U.S. patent application number 17/530950 was filed with the patent office on 2022-05-26 for delayed harvest of short stature corn plants.
This patent application is currently assigned to MONSANTO TECHNOLOGY LLC. The applicant listed for this patent is MONSANTO TECHNOLOGY LLC. Invention is credited to Ty J. BARTEN, Edward J. CARGILL, Bryce LEMKE.
Application Number | 20220162632 17/530950 |
Document ID | / |
Family ID | |
Filed Date | 2022-05-26 |
United States Patent
Application |
20220162632 |
Kind Code |
A1 |
BARTEN; Ty J. ; et
al. |
May 26, 2022 |
DELAYED HARVEST OF SHORT STATURE CORN PLANTS
Abstract
Methods for delayed harvesting of corn fields are provided
herein. These methods provide an extended, flexible period of time
to harvest corn. The methods allow growers to harvest their corn at
the optimal time for drying down or accessing seed, without
increasing the risk of losing yield to lodging.
Inventors: |
BARTEN; Ty J.; (Ankeny,
IA) ; CARGILL; Edward J.; (Chesterfield, MO) ;
LEMKE; Bryce; (Nevada, IA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MONSANTO TECHNOLOGY LLC |
St. Louis |
MO |
US |
|
|
Assignee: |
MONSANTO TECHNOLOGY LLC
St. Louis
MO
|
Appl. No.: |
17/530950 |
Filed: |
November 19, 2021 |
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International
Class: |
C12N 15/82 20060101
C12N015/82; C12N 9/02 20060101 C12N009/02; C07K 14/415 20060101
C07K014/415; A01D 91/04 20060101 A01D091/04 |
Claims
1. A method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises: (a) a mutant allele of an endogenous GA20 oxidase_3
locus, wherein the mutant allele comprises a first DNA segment
inserted into the endogenous GA20 oxidase_3 locus, wherein the
first DNA segment encodes an antisense RNA sequence that is at
least 70% complementary to at least 20 consecutive nucleotides of
one or more of SEQ ID NOs: 182-184 and 186-188, and wherein the
mutant allele of the endogenous GA20 oxidase_3 locus produces a RNA
transcript comprising the antisense RNA sequence; or (b) a first
mutant allele of an endogenous GA20 oxidase_5 locus, wherein the
first mutant allele of the endogenous GA20 oxidase_5 locus
comprises a second DNA segment inserted into the endogenous GA20
oxidase_5 locus, wherein the second DNA segment encodes an
antisense RNA sequence that is at least 70% complementary to at
least 20 consecutive nucleotides of one or more of SEQ ID NOs:
182-184 and 186-188, and wherein the mutant allele of the
endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence; or (c) a first mutant allele
of an endogenous Brachytic2 (br2) locus, wherein the first mutant
allele of an endogenous br2 locus comprises a third DNA segment
inserted into the endogenous br2 locus, wherein the third DNA
segment encodes an antisense RNA that is at least 70% complementary
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180,
and wherein the mutant allele of the endogenous br2 locus produces
an RNA transcript comprising the antisense RNA sequence; or (d) a
second mutant allele of an endogenous br2 locus, wherein the second
mutant allele of an endogenous br2 locus comprises a deletion of at
least one nucleotide from an endogenous br2 locus as compared to
SEQ ID NO: 132; or (e) a dominant or semi-dominant transgene or
mutant allele of a gene, and wherein the transgene or mutant allele
causes a short stature phenotype in the at least one corn plant; or
(f) a premature stop codon within a nucleic acid sequence encoding
a Brachytic2 protein as compared to a control corn plant; or (g) a
second mutant allele of an endogenous GA20 oxidase_5 locus, wherein
the second mutant allele of the endogenous GA20 oxidase_5 locus
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene; or (h) a
third mutant allele of an endogenous GA20 oxidase_5 locus, wherein
the third mutant allele of the endogenous GA20 oxidase_5 locus
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene; or (i) a genome modification comprising a deletion of at
least a portion of one or more of the following: 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3' UTR, and any portion thereof, and the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any portion
thereof, of the endogenous Zm.SAMT gene; or (j) a fourth mutant
allele of an endogenous GA20 oxidase_5 locus, wherein the fourth
mutant allele of the endogenous GA20 oxidase_5 locus comprises a
genome modification which results in the transcription of an
antisense strand of at least an exon, an intron, or an untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof; or (k) a fifth mutant allele of an endogenous GA20
oxidase_5 locus, wherein the fifth mutant allele of the endogenous
GA20 oxidase_5 locus comprises the Zm.SAMT gene promoter, or a
functional part thereof, operably linked to at least one
transcribable antisense sequence of at least an exon, intron or
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof; or (l) a sixth mutant allele of an endogenous
GA20 oxidase_5 locus, wherein the sixth mutant allele of the
endogenous GA20 oxidase_5 locus comprises a sequence selected from
the group consisting of SEQ ID NOs: 87-105; or (m) a seventh mutant
allele of an endogenous GA20 oxidase_5 locus, wherein the seventh
mutant allele of the endogenous GA20 oxidase_5 locus comprises a
first sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides; or (n) an eighth mutant
allele of an endogenous GA20 oxidase_5 locus, wherein the eighth
mutant allele of the endogenous GA20 oxidase_5 locus comprises a
genomic deletion relative to a wild type allele of the endogenous
GA20 oxidase_5 locus, wherein the genomic deletion is flanked by a
first sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; or (o) a genomic sequence comprising a
first sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length; or (p) a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
2. A method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises: (a) a mutant
allele of an endogenous GA20 oxidase_3 locus, wherein the mutant
allele comprises a first DNA segment inserted into the endogenous
GA20 oxidase_3 locus, wherein the first DNA segment encodes an
antisense RNA sequence that is at least 70% complementary to at
least 20 consecutive nucleotides of one or more of SEQ ID NOs:
182-184 and 186-188, and wherein the mutant allele of the
endogenous GA20 oxidase_3 locus produces a RNA transcript
comprising the antisense RNA sequence; or (b) a first mutant allele
of an endogenous GA20 oxidase_5 locus, wherein the first mutant
allele of the endogenous GA20 oxidase_5 locus comprises a second
DNA segment inserted into the endogenous GA20 oxidase_5 locus,
wherein the second DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA sequence; or
(c) a first mutant allele of an endogenous Brachytic2 (br2) locus,
wherein the first mutant allele of the endogenous br2 locus
comprises a third DNA segment inserted into the endogenous br2
locus, wherein the third DNA segment encodes an antisense RNA that
is at least 70% complementary to at least 20 consecutive
nucleotides of SEQ ID NO: 132 or 180, and wherein the mutant allele
of the endogenous br2 locus produces an RNA transcript comprising
the antisense RNA sequence; or (d) a second mutant allele of an
endogenous br2 locus, wherein the second mutant allele of the
endogenous br2 locus comprises a deletion of at least one
nucleotide from an endogenous br2 locus as compared to SEQ ID NO:
132; or (e) a dominant or semi-dominant transgene or mutant allele
of a gene, and wherein the transgene or mutant allele causes a
short stature phenotype in the at least one corn plant; or (f) a
premature stop codon within a nucleic acid sequence encoding a
Brachytic2 protein as compared to a control corn plant; or (g) a
second mutant allele of an endogenous GA20 oxidase_5 locus, wherein
the second mutant allele of the endogenous GA20 oxidase_5 locus
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene; or (h) a
third mutant allele of an endogenous GA20 oxidase_5 locus, wherein
the third mutant allele of the endogenous GA20 oxidase_5 locus
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene; or (i) a genome modification comprising a deletion of at
least a portion of one or more of the following: 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3' UTR, and any portion thereof, and the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any portion
thereof, of the endogenous Zm.SAMT gene; or (j) a fourth mutant
allele of an endogenous GA20 oxidase_5 locus, wherein the fourth
mutant allele of the endogenous GA20 oxidase_5 locus comprises a
genome modification which results in the transcription of an
antisense strand of at least an exon, an intron, or an untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof; or (k) a fifth mutant allele of an endogenous GA20
oxidase_5 locus, wherein the fifth mutant allele of the endogenous
GA20 oxidase_5 locus comprises the Zm.SAMT gene promoter, or a
functional part thereof, operably linked to at least one
transcribable antisense sequence of at least an exon, intron or
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof; or (l) a sixth mutant allele of an endogenous
GA20 oxidase_5 locus, wherein the sixth mutant allele of the
endogenous GA20 oxidase_5 locus comprises a sequence selected from
the group consisting of SEQ ID NOs: 87-105; or (m) an seventh
mutant allele of an endogenous GA20 oxidase_5 locus, wherein the
seventh mutant allele of the endogenous GA20 oxidase_5 locus
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides; or (n) a eighth mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the eighth
mutant allele of the endogenous GA20 oxidase_5 locus comprises a
genomic deletion relative to a wild type allele of the endogenous
GA20 oxidase_5 locus, wherein the genomic deletion is flanked by a
first sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; or (o) a genomic sequence comprising a
first sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length; or (p) a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
3. The method of claim 1, wherein the average kernel moisture
content is less than or equal to 30%
4. The method of claim 2, wherein average kernel moisture content
is less than or equal to 30%
5. The method of claim 1, wherein the average yield of said field
is at least 170 bushels per acre.
6. The method of claim 2, wherein the average yield of said field
is at least 170 bushels per acre.
7. A method comprising harvesting a plurality of corn plants from a
field at least 1 day after the average kernel moisture content of
at least 50% of said plurality of corn plants is between 10% and
30%, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises: (a) a mutant allele of
an endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a first DNA segment inserted into the endogenous GA20
oxidase_3 locus, wherein the first DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_3 locus produces a RNA transcript comprising the antisense
RNA sequence; or (b) a first mutant allele of an endogenous GA20
oxidase_5 locus, wherein the first mutant allele of the endogenous
GA20 oxidase_5 locus comprises a second DNA segment inserted into
the endogenous GA20 oxidase_5 locus, wherein the second DNA segment
encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence; or (c) a first mutant allele
of an endogenous Brachytic2 (br2) locus, wherein the first mutant
allele of the endogenous br2 locus comprises a third DNA segment
inserted into the endogenous br2 locus, wherein the third DNA
segment encodes an antisense RNA that is at least 70% complementary
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180,
and wherein the mutant allele of the endogenous br2 locus produces
an RNA transcript comprising the antisense RNA sequence; or (d) a
second mutant allele of an endogenous br2 locus, wherein the second
mutant allele comprises a deletion of at least one nucleotide from
an endogenous br2 locus as compared to SEQ ID NO: 132; or (e) a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant; or (f) a premature stop
codon within a nucleic acid sequence encoding a Brachytic2 protein
as compared to a control corn plant; or (g) a second mutant allele
of an endogenous GA20 oxidase_5 locus, wherein the second mutant
allele of the endogenous GA20 oxidase_5 locus comprises a genome
modification comprising a deletion of at least a portion of the
transcription termination sequence of the endogenous Zm.SAMT gene,
and wherein the mutant allele produces a RNA molecule comprising an
antisense sequence complementary to all or part of the sense strand
of the endogenous GA20 oxidase_5 gene; or (h) a third mutant allele
of an endogenous GA20 oxidase_5 locus, wherein the third mutant
allele of the endogenous GA20 oxidase_5 locus comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene; or (i) a
genome modification comprising a deletion of at least a portion of
one or more of the following: 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any portion thereof, and the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3.sup.rd
intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th
intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th
intron, 8.sup.th exon, 3' UTR, and any portion thereof, of the
endogenous Zm.SAMT gene; or (j) a fourth mutant allele of an
endogenous GA20 oxidase_5 locus, wherein the fourth mutant allele
of the endogenous GA20 oxidase_5 locus comprises a genome
modification which results in the transcription of an antisense
strand of at least an exon, an intron, or an untranslated region
(UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof; or (k) a fifth mutant allele of an endogenous GA20
oxidase_5 locus, wherein the fifth mutant allele of the endogenous
GA20 oxidase_5 locus comprises the Zm.SAMT gene promoter, or a
functional part thereof, operably linked to at least one
transcribable antisense sequence of at least an exon, intron or
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof; or (l) a sixth mutant allele of an endogenous
GA20 oxidase_5 locus, wherein the sixth mutant allele of the
endogenous GA20 oxidase_5 locus comprises a sequence selected from
the group consisting of SEQ ID NOs: 87-105; or (m) an seventh
mutant allele of an endogenous GA20 oxidase_5 locus, wherein the
seventh mutant allele of the endogenous GA20 oxidase_5 locus
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides; or (n) an eighth mutant
allele of an endogenous GA20 oxidase_5 locus, wherein the eighth
mutant allele of the endogenous GA20 oxidase_5 locus comprises a
genomic deletion relative to a wild type allele of the endogenous
GA20 oxidase_5 locus, wherein the genomic deletion is flanked by a
first sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; or (o) a genomic sequence comprising a
first sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length; or (p) a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
8. The method of claim 1, wherein (i) at least 50% of said corn
plants are inbred corn plants or (ii) wherein at least 50% of said
corn plants are hybrid corn plants.
9. The method of claim 1, wherein (i) at least 50% of said corn
plants are semi-dwarf corn plants; (ii) at least 50% of said corn
plants are dwarf corn plants; or (iii) at least 50% of said corn
plants are brachytic corn plants.
10. The method of claim 1, wherein said field comprises a planting
density of at least 10,000 corn plants per acre.
11. The method of claim 1, wherein (i) the mutant allele of the
endogenous GA20 oxidase_3 locus suppresses the expression of a
wild-type allele of the endogenous GA20 oxidase_3 locus, a
wild-type allele of the endogenous GA20 oxidase_5 locus, or both;
or (ii) the first mutant allele of an endogenous GA20 oxidase_5
locus suppresses the expression of a wild-type allele of the
endogenous GA20 oxidase_3 locus, a wild-type allele of the
endogenous GA20 oxidase_5 locus, or both.
12. The method of claim 1, wherein the first DNA segment (i)
comprises a nucleotide sequence originating from the endogenous
GA20 oxidase_3 locus; (ii) corresponds to an inverted genomic
fragment of the endogenous GA20 oxidase_3 locus; or (iii) comprises
a nucleotide sequence originating from the endogenous GA20
oxidase_5 locus.
13. The method of claim 1, wherein the first DNA segment comprises
a sequence having at least at least 70% identity to one or more of
SEQ ID Nos: 194, 195, 207, 209, 211, 213, and 217.
14. The method of claim 1, wherein the second DNA segment comprises
a sequence having at least at least 70% identity to one or more of
SEQ ID Nos: 194, 195, 207, 209, 211, 213, and 217.
15. The method of claim 1, wherein the level of one or more active
GAs in at least one internode tissue of the stem or stalk of the
modified corn plant is lower than the same internode tissue of an
unmodified control plant.
16. The method of claim 1, wherein the second mutant allele of an
endogenous GA20 oxidase_5 gene comprises the endogenous Zm.SAMT
gene promoter, or a portion thereof, operably linked to a
transcribable DNA sequence encoding: (i) a RNA molecule that causes
suppression of one or both of the endogenous GA20 oxidase_3 gene
and the endogenous GA20 oxidase_5 gene; (ii) a RNA molecule
comprising an antisense sequence that is at least 80% complementary
to all or part of the endogenous GA20 oxidase_3 or GA20 oxidase_5
gene; or (iii) both (i) and (ii).
17. The method of claim 1, wherein the at least one transcribable
antisense sequence is at least 80% complementary to at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 218-220,
222-224, 226, and 228-255.
18. The method of claim 1, wherein the first mutant allele of the
endogenous br2 locus suppresses the expression of a wild-type
allele of the endogenous br2 locus.
19. The method of claim 1, wherein the third DNA segment comprises
a sequence having at least at least 70% identity to one or more of
SEQ ID Nos: 132 and 180.
20. The method claim 1, wherein the second mutant allele of an
endogenous br2 locus comprises the deletion of (i) at least one
nucleotide of at least one exon of the endogenous br2 locus as
compared to SEQ ID NO: 132; and/or (ii) a deletion of at least one
nucleotide from at least one intron of the endogenous br2
locus.
21. The method of claim 1, wherein the second mutant allele of an
endogenous br2 locus encodes a truncated protein as compared to SEQ
ID NO: 181.
22. The method of claim 1, wherein the at least one corn plant has
improved lodging resistance relative to an unmodified control
plant.
23. The method of claim 1, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80% identical to one or more
of SEQ ID NOs: 325, 327, 329, 331, 333, 335, 337, 339, 341, 343,
345, 347, 349, 351, 353, 355, 357, 359, 361, 363, 365, 367, 369,
371, 373, 375, 377, 379, 381, 383, 385, 387, 389, 391, 393, 395,
397, 399, 401, 403, 405, 407, 409, 411, 413, 415, 417, 419, 421,
423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443, 445, 447,
449, 451, 453, 455, 457, 459, 461, 463, 465, 467, 469, 471, 473,
475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495, 497, 499,
501, 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525,
527, 529, 531, 533, 535, 537, 539, 541, 543, 545, 547, 549, 551,
553, 555, 557, 559, 561, 563, 565, 567, 569, 571, 573, 575, 577,
579, 581, 583, 585, 587, 589, 591, 593, 595, 597, 599, 601, 603,
605, 607, 609, 611, 613, 615, 617, 619, 621, 623, 625, 627, 629,
631, 633, 635, 637, 639, 641, 643, 645, 647, 649, 651, 653, and/or
655.
24. The method of claim 1, wherein the transcribable DNA sequence
is, or comprises a sequence that is, at least 80% identical to one
or more of SEQ ID NOs: 324, 326, 328, 330, 332, 334, 336, 338, 340,
342, 344, 346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366,
368, 370, 372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392,
394, 396, 398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418,
420, 421, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444,
446, 448, 450, 452, 454, 456, 458, 460, 462, 464, 466, 468, 470,
472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494, 496,
498, 500, 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522,
524, 526, 528, 530, 532, 534, 536, 538, 540, 542, 544, 546, 548,
550, 552, 554, 556, 558, 560, 562, 564, 566, 568, 570, 572, 574,
576, 578, 580, 582, 584, 586, 588, 590, 592, 594, 596, 598, 600,
602, 604, 606, 608, 610, 612, 614, 616, 618, 620, 622, 624, 626,
628, 630, 632, 634, 636, 638, 640, 642, 644, 646, 648, 650, 652,
and/or 655.
25. The method of claim 1, wherein the plant-expressible promoter
is a (i) vascular promoter; (ii) a leaf promoter; or (iii) a
constitutive promoter.
26. The method of claim 1, wherein the plant-expressible promoter
is a rice tungro bacilliform virus (RTBV) promoter.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit of U.S. Provisional
Application No. 63/117,231, filed Nov. 23, 2020; U.S. Provisional
Application No. 63/117,237, filed Nov. 23, 2020; U.S. Provisional
Application No. 63/117,247, filed Nov. 23, 2020; U.S. Provisional
Application No. 63/117,225, filed Nov. 23, 2020; and U.S.
Provisional Application No. 63/125,752, filed Dec. 15, 2020; and
U.S. Provisional Application No. 63/180,344, filed Apr. 27, 2021,
all of which are incorporated by reference in their entireties
herein.
INCORPORATION OF SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted electronically in ASCII format and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Nov. 19, 2021, is named P35026US01_SL.txt and is 1,161,026 bytes
in size.
FIELD
[0003] The present disclosure relates to methods of delayed harvest
of corn fields.
BACKGROUND
[0004] Corn can be harvested after fertilization, grain fill and
maturity, but typically after drying down to a desired moisture
content for storage. Growers have to balance product value, plant
health, kernel moisture content, and standability (e.g., due to the
propensity of corn to lodge) of corn plants when determining the
optimum time for harvesting. If a grower harvests corn before it
reaches its optimal kernel moisture content, the grower may have to
use artificial drying methods to further reduce the kernel moisture
content before storage. Conversely, if a grower waits to harvest
corn (or cannot harvest due to physical weather barriers such as
rain or snow) until it reaches or passes optimal kernel moisture,
then the longer the amount of time the crop remains in the field,
the greater the risk of lodging from weather events (e.g., strong
winds) and/or plant senescence (i.e., deterioration from age).
Thus, there is a need for farmers to have greater flexibility to
leave corn plants in the field for later harvest to allow for
greater access and/or dry down of kernels.
SUMMARY
[0005] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0006] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0007] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence.
[0008] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132.
[0009] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant.
[0010] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a premature stop
codon within a nucleic acid sequence encoding a Brachytic2 protein
as compared to a control corn plant.
[0011] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0012] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0013] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0014] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0015] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0016] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0017] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0018] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0019] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0020] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0021] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0022] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a DNA
segment inserted into the endogenous br2 locus, wherein the DNA
segment encodes an antisense RNA that is at least 70% complementary
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180,
and wherein the mutant allele of the endogenous br2 locus produces
an RNA transcript comprising the antisense RNA sequence.
[0023] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a
deletion of at least one nucleotide from an endogenous br2 locus as
compared to SEQ ID NO: 132.
[0024] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a dominant or semi-dominant
transgene or mutant allele of a gene, and wherein the transgene or
mutant allele causes a short stature phenotype in the at least one
corn plant.
[0025] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a premature stop codon within a
nucleic acid sequence encoding a Brachytic2 protein as compared to
a control corn plant.
[0026] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0027] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0028] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0029] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0030] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0031] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0032] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0033] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0034] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0035] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_3 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_3 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_3 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0036] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_5 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_5 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0037] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
br2 locus, wherein the DNA segment encodes an antisense RNA that is
at least 70% complementary to at least 20 consecutive nucleotides
of SEQ ID NO: 132 or 180, and wherein the mutant allele of the
endogenous br2 locus produces an RNA transcript comprising the
antisense RNA sequence.
[0038] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a deletion of at least one nucleotide from
an endogenous br2 locus as compared to SEQ ID NO: 132.
[0039] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant.
[0040] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
premature stop codon within a nucleic acid sequence encoding a
Brachytic2 protein as compared to a control corn plant.
[0041] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the transcription termination sequence of
the endogenous Zm.SAMT gene, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0042] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the intergenic region between the
endogenous GA20 oxidase_5 and Zm.SAMT genes, and wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
complementary to all or part of the sense strand of the endogenous
GA20 oxidase_5 gene.
[0043] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of one or more of the following: 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon,
2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon,
5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon,
3' UTR, and any portion thereof, of the endogenous Zm.SAMT
gene.
[0044] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification which results in the
transcription of an antisense strand of at least an exon, an
intron, or an untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof.
[0045] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises the Zm.SAMT gene promoter, or a functional
part thereof, operably linked to at least one transcribable
antisense sequence of at least an exon, intron or untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0046] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a sequence selected from the group
consisting of SEQ ID NOs: 87-105.
[0047] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0048] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic deletion relative to a wild type
allele of the endogenous GA20 oxidase_5 locus, wherein the genomic
deletion is flanked by a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0049] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic sequence comprising a first
sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length.
[0050] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0051] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0052] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence.
[0053] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132.
[0054] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant.
[0055] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a premature stop
codon within a nucleic acid sequence encoding a Brachytic2 protein
as compared to a control corn plant.
[0056] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0057] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0058] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0059] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0060] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0061] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0062] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0063] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0064] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0065] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_3 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_3 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_3 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0066] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_5 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_5 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0067] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
br2 locus, wherein the DNA segment encodes an antisense RNA that is
at least 70% complementary to at least 20 consecutive nucleotides
of SEQ ID NO: 132 or 180, and wherein the mutant allele of the
endogenous br2 locus produces an RNA transcript comprising the
antisense RNA sequence.
[0068] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a deletion of at least one nucleotide from
an endogenous br2 locus as compared to SEQ ID NO: 132.
[0069] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant.
[0070] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
premature stop codon within a nucleic acid sequence encoding a
Brachytic2 protein as compared to a control corn plant.
[0071] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the transcription termination sequence of
the endogenous Zm.SAMT gene, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0072] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the intergenic region between the
endogenous GA20 oxidase_5 and Zm.SAMT genes, and wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
complementary to all or part of the sense strand of the endogenous
GA20 oxidase_5 gene.
[0073] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of one or more of the following: 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon,
2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon,
5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon,
3' UTR, and any portion thereof, of the endogenous Zm.SAMT
gene.
[0074] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification which results in the
transcription of an antisense strand of at least an exon, an
intron, or an untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof.
[0075] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises the Zm.SAMT gene promoter, or a functional
part thereof, operably linked to at least one transcribable
antisense sequence of at least an exon, intron or untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0076] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a sequence selected from the group
consisting of SEQ ID NOs: 87-105.
[0077] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0078] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic deletion relative to a wild type
allele of the endogenous GA20 oxidase_5 locus, wherein the genomic
deletion is flanked by a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0079] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic sequence comprising a first
sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length.
[0080] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0081] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0082] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of an
endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence.
[0083] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of an
endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132.
[0084] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant.
[0085] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a premature stop codon
within a nucleic acid sequence encoding a Brachytic2 protein as
compared to a control corn plant.
[0086] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0087] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0088] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0089] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0090] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0091] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0092] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0093] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0094] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0095] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_3 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_3 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_3 locus produces a RNA transcript
comprising the antisense RNA sequence.
[0096] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_5 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence.
[0097] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous br2 locus, wherein the DNA segment
encodes an antisense RNA that is at least 70% complementary to at
least 20 consecutive nucleotides of SEQ ID NO: 132 or 180, and
wherein the mutant allele of the endogenous br2 locus produces an
RNA transcript comprising the antisense RNA sequence.
[0098] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a deletion of at
least one nucleotide from an endogenous br2 locus as compared to
SEQ ID NO: 132.
[0099] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a dominant or semi-dominant transgene or
mutant allele of a gene, and wherein the transgene or mutant allele
causes a short stature phenotype in the at least one corn
plant.
[0100] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a premature stop codon within a nucleic acid
sequence encoding a Brachytic2 protein as compared to a control
corn plant.
[0101] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
transcription termination sequence of the endogenous Zm.SAMT gene,
and wherein the mutant allele produces a RNA molecule comprising an
antisense sequence complementary to all or part of the sense strand
of the endogenous GA20 oxidase_5 gene.
[0102] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0103] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of one or
more of the following: 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3' UTR, and any portion thereof, and the 5' UTR,
1st exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3rd intron,
4th exon, 4th intron, 5th exon, 5th intron, 6th exon, 6th intron,
7th exon, 7th intron, 8th exon, 3' UTR, and any portion thereof, of
the endogenous Zm.SAMT gene.
[0104] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification which results in the transcription of an antisense
strand of at least an exon, an intron, or an untranslated region
(UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0105] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises the Zm.SAMT
gene promoter, or a functional part thereof, operably linked to at
least one transcribable antisense sequence of at least an exon,
intron or untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof.
[0106] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a sequence
selected from the group consisting of SEQ ID NOs: 87-105.
[0107] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3' UTR, and any complementary sequence
thereof, and any portion of the foregoing, of the endogenous
Zm.GA20 oxidase_5 gene; and wherein the second sequence comprises
one or more of the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene; wherein the first
sequence and the second sequence are contiguous or separated only
by an intervening sequence of fewer than 555 nucleotides.
[0108] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
deletion relative to a wild type allele of the endogenous GA20
oxidase_5 locus, wherein the genomic deletion is flanked by a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3' UTR, and any complementary sequence
thereof, and any portion of the foregoing, of the endogenous
Zm.GA20 oxidase_5 gene; and wherein the second sequence comprises
one or more of the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene.
[0109] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
sequence comprising a first sequence and a second sequence; wherein
the first sequence comprises at least 15 consecutive nucleotides of
one or more of SEQ ID NOs: 228-235 and 276-283; wherein the second
sequence comprises at least 15 consecutive nucleotides of one or
more of SEQ ID NOs: 235-276; and wherein the genomic sequence is at
least 50 consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length.
[0110] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0111] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible
promoter.
[0112] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
recombinant DNA construct comprising a transcribable DNA sequence
encoding a GA2 oxidase protein and a plant-expressible promoter,
wherein the transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0113] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0114] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
recombinant DNA construct comprising a transcribable DNA sequence
encoding a GA2 oxidase protein and a plant-expressible promoter,
wherein the transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0115] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0116] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter.
BRIEF DESCRIPTION OF DRAWINGS
[0117] FIG. 1 depicts the stalk health ratings of short and tall
corn plants at normal and late harvest times.
[0118] FIG. 2 depicts an example of a planting, maturation, and
harvesting schedule for corn.
DETAILED DESCRIPTION
[0119] Unless defined otherwise, all technical and scientific terms
used have the same meaning as commonly understood by one of
ordinary skill in the art to which this disclosure belongs. Unless
otherwise provided, where a term is provided in the singular, this
disclosure also contemplates the plural of that term. Where there
are discrepancies in terms and definitions used in references that
are incorporated by reference, the terms used in this application
shall have the definitions given herein. Other technical terms used
have their ordinary meaning in the art in which they are used, as
exemplified by various art-specific dictionaries, for example, "The
American Heritage.RTM. Science Dictionary" (Editors of the American
Heritage Dictionaries, 2011, Houghton Mifflin Harcourt, Boston and
New York), the "McGraw-Hill Dictionary of Scientific and Technical
Terms" (6th edition, 2002, McGraw-Hill, New York), or the "Oxford
Dictionary of Biology" (6th edition, 2008, Oxford University Press,
Oxford and New York). Any references cited herein, including, e.g.,
all patents, published patent applications, and non-patent
publications, are incorporated herein by reference in their
entirety.
[0120] When a grouping of alternatives is presented, any and all
combinations of the members that make up that grouping of
alternatives is specifically envisioned. For example, if an item is
selected from a group consisting of A, B, C, and D, the inventors
specifically envision each alternative individually (e.g., A alone,
B alone, etc.), as well as combinations such as A, B, and D; A, C,
and D; A, B, and C; A and C; B and C; A and B; etc. The term
"and/or" when used in a list of two or more items means any one of
the listed items by itself or in combination with any one or more
of the other listed items. For example, the expression "A and/or B"
is intended to mean either or both of A and B--i.e., A alone, B
alone, or A and B in combination. The expression "A, B and/or C" is
intended to mean A alone, B alone, C alone, A and B in combination,
A and C in combination, B and C in combination, or A, B, and C in
combination.
[0121] As well understood in the art, metric measurement values
provided herein can be easily converted to standard (S.I.) units
where relevant, and vice versa.
[0122] As used herein, a "plant" includes an explant, plant part,
seedling, plantlet or whole plant at any stage of regeneration or
development. As commonly understood, a "corn plant" or "maize
plant" refers to any plant of species Zea mays and includes all
plant varieties that can be bred with corn, including wild maize
species.
[0123] As used herein, a "plant part" can refer to any organ or
intact tissue of a plant, such as a meristem, shoot organ/structure
(e.g., leaf, stem or node), root, flower or floral organ/structure
(e.g., bract, sepal, petal, stamen, carpel, anther and ovule),
seed, embryo, endosperm, seed coat, fruit, the mature ovary,
propagule, or other plant tissues (e.g., vascular tissue, dermal
tissue, ground tissue, and the like), or any portion thereof. Plant
parts of the present disclosure can be viable, nonviable,
regenerable, and/or non-regenerable. A "propagule" can include any
plant part that can grow into an entire plant.
[0124] As used herein, a "locus" is a chromosomal locus or region
where a polymorphic nucleic acid, trait determinant, gene, or
marker is located. A "locus" can be shared by two homologous
chromosomes to refer to their corresponding locus or region.
Without being limiting, a locus can comprise a polynucleotide that
encodes a protein or an RNA. A locus can also comprise a non-coding
RNA. A locus can comprise a gene. A locus can comprise a promoter,
a 5'-untranslated region (UTR), an exon, an intron, a 3'-UTR, or
any combination thereof. A locus can comprise a coding region.
[0125] As used herein, an "allele" refers to an alternative (e.g.,
variant) nucleic acid sequence of a gene or at a particular locus
(e.g., a nucleic acid sequence of a gene or locus that is different
than other alleles for the same gene or locus). Such an allele can
be considered (i) wild-type or (ii) mutant if one or more mutations
or edits are present in the nucleic acid sequence of the mutant
allele relative to the wild-type allele. A mutant or edited allele
for a gene may have a reduced or eliminated activity or expression
level for the gene relative to the wild-type allele. According to
present embodiments, a mutant or edited allele for a gene may have
an inversion or antisense sequence that may be complementary to
another portion of the gene and/or a coding sequence of another
copy or allele of the gene and/or another gene. For diploid
organisms such as corn, a first allele can occur on one chromosome,
and a second allele can occur at the same locus on a second
homologous chromosome. If one allele at a locus on one chromosome
of a plant is a mutant or edited allele and the other corresponding
allele on the homologous chromosome of the plant is wild-type, then
the plant is described as being heterozygous for the mutant or
edited allele. However, if both alleles at a locus are mutant or
edited alleles, then the plant is described as being homozygous for
the mutant or edited alleles. A plant homozygous for mutant alleles
at a locus may comprise the same mutant or edited allele or
different mutant or edited alleles if heteroallelic or
biallelic.
[0126] As used herein, an "endogenous locus" refers to a locus at
its natural and original chromosomal location. As used herein, the
"endogenous GA20 oxidase_3 locus" refers to the GA20 oxidase_3
genic locus at its original chromosomal location. As used herein,
the "endogenous GA20 oxidase_5 locus" refers to the GA20 oxidase_5
genic locus at its original chromosomal location. As used herein,
the "endogenous GA3 oxidase locus" refers to the GA3 oxidase genic
locus at its original chromosomal location. As used herein, the
"endogenous br2 locus" refers to the br2 genic locus at its
original chromosomal location.
[0127] As used herein, a "female" corn plant refers to a corn plant
that comprises one or more female reproductive structures that are
capable of producing corn ear(s) and kernels (seed). In an aspect,
a corn plant provided herein is a female corn plant. In an aspect,
a female corn plant is male sterile. In another aspect, a female
corn plant is detasseled. In an aspect, the male reproductive
organs or flowers (e.g., tassels) of a female corn plant are
chemically sterilized (e.g., by application of an herbicide to
plants lacking tolerance to the herbicide in those male
reproductive organs, flowers or tassels), such as with a
Roundup.RTM. Hybridization System (RHS). In another aspect, the
male reproductive organs or flowers (e.g., tassels) of a female
corn plant are sterilized due to cytoplasmic male sterility (or
CMS). It is appreciated in the art that a corn plant is monoecious
and can be considered both a male corn plant and a female corn
plant. In an aspect, a female corn plant is capable of producing
pollen. For purposes of the present disclosure, a corn plant having
one or more male reproductive organ or structure, such as tassels,
and/or capable of producing pollen, is considered a "female" plant
if used to generate a corn ear(s) and/or corn seed (i.e., kernels)
for production and harvest. A corn plant lacking a male
reproductive organ or structure, such as tassels, having a
sterilized male reproductive organ or structure, and/or incapable
of producing pollen, is also considered a "female" plant if used to
generate a corn ear(s) and/or seed (i.e., kernels) for production
and harvest. A "female" corn plant may include any pollen-receiving
corn plant that produces an ear or female reproductive organ, which
can receive pollen from a pollen-bearing corn plant.
[0128] As used herein, a "male" corn plant refers to a corn plant
that is capable of producing pollen (e.g., form one or more
tassels) and is used to pollinate and/or fertilize one or more
female corn plant(s) for seed production and harvest, even if the
male plant further has a female reproductive structure(s) that
is/are capable of producing a corn ear and kernels (seed), which
may or may not be harvested. A "male" corn plant may include any
pollen-bearing (or pollen-producing) corn plant, which can provide
its pollen to a pollen-receiving corn plant.
[0129] As used herein, the phrase "at least one" in reference to
something (e.g., any object, method step, etc.) means one or more
of that something. For example, "at least one plant" or "at least
one plants" each means one or more plants. Accordingly, "at least
one" can include one or a plurality. Thus, where the present
disclosure provides "one or more" of something or "at least one" of
something, then the description further supports a plurality of
that something.
[0130] In an aspect, any mutant allele, mutation (e.g., without
being limiting, a deletion, inversion, insertion, or combinations
thereof) or transgene provided herein is present in the genome of a
female plant. In an aspect, any mutant allele, mutation (e.g.,
without being limiting, a deletion, inversion, insertion, or
combinations thereof) or transgene provided herein is present in
the genome of a male plant.
[0131] In an aspect, this disclosure provides corn plants
comprising a dominant mutant allele. In an aspect, this disclosure
provides corn plants comprising a semi-dominant mutant allele.
Dominant alleles are alleles that mask the contribution of a second
allele at the same locus. A dominant allele can be a "dominant
negative allele" or a "dominant positive allele." Dominant negative
alleles, or antimorphs, are alleles that act in opposition to
normal allelic function. A dominant negative allele typically does
not function normally and either directly inhibits the activity of
a wild-type protein (e.g., through dimerization) or inhibits the
activity of a second protein that is required for the normal
function of the wild-type protein (e.g., an activator or a
downstream component of a pathway). For example, a dominant
negative allele abrogates or reduces the normal function of an
allele in a heterozygous or homozygous state. Dominant positive
alleles can increase normal gene function (e.g., a hypermorph) or
provide new functions for a gene (e.g., a neomorph). A
semi-dominant allele occurs when penetrance of a linked phenotype
in individuals heterozygous for the allele is less than that which
is observed in individuals homozygous for the allele.
[0132] Creation of dominant alleles that work in a heterozygous
state, can speed up effective trait development, deployment, and
launch of gene editing-derived products in hybrid crops such as
corn. Dominant negative alleles have the potential advantage of
providing a positive or beneficial plant trait in a heterozygous
state--e.g., when present in a single copy. As a result, a dominant
negative mutant allele can be introduced through crossing into a
progeny plant from a single parent without having to introduce the
allele from both parent plants as with a recessive allele.
[0133] In an aspect, a dominant mutant allele is a dominant
negative allele. In an aspect, a dominant mutant allele is a
dominant positive allele.
[0134] In an aspect, a dominant mutant allele or a semi-dominant
mutant allele comprises an insertion, an inversion, a deletion, or
any combination thereof as compared to a wildtype allele of a
gene.
[0135] In an aspect, a mutant allele provided herein is a dominant
mutant allele. In an aspect, a mutant allele provided herein is a
semi-dominant mutant allele. In an aspect, a mutant allele provided
herein is a dominant negative mutant allele.
[0136] In an aspect, a female corn plant is a modified corn plant.
In another aspect, a female inbred corn plant is a modified corn
plant. In an aspect, a modified plant provided herein is homozygous
(or biallelic) for a dominant mutant allele(s) or transgene. In an
aspect, a modified plant provided herein is homozygous (or
biallelic) for a semi-dominant mutant allele(s). In an aspect, a
modified plant provided herein is homozygous (or biallelic) for a
dominant negative mutant allele. In an aspect, a modified plant
provided herein is heterozygous or hemizygous for a dominant mutant
allele or transgene. In an aspect, a modified plant provided herein
is heterozygous or hemizygous for a semi-dominant mutant allele or
transgene. In an aspect, a modified plant provided herein is
heterozygous for a dominant negative mutant allele. As used herein,
"modified", in the context of plants, seeds, plant components,
plant cells, and plant genomes, refers to a state containing
changes or variations from their natural or native state. According
to an aspect, a modified corn plant, which may be a female corn
plant, has a shorter plant height as compared to a control plant
and/or a male corn plant.
[0137] As used herein, the term "control plant" (or likewise a
"control" plant seed, plant part, plant cell and/or plant genome)
refers to a plant (or plant seed, plant part, plant cell and/or
plant genome) that is used for comparison to a modified plant (or
modified plant seed, plant part, plant cell and/or plant genome)
and has the same or similar genetic background (e.g., same parental
lines, hybrid cross, inbred line, testers, etc.) as the modified
plant (or plant seed, plant part, plant cell and/or plant genome),
except for a transgenic event and/or genome edit(s) (e.g., an
inversion or antisense insertion) affecting one or more genes. For
example, a control plant may be an inbred line that is the same as
the inbred line used to make the modified plant, or a control plant
may be the product of the same hybrid cross of inbred parental
lines as the modified plant, except for the absence in the control
plant of any transgenic or genome edit(s) affecting one or more GA
oxidase or br2 genes. Similarly, an unmodified control plant refers
to a plant that shares a substantially similar or essentially
identical genetic background as a modified plant, but without the
one or more engineered changes to the genome (e.g., transgene,
mutation or edit) of the modified plant. For purposes of comparison
to a modified plant, plant seed, plant part, plant cell and/or
plant genome, a "wild-type plant" (or likewise a "wild-type" plant
seed, plant part, plant cell and/or plant genome) refers to a
non-transgenic and non-genome edited control plant, plant seed,
plant part, plant cell and/or plant genome. As used herein, a
"control" plant, plant seed, plant part, plant cell and/or plant
genome may also be a plant, plant seed, plant part, plant cell
and/or plant genome having a similar (but not the same or
identical) genetic background to a modified plant, plant seed,
plant part, plant cell and/or plant genome, if deemed sufficiently
similar for comparison of the characteristics or traits to be
analyzed.
[0138] In an aspect, this disclosure provides a dominant mutant
allele in a GA20 oxidase_3 gene. In an aspect, this disclosure
provides a dominant mutant allele in a GA20 oxidase_5 gene. In an
aspect, this disclosure provides a dominant mutant allele in a GA3
oxidase gene. In an aspect, this disclosure provides a dominant
mutant allele in a brachytic2 gene.
[0139] In an aspect, this disclosure provides a semi-dominant
mutant allele in a GA20 oxidase_3 gene. In an aspect, this
disclosure provides a semi-dominant mutant allele in a GA20
oxidase_5 gene. In an aspect, this disclosure provides a
semi-dominant mutant allele in a GA3 oxidase gene. In an aspect,
this disclosure provides a semi-dominant mutant allele in a
brachytic2 gene.
[0140] In an aspect, this disclosure provides a dominant negative
mutant allele in a GA20 oxidase_3 gene. In an aspect, this
disclosure provides a dominant negative mutant allele in a GA20
oxidase_5 gene. In an aspect, this disclosure provides a dominant
negative mutant allele in a GA3 oxidase gene. In an aspect, this
disclosure provides a dominant negative mutant allele in a
brachytic2 gene.
[0141] In an aspect, a female corn plant is homozygous (or
biallelic) for a mutant allele or transgene provided herein. In an
aspect, a female corn plant is heterozygous for a mutant allele
provided herein. In an aspect, a male corn plant is homozygous (or
biallelic) for a mutant allele or transgene provided herein. In an
aspect, a male corn plant is heterozygous for a mutant allele
provided herein. In an aspect, a corn plant is homozygous (or
biallelic) for a mutant allele or transgene provided herein. In an
aspect, a corn plant is heterozygous for a mutant allele provided
herein.
[0142] In an aspect, a dominant negative mutant allele generates an
antisense RNA transcript capable of triggering suppression of an
unmodified or wildtype allele of the gene. In an aspect, a dominant
negative mutant allele encodes a truncated protein as compared to
an unmodified allele of the gene. In an aspect, a dominant negative
mutant allele generates at least one RNA transcript capable of
forming a hairpin-loop secondary structure. In an aspect, the
coding sequence of a dominant negative mutant allele is operably
linked to a promoter of the native copy of the gene. In an aspect,
a dominant negative mutant allele comprises a heterologous
non-coding RNA target site in the endogenous locus of the gene. In
an aspect, a dominant negative mutant allele comprises an inverted
copy of the gene, or a portion thereof, adjacent to a wildtype copy
of the gene at the endogenous locus of the gene. As used herein, a
"portion" of a gene refers to at least 10, at least 20, at least
30, at least 40, at least 50, at least 100, at least 250, at least
500, at least 1000, or at least 2500 consecutive nucleotides of the
gene. As used herein, "adjacent" refers to a nucleic acid sequence
that is in close proximity, or next to another nucleic acid
sequence. In one aspect, adjacent nucleic acid sequences are
physically linked. In another aspect, adjacent nucleic acid
sequences or genes are immediately next to each other such that
there are no intervening nucleotides between the end of a first
nucleic acid sequence and the start of a second nucleic acid
sequence. In an aspect, a first gene and a second gene are adjacent
to each other if they are separated by less than 50,000, less than
25,000, less than 10,000, less than 9000, less than 8000, less than
7000, less than 6000, less than 5000, less than 4000, less than
3000, less than 2500, less than 2000, less than 1750, less than
1500, less than 1250, less than 1000, less than 900, less than 800,
less than 700, less than 600, less than 500, less than 400, less
than 300, less than 200, less than 100, less than 75, less than 50,
less than 25, less than 20, less than 10, less than 5, less than 4,
less than 3, less than 2, or less than 1 nucleotides.
[0143] In an aspect, a dominant negative mutant allele comprises a
deletion of a portion of a chromosome between a first region of the
gene and a second region of the gene, wherein an antisense RNA
transcript of the first region of the gene is generated following
the deletion of the portion of the chromosome. In an aspect, a
dominant negative mutant allele comprises a first promoter and a
second promoter separated by an intervening region, wherein the
first promoter and the second promoter are positioned in opposite
orientations, wherein the second promoter generates at least one
antisense RNA transcript, and wherein expression of the gene is
reduced as compared to a control corn plant that lacks the dominant
negative mutant allele. In an aspect, a dominant negative mutant
allele comprises a tissue-specific or tissue-preferred promoter
inserted into the gene in reverse orientation as compared to the
native promoter of the gene, wherein the tissue-specific or
tissue-preferred promoter generates at least one antisense RNA
transcript, and wherein expression of the gene is reduced as
compared to a control corn plant that lacks the dominant negative
mutant allele.
[0144] In an aspect, this disclosure provides corn plants
comprising a dominant or semi-dominant transgene or mutant allele
of a gene that causes a short stature phenotype. As used herein, a
"short stature phenotype" refers to a dwarf corn plant, a
semi-dwarf corn plant, or a brachytic corn plant. In an aspect, a
plant is homozygous (or biallelic) for a transgene. In an aspect, a
plant is heterozygous for a transgene. In an aspect, a plant is
hemizygous for a transgene. In an aspect, a corn plant is
homozygous (or biallelic) for a transgene provided herein. In an
aspect, a corn plant is heterozygous for a transgene provided
herein. In an aspect, a corn plant is hemizygous for a transgene
provided herein.
[0145] In an aspect, a transgene comprises a recombinant
polynucleotide encoding an RNA molecule that suppresses expression
of an endogenous GA20 oxidase gene. In an aspect, a transgene
comprises a recombinant polynucleotide encoding an RNA molecule
that suppresses expression of an endogenous GA20 oxidase_3 gene. In
an aspect, a transgene comprises a recombinant polynucleotide
encoding an RNA molecule that suppresses expression of an
endogenous GA20 oxidase_5 gene. In an aspect, a transgene comprises
a recombinant polynucleotide encoding an RNA molecule that
suppresses expression of an endogenous GA3 oxidase gene. In an
aspect, a transgene comprises a recombinant polynucleotide encoding
an RNA molecule that suppresses expression of an endogenous br2
gene. In an aspect, a recombinant polynucleotide is operably linked
to a promoter. In an aspect, a transgene comprises a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein.
[0146] As used herein, the term "plurality" in reference to an item
means two or more of such items. For example, a "plurality of
plants" means two or more plants.
[0147] In an aspect, corn plants disclosed herein are selected from
the subspecies Zea mays L. ssp. mays. In an additional aspect, corn
plants disclosed herein are selected from the group Zea mays L.
subsp. mays Indentata, otherwise known as dent corn. In another
aspect, corn plants disclosed herein are selected from the group
Zea mays L. subsp. mays Indurata, otherwise known as flint corn. In
an aspect, corn plants disclosed herein are selected from the group
Zea mays L. subsp. mays Saccharata, otherwise known as sweet corn.
In another aspect, corn plants disclosed herein are selected from
the group Zea mays L. subsp. mays Amylacea, otherwise known as
flour corn. In a further aspect, corn plants disclosed herein are
selected from the group Zea mays L. subsp. mays Everta, otherwise
known as popcorn. Plants disclosed herein also include hybrids,
inbreds, partial inbreds, or members of defined or undefined
populations.
[0148] Growers must balance crop prices, standability, plant
health, and kernel moisture content when determining when to
harvest a corn field. As provided herein, corn plants with better
standability, such as dwarf corn plants, semi-dwarf corn plants,
and brachytic corn plants, are resistant to lodging and thus can
remain in the field for a longer period of time prior to harvest
without significant loss of yield, or with improved yield relative
to taller corn plants (especially when compared to corn plants that
have lodged). According to aspects of the present disclosure, the
improved standability of short stature corn plants provides growers
and seed producers with more flexibility on when to harvest, allows
more time for drying down seed or grain prior to harvest, and/or
enables or improves direct harvest applications, particularly in
corn seed production operations. As used herein, "direct
harvesting" refers to the harvesting of crop seeds from plants with
a combine harvester in the field with little or no further drying
or other processing or desiccation steps prior to seed storage. As
used herein, "standability" refers to the ability of a plant or a
plurality, population or field of plants, such as a corn plant or a
plurality, population or field of corn plants, to stand upright in
a position that enables the plant(s) to be harvested by standard
farm equipment (e.g., a combine harvester). As used herein,
"lodging" can refer to either "stalk lodging" or "root lodging."
Stalk lodging occurs when the corn plant stalk is severely bent or
broken below the ear. Root lodging occurs when the corn plant is
leaning at an angle (e.g., greater than or equal to 45.degree.
relative to perpendicular from the ground, or at an angle less than
45.degree. relative to the ground). Lodged corn plants, whether
stalk lodged and/or root lodged, severely limit harvestability by
standard farm equipment (e.g., a combine harvester) resulting in up
to 100% yield loss of the lodged corn plants.
[0149] In an aspect, a modified corn plant provided herein has
improved lodging resistance relative to an unmodified control
plant.
[0150] Growers will leave harvestable corn standing in a field to
reduce the kernel moisture content of the grain. Optimal kernel
moisture content can vary by growing region and by individual
grower. Typically, kernel moisture content decreases the longer the
corn plants are left in the field (e.g., the longer the period of
time between fertilization or reaching maturity and harvest).
However, extending the period of time between fertilization (or
reaching maturity or some other developmental stage) and harvesting
can increase the chance that plants will lodge, which can result in
significant decreases in yield (even up to 100%). As provided
herein, by providing plants with reduced heights that have high
standability performance (i.e., resistance to lodging), growers are
enabled to allow for greater periods of time until harvest without
increasing (or significantly or substantially increasing) their
risk of yield loss due to lodging. Typical grain moisture contents
for harvesting corn are between 15% and 25%, although wider ranges
of 13-30% or higher are possible. According to present embodiments,
corn plants may be left in the field for a longer period of time
after reaching a given grain moisture content percentage.
[0151] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field at least 30 days, at least
40 days, at least 50 days, at least 60 days, at least 70 days, at
least 80 days, at least 90 days, at least 100 days, or at least 110
days after at least 50%, at least 60%, at least 70%, at least 80%,
at least 90% or 100% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest. In another aspect, a method comprises
harvesting a plurality of corn plants from a field at least 30
days, at least 40 days, at least 50 days, at least 60 days, at
least 70 days, at least 80 days, at least 90 days, at least 100
days, or at least 110 days after at least 50%, at least 60%, at
least 70%, at least 80%, at least 90% or 100% of said corn plants
have reached R3 stage, wherein average kernel moisture content is
less than or equal to 30% or the kernel moisture content of a corn
plant of the plurality of corn plants is less than or equal to 30%,
and wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest. In another aspect, a method
comprising harvesting a plurality of corn plants from a field at
least 30 days, at least 40 days, at least 50 days, at least 60
days, at least 70 days, at least 80 days, at least 90 days, at
least 100 days, or at least 110 days after at least 50%, at least
60%, at least 70%, at least 80%, at least 90% or 100% of said corn
plants have reached R3 stage, wherein the average yield of said
field is at least 170 bushels per acre, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest.
[0152] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field at least 30 days, at least
40 days, at least 50 days, at least 60 days, at least 70 days, at
least 80 days at least 90 days, at least 100 days, or at least 110
days after fertilization or silking of said plurality of corn
plants, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest. In another aspect, a method provided
herein comprises harvesting a plurality of corn plants from a field
at least 30 days, at least 40 days, at least 50 days, at least 60
days, at least 70 days, at least 80 days at least 90 days, at least
100 days, or at least 110 days after fertilization or silking of
said plurality of corn plants, wherein the average kernel moisture
content is less than or equal to 30% or the kernel moisture content
of a corn plant of the plurality of corn plants is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest. In another aspect, a
method provided herein comprises harvesting a plurality of corn
plants from a field at least 30 days, at least 40 days, at least 50
days, at least 60 days, at least 70 days, at least 80 days at least
90 days, at least 100 days, or at least 110 days after at
fertilization or silking of said plurality of corn plants, wherein
the average yield of said field is at least 170 bushels per acre,
and wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest.
[0153] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field when, or at least 1 day
after, the average kernel moisture content of said plurality of
corn plants is between 10% and 30%, or less than or equal to 30%,
or the kernel moisture content of a corn plant of the plurality of
corn plants is between 10% and 30%, or less than or equal to 30%,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest. In an aspect, methods provided herein
comprise harvesting a plurality of corn plants from a field when,
or at least 1 day after, the average kernel moisture content of
said plurality of corn plants is between 15% and 25%, or less than
or equal to 25%, or less than or equal to 20%, or less than or
equal to 15%, or the kernel moisture content of a corn plant of the
plurality of corn plants is between 15% and 25%, or less than or
equal to 25%, or less than or equal to 20%, or less than or equal
to 15%, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest. In each of these aspects, the
average yield of said plants in a field may be at least 170 bushels
per acre.
[0154] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field at least 30 days, at least
40 days, at least 50 days, at least 60 days, at least 70 days, at
least 80 days, at least 90 days, at least 100 days, or at least 110
days after fertilization or silking of at least 50%, at least 60%,
at least 70%, at least 80%, at least 85%, at least 90%, or 100% of
said plurality of corn plants, wherein fewer than or equal to 50%
of said corn plants have lodged at the time of harvest. In another
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a field at least 30 days, at least 40 days, at
least 50 days, at least 60 days, at least 70 days, at least 80
days, at least 90 days, at least 100 days, or at least 110 days
after fertilization or silking of at least 50%, at least 60%, at
least 70%, at least 80%, at least 85%, at least 90%, or 100% of
said plurality of corn plants, wherein the average kernel moisture
content is less than or equal to 30% or the kernel moisture content
of a corn plant of the plurality of corn plants is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest. In another aspect, a
method provided herein comprises harvesting a plurality of corn
plants from a field at least 30 days, at least 40 days, at least 50
days, at least 60 days, at least 70 days, at least 80 days, at
least 90 days, at least 100 days, or at least 110 days after at
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 85%, at least 90%, or 100% of said
plurality of corn plants, wherein the average yield of said field
is at least 170 bushels per acre, and wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest.
[0155] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field when, or at least 1 day
after, the average kernel moisture content of at least 50%, at
least 60%, at least 70%, at least 80%, at least 85%, at least 90%,
or 100% of said plurality of corn plants is between 10% and 30%, or
less than or equal to 30%, or the kernel moisture content of a corn
plant of the plurality of corn plants is between 10% and 30%, or
less than or equal to 30%, wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest. In an aspect,
methods provided herein comprise harvesting a plurality of corn
plants from a field when, or at least 1 day after, the average
kernel moisture content of at least 50%, at least 60%, at least
70%, at least 80%, at least 85%, at least 90%, or 100% of said
plurality of corn plants is between 15% and 25%, or less than or
equal to 25%, or less than or equal to 20%, or less than or equal
to 15%, or the kernel moisture content of a corn plant of the
plurality of corn plants is between 15% and 25%, or less than or
equal to 25%, or less than or equal to 20%, or less than or equal
to 15%, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest. In each of these aspects, the
average yield of said plants in a field may be at least 170 bushels
per acre.
[0156] In an aspect, a method provided herein further comprises
growing a plurality of corn plants in a corn field prior to
harvesting the plurality of corn plants.
[0157] In an aspect, corn plants provided herein are inbred corn
plants. As used herein, the term "inbred" means a line that has
been bred for genetic homogeneity. In another aspect, corn plants
provided herein are hybrid corn plants. As used herein, the term
"hybrid" means a progeny of mating between at least two genetically
dissimilar parents or inbreds. In an aspect, corn plants provided
herein are transgenic, mutant and/or edited corn plants.
[0158] In an aspect, at least 10% of the corn plants in a field are
inbred corn plants. In an aspect, at least 20% of the corn plants
in a field are inbred corn plants. In an aspect, at least 30% of
the corn plants in a field are inbred corn plants. In an aspect, at
least 40% of the corn plants in a field are inbred corn plants. In
an aspect, at least 50% of the corn plants in a field are inbred
corn plants. In an aspect, at least 60% of the corn plants in a
field are inbred corn plants. In an aspect, at least 70% of the
corn plants in a field are inbred corn plants. In an aspect, at
least 80% of the corn plants in a field are inbred corn plants. In
an aspect, at least 90% of the corn plants in a field are inbred
corn plants. In an aspect, 100% of the corn plants in a field are
inbred corn plants.
[0159] In an aspect, between 1% and 100% of the corn plants in a
field are inbred corn plants. In an aspect, between 10% and 100% of
the corn plants in a field are inbred corn plants. In an aspect,
between 20% and 100% of the corn plants in a field are inbred corn
plants. In an aspect, between 30% and 100% of the corn plants in a
field are inbred corn plants. In an aspect, between 40% and 100% of
the corn plants in a field are inbred corn plants. In an aspect,
between 50% and 100% of the corn plants in a field are inbred corn
plants. In an aspect, between 60% and 100% of the corn plants in a
field are inbred corn plants. In an aspect, between 70% and 100% of
the corn plants in a field are inbred corn plants. In an aspect,
between 80% and 100% of the corn plants in a field are inbred corn
plants. In an aspect, between 90% and 100% of the corn plants in a
field are inbred corn plants.
[0160] In an aspect, at least 10% of the corn plants in a field are
hybrid corn plants. In an aspect, at least 20% of the corn plants
in a field are hybrid corn plants. In an aspect, at least 30% of
the corn plants in a field are hybrid corn plants. In an aspect, at
least 40% of the corn plants in a field are hybrid corn plants. In
an aspect, at least 50% of the corn plants in a field are hybrid
corn plants. In an aspect, at least 60% of the corn plants in a
field are hybrid corn plants. In an aspect, at least 70% of the
corn plants in a field are hybrid corn plants. In an aspect, at
least 80% of the corn plants in a field are hybrid corn plants. In
an aspect, at least 90% of the corn plants in a field are hybrid
corn plants. In an aspect, 100% of the corn plants in a field are
hybrid corn plants.
[0161] In an aspect, between 1% and 100% of the corn plants in a
field are hybrid corn plants. In an aspect, between 10% and 100% of
the corn plants in a field are hybrid corn plants. In an aspect,
between 20% and 100% of the corn plants in a field are hybrid corn
plants. In an aspect, between 30% and 100% of the corn plants in a
field are hybrid corn plants. In an aspect, between 40% and 100% of
the corn plants in a field are hybrid corn plants. In an aspect,
between 50% and 100% of the corn plants in a field are hybrid corn
plants. In an aspect, between 60% and 100% of the corn plants in a
field are hybrid corn plants. In an aspect, between 70% and 100% of
the corn plants in a field are hybrid corn plants. In an aspect,
between 80% and 100% of the corn plants in a field are hybrid corn
plants. In an aspect, between 90% and 100% of the corn plants in a
field are hybrid corn plants.
[0162] In another aspect, a corn plant provided herein is a
semi-dwarf corn plant. As used herein, a "semi-dwarf plant" refers
to a plant having a stature or height that is reduced relative to a
control wild-type plant by at least 5%, at least 10%, at least 15%,
at least 20%, at least 25%, at least 30%, at least 35%, at least
40%, at least 45%, or at least 50%. Such a semi-dwarf plant can be
characterized by a reduced stem, stalk, or trunk length when
compared to the control wild-type plant under comparable growth
conditions, which can result from fewer internodes or shorter
average internode length. As used herein, an "internode" refers to
the region between two nodes on a corn stalk, and a "node" refers
to the point on the corn stalk (e.g., stem) where leaves and/or
ears originate.
[0163] In an aspect, at least 10% of the corn plants in a field are
semi-dwarf corn plants. In an aspect, at least 20% of the corn
plants in a field are semi-dwarf corn plants. In an aspect, at
least 30% of the corn plants in a field are semi-dwarf corn plants.
In an aspect, at least 40% of the corn plants in a field are
semi-dwarf corn plants. In an aspect, at least 50% of the corn
plants in a field are semi-dwarf corn plants. In an aspect, at
least 60% of the corn plants in a field are semi-dwarf corn plants.
In an aspect, at least 70% of the corn plants in a field are
semi-dwarf corn plants. In an aspect, at least 80% of the corn
plants in a field are semi-dwarf corn plants. In an aspect, at
least 90% of the corn plants in a field are semi-dwarf corn plants.
In an aspect, 100% of the corn plants in a field are semi-dwarf
corn plants.
[0164] In an aspect, between 1% and 100% of the corn plants in a
field are semi-dwarf corn plants. In an aspect, between 10% and
100% of the corn plants in a field are semi-dwarf corn plants. In
an aspect, between 20% and 100% of the corn plants in a field are
semi-dwarf corn plants. In an aspect, between 30% and 100% of the
corn plants in a field are semi-dwarf corn plants. In an aspect,
between 40% and 100% of the corn plants in a field are semi-dwarf
corn plants. In an aspect, between 50% and 100% of the corn plants
in a field are semi-dwarf corn plants. In an aspect, between 60%
and 100% of the corn plants in a field are semi-dwarf corn plants.
In an aspect, between 70% and 100% of the corn plants in a field
are semi-dwarf corn plants. In an aspect, between 80% and 100% of
the corn plants in a field are semi-dwarf corn plants. In an
aspect, between 90% and 100% of the corn plants in a field are
semi-dwarf corn plants.
[0165] In an aspect, a corn plant provided herein is a dwarf corn
plant. As used herein, a "dwarf" plant refers to an atypically
small plant. Generally, such a "dwarf plant" has a stature or
height that is reduced from that of a control wild-type plant
(e.g., a sibling plant comprising all other traits except the dwarf
trait) by at least 30%, at least 35%, at least 40%, at least 45%,
at least 50%, at least 55%, at least 60%, or at least 75%.
[0166] In an aspect, at least 10% of the corn plants in a field are
dwarf corn plants. In an aspect, at least 20% of the corn plants in
a field are dwarf corn plants. In an aspect, at least 30% of the
corn plants in a field are dwarf corn plants. In an aspect, at
least 40% of the corn plants in a field are dwarf corn plants. In
an aspect, at least 50% of the corn plants in a field are dwarf
corn plants. In an aspect, at least 60% of the corn plants in a
field are dwarf corn plants. In an aspect, at least 70% of the corn
plants in a field are dwarf corn plants. In an aspect, at least 80%
of the corn plants in a field are dwarf corn plants. In an aspect,
at least 90% of the corn plants in a field are dwarf corn plants.
In an aspect, 100% of the corn plants in a field are dwarf corn
plants.
[0167] In an aspect, between 1% and 100% of the corn plants in a
field are dwarf corn plants. In an aspect, between 10% and 100% of
the corn plants in a field are dwarf corn plants. In an aspect,
between 20% and 100% of the corn plants in a field are dwarf corn
plants. In an aspect, between 30% and 100% of the corn plants in a
field are dwarf corn plants. In an aspect, between 40% and 100% of
the corn plants in a field are dwarf corn plants. In an aspect,
between 50% and 100% of the corn plants in a field are dwarf corn
plants. In an aspect, between 60% and 100% of the corn plants in a
field are dwarf corn plants. In an aspect, between 70% and 100% of
the corn plants in a field are dwarf corn plants. In an aspect,
between 80% and 100% of the corn plants in a field are dwarf corn
plants. In an aspect, between 90% and 100% of the corn plants in a
field are dwarf corn plants.
[0168] As used herein, the term "polynucleotide" refers to a
nucleic acid molecule containing multiple nucleotides and generally
comprises at least 2, at least 5, at least 10, at least 20, at
least 30, at least 40, at least 50, at least 100, at least 250, at
least 500, at least 1,000, at least 1,500, at least 2,000, at least
2,500, at least 3,000, at least 5,000, or at least 10,000
nucleotide bases. As an example, a polynucleotide provided herein
can be a plasmid. The use of the terms "polynucleotide" or "nucleic
acid molecule" is not intended to limit the present disclosure to
polynucleotides comprising deoxyribonucleic acid (DNA). For
example, ribonucleic acid (RNA) molecules are also envisioned.
Those of ordinary skill in the art will recognize that
polynucleotides and nucleic acid molecules can comprise
ribonucleotides and combinations of ribonucleotides and
deoxyribonucleotides. Such deoxyribonucleotides and ribonucleotides
include both naturally occurring molecules and synthetic analogues.
The polynucleotides of the present disclosure also encompass all
forms of sequences including, but not limited to, single-stranded
forms, double-stranded forms, hairpins, stem-and-loop structures,
and the like. In an aspect, a nucleic acid molecule provided herein
is a DNA molecule. In another aspect, a nucleic acid molecule
provided herein is an RNA molecule. In an aspect, a nucleic acid
molecule provided herein is single-stranded. In another aspect, a
nucleic acid molecule provided herein is double-stranded. In an
aspect, a polynucleotide provided herein is single-stranded. In
another aspect, a polynucleotide provided herein is
double-stranded.
[0169] A non-coding RNA molecule can act as a suppression element
that targets one or more gene(s) in a plant cell, such as one or
more endogenous br2, GA20 or GA3 oxidase gene(s), or as a RNA
molecule, such as a guide RNA, etc., that guides a
sequence-specific nuclease to cut and trigger a genome editing
event at a target site in the genome. Non-limiting examples of
non-coding RNA molecules include a microRNA (miRNA), a miRNA
precursor (pre-miRNA), a small interfering RNA (siRNA), a small RNA
(18-26 nt in length) and precursor encoding same, a heterochromatic
siRNA (hc-siRNA), a Piwi-interacting RNA (piRNA), a hairpin double
strand RNA (hairpin dsRNA), a trans-acting siRNA (ta-siRNA), a
naturally occurring antisense siRNA (nat-siRNA), a CRISPR RNA
(crRNA), a tracer RNA (tracrRNA), a guide RNA (gRNA), and a
single-guide RNA (sgRNA). In an aspect, a non-coding RNA provided
herein is selected from the group consisting of a microRNA, a small
interfering RNA, a secondary small interfering RNA, a transfer RNA,
a ribosomal RNA, a trans-acting small interfering RNA, a naturally
occurring antisense small interfering RNA, a heterochromatic small
interfering RNA, and precursors thereof. In another aspect, a
non-coding RNA provided herein is selected from the group
consisting of a miRNA, a pre-miRNA, a siRNA, a hc-siRNA, a piRNA, a
hairpin dsRNA, a ta-siRNA, a nat-siRNA, a crRNA, a tracrRNA, a
gRNA, and a sgRNA. In another aspect, a non-coding RNA provided
herein is a miRNA. In another aspect, a non-coding RNA provided
herein is a siRNA.
[0170] As used herein, a "stem-loop structure" refers to a
secondary structure in a RNA molecule having a double stranded
region (e.g., stem) made up by two annealing RNA strands, sequences
or segments of the RNA molecule, connected by a single stranded
intervening RNA sequence of the RNA molecule (e.g., a loop or
hairpin). A "stem-loop structure" of a RNA molecule can have a more
complicated secondary RNA structure, for example, comprising
self-annealing double stranded RNA sequences having internal
mismatches, bulges and/or loops.
[0171] As used herein, a "native sequence" refers to a nucleic acid
sequence naturally present in its original chromosomal
location.
[0172] As used herein, a "wild-type gene" or "wild-type allele"
refers to a gene or allele having a sequence or genotype that is
most common in a particular plant species or another sequence or
genotype having only natural variations, polymorphisms, or other
silent mutations relative to the most common sequence or genotype
that do not significantly impact the expression and activity of the
gene or allele. Indeed, a "wild-type" gene or allele contains no
variation, polymorphism, or any other type of mutation that
substantially affects the normal function, activity, expression, or
phenotypic consequence of the gene or allele relative to the most
common sequence or genotype.
[0173] The terms "percent identity" or "percent identical" as used
herein in reference to two or more nucleotide or protein sequences
is calculated by (i) comparing two optimally aligned sequences
(nucleotide or protein) over a window of comparison, (ii)
determining the number of positions at which the identical nucleic
acid base (for nucleotide sequences) or amino acid residue (for
proteins) occurs in both sequences to yield the number of matched
positions, (iii) dividing the number of matched positions by the
total number of positions in the window of comparison, and then
(iv) multiplying this quotient by 100% to yield the percent
identity. For purposes of calculating "percent identity" between
DNA and RNA sequences, a uracil (U) of a RNA sequence is considered
identical to a thymine (T) of a DNA sequence. If the window of
comparison is defined as a region of alignment between two or more
sequences (i.e., excluding nucleotides at the 5' and 3' ends of
aligned polynucleotide sequences, or amino acids at the N-terminus
and C-terminus of aligned protein sequences, that are not identical
between the compared sequences), then the "percent identity" may
also be referred to as a "percent alignment identity". If the
"percent identity" is being calculated in relation to a reference
sequence without a particular comparison window being specified,
then the percent identity is determined by dividing the number of
matched positions over the region of alignment by the total length
of the reference sequence. Accordingly, for purposes of the present
disclosure, when two sequences (query and subject) are optimally
aligned (with allowance for gaps in their alignment), the "percent
identity" for the query sequence is equal to the number of
identical positions between the two sequences divided by the total
number of positions in the query sequence over its length (or a
comparison window), which is then multiplied by 100%.
[0174] For optimal alignment of sequences to calculate their
percent identity, various pair-wise or multiple sequence alignment
algorithms and programs are known in the art, such as ClustalW, or
Basic Local Alignment Search Tool.RTM. (BLAST.RTM.), etc., that may
be used to compare the sequence identity or similarity between two
or more nucleotide or protein sequences. Although other alignment
and comparison methods are known in the art, the alignment between
two sequences (including the percent identity ranges described
above) may be as determined by the ClustalW or BLAST.RTM.
algorithm, see, e.g., Chenna R. et al., "Multiple sequence
alignment with the Clustal series of programs," Nucleic Acids
Research 31: 3497-3500 (2003); Thompson J D et al., "Clustal W:
Improving the sensitivity of progressive multiple sequence
alignment through sequence weighting, position-specific gap
penalties and weight matrix choice," Nucleic Acids Research 22:
4673-4680 (1994); and Larkin M A et al., "Clustal W and Clustal X
version 2.0," Bioinformatics 23: 2947-48 (2007); and Altschul, S.
F., Gish, W., Miller, W., Myers, E. W. & Lipman, D. J. (1990)
"Basic local alignment search tool." J. Mol. Biol. 215:403-410
(1990), the entire contents and disclosures of which are
incorporated herein by reference.
[0175] The terms "percent complementarity" or "percent
complementary", as used herein in reference to two nucleotide
sequences, is similar to the concept of percent identity but refers
to the percentage of nucleotides of a query sequence that optimally
base-pair or hybridize to nucleotides of a subject sequence when
the query and subject sequences are linearly arranged and optimally
base paired without secondary folding structures, such as loops,
stems or hairpins. Such a percent complementarity may be between
two DNA strands, two RNA strands, or a DNA strand and a RNA strand.
The "percent complementarity" is calculated by (i) optimally
base-pairing or hybridizing the two nucleotide sequences in a
linear and fully extended arrangement (i.e., without folding or
secondary structures) over a window of comparison, (ii) determining
the number of positions that base-pair between the two sequences
over the window of comparison to yield the number of complementary
positions, (iii) dividing the number of complementary positions by
the total number of positions in the window of comparison, and (iv)
multiplying this quotient by 100% to yield the percent
complementarity of the two sequences. Optimal base pairing of two
sequences may be determined based on the known pairings of
nucleotide bases, such as G-C, A-T, and A-U, through hydrogen
bonding. If the "percent complementarity" is being calculated in
relation to a reference sequence without specifying a particular
comparison window, then the percent identity is determined by
dividing the number of complementary positions between the two
linear sequences by the total length of the reference sequence.
Thus, for purposes of the present disclosure, when two sequences
(query and subject) are optimally base-paired (with allowance for
mismatches or non-base-paired nucleotides but without folding or
secondary structures), the "percent complementarity" for the query
sequence is equal to the number of base-paired positions between
the two sequences divided by the total number of positions in the
query sequence over its length (or by the number of positions in
the query sequence over a comparison window), which is then
multiplied by 100%.
[0176] As used herein, with respective to a given sequence, a
"complement", a "complementary sequence" and a "reverse complement"
are used interchangeably. All three terms refer to the inversely
complementary sequence of a nucleotide sequence, i.e. to a sequence
complementary to a given sequence in reverse order of the
nucleotides. As an example, the reverse complement of a nucleotide
sequence having the sequence 5'-atggttc-3' is 5'-gaaccat-3'.
[0177] As used herein, the term "antisense" refers to DNA or RNA
sequences that are complementary to a specific DNA or RNA sequence.
Antisense RNA molecules are single-stranded nucleic acids which can
combine with a sense RNA strand or sequence or mRNA to form
duplexes due to complementarity of the sequences. The term
"antisense strand" refers to a nucleic acid strand that is
complementary to the "sense" strand. The "sense strand" of a gene
or locus is the strand of DNA or RNA that has the same sequence as
a RNA molecule transcribed from the gene or locus (with the
exception of Uracil in RNA and Thymine in DNA).
[0178] As used herein, an "inverted genomic fragment" refers to a
genomic segment that is inverted in the genome such that the
original sense strand and anti sense strand sequences are reversed
or switched in the opposite orientation for the entire genomic
segment.
[0179] As used herein, in the context of a "corresponding
endogenous sequence" or a "corresponding endogenous DNA segment,"
an endogenous sequence or endogenous DNA segment is considered to
correspond to another sequence or DNA segment (e.g., an
non-endogenous, introduced or inserted sequence or DNA segment)
when the sequences or DNA segments share sufficient sequence
homology, identity, or complementarity.
[0180] The term "operably linked" refers to a functional linkage
between a promoter or other regulatory element and an associated
transcribable DNA sequence or coding sequence of a gene (or
transgene), such that the promoter, etc., operates or functions to
initiate, assist, affect, cause, and/or promote the transcription
and expression of the associated transcribable DNA sequence or
coding sequence, at least in certain cell(s), tissue(s),
developmental stage(s), and/or condition(s). Two transcribable DNA
sequences can also be "operably linked" to each other if their
transcription is subject to the control of a common promoter or
other regulatory element. In an aspect, a promoter is selected from
the group consisting of a constitutive promoter, an inducible
promoter, a tissue-specific promoter, and a tissue-preferred
promoter. In an aspect, a promoter is a native promoter.
[0181] As used herein, an "encoding region" or "coding region"
refers to a portion of a polynucleotide that encodes a functional
unit or molecule (e.g., without being limiting, a mRNA, protein, or
non-coding RNA sequence or molecule). An "encoding region" or
"coding region" can contain, for example, one or more exons, one or
more introns, a 5'-UTR, a 3'-UTR, or any combination thereof.
[0182] As used herein, an "intervening region" or "intervening
sequence" refers to a polynucleotide sequence between a physically
linked first polynucleotide sequence and second polynucleotide
sequence. The intervening sequence may form a loop, and the first
and second sequences may hybridize to form a stem, of a stem-loop
structure. In one aspect, an intervening region or intervening
sequence comprises at least 1, at least 2, at least 3, at least 4,
at least 5, at least 10, at least 25, at least 50, at least 100, at
least 150, at least 200, at least 250, at least 500, at least 1000,
at least 1250, at least 1500, at least 1750, at least 2000, at
least 2500, at least 3000, at least 4000, at least 5000, at least
6000, at least 7000, at least 8000, at least 9000, at least 10,000,
at least 15,000, at least 20,000, at least 25,000, or at least
50,000 nucleotides. In one aspect, an intervening region or
intervening sequence comprises a DNA sequence. In one aspect, an
intervening region or intervening sequence comprises an RNA
sequence. In one aspect, an intervening region or intervening
sequences comprises an endogenous or native nucleic acid sequence.
In another aspect, an intervening region or intervening sequences
comprises a transgenic or exogenous nucleic acid sequence. In one
aspect, an intervening region or intervening sequences comprises an
endogenous or native nucleic acid sequence and a transgenic or
exogenous nucleic acid sequence.
[0183] The term "recombinant" in reference to a polynucleotide (DNA
or RNA) molecule, protein, construct, vector, etc., refers to a
polynucleotide or protein molecule or sequence that is man-made and
not normally found in nature, and/or is present in a context in
which it is not normally found in nature, including a
polynucleotide (DNA or RNA) molecule, protein, construct, etc.,
comprising a combination of two or more polynucleotide or protein
sequences that would not naturally occur together in the same
manner without human intervention, such as a polynucleotide
molecule, protein, construct, etc., comprising at least two
polynucleotide or protein sequences that are operably linked but
heterologous with respect to each other. For example, the term
"recombinant" can refer to any combination of two or more DNA or
protein sequences in the same molecule (e.g., a plasmid, construct,
vector, chromosome, protein, etc.) where such a combination is
man-made and not normally found in nature. As used in this
definition, the phrase "not normally found in nature" means not
found in nature without human introduction. A recombinant
polynucleotide or protein molecule, construct, etc., can comprise
polynucleotide or protein sequence(s) that is/are (i) separated
from other polynucleotide or protein sequence(s) that exist in
proximity to each other in nature, and/or (ii) adjacent to (or
contiguous with) other polynucleotide or protein sequence(s) that
are not naturally in proximity with each other. Such a recombinant
polynucleotide molecule, protein, construct, etc., can also refer
to a polynucleotide or protein molecule or sequence that has been
genetically engineered and/or constructed outside of a cell. For
example, a recombinant DNA molecule can comprise any engineered or
man-made plasmid, vector, etc., and can include a linear or
circular DNA molecule. Such plasmids, vectors, etc., can contain
various maintenance elements including a prokaryotic origin of
replication and selectable marker, as well as one or more
transgenes or expression cassettes perhaps in addition to a plant
selectable marker gene, etc.
[0184] As used herein, the term "transgene" refers to a recombinant
DNA molecule, construct, or sequence comprising a gene and/or
transcribable DNA sequence and integrated or inserted into a plant
genome.
[0185] As used herein, a "transgenic plant" refers to a plant whose
genome has been altered by the integration or insertion of a
recombinant DNA molecule, construct, cassette or sequence for
expression of a non-coding RNA molecule, mRNA and/or protein in the
plant. A transgenic plant includes an R.sub.0 plant developed or
regenerated from an originally transformed plant cell(s) as well as
progeny transgenic plants in later generations or crosses from the
R.sub.0 transgenic plant that comprise the recombinant DNA
molecule, construct, cassette or sequence. A plant having an
integrated or inserted recombinant DNA molecule, construct,
cassette or sequence is considered a transgenic plant even if the
plant also has other mutation(s) or edit(s) that would not
themselves be considered transgenic.
[0186] As used herein, the term "heterologous" can refer broadly to
a combination of two or more DNA molecules or sequences, such as a
promoter and an associated transcribable DNA sequence, coding
sequence, or gene, when such a combination is man-made and not
normally found in nature. The term "heterologous" in reference to a
promoter or other regulatory sequence in relation to an associated
polynucleotide sequence (e.g., a transcribable DNA sequence or
coding sequence or gene) is a promoter or regulatory sequence that
is not operably linked to such associated polynucleotide sequence
in nature--e.g., the promoter or regulatory sequence has a
different origin relative to the associated polynucleotide sequence
and/or the promoter or regulatory sequence is not naturally
occurring in a plant species to be transformed with the promoter or
regulatory sequence. For example, a transcribable DNA sequence
encoding a non-coding RNA molecule that targets one or more GA
oxidase gene(s) for suppression can be operably linked to a
heterologous plant-expressible promoter.
[0187] As used herein, the term "expression" refers to the process
for converting the genetic information of a gene into a functional
unit (without being limiting, for example, a mRNA and/or protein or
a non-coding RNA molecule).
[0188] As used herein, the terms "suppress," "suppression,"
"inhibit," "inhibition," "inhibiting," and "downregulation" refer
to a lowering, reduction or elimination of the expression level of
a mRNA and/or protein encoded by a gene in a plant, plant cell, or
plant tissue at one or more stage(s) of plant development, as
compared to the expression level of such mRNA and/or protein in a
wild-type or control plant, cell, or tissue at the same stage(s) of
plant development. In an aspect, a polynucleotide provided herein
can suppress the expression of a complementary target gene. In
another aspect, a non-coding RNA molecule can suppress the
expression of a complementary target gene.
[0189] As used herein, a "mutation" refers to an insertion,
deletion, substitution, duplication, or inversion of one or more
nucleotides and/or encoded amino acids as compared to a reference
or wild-type nucleotide and/or amino acid sequence, which can be
introduced by any suitable mutagenesis or gene editing
technique.
[0190] There are different ways in which a corn plant can be made
to have a shorter semi-dwarf plant height. According to many
aspects, a corn plant can be made shorter or semi-dwarf relative to
a control plant by lowering the level(s) of active GAs in one or
more tissue(s) of the plant, such as by suppressing, mutating or
editing a GA oxidase gene in the corn plant. In an aspect, a corn
plant provided herein comprises a recombinant polynucleotide
capable of suppressing expression of one or more GA20 oxidase
and/or GA3 oxidase gene(s) and/or mRNA(s) transcribed therefrom.
Alternatively, a corn plant provided herein comprises one or more
mutation(s) or edit(s) in one or more GA20 oxidase and/or GA3
oxidase gene(s). According to other aspects, corn plants can have a
mutation or edit in an auxin, brassinosteroid, jasmonic acid, cell
cycle regulation, and/or other pathway gene(s) that are shown to
affect plant height. According to yet further embodiments, corn
plants can be made shorter by application of one or more
chemistries shown to affect plant height. According to another
aspect, a corn plant or plurality of corn plants provided herein
can comprise a mutation or edit in one or more loci or genes, or a
transgene targeting such one or more loci or genes, that have been
associated with a short stature phenotype in corn, such as one or
more of the following: anther ear 1 (An1), brachytic 1 (Br1),
brevis plant 1 (Bv1) or brachytic 3 (br3), crinkly 4 (Cr4), compact
plant 2 (Ct2), dwarf plant 1 (d1), dwarf plant 8 (d8), dwarf plant
9 (d9), nana plant 1 (Na1), nana plant 2 (Na2), non-chromosomal
stripe 3 (Nsc3), narrow leaf dwarf/(Nld1), reduced plant 1 (Rd1),
semi-dwarf/(Sdw1), semi-dwarf 2 (Sdw2), tangled 1 (Tan1), terminal
ear 1 (Te1), and vanishing tassel 2 (Vt2). As used herein, a
"mutation" includes an edit--i.e., a mutation introduced via a
genome editing technique.
[0191] In an aspect, a corn plant(s) is homozygous for one or more
mutation(s) and/or edit(s) in one of the foregoing native corn
genes. In an aspect, a corn plant is biallelic for a first mutation
and/or edit and a second mutation and/or edit in one of the
foregoing native corn genes.
[0192] As used herein, a "brachytic plant" refers to a plant having
a mutated, edited or suppressed brachytic gene and a short
semi-dwarf height and stature relative to a control plant (e.g., a
wild-type sibling plant comprising all other traits except the
brachytic trait) due to a shortening of the average internode
length. Such a brachytic mutant plant can have a short semi-dwarf
height and stature due to a shortening of the average internode
length. As used herein, a "brachytic gene", "Br gene" or "br gene",
or "Br gene" refers to any brachytic gene in a corn plant that when
suppressed, mutated or edited to reduce its expression or function
can result in a shorter, semi-dwarf corn plant and phenotype.
[0193] Certain mutations of brachytic genes have been shown to
result in a short stature, semi-dwarf phenotype. In an aspect of
the present disclosure, a corn plant is provided having a
non-silent mutation or edit in a brachytic gene. See, e.g., PCT
Application No. PCT/US2016/029492 and PCT/US2017/067888, the entire
contents and disclosures of which are incorporated herein by
reference. Thus, a shorter corn plant can comprise a mutation (or
edit) in a brachytic gene, and can be homozygous (or biallelic) for
a mutation (or edit) in a brachytic gene. As used herein, a
"brachytic mutant plant" refers to a plant having a short
semi-dwarf height and stature relative to a control plant (e.g., a
wild-type sibling plant comprising all other traits except the
brachytic trait) due to a shortening of the average internode
length. Such a brachytic mutant plant can have a short semi-dwarf
height and stature due to a shortening of the average internode
length. As used herein, a "brachytic gene", "BR gene" or "br gene",
or "Br gene" refers to any brachytic gene in a corn plant that when
mutated or edited to reduce its expression or function can result
in a shorter, semi-dwarf corn plant and phenotype. In an aspect, an
inbred corn plant or plurality of inbred corn plants provided
herein each has a non-silent mutation or edit in a brachytic gene.
In an aspect, the brachytic gene is a br1 mutant gene. In another
aspect, the brachytic gene is a br2 mutant gene. In yet another
aspect, the brachytic gene is a br3 mutant gene.
[0194] In maize, brachytic mutants have a short stature due to a
shortening of the internode length without a corresponding
reduction in the number of internodes or the number and size of
other organs, including the leaves, ear and tassel. See Kempton J.
Hered. 11:111-115(1920); Pilu et al., Molecular Breeding,
20:83-91(2007). Three brachytic mutants have been isolated in maize
to date: brachytic1 (br1), brachytic2 (br2) and brachytic3 (br3).
Both br1 and br3 mutations cause a reduction in corn plant height,
which has been thought too severe for commercial exploitation due
to potential impacts on yield. In contrast, the br2 mutant has
particular agronomic potential because of shortening of the
internodes of the lower stalk without an obvious reduction in other
plant organs. In addition, br2 lines exhibit an unusual stalk
strength and tolerance to wind lodging, while the leaves are often
darker and persist longer in the active green than those of the
wild-type plants. The br2 phenotype is insensitive to treatment
with gibberellins, auxins, brassinosteroids and cytokinins,
suggesting that the biosynthesis of these hormones is not modified
by the br2 mutation. Multani et al. identified the genomic sequence
of the br2 gene (SEQ ID NO: 58) and deposited it under GenBank
Accession No. AY366085. See Multani et al., Science, 302(5642)81-84
(2003). br2 was annotated to encode a putative protein similar to
adenosine triphosphate (ATP)-binding cassette transporters of the
multidrug resistant (MDR) class of P-glycoproteins (PGPs). Pilu et
al. reported a br2-23 allele having an 8-bp deletion in the 3' end
of the br2 gene and claimed a direct relationship between this
deletion and the brachytic phenotype in their br2-23 plants. See
Pilu et al., Molecular Breeding, 20:83-91(2007). Nevertheless, the
use of brachytic mutations in corn has not been exploited
commercially partly because of the severity of the available
brachytic mutant alleles.
[0195] A wild-type genomic DNA sequence of the br2 locus from a
reference genome is provided in SEQ ID NO: 132. A wild-type cDNA
sequence of the br2 locus from a reference genome is provided in
SEQ ID NO: 180. A wild-type amino acid sequence encoded by SEQ ID
NO: 180 is provided in SEQ ID NO: 181.
[0196] For the br2 gene, SEQ ID NO: 132 provides 954 nucleotides
upstream of the br2 5'-UTR; nucleotides 955-1000 correspond to the
5'-UTR; nucleotides 1001-1604 correspond to the first exon;
nucleotides 1605-1747 correspond to the first intron; nucleotides
1748-2384 correspond to the second exon; nucleotides 2385-2473
correspond to the second intron; nucleotides 2474-2784 correspond
to the third exon; nucleotides 2785-3410 correspond to the third
intron; nucleotides 3411-3640 correspond to the fourth exon;
nucleotides 3641-5309 correspond to the fourth intron; nucleotides
5310-7667 correspond to the fifth exon; and nucleotides 7668-8029
correspond to the 3'-UTR. SEQ ID NO: 132 also provides 638
nucleotides downstream of the end of the 3'-UTR (nucleotides
8030-8667).
[0197] As used herein, a "brachytic allele" is an allele at a
particular genomic locus that confers, or contributes to, a
brachytic or semi-dwarf phenotype, such as an allele of a brachytic
gene that causes a brachytic or semi-dwarf phenotype, or
alternatively, is an allele that allows for the identification of
plants that comprise a brachytic phenotype or plants that can give
rise to progenies with a brachytic phenotype. For example, a
brachytic allele of a marker can be a marker allele that segregates
with a brachytic phenotype.
[0198] In some aspects, a brachytic, dwarf, or semi-dwarf corn
plant comprises a reduced level of br2 mRNA and/or protein, as
compared to a control corn plant not having the brachytic allele.
In other aspects, the corn plants or seeds comprise reduced Br2
protein activity compared to a control plant not having the
brachytic allele. In some aspects, the height of a brachytic,
dwarf, or semi-dwarf plant comprising a brachytic allele at
maturity is reduced by at least 10%, at least 20%, at least 30%, at
least 40%, at least 50%, at least 60%, or at least 70% compared to
a control plant not having a brachytic allele. In another aspect,
the yield of a brachytic, dwarf, or semi-dwarf corn plant
comprising a brachytic allele is equal to or more than the yield of
a control plant not having the brachytic allele. In an aspect, a
brachytic, dwarf, or semi-dwarf corn plant comprising a brachytic
allele requires about 5%, 10%, 15%, 20%, or 25% fewer heat units
than a control plant not having the brachytic allele to reach
anthesis. In an aspect, a brachytic, dwarf, or semi-dwarf corn
plant is homozygous for a brachytic allele. In another aspect, a
brachytic, dwarf, or semi-dwarf corn plant is heterozygous for a
brachytic allele. In another aspect, a brachytic, dwarf, or
semi-dwarf corn plant is a hybrid. In another aspect, a brachytic,
dwarf, or semi-dwarf corn plant is an inbred, such as a female
inbred.
[0199] In an aspect, this disclosure provides brachytic, dwarf, or
semi-dwarf corn plants comprising a brachytic allele comprising one
or more sequences selected from the group consisting of SEQ ID NOs:
59-85. In another aspect, a brachytic, dwarf, or semi-dwarf corn
plant comprises a single gene conversion of the br2 genomic
region.
[0200] In an aspect, a brachytic, dwarf, or semi-dwarf corn plant
comprises a brachytic allele at a polymorphic locus, wherein the
polymorphic locus is associated with, or linked to, a marker
selected from the group consisting of SEQ ID NOs: 86-131. In
another aspect, a brachytic allele at a polymorphic locus is within
20 cM (centimorgans), within 10 cM, within 5 cM, within 1 cM, or
within 0.5 cM of a marker selected from the group consisting of SEQ
ID NOs: 86-131. In another aspect, a brachytic allele is at a
polymorphic locus within 20 cM, within 10 cM, within 5 cM, within 1
cM, or within 0.5 cM of a marker selected from the group consisting
of SEQ ID NOs: 90-117. In another aspect, a brachytic allele is at
a polymorphic locus within 20 cM, within 10 cM, within 5 cM, within
1 cM, or within 0.5 cM of a marker selected from the group
consisting of SEQ ID NOs: 92 and 117.
[0201] In an aspect, a corn plant or plurality of corn plants
provided herein, such as a female corn plant or inbred or a
plurality or population of female corn plants, can comprise at
least one non-natural brachytic mutation, where the corn plant
exhibits a semi-dwarf phenotype compared to a control corn plant
not comprising the at least one non-natural brachytic mutation when
grown under comparable conditions. In another aspect, a corn plant
provided herein can comprise at least one non-natural brachytic
mutation. In another aspect, a corn plant provided herein can
comprise at least one non-natural brachytic mutant allele. In
another aspect, a corn plant provided herein can comprise at least
one non-natural brachytic mutation and exhibits a semi-dwarf
phenotype. In another aspect, a corn plant provided herein can
comprise at least one non-natural brachytic mutant allele and
exhibit a semi-dwarf phenotype. In another aspect, a corn plant
provided herein can comprise a non-naturally occurring mutation in
a br gene reducing the activity of the br gene, where the mutation
is not introduced via a transposon. In another aspect, a corn plant
provided herein can comprise a mutation in a br2 locus or gene as
compared to a wildtype br2 locus or gene. In an aspect, a corn
plant provided herein is homozygous (or biallelic) for a mutation
in a br2 locus or gene as compared to a wildtype br2 locus or gene.
In another aspect, a corn plant provided herein is heterozygous for
a mutation in a br2 locus or gene as compared to a wildtype br2
locus or gene. In another aspect, a corn plant provided herein can
comprise a modified br2 gene with reduced activity, where the corn
plant does not comprise a br2-23 brachytic allele or SNP5259. In
another aspect, a corn plant provided herein can comprise a
synthetic mutation in a br gene, reducing the activity of the br
gene.
[0202] In an aspect, a corn plant or plurality of corn plants
provided herein, such as a female corn plant or inbred or a
plurality or population of female corn plants, can each comprise a
non-transgene or non-transposon mediated mutation in a br gene
reducing the activity of the br gene. In another aspect, a corn
plant provided herein can comprise a recessive, non-transgenic br
mutant allele. In another aspect, a corn plant provided herein can
comprise a heterologous polynucleotide capable of suppressing
expression of a br gene or an mRNA transcribed therefrom. In
another aspect, a corn plant provided herein can comprise a
heterologous polynucleotide capable of suppressing expression of a
br1 gene or an mRNA transcribed therefrom. In another aspect, a
corn plant provided herein can comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In another aspect, a corn plant
provided herein can comprise a heterologous polynucleotide capable
of suppressing expression of a br3 gene or an mRNA transcribed
therefrom. Additional details about altering the expression of br
genes can be found in PCT Application No. PCT/US2016/029492 and
PCT/US2017/067888, the entire contents and disclosure of which are
incorporated herein by reference.
[0203] In an aspect, this disclosure provides a mutant allele of an
endogenous br2 locus, where the mutant allele comprises a DNA
segment inserted into the endogenous br2 locus, wherein the DNA
segment encodes an antisense RNA that is at least 70%, at least
80%, at least 90%, at least 91%, at least 92%, at least 93%, at
least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, or 100% complementary to at least 20, at least 30, at
least 40, at least 50, at least 100, at least 200, at least 300, at
least 400, at least 500, at least 600, at least 700, at least 800,
at least 900, or at least 1000 consecutive nucleotides of SEQ ID
NOs: 132 or 180, and wherein the mutant allele of the endogenous
br2 locus produces an RNA transcript comprising the antisense RNA
sequence. In an aspect, a mutant allele of an endogenous br2 locus
further comprises deletion of at least one portion of the
endogenous br2 locus. In an aspect, a "portion" of an endogenous
br2 locus refers to at least 1 nucleotide.
[0204] In an aspect, a modified corn plant, or plant part thereof,
is homozygous for a deletion within an endogenous br2 locus. In an
aspect, a modified corn plant, or plant part thereof, is biallelic
for a first mutant allele and a second mutant allele each within an
endogenous br2 locus. In an aspect, a first mutant allele comprises
a deletion and/or an inversion or antisense sequence. In an aspect,
a second mutant allele comprises a deletion and/or an inversion or
antisense sequence. In an aspect, a modified corn plant, or plant
part thereof, is heterozygous for a deletion and/or an inversion or
antisense sequence within an endogenous br2 locus.
[0205] In an aspect, a modified corn plant, or plant part thereof,
is homozygous for a mutant allele at an endogenous br2 locus. In an
aspect, a modified corn plant, or plant part thereof, is biallelic
for a first mutant allele and a second mutant allele at an
endogenous br2 locus. In an aspect, a modified corn plant, or plant
part thereof, is heterozygous for a mutant allele at an endogenous
br2 locus.
[0206] In an aspect, a deletion within an endogenous br2 locus
comprises between 1 nucleotide and 8667 nucleotides, between 1
nucleotide and 8000 nucleotides, between 1 nucleotide and 7000
nucleotides, between 1 nucleotide and 6000 nucleotides, between 1
nucleotide and 5000 nucleotides, between 1 nucleotide and 4000
nucleotides, between 1 nucleotide and 3000 nucleotides, between 1
nucleotide and 2000 nucleotides, between 1 nucleotide and 1000
nucleotides, between 1 nucleotide and 750 nucleotides, between 1
nucleotide and 500 nucleotides, between 1 nucleotide and 250
nucleotides, between 1 nucleotide and 100 nucleotides, between 1
nucleotide and 50 nucleotides, between 10 nucleotide and 8000
nucleotides, between 10 nucleotide and 5000 nucleotides, between 10
nucleotide and 2500 nucleotides, between 10 nucleotide and 1000
nucleotides, between 10 nucleotide and 100 nucleotides, between 100
nucleotide and 8000 nucleotides, between 100 nucleotide and 5000
nucleotides, between 100 nucleotide and 2500 nucleotides, between
100 nucleotide and 1000 nucleotides, or between 100 nucleotide and
500 nucleotides. In an aspect, a deletion within an endogenous br2
locus comprises at least 1 nucleotide. In an aspect, a deletion
within an endogenous br2 locus comprises at least 2 nucleotides. In
an aspect, a deletion within an endogenous br2 locus comprises at
least 5 nucleotides. In an aspect, a deletion within an endogenous
br2 locus comprises at least 10 nucleotides. In an aspect, a
deletion within an endogenous br2 locus comprises at least 20
nucleotides. In an aspect, a deletion within an endogenous br2
locus comprises at least 30 nucleotides. In an aspect, a deletion
within an endogenous br2 locus comprises at least 40 nucleotides.
In an aspect, a deletion within an endogenous br2 locus comprises
at least 50 nucleotides. In an aspect, a deletion within an
endogenous br2 locus comprises at least 100 nucleotides. In an
aspect, a deletion within an endogenous br2 locus comprises at
least 200 nucleotides. In an aspect, a deletion within an
endogenous br2 locus comprises at least 300 nucleotides. In an
aspect, a deletion within an endogenous br2 locus comprises at
least 400 nucleotides. In an aspect, a deletion within an
endogenous br2 locus comprises at least 500 nucleotides.
[0207] In an aspect, this disclosure provides a mutant allele of an
endogenous br2 locus, where the mutant allele comprises a deletion
of at least one nucleotide from at least one exon of an endogenous
br2 locus as compared to SEQ ID NO: 132. In an aspect, a deletion
further comprises the deletion of at least one exon of an
endogenous br2 locus as compared to SEQ ID NO: 132. In an aspect, a
deletion comprises the deletion of an endogenous br2 locus. In an
aspect, a deletion comprises the deletion of at least two exons
from an endogenous br2 locus. In an aspect, two deleted exons from
an endogenous br2 locus are contiguous. In an aspect, two deleted
exons from an endogenous br2 locus are not contiguous. In an
aspect, the first exon of an endogenous br2 locus is deleted. In an
aspect, the second exon of an endogenous br2 locus is deleted. In
an aspect, the third exon of an endogenous br2 locus is deleted. In
an aspect, the fourth exon of an endogenous br2 locus is deleted.
In an aspect, the fifth exon of an endogenous br2 locus is deleted.
In an aspect, a deletion further comprises the deletion of at least
one nucleotide from at least one intron of an endogenous br2 locus.
In an aspect, a deletion further comprises the deletion of at least
one nucleotide from at least one intron of an endogenous br2 locus.
In an aspect, a deletion comprises the deletion of at least one
intron of an endogenous br2 locus. In an aspect, a deletion
comprises the deletion of at least one nucleotide of the
5'-untranslated region of the endogenous br2 locus. In an aspect, a
deletion comprises the deletion of at least one nucleotide of the
3'-untranslated region of the endogenous br2 locus.
[0208] In an aspect, a deletion comprises deletion of at least one
nucleotide of the first exon of an endogenous br2 locus. In an
aspect, a deletion comprises deletion of at least one nucleotide of
the second exon of an endogenous br2 locus. In an aspect, a
deletion comprises deletion of at least one nucleotide of the third
exon of an endogenous br2 locus. In an aspect, a deletion comprises
deletion of at least one nucleotide of the fourth exon of an
endogenous br2 locus. In an aspect, a deletion comprises deletion
of at least one nucleotide of the fifth exon of an endogenous br2
locus.
[0209] In an aspect, a deletion comprises deletion of the first
exon of an endogenous br2 locus. In an aspect, a deletion comprises
deletion of the second exon of an endogenous br2 locus. In an
aspect, a deletion comprises deletion of the third exon of an
endogenous br2 locus. In an aspect, a deletion comprises deletion
of the fourth exon of an endogenous br2 locus. In an aspect, a
deletion comprises deletion of the fifth exon of an endogenous br2
locus.
[0210] In an aspect, a deletion comprises deletion of at least one
nucleotide of at least one intron of an endogenous br2 locus. In an
aspect, a deletion comprises deletion of at least one intron of an
endogenous br2 locus. In an aspect, a deletion comprises deletion
of at least one nucleotide of the 5'-untranslated region of an
endogenous br2 locus. In an aspect, a deletion comprises deletion
of the 5'-untranslated region of an endogenous br2 locus. In an
aspect, a deletion comprises deletion of at least one nucleotide of
the 3'-untranslated region of an endogenous br2 locus. In an
aspect, a deletion comprises deletion of the 3'-untranslated region
of an endogenous br2 locus.
[0211] In an aspect, a deletion comprises a deletion of at least
one nucleotide of at least one intron, a deletion of at least one
nucleotide of at least one exon, at least one nucleotide of a
5'-untranslated region (UTR), at least one nucleotide of a 3'-UTR,
or any combination thereof of an endogenous br2 locus.
[0212] In an aspect, a deletion comprises deletion of at least one
nucleotide from a first exon and at least one nucleotide from a
second exon of an endogenous br2 locus. In an aspect, a deletion
comprises deletion of at least one nucleotide from a first exon, at
least one nucleotide from a second exon, and at least one
nucleotide from a third exon of an endogenous br2 locus. In an
aspect, a deletion comprises deletion of at least one nucleotide
from a first exon, at least one nucleotide from a second exon, at
least one nucleotide from a third exon, and at least one nucleotide
from a fourth exon of an endogenous br2 locus. In an aspect, a
deletion comprises deletion of at least one nucleotide from a first
exon, at least one nucleotide from a second exon, at least one
nucleotide from a third exon, at least one nucleotide from a fourth
exon, and at least one nucleotide from a fifth exon of an
endogenous br2 locus.
[0213] In an aspect, a deletion comprises a deletion of a first
exon and a second exon from an endogenous br2 locus. In an aspect,
a first deleted exon and a second deleted exon are contiguous. In
an aspect, a first deleted exon and a second deleted exon are not
contiguous. In an aspect, a deletion comprises deletion of a first
exon and a second exon from an endogenous br2 locus. In an aspect,
a deletion comprises deletion of a first exon, a second exon, and a
third exon from an endogenous br2 locus. In an aspect, a deletion
comprises deletion of a first exon, a second exon, a third exon,
and a fourth exon from an endogenous br2 locus. In an aspect, a
deletion comprises deletion of a first exon, a second exon, a third
exon, a fourth exon, and a fifth exon from an endogenous br2
locus.
[0214] In an aspect, a deletion in an endogenous br2 locus results
in a premature stop codon within an mRNA transcript encoding a Br2
protein. In an aspect, this disclosure provides a modified corn
plant, or plant part thereof, comprising a premature stop codon
within a nucleic acid sequence encoding a Brachytic2 protein as
compared to a control corn plant or plant part thereof. In an
aspect, a mutant allele encodes an mRNA transcript comprising a
premature stop codon as compared to SEQ ID NO: 180.
[0215] According to some embodiments, an endogenous gene can be
edited or engineered to express a truncated protein relative to a
wild type protein by the introduction of a premature stop codon
into the coding sequence and the encoded mRNA transcript of the
endogenous gene. Without being bound by theory, a truncated Br2
protein expressed from an edited endogenous br2 gene comprising a
premature stop codon may not only be non-functional or have reduced
function, but also interfere with the functioning of a wild type
Br2 protein to act in a dominant or semi-dominant manner. In an
aspect, a premature stop codon within an mRNA transcript results in
translation of a truncated protein as compared to a control mRNA
transcript that lacks the premature stop codon. As used herein, a
"stop codon" refers to a nucleotide triplet within an mRNA
transcript that signals a termination of protein translation. A
"premature stop codon" refers to a stop codon positioned earlier
(e.g., on the 5'-side) than the normal stop codon position in an
endogenous mRNA transcript. A stop codon is a nucleotide triplet in
a mRNA that signals the termination of protein translation from the
mRNA. Without being limiting, several stop codons are known in the
art, including "UAG," "UAA," "UGA," "TAG," "TAA," and "TGA." In an
aspect, a premature stop codon can arise from a frameshift
mutation. Frameshift mutations can be caused by the insertion or
deletion of one or more nucleotides in a protein-coding sequence.
In an aspect, a premature stop codon can arise from a substitution,
missense or nonsense mutation. In an aspect, a nonsense, missense
or frameshift mutation provided herein is located in an exon of a
br2 gene. In an aspect, a substitution, insertion or deletion
provided herein is located in a gene element selected from the
group consisting of an exon and an intron/exon splice site. A
substitution, insertion or deletion provided herein can generate a
protein with one or more, two or more, three or more, four or more,
five or more, six or more, seven or more, eight or more, nine or
more, or ten or more nonsense mutations. According to present
embodiments, a premature stop codon may be introduced into the
coding sequence of an endogenous br2 gene via a targeted editing
technique and/or site-directed integration. The premature stop
codon may be generated via imperfect DNA repair following a double
strand break introduced into a br2 gene, or via template-assisted
repair following introduction of the double strand break using a
DNA donor template comprising the premature stop codon. Such a DNA
donor template may further comprise one or more flanking homologous
arms or sequences that are identical, homologous or complementary
to a corresponding sequence of the endogenous br2 gene to help
promote recombination between the donor template and the target
site in the endogenous br2 gene for insertion of a sequence
comprising the premature stop codon at the desired target site.
[0216] In an aspect, a premature stop codon is positioned within
the first exon of an endogenous br2 locus. In an aspect, a
premature stop codon is positioned within the second exon of an
endogenous br2 locus. In an aspect, a premature stop codon is
positioned within the third exon of an endogenous br2 locus. In an
aspect, a premature stop codon is positioned within the fourth exon
of an endogenous br2 locus. In an aspect, a premature stop codon is
positioned within the fifth exon of an endogenous br2 locus.
[0217] In an aspect, a mutant allele provided herein encodes a
truncated protein as compared to SEQ ID NO: 181. As used herein, a
"truncated" protein or polypeptide comprises at least one fewer
amino acid as compared to an endogenous control protein or
polypeptide. For example, if endogenous Protein A comprises 100
amino acids, a truncated version of Protein A can comprise between
1 and 99 amino acids.
[0218] In an aspect, this disclosure provides a modified corn
plant, or plant part thereof, comprising a premature stop codon
within a nucleic acid sequence encoding a Brachytic2 protein as
compared to a nucleic acid sequence of a control corn plant or
plant part thereof. In an aspect, this disclosure provides a
modified corn plant, or plant part thereof, comprising a premature
stop codon within a nucleic acid sequence encoding a Brachytic2
protein. In an aspect, this disclosure provides a modified corn
plant, or plant part thereof, comprising a truncated Brachytic2
protein encoded by a nucleic acid sequence comprising a premature
stop codon as compared to a wildtype or control nucleic acid
sequence. In an aspect, this disclosure provides a modified corn
plant, or plant part thereof, comprising a premature stop codon in
a nucleic acid sequence as compared to SEQ ID NO: 180.
[0219] In an aspect, a premature stop codon is positioned within a
region of a br2 mRNA transcript selected from the group consisting
of the first exon, the second exon, the third exon, the fourth
exon, and the fifth exon.
[0220] In an aspect, a truncated Br2 protein sequence comprises
fewer than 1378 amino acids. In an aspect, a truncated Br2 protein
sequence comprises fewer than 1375 amino acids. In an aspect, a
truncated Br2 protein sequence comprises fewer than 1350 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
fewer than 1300 amino acids. In an aspect, a truncated Br2 protein
sequence comprises fewer than 1200 amino acids. In an aspect, a
truncated Br2 protein sequence comprises fewer than 1100 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
fewer than 1000 amino acids. In an aspect, a truncated Br2 protein
sequence comprises fewer than 900 amino acids. In an aspect, a
truncated Br2 protein sequence comprises fewer than 800 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
fewer than 700 amino acids. In an aspect, a truncated Br2 protein
sequence comprises fewer than 600 amino acids. In an aspect, a
truncated Br2 protein sequence comprises fewer than 500 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
fewer than 400 amino acids. In an aspect, a truncated Br2 protein
sequence comprises fewer than 300 amino acids. In an aspect, a
truncated Br2 protein sequence comprises fewer than 200 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
fewer than 100 amino acids. In an aspect, a truncated Br2 protein
sequence comprises fewer than 50 amino acids.
[0221] In an aspect, a truncated Br2 protein sequence comprises
between 1 amino acid and 1378 amino acids. In an aspect, a
truncated Br2 protein sequence comprises between 25 amino acids and
1378 amino acids. In an aspect, a truncated Br2 protein sequence
comprises between 50 amino acids and 1378 amino acids. In an
aspect, a truncated Br2 protein sequence comprises between 100
amino acids and 1378 amino acids. In an aspect, a truncated Br2
protein sequence comprises between 250 amino acids and 1378 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
between 500 amino acids and 1378 amino acids. In an aspect, a
truncated Br2 protein sequence comprises between 750 amino acids
and 1378 amino acids. In an aspect, a truncated Br2 protein
sequence comprises between 1000 amino acids and 1378 amino acids.
In an aspect, a truncated Br2 protein sequence comprises between
1250 amino acids and 1378 amino acids. In an aspect, a truncated
Br2 protein sequence comprises between 100 amino acids and 1000
amino acids. In an aspect, a truncated Br2 protein sequence
comprises between 250 amino acids and 1000 amino acids. In an
aspect, a truncated Br2 protein sequence comprises between 500
amino acids and 1000 amino acids. In an aspect, a truncated Br2
protein sequence comprises between 750 amino acids and 1000 amino
acids. In an aspect, a truncated Br2 protein sequence comprises
between 1000 amino acids and 1378 amino acids.
[0222] In an aspect, a mutant allele of an endogenous br2 locus
suppresses the expression of a wild-type allele of the endogenous
br2 locus.
[0223] In an aspect, an RNA transcript comprises one or more
sequence elements of the endogenous br2 locus selected from the
group consisting of 5'-UTR, first exon, first intron, second exon,
second intron, third exon, third intron, fourth exon, fourth
intron, fifth exon, 3'-UTR, and any portion thereof. In an aspect,
an endogenous sequence of an RNA transcript is at least 85%, at
least 90%, at least 91%, at least 92%, at least 93%, at least 94%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, or 100% identical to at least 20, at least 30, at least 40, at
least 50, at least 100, at least 200, at least 300, at least 400,
at least 500, at least 600, at least 700, at least 800, at least
900, or at least 1000 consecutive nucleotides of SEQ ID NOs: 132 or
180.
[0224] In an aspect, a DNA segment comprises a nucleotide sequence
originating from the endogenous br2 locus. In an aspect, a DNA
segment comprises an inverted genomic fragment of the endogenous
br2 locus. In an aspect, a DNA segment is inserted near or adjacent
to a corresponding endogenous DNA segment of an endogenous br2
locus. In an aspect, a DNA segment is inserted within a region
selected from the group consisting of the 5' untranslated region
(UTR), first exon, first intron, second exon, second intron, third
exon, third intron, fourth exon, fourth intron, fifth exon, and 3'
UTR of an endogenous br2 locus, and a combination thereof. In an
aspect, the sense strand of a DNA segment comprises a sequence that
is at least 70%, at least 80%, at least 85%, at least 90%, at least
91%, at least 92%, at least 93%, at least 94%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, or 100%
identical or complementary to an exon sequence of an endogenous br2
locus. In an aspect, the sense strand of a DNA segment comprises a
sequence at least 70%, at least 80%, at least 85%, at least 90%, at
least 91%, at least 92%, at least 93%, at least 94%, at least 95%,
at least 96%, at least 97%, at least 98%, at least 99%, or 100%
identical or complementary to an untranslated region (UTR) sequence
of the endogenous br2 locus. In an aspect, the sense strand of a
DNA segment comprises a sequence at least 70%, at least 80%, at
least 85%, at least 90%, at least 91%, at least 92%, at least 93%,
at least 94%, at least 95%, at least 96%, at least 97%, at least
98%, at least 99%, or 100% identical or complementary to an exon
sequence and an intron sequence of the endogenous br2 locus, the
exon sequence and the intron sequence being contiguous within the
endogenous locus. In an aspect, a DNA segment comprises a sequence
having at least 70%, at least 80%, at least 85%, at least 90%, at
least 91%, at least 92%, at least 93%, at least 94%, at least 95%,
at least 96%, at least 97%, at least 98%, at least 99%, or 100%
identical or complementary to SEQ ID NOs: 132 or 180.
[0225] In an aspect, a mutant allele encodes a truncated Br2
protein as compared to SEQ ID NO: 181. In an aspect, a truncated
Br2 protein is at least 1, at least 2, at least 3, at least 4, at
least 5, at least 10, at least 20, at least 30, at least 40, at
least 50, at least 60, at least 70, at least 80, at least 90, at
least 100, or at least 150 amino acids shorter than the amino acid
sequence of SEQ ID NO: 181.
[0226] In an aspect, an intervening DNA sequence comprises a native
sequence of an endogenous br2 locus. In an aspect, an intervening
DNA sequence comprises an exogenous sequence inserted into an
endogenous br2 locus.
[0227] Corn plants or seeds can be screened and/or selected for the
presence of mutations, edits, or transgenes using any methodologies
known to those having ordinary skill in the art. Examples of
screening and selection methodologies include, but are not limited
to, Southern analysis, PCR amplification for detection of a
polynucleotide, Northern blots, RNase protection, primer-extension,
RT-PCR amplification for detecting RNA transcripts, Sanger
sequencing, Next Generation sequencing technologies (e.g.,
Illumina, PacBio, Ion Torrent, 454) enzymatic assays for detecting
enzyme or ribozyme activity of polypeptides and polynucleotides,
and protein gel electrophoresis, Western blots,
immunoprecipitation, and enzyme-linked immunoassays to detect
polypeptides. Other techniques such as in situ hybridization,
enzyme staining, and immunostaining also can be used to detect the
presence or expression of polypeptides and/or polynucleotides.
Methods for performing all of the referenced techniques are
known.
[0228] In an aspect, selecting is performed following genotyping
seeds. In an aspect, selecting is performed following phenotypic
analysis. In an aspect, selecting is performed following
germination of the seeds. In an aspect, selecting is performed
after determining the zygosity of the seed. In an aspect, selecting
comprises a visual assay of the seed. In an aspect, selecting
comprises separating seeds. In an aspect, selecting comprises
placing selected seeds in a container or packet.
[0229] Without being limiting, this disclosure provides several
methods related to the harvesting of corn plants.
[0230] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0231] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0232] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence.
[0233] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132.
[0234] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant.
[0235] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a premature stop
codon within a nucleic acid sequence encoding a Brachytic2 protein
as compared to a control corn plant.
[0236] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0237] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0238] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0239] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0240] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0241] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0242] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0243] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0244] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0245] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0246] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0247] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a DNA
segment inserted into the endogenous br2 locus, wherein the DNA
segment encodes an antisense RNA that is at least 70% complementary
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180,
and wherein the mutant allele of the endogenous br2 locus produces
an RNA transcript comprising the antisense RNA sequence.
[0248] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a
deletion of at least one nucleotide from an endogenous br2 locus as
compared to SEQ ID NO: 132.
[0249] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a dominant or semi-dominant
transgene or mutant allele of a gene, and wherein the transgene or
mutant allele causes a short stature phenotype in the at least one
corn plant.
[0250] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a premature stop codon within a
nucleic acid sequence encoding a Brachytic2 protein as compared to
a control corn plant.
[0251] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0252] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0253] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0254] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0255] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0256] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0257] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0258] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0259] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0260] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_3 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_3 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_3 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0261] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_5 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_5 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0262] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
br2 locus, wherein the DNA segment encodes an antisense RNA that is
at least 70% complementary to at least 20 consecutive nucleotides
of SEQ ID NO: 132 or 180, and wherein the mutant allele of the
endogenous br2 locus produces an RNA transcript comprising the
antisense RNA sequence.
[0263] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a deletion of at least one nucleotide from
an endogenous br2 locus as compared to SEQ ID NO: 132.
[0264] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant.
[0265] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
premature stop codon within a nucleic acid sequence encoding a
Brachytic2 protein as compared to a control corn plant.
[0266] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the transcription termination sequence of
the endogenous Zm.SAMT gene, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0267] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the intergenic region between the
endogenous GA20 oxidase_5 and Zm.SAMT genes, and wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
complementary to all or part of the sense strand of the endogenous
GA20 oxidase_5 gene.
[0268] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of one or more of the following: 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon,
2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon,
5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon,
3' UTR, and any portion thereof, of the endogenous Zm.SAMT
gene.
[0269] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification which results in the
transcription of an antisense strand of at least an exon, an
intron, or an untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof.
[0270] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises the Zm.SAMT gene promoter, or a functional
part thereof, operably linked to at least one transcribable
antisense sequence of at least an exon, intron or untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0271] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a sequence selected from the group
consisting of SEQ ID NOs: 87-105.
[0272] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0273] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic deletion relative to a wild type
allele of the endogenous GA20 oxidase_5 locus, wherein the genomic
deletion is flanked by a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0274] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein the average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic sequence comprising a first
sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length.
[0275] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0276] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0277] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence.
[0278] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132.
[0279] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant.
[0280] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a premature stop
codon within a nucleic acid sequence encoding a Brachytic2 protein
as compared to a control corn plant.
[0281] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0282] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0283] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0284] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0285] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0286] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0287] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0288] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0289] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein average kernel moisture content is less than or equal to
30%, and wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0290] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_3 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_3 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_3 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0291] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_5 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_5 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0292] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
br2 locus, wherein the DNA segment encodes an antisense RNA that is
at least 70% complementary to at least 20 consecutive nucleotides
of SEQ ID NO: 132 or 180, and wherein the mutant allele of the
endogenous br2 locus produces an RNA transcript comprising the
antisense RNA sequence.
[0293] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a deletion of at least one nucleotide from
an endogenous br2 locus as compared to SEQ ID NO: 132.
[0294] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant.
[0295] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
premature stop codon within a nucleic acid sequence encoding a
Brachytic2 protein as compared to a control corn plant.
[0296] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the transcription termination sequence of
the endogenous Zm.SAMT gene, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0297] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the intergenic region between the
endogenous GA20 oxidase_5 and Zm.SAMT genes, and wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
complementary to all or part of the sense strand of the endogenous
GA20 oxidase_5 gene.
[0298] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of one or more of the following: 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon,
2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon,
5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon,
3' UTR, and any portion thereof, of the endogenous Zm.SAMT
gene.
[0299] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification which results in the
transcription of an antisense strand of at least an exon, an
intron, or an untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof.
[0300] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises the Zm.SAMT gene promoter, or a functional
part thereof, operably linked to at least one transcribable
antisense sequence of at least an exon, intron or untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0301] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a sequence selected from the group
consisting of SEQ ID NOs: 87-105.
[0302] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0303] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic deletion relative to a wild type
allele of the endogenous GA20 oxidase_5 locus, wherein the genomic
deletion is flanked by a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR, 1st
exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0304] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genomic sequence comprising a first
sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length.
[0305] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0306] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA
sequence.
[0307] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of an
endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence.
[0308] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of an
endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132.
[0309] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant.
[0310] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a premature stop codon
within a nucleic acid sequence encoding a Brachytic2 protein as
compared to a control corn plant.
[0311] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0312] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene.
[0313] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any portion
thereof, and the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0314] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0315] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof.
[0316] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a sequence selected from the group consisting of SEQ ID
NOs: 87-105.
[0317] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th
exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene; wherein
the first sequence and the second sequence are contiguous or
separated only by an intervening sequence of fewer than 555
nucleotides.
[0318] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1st exon, 1st
intron, 2nd exon, 2nd intron, 3rd exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.GA20 oxidase_5 gene; and wherein the second
sequence comprises one or more of the 5' UTR, 1st exon, 1st intron,
2nd exon, 2nd intron, 3rd exon, 3rd intron, 4th exon, 4th intron,
5th exon, 5th intron, 6th exon, 6th intron, 7th exon, 7th intron,
8th exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene.
[0319] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length.
[0320] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_3 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_3 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_3 locus produces a RNA transcript
comprising the antisense RNA sequence.
[0321] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_5 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence.
[0322] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous br2 locus, wherein the DNA segment
encodes an antisense RNA that is at least 70% complementary to at
least 20 consecutive nucleotides of SEQ ID NO: 132 or 180, and
wherein the mutant allele of the endogenous br2 locus produces an
RNA transcript comprising the antisense RNA sequence.
[0323] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a deletion of at
least one nucleotide from an endogenous br2 locus as compared to
SEQ ID NO: 132.
[0324] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a dominant or semi-dominant transgene or
mutant allele of a gene, and wherein the transgene or mutant allele
causes a short stature phenotype in the at least one corn
plant.
[0325] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a premature stop codon within a nucleic acid
sequence encoding a Brachytic2 protein as compared to a control
corn plant.
[0326] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
transcription termination sequence of the endogenous Zm.SAMT gene,
and wherein the mutant allele produces a RNA molecule comprising an
antisense sequence complementary to all or part of the sense strand
of the endogenous GA20 oxidase_5 gene.
[0327] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0328] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of one or
more of the following: 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3' UTR, and any portion thereof, and the 5' UTR,
1st exon, 1st intron, 2nd exon, 2nd intron, 3rd exon, 3rd intron,
4th exon, 4th intron, 5th exon, 5th intron, 6th exon, 6th intron,
7th exon, 7th intron, 8th exon, 3' UTR, and any portion thereof, of
the endogenous Zm.SAMT gene.
[0329] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification which results in the transcription of an antisense
strand of at least an exon, an intron, or an untranslated region
(UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0330] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises the Zm.SAMT
gene promoter, or a functional part thereof, operably linked to at
least one transcribable antisense sequence of at least an exon,
intron or untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof.
[0331] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a sequence
selected from the group consisting of SEQ ID NOs: 87-105.
[0332] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3' UTR, and any complementary sequence
thereof, and any portion of the foregoing, of the endogenous
Zm.GA20 oxidase_5 gene; and wherein the second sequence comprises
one or more of the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene; wherein the first
sequence and the second sequence are contiguous or separated only
by an intervening sequence of fewer than 555 nucleotides.
[0333] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
deletion relative to a wild type allele of the endogenous GA20
oxidase_5 locus, wherein the genomic deletion is flanked by a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1st exon, 1st intron, 2nd
exon, 2nd intron, 3rd exon, 3' UTR, and any complementary sequence
thereof, and any portion of the foregoing, of the endogenous
Zm.GA20 oxidase_5 gene; and wherein the second sequence comprises
one or more of the 5' UTR, 1st exon, 1st intron, 2nd exon, 2nd
intron, 3rd exon, 3rd intron, 4th exon, 4th intron, 5th exon, 5th
intron, 6th exon, 6th intron, 7th exon, 7th intron, 8th exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene.
[0334] In an aspect, this disclosure provides a method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
sequence comprising a first sequence and a second sequence; wherein
the first sequence comprises at least 15 consecutive nucleotides of
one or more of SEQ ID NOs: 228-235 and 276-283; wherein the second
sequence comprises at least 15 consecutive nucleotides of one or
more of SEQ ID NOs: 235-276; and wherein the genomic sequence is at
least 50 consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length.
[0335] In an aspect, at least 10% of the corn plants in a field are
brachytic corn plants. In an aspect, at least 20% of the corn
plants in a field are brachytic corn plants. In an aspect, at least
30% of the corn plants in a field are brachytic corn plants. In an
aspect, at least 40% of the corn plants in a field are brachytic
corn plants. In an aspect, at least 50% of the corn plants in a
field are brachytic corn plants. In an aspect, at least 60% of the
corn plants in a field are brachytic corn plants. In an aspect, at
least 70% of the corn plants in a field are brachytic corn plants.
In an aspect, at least 80% of the corn plants in a field are
brachytic corn plants. In an aspect, at least 90% of the corn
plants in a field are brachytic corn plants. In an aspect, 100% of
the corn plants in a field are brachytic corn plants.
[0336] In an aspect, between 1% and 100% of the corn plants in a
field are brachytic corn plants. In an aspect, between 10% and 100%
of the corn plants in a field are brachytic corn plants. In an
aspect, between 20% and 100% of the corn plants in a field are
brachytic corn plants. In an aspect, between 30% and 100% of the
corn plants in a field are brachytic corn plants. In an aspect,
between 40% and 100% of the corn plants in a field are brachytic
corn plants. In an aspect, between 50% and 100% of the corn plants
in a field are brachytic corn plants. In an aspect, between 60% and
100% of the corn plants in a field are brachytic corn plants. In an
aspect, between 70% and 100% of the corn plants in a field are
brachytic corn plants. In an aspect, between 80% and 100% of the
corn plants in a field are brachytic corn plants. In an aspect,
between 90% and 100% of the corn plants in a field are brachytic
corn plants.
[0337] It will be appreciated in the art that dwarf, semi-dwarf,
and brachytic plants can be inbred or hybrid plants.
[0338] Three brachytic mutants have been isolated in maize to date:
brachytic1 (br1), brachytic2 (br2) and brachytic3 (br3). brachytic3
is also known as brevis plant 1 (Bv1). Both br1 and br3 mutations
cause a reduction in corn plant height which has been thought too
severe for commercial exploitation due to potential impacts on
yield. In contrast, the br2 mutant has particular agronomic
potential because of shortening of the internodes of the lower
stalk without an obvious reduction in other plant organs. In
addition, br2 lines exhibit an unusual stalk strength and tolerance
to wind lodging, while the leaves are often darker and persist
longer in the active green than those of the wild-type plants. The
br2 phenotype is insensitive to treatment with Gibberellins,
auxins, brassinosteroids and cytokinins, suggesting that the
biosynthesis of these hormones is not modified by the br2 mutation.
Multani et al. identified the genomic sequence of the br2 gene and
deposited it under GenBank Accession No. AY366085. See Multani et
al., Science, 302:81-84 (2003). Br2 was annotated to encode a
putative protein similar to adenosine triphosphate (ATP)-binding
cassette transporters of the multidrug resistant (MDR) class of
P-glycoproteins (PGPs). Pilu et al. reported a br2-23 allele having
an 8-bp deletion in the 3' end of the br2 gene and claimed a direct
relationship between this deletion and the brachytic phenotype in
their br2-23 plants. See Pilu et al., Molecular Breeding,
20:83-91(2007). Other brachytic gene alleles are known in the art
which may be used according to embodiments of the present
disclosure.
[0339] In some aspects, a brachytic, dwarf, or semi-dwarf corn
plant comprises a reduced level of Br2 mRNA and/or protein compared
to a control corn plant not having the brachytic allele. In other
aspects, the corn plants or seeds comprise reduced Br2 protein
activity compared to a control plant not having the brachytic
allele. In some aspects, the height of a brachytic, dwarf, or
semi-dwarf plant comprising a brachytic allele at maturity is
reduced by at least 5%, at least 10%, at least 20%, at least 30%,
at least 40%, at least 50%, at least 60%, or at least 70% compared
to a control plant not having a brachytic allele. In another
aspect, the yield of a brachytic, dwarf, or semi-dwarf corn plant
comprising a brachytic allele is equal to or more than the yield of
a control plant not having the brachytic allele.
[0340] In an aspect, at least 10% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 20% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 30% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 40% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 50% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 60% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 70% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 80% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, at least 90% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, 100% of the corn plants in a field comprise a
mutation in a br2 locus as compared to a wildtype br2 locus.
[0341] In an aspect, between 1% and 100% of the corn plants in a
field comprise a mutation in a br2 locus as compared to a wildtype
br2 locus. In an aspect, between 10% and 100% of the corn plants in
a field comprise a mutation in a br2 locus as compared to a
wildtype br2 locus. In an aspect, between 20% and 100% of the corn
plants in a field comprise a mutation in a br2 locus as compared to
a wildtype br2 locus. In an aspect, between 30% and 100% of the
corn plants in a field comprise a mutation in a br2 locus as
compared to a wildtype br2 locus. In an aspect, between 40% and
100% of the corn plants in a field comprise a mutation in a br2
locus as compared to a wildtype br2 locus. In an aspect, between
50% and 100% of the corn plants in a field comprise a mutation in a
br2 locus as compared to a wildtype br2 locus. In an aspect,
between 60% and 100% of the corn plants in a field comprise a
mutation in a br2 locus as compared to a wildtype br2 locus. In an
aspect, between 70% and 100% of the corn plants in a field comprise
a mutation in a br2 locus as compared to a wildtype br2 locus. In
an aspect, between 80% and 100% of the corn plants in a field
comprise a mutation in a br2 locus as compared to a wildtype br2
locus. In an aspect, between 90% and 100% of the corn plants in a
field comprise a mutation in a br2 locus as compared to a wildtype
br2 locus.
[0342] In another aspect, a corn plant provided herein comprises a
non-transgene or non-transposon mediated mutation in a Br gene
reducing the activity of the Br gene. In a further aspect, a corn
plant provided herein comprises a recessive, non-transgenic Br
mutant allele. In another aspect, a corn plant provided herein
comprises a heterologous polynucleotide capable of suppressing
expression of a Br gene or an mRNA transcribed therefrom. In
another aspect, a corn plant provided herein comprises a
heterologous polynucleotide capable of suppressing expression of a
Br1 gene or an mRNA transcribed therefrom. In another aspect, a
corn plant provided herein comprises a heterologous polynucleotide
capable of suppressing expression of a Br2 gene or an mRNA
transcribed therefrom. In another aspect, a corn plant provided
herein comprises a heterologous polynucleotide capable of
suppressing expression of a Br3 gene or an mRNA transcribed
therefrom. Additional details about corn plants and altering the
expression of Br genes can be found in PCT Application No.
PCT/US2016/029492 and PCT/US2017/067888, which are incorporated
herein by reference in their entirety.
[0343] In an aspect, a mutant allele of an endogenous br2 locus
suppresses the expression of a wild-type allele of the endogenous
br2 locus. In an aspect, a mutant allele product of an endogenous
br2 locus disrupts the function of a wild-type allele product of
the endogenous br2 locus. In an aspect, a "product" of a mutant
allele is a mRNA transcript. In an aspect, a "product" of a mutant
allele comprises an antisense RNA. In an aspect, a "product" of a
mutant allele is a protein.
[0344] In an aspect, at least 10% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a br2 gene or an mRNA transcribed therefrom. In an
aspect, at least 20% of the corn plants in a field comprise a
heterologous polynucleotide capable of suppressing expression of a
br2 gene or an mRNA transcribed therefrom. In an aspect, at least
30% of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, at least 40% of the
corn plants in a field comprise a heterologous polynucleotide
capable of suppressing expression of a br2 gene or an mRNA
transcribed therefrom. In an aspect, at least 50% of the corn
plants in a field comprise a heterologous polynucleotide capable of
suppressing expression of a br2 gene or an mRNA transcribed
therefrom. In an aspect, at least 60% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a br2 gene or an mRNA transcribed therefrom. In an
aspect, at least 70% of the corn plants in a field comprise a
heterologous polynucleotide capable of suppressing expression of a
br2 gene or an mRNA transcribed therefrom. In an aspect, at least
80% of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, at least 90% of the
corn plants in a field comprise a heterologous polynucleotide
capable of suppressing expression of a br2 gene or an mRNA
transcribed therefrom. In an aspect, 100% of the corn plants in a
field comprise a heterologous polynucleotide capable of suppressing
expression of a br2 gene or an mRNA transcribed therefrom.
[0345] In an aspect, between 1% and 100% of the corn plants in a
field comprise a heterologous polynucleotide capable of suppressing
expression of a br2 gene or an mRNA transcribed therefrom. In an
aspect, between 10% and 100% of the corn plants in a field comprise
a heterologous polynucleotide capable of suppressing expression of
a br2 gene or an mRNA transcribed therefrom. In an aspect, between
20% and 100% of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 30% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 40% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 50% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 60% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 70% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 80% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom. In an aspect, between 90% and 100%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a br2 gene or
an mRNA transcribed therefrom.
[0346] Active or bioactive gibberellic acids (i.e., "active
gibberellins" or "active GAs") are known in the art for a given
plant species, as distinguished from inactive GAs. For example,
active GAs in corn and higher plants include the following: GA1,
GA3, GA4, and GA7. Thus, an "active GA-producing tissue" is a plant
tissue that produces one or more active GAs. In an aspect, a
modified corn plant comprises a level of one or more active GAs in
at least one internode tissue of a stem or stalk that is at least
1%, at least 2%, at least 2.5%, at least 3%, at least 4%, at least
5%, at least 6%, at least 7%, at least 8%, at least 9%, at least
10%, at least 11%, at least 12%, at least 13%, at least 14%, at
least 15%, at least 20%, at least 25%, at least 30%, at least 35%,
at least 40%, at least 45%, or at least 50% lower than the level of
the one or more active GAs in the same internode tissue of an
unmodified control plant. In an aspect, a modified corn plant
provided herein comprises a lower level of one or more active GAs
in at least one internode tissue of a stem or stalk as compared to
the same internode tissue of an unmodified control plant.
[0347] Certain biosynthetic enzymes (e.g., GA20 oxidase and GA3
oxidase) and catabolic enzymes (e.g., GA2 oxidase) in the GA
pathway participate in GA synthesis and degradation, respectively,
to affect active GA levels in plant tissues. Thus, in addition to
suppression of certain GA20 oxidase genes, it is further proposed
that suppression of a GA3 oxidase gene in a constitutive or
tissue-specific or tissue-preferred manner can also produce corn
plants having a short stature phenotype and increased lodging
resistance, with possible increased yield, but without off-types in
the ear.
[0348] Without being bound by theory, it is proposed that
suppression of GA20 or GA3 oxidase gene(s) and/or targeting of a
subset of one or more GA oxidase gene(s) can be effective in
achieving a short stature, semi-dwarf phenotype with increased
resistance to lodging, but without reproductive off-types in the
ear. It is further proposed, without being limited by theory, that
suppression of GA20 and/or GA3 oxidase gene(s) through constitutive
expression or in active GA-producing tissues, such as the vascular
and/or leaf tissues of the plant, can produce a short-stature plant
with increased lodging resistance, but without significant
off-types in reproductive tissues. Expression of a GA20 or GA3
oxidase suppression element using a constitutive, vascular and/or
leaf promoter can be sufficient and effective at producing plants
with the short stature phenotype, while avoiding potential
off-types in reproductive tissues that were previously observed
with GA mutants in corn. For example, GA20 and/or GA3 oxidase
gene(s) can be targeted for suppression using a vascular promoter,
such as a rice tungro bacilliform virus (RTBV) promoter, that
drives expression in vascular tissues of plants. The expression
pattern of the RTBV promoter is enriched in vascular tissues of
corn plants relative to non-vascular tissues, which is sufficient
to produce a semi-dwarf phenotype in corn plants when operably
linked to a suppression element targeting GA20 and GA3 oxidase
gene(s). Lowering of active GA levels in tissue(s) of a corn plant,
such as in the stalk, stem, or internode(s) of corn plant, that
produce active GAs can reduce plant height and increase lodging
resistance, and avoid off-types in the reproductive tissues of the
plant, such as in the female (ear) or male (tassel) tissues of the
plant.
[0349] Without being limited by theory, it is proposed that short
stature, semi-dwarf phenotypes in corn plants can result from a
sufficient level of expression of a suppression construct targeting
certain GA oxidase gene(s) in active GA-producing tissue(s) of the
plant. For targeted suppression of certain GA20 oxidase genes in
corn, restricting the pattern of expression to avoid reproductive
ear tissues may not be necessary to avoid reproductive off-types in
the developing ear. However, expression of a GA20 oxidase
suppression construct at low levels, and/or in a limited number of
plant tissues, can be insufficient to cause a significant short
stature, semi-dwarf phenotype. Given that the observed semi-dwarf
phenotype with targeted GA20 oxidase suppression is the result of
shortening the stem internodes of the plant, suppression of GA20
oxidase genes in at least some stem tissues was not sufficient to
cause shortening of the internodes and reduced plant height.
Without being bound by theory, it is proposed that suppression of
certain GA oxidase gene(s) in tissue(s) and/or cell(s) of the plant
where active GAs are produced, and not necessarily in stem or
internode tissue(s), can be sufficient to produce semi-dwarf
plants, even though the short stature trait is due to shortening of
the stem internodes. Given that GAs can migrate through the
vasculature of the plant, manipulating GA oxidase genes in plant
tissue(s) where active GAs are produced can result in a short
stature, semi-dwarf plant, even though this may be largely achieved
by suppressing the level of active GAs produced in non-stem tissues
(i.e., away from the site of action in the stem where reduced
internode elongation leads to the semi-dwarf phenotype). Indeed,
suppression of certain GA20 oxidase genes in leaf tissues causes a
moderate semi-dwarf phenotype in corn plants. Given that expression
of a GA20 oxidase suppression construct with several different
"stem" promoters did not produce the semi-dwarf phenotype in corn,
it is noteworthy that expression of the same GA20 oxidase
suppression construct with a vascular promoter was effective at
consistently producing the semi-dwarf phenotype with a high degree
of penetrance across events and germplasms. A semi-dwarf phenotype
was also observed with expression of the same GA20 oxidase
suppression construct using other vascular promoters and with
various constitutive promoters without any observable off-types.
Additional details about corn plants and altering the expression of
GA20 oxidase and GA3 oxidase genes can be found in PCT Application
No. PCT/US2017/047405 and PCT/US2019/018133, which are incorporated
herein by reference in their entirety.
[0350] In an aspect, a corn plant or plurality of corn plants
provided herein can each comprise a recombinant DNA construct or
polynucleotide sequence, where the recombinant DNA construct or
polynucleotide sequence comprises a transcribable DNA sequence
encoding a non-coding RNA molecule that targets at least one
endogenous GA20 or GA3 oxidase gene for suppression. In another
aspect, a corn plant provided herein can comprise suppressed GA3
oxidase gene expression in one or more tissues as compared to a
wild-type control plant. In another aspect, a corn plant provided
herein can comprise suppressed GA20 oxidase gene expression in one
or more tissues as compared to a wild-type control plant. In
another aspect, a corn plant provided herein can comprise a
mutation at or near an endogenous GA oxidase gene, where the
expression level of the endogenous GA oxidase gene is reduced or
eliminated in the corn plant, and where the corn plant has a
shorter plant height as compared to a wild-type control plant. In
an aspect, a corn plant provided herein can comprise a recombinant
polynucleotide capable of suppressing expression of one or more
GA20 oxidase and/or GA3 oxidase gene(s) and/or mRNA(s) transcribed
therefrom. Alternatively, a corn plant provided herein can comprise
one or more mutation(s) or edit(s) in one or more GA20 oxidase
and/or GA3 oxidase gene(s). In an aspect, a corn plant provided
herein can comprise a mutation in a GA20 oxidase locus or gene as
compared to a wildtype GA20 oxidase locus or gene. In an aspect, a
corn plant provided herein is homozygous (or biallelic) for a
mutation or an edit in one or more GA20 oxidase loci or genes as
compared to a wildtype GA20 oxidase locus or gene. In an aspect, a
corn plant provided herein is homozygous (or biallelic) for a
mutation or an edit in a GA20 oxidase_3 gene as compared to a
wildtype GA20 oxidase_3 gene. In an aspect, a corn plant provided
herein is homozygous (or biallelic) for a mutation or an edit in a
GA20 oxidase_5 gene as compared to a wildtype GA20 oxidase_5 gene.
In another aspect, a corn plant provided herein is heterozygous for
a mutation or an edit in one or more GA20 oxidase loci or genes as
compared to a wildtype GA20 oxidase locus or gene. In an aspect, a
corn plant provided herein is heterozygous for a mutation or an
edit in a GA20 oxidase_3 gene as compared to a wildtype GA20
oxidase_3 gene. In an aspect, a corn plant provided herein is
heterozygous for a mutation or an edit in a GA20 oxidase_5 gene as
compared to a wildtype GA20 oxidase_5 gene. In another aspect, a
corn plant provided herein can comprise a mutation in a GA3 oxidase
locus or gene as compared to a wildtype GA3 oxidase locus or gene.
In an aspect, a corn plant provided herein is homozygous (or
biallelic) for a mutation or an edit in one or more GA3 oxidase
loci or genes as compared to a wildtype GA3 oxidase locus or gene.
In another aspect, a corn plant provided herein is heterozygous for
a mutation or an edit in a one or more GA3 oxidase loci or genes as
compared to a wildtype GA3 oxidase locus or gene. In another
aspect, a corn plant provided herein can comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. Additional details about
altering the expression of GA20 and/or GA3 oxidase gene(s) through
suppression, mutation, or editing of those gene(s) can be found in
PCT Application No. PCT/US2017/047405, the entire contents and
disclosure of which is incorporated herein by reference.
[0351] In an aspect, a modified corn plant, or plant part thereof,
is homozygous for a mutant allele at an endogenous GA oxidase
locus. In an aspect, a modified corn plant, or plant part thereof,
is biallelic for a first mutant allele and a second mutant allele
at an endogenous GA oxidase locus. In an aspect, a modified corn
plant, or plant part thereof, is heterozygous for a mutant allele
at an endogenous GA oxidase locus. In an aspect, a modified corn
plant, or plant part thereof, is homozygous for a mutant allele at
an endogenous GA20 oxidase locus. In an aspect, a modified corn
plant, or plant part thereof, is biallelic for a first mutant
allele and a second mutant allele at an endogenous GA20 oxidase
locus. In an aspect, a modified corn plant, or plant part thereof,
is heterozygous for a mutant allele at an endogenous GA20 oxidase
locus. In an aspect, a modified corn plant, or plant part thereof,
is homozygous for a mutant allele at an endogenous GA20 oxidase_5
locus. In an aspect, a modified corn plant, or plant part thereof,
is biallelic for a first mutant allele and a second mutant allele
at an endogenous GA20 oxidase_5 locus. In an aspect, a modified
corn plant, or plant part thereof, is heterozygous for a mutant
allele at an endogenous GA20 oxidase_5 locus. In an aspect, a
modified corn plant, or plant part thereof, is homozygous for a
mutant allele at an endogenous GA20 oxidase_3 locus. In an aspect,
a modified corn plant, or plant part thereof, is biallelic for a
first mutant allele and a second mutant allele at an endogenous
GA20 oxidase_3 locus. In an aspect, a modified corn plant, or plant
part thereof, is heterozygous for a mutant allele at an endogenous
GA20 oxidase_3 locus. In an aspect, a modified corn plant, or plant
part thereof, is homozygous for a mutant allele at an endogenous
GA3 oxidase locus. In an aspect, a modified corn plant, or plant
part thereof, is biallelic for a first mutant allele and a second
mutant allele at an endogenous GA3 oxidase locus. In an aspect, a
modified corn plant, or plant part thereof, is heterozygous for a
mutant allele at an endogenous GA3 oxidase locus.
[0352] By targeting a subset of one or more endogenous GA3 or GA20
oxidase genes for suppression within a plant, a more pervasive
pattern of expression (e.g., with a constitutive promoter) can be
used to produce semi-dwarf plants without significant reproductive
off-types and/or other undesirable traits in the plant, even with
expression of the suppression construct in reproductive tissue(s).
Indeed, suppression elements and constructs are provided herein
that selectively target the GA20 oxidase_3 and/or GA20 oxidase_5
genes for suppression, which can be operably linked to a vascular,
leaf and/or constitutive promoter.
[0353] As introduced above, instead of suppressing one or more GA
oxidase gene(s), active GA levels can also be reduced in a corn
plant by mutation or editing of one or more GA20 and/or GA3 oxidase
gene(s).
[0354] Corn has a family of at least nine GA20 oxidase genes that
includes GA20 oxidase_1, GA20 oxidase_2, GA20 oxidase_3, GA20
oxidase_4, GA20 oxidase_5, GA20 oxidase_6, GA20 oxidase_7, GA20
oxidase_8, and GA20 oxidase_9. However, there are only two GA3
oxidases in corn, GA3 oxidase_1 and GA3 oxidase_2. The DNA and
protein sequences by SEQ ID NOs for each of these GA20 oxidase
genes are provided in Table 1, and the DNA and protein sequences by
SEQ ID NOs for each of these GA3 oxidase genes are provided in
Table 2.
[0355] In an aspect, a corn plant provided herein is homozygous (or
biallelic) for a mutation or an edit in a GA20 oxidase_5 locus or
gene as compared to a wildtype GA20 oxidase_5 locus or gene and
homozygous (or biallelic) for a mutation or an edit in a GA20
oxidase_3 locus or gene as compared to a wildtype GA20 oxidase_3
locus or gene. In an aspect, a corn plant provided herein is
homozygous (or biallelic) for a mutation or an edit in a GA20
oxidase_5 locus or gene as compared to a wildtype GA20 oxidase_5
locus or gene and heterozygous for a mutation or an edit in a GA20
oxidase_3 locus or gene as compared to a wildtype GA20 oxidase_3
locus or gene. In an aspect, a corn plant provided herein is
heterozygous for a mutation or an edit in a GA20 oxidase_5 locus or
gene as compared to a wildtype GA20 oxidase_5 locus or gene and
homozygous (or biallelic) for a mutation or an edit in a GA20
oxidase_3 locus or gene as compared to a wildtype GA20 oxidase_3
locus or gene. See, e.g., U.S. Provisional Patent Application Nos.
62/631,412; 62/631,416; and 62/710,302; the contents and
disclosures of which are incorporated herein by reference in their
entireties.
TABLE-US-00001 TABLE 1 DNA and protein sequences by sequence
identifier for GA20 oxidase genes in corn. GA20 Coding Sequence
oxidase Gene cDNA (CDS) Protein GA20 oxidase_1 SEQ ID NO: 1 SEQ ID
NO: 2 SEQ ID NO: 3 GA20 oxidase_2 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID
NO: 6 GA20 oxidase_3 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 9 GA20
oxidase_4 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 GA20 oxidase_5
SEQ ID NO: 13 SEQ ID NO: 14 SEQ ID NO: 15 GA20 oxidase_6 SEQ ID NO:
16 SEQ ID NO: 17 SEQ ID NO: 18 GA20 oxidase_7 SEQ ID NO: 19 SEQ ID
NO: 20 SEQ ID NO: 21 GA20 oxidase_8 SEQ ID NO: 22 SEQ ID NO: 23 SEQ
ID NO: 24 GA20 oxidase_9 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO:
27
TABLE-US-00002 TABLE 2 DNA and protein sequences by sequence
identifier for GA3 oxidase genes in corn. Coding GA3 oxidase
Sequence Gene cDNA (CDS) Protein GA3 oxidase_1 SEQ ID NO: 28 SEQ ID
NO: 29 SEQ ID NO: 30 GA3 oxidase_2 SEQ ID NO: 31 SEQ ID NO: 32 SEQ
ID NO: 33
[0356] The genomic DNA sequence of GA20 oxidase_3 is provided in
SEQ ID NO: 34, and the genomic DNA sequence of GA20 oxidase_5 is
provided in SEQ ID NO: 35. For the GA20 oxidase_3 gene, SEQ ID NO:
34 provides 3000 nucleotides upstream of the GA20 oxidase_3 5'-UTR;
nucleotides 3001-3096 correspond to the 5'-UTR; nucleotides
3097-3665 correspond to the first exon; nucleotides 3666-3775
correspond to the first intron; nucleotides 3776-4097 correspond to
the second exon; nucleotides 4098-5314 correspond to the second
intron; nucleotides 5315-5584 correspond to the third exon; and
nucleotides 5585-5800 correspond to the 3'-UTR. SEQ ID NO: 34 also
provides 3000 nucleotides downstream of the end of the 3'-UTR
(nucleotides 5801-8800).
[0357] For the GA20 oxidase_5 gene, SEQ ID NO: 35 provides 3000
nucleotides upstream of the GA20 oxidase_5 start codon (nucleotides
1-3000); nucleotides 3001-3791 correspond to the first exon;
nucleotides 3792-3906 correspond to the first intron; nucleotides
3907-4475 correspond to the second exon; nucleotides 4476-5197
correspond to the second intron; nucleotides 5198-5473 correspond
to the third exon; and nucleotides 5474-5859 correspond to the
3'-UTR. SEQ ID NO: 35 also provides 3000 nucleotides downstream of
the end of the 3'-UTR (nucleotides 5860-8859).
[0358] The genomic DNA sequence of GA3 oxidase_1 is provided in SEQ
ID NO: 36, and the genomic DNA sequence of GA3 oxidase_2 is
provided in SEQ ID NO: 37. For the GA3 oxidase_1 gene, nucleotides
1-29 of SEQ ID NO: 36 correspond to the 5'-UTR; nucleotides 30-514
of SEQ ID NO: 36 correspond to the first exon; nucleotides 515-879
of SEQ ID NO: 36 correspond to the first intron; nucleotides
880-1038 of SEQ ID NO: 36 correspond to the second exon;
nucleotides 1039-1158 of SEQ ID NO: 36 correspond to the second
intron; nucleotides 1159-1663 of SEQ ID NO: 36 correspond to the
third exon; and nucleotides 1664-1788 of SEQ ID NO: 36 correspond
to the 3'-UTR. For the GA3 oxidase_2 gene, nucleotides 1-38 of SEQ
ID NO: 37 correspond to the 5-UTR; nucleotides 39-532 of SEQ ID NO:
37 correspond to the first exon; nucleotides 533-692 of SEQ ID NO:
37 correspond to the first intron; nucleotides 693-851 of SEQ ID
NO: 37 correspond to the second exon; nucleotides 852-982 of SEQ ID
NO: 37 correspond to the second intron; nucleotides 983-1445 of SEQ
ID NO: 37 correspond to the third exon; and nucleotides 1446-1698
of SEQ ID NO: 37 correspond to the 3'-UTR.
[0359] In addition to phenotypic observations with targeting the
GA20 oxidase_3 and/or GA20 oxidase_5 gene(s), or the GA3 oxidase_1
and/or GA3 oxidase_2 gene(s), for suppression, a semi-dwarf
phenotype is also observed with suppression of the GA20 oxidase_4
gene. The genomic DNA sequence of GA20 oxidase_4 is provided in SEQ
ID NO: 38. For the GA oxidase_4 gene, SEQ ID NO: 38 provides
nucleotides 1-1416 upstream of the 5'-UTR; nucleotides 1417-1543 of
SEQ ID NO: 38 correspond to the 5'-UTR; nucleotides 1544-1995 of
SEQ ID NO: 38 correspond to the first exon; nucleotides 1996-2083
of SEQ ID NO: 38 correspond to the first intron; nucleotides
2084-2411 of SEQ ID NO: 38 correspond to the second exon;
nucleotides 2412-2516 of SEQ ID NO: 38 correspond to the second
intron; nucleotides 2517-2852 of SEQ ID NO: 38 correspond to the
third exon; nucleotides 2853-3066 of SEQ ID NO: 38 correspond to
the 3'-UTR; and nucleotides 3067-4465 of SEQ ID NO: 38 corresponds
to genomic sequence downstream of to the 3'-UTR.
[0360] In an aspect, the present disclosure provides a corn plant
or plurality of corn plants each comprising a recombinant DNA
construct or polynucleotide sequence comprising a transcribable DNA
sequence encoding a non-coding RNA molecule, wherein the non-coding
RNA molecule comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% complementary to
at least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, or at least 27 consecutive nucleotides of a
mRNA molecule encoding an endogenous GA oxidase protein in a corn
plant or corn cell, the endogenous GA oxidase protein being at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 9, 12, 15, 30, and/or 33, and wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter, which can be heterologous with respect
to the transcribable DNA sequence and/or the corn plant.
[0361] Recombinant DNA constructs and transgenic corn plants are
provided herein comprising a GA20 or GA3 oxidase suppression
element or sequence operably linked to a plant expressible
promoter, which can be a constitutive or tissue-specific or
tissue-preferred promoter. Such a tissue-specific or
tissue-preferred promoter can drive expression of its associated GA
oxidase suppression element or sequence in one or more active
GA-producing tissue(s) of the plant to suppress or reduce the level
of active GAs produced in those tissue(s). Such a tissue-specific
or tissue-preferred promoter can drive expression of its associated
GA oxidase suppression construct or transgene during one or more
vegetative stage(s) of development. Such a tissue-specific or
tissue-preferred promoter can also have little or no expression in
one or more cell(s) or tissue(s) of the developing female organ or
ear of the plant to avoid the possibility of off-types in those
reproductive tissues.
[0362] As used herein, a "plant-expressible promoter" refers to a
promoter that drives, causes, or initiates expression of a
transcribable DNA sequence or transgene operably linked to such
promoter in one or more plant cells or tissues, such as one or more
cells or tissues of a corn plant. In an aspect, a plant-expressible
promoter is a constitutive promoter. In another aspect, a
plant-expressible promoter is a vascular promoter. As used herein,
a "vascular promoter" refers to a plant-expressible promoter that
drives, causes or initiates expression of a transcribable DNA
sequence or transgene operably linked to such promoter in one or
more vascular tissue(s) of the plant, even if the promoter is also
expressed in other non-vascular plant cell(s) or tissue(s). Such
vascular tissue(s) can comprise one or more of the phloem, vascular
parenchymal, and/or bundle sheath cell(s) or tissue(s) of the
plant. A "vascular promoter" is distinguished from a constitutive
promoter in that it has a regulated and relatively more limited
pattern of expression that includes one or more vascular tissue(s)
of the plant. A vascular promoter includes both vascular-specific
promoters and vascular-preferred promoters. In another aspect, a
plant-expressible promoter is a leaf promoter. As used herein, a
"leaf promoter" refers to a plant-expressible promoter that drives,
causes or initiates expression of a transcribable DNA sequence or
transgene operably linked to such promoter in one or more leaf
tissue(s) of the plant, even if the promoter is also expressed in
other non-leaf plant cell(s) or tissue(s). A leaf promoter includes
both leaf-specific promoters and leaf-preferred promoters. A "leaf
promoter" is distinguished from a vascular promoter in that it is
expressed more predominantly or exclusively in leaf tissue(s) of
the plant relative to other plant tissues, whereas a vascular
promoter is expressed in vascular tissue(s) more generally
including vascular tissue(s) outside of the leaf, such as the
vascular tissue(s) of the stem, or stem and leaves, of the
plant.
[0363] Promoters that drive enhanced expression in certain tissues
of the plant relative to other plant tissues are referred to as
"tissue-enhanced" or "tissue-preferred" promoters. Thus, a
"tissue-preferred" promoter causes relatively higher or
preferential or predominant expression in a specific tissue(s) of
the plant, but with lower levels of expression in other tissue(s)
of the plant. Promoters that express within a specific tissue(s) of
the plant, with little or no expression in other plant tissues, are
referred to as "tissue-specific" promoters.
[0364] A non-limiting exemplary plant-expressible promoter is the
RTBV promoter. In an aspect, a plant-expressible promoter is an
RTBV promoter. In another aspect, a plant expressible promoter
comprises a DNA sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5% or 100% identical to one or more of
SEQ ID NO: 39, SEQ ID NO: 40, or a functional portion thereof.
[0365] Non-limiting exemplary vascular promoters include a sucrose
synthase promoter, a sucrose transporter promoter, a Sh1 promoter,
Commelina yellow mottle virus (CoYMV) promoter, a wheat dwarf
geminivirus (WDV) large intergenic region (LIR) promoter, a maize
streak geminivirus (MSV) coat protein (CP) promoter, a rice yellow
stripe 1 (YS1)-like promoter, and a rice yellow stripe 2 (OsYSL2)
promoter. In an aspect, a vascular promoter is selected from the
group consisting of a sucrose synthase promoter, a sucrose
transporter promoter, a Sh1 promoter, Commelina yellow mottle virus
(CoYMV) promoter, a wheat dwarf geminivirus (WDV) large intergenic
region (LIR) promoter, a maize streak geminivirus (MSV) coat
protein (CP) promoter, a rice yellow stripe 1 (YS1)-like promoter,
a rice yellow stripe 2 (OsYSL2) promoter, and functional portions
thereof. In an aspect, a vascular promoter comprises a DNA sequence
that is at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%
or 100% identical to one or more of SEQ ID NO: 41, SEQ ID NO: 42,
SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, or a functional
portion thereof.
[0366] Non-limiting exemplary leaf promoters include a RuBisCO
promoter, a PPDK promoter, a FDA promoter, a Nadh-Gogat promoter, a
chlorophyll a/b binding protein gene promoter, a
phosphoenolpyruvate carboxylase (PEPC) promoter, and a Myb gene
promoter. In an aspect, a leaf promoter is selected from the group
consisting of a RuBisCO promoter, a PPDK promoter, a FDA promoter,
a Nadh-Gogat promoter, a chlorophyll a/b binding protein gene
promoter, a phosphoenolpyruvate carboxylase (PEPC) promoter, a Myb
gene promoter, and functional portions thereof. In an aspect, a
leaf promoter comprises a DNA sequence that is at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5% or 100% identical to one
or more of SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, or a
functional portion thereof.
[0367] Non-limiting exemplary constitutive promoters include an
actin promoter, a CaMV 35S or 19S promoter, a plant ubiquitin
promoter, a plant Gos2 promoter, a FMV promoter, a CMV promoter, a
MMV promoter, a PCLSV promoter, an Emu promoter, a tubulin
promoter, a nopaline synthase promoter, an octopine synthase
promoter, a mannopine synthase promoter, and a maize alcohol
dehydrogenase. In an aspect, a constitutive promoter is selected
from the group consisting of an actin promoter, a CaMV 35S or 19S
promoter, a plant ubiquitin promoter, a plant Gos2 promoter, a FMV
promoter, a CMV promoter, a MMV promoter, a PCLSV promoter, an Emu
promoter, a tubulin promoter, a nopaline synthase promoter, an
octopine synthase promoter, a mannopine synthase promoter, a maize
alcohol dehydrogenase, or functional portions thereof. In an
aspect, a constitutive promoter comprises a DNA sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5% or 100%
identical to one or more of SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID
NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55,
SEQ ID NO: 56, SEQ ID NO: 57, or a functional portion thereof.
[0368] In another aspect, the present disclosure provides a corn
plant or plurality of corn plants each comprising a recombinant DNA
construct or polynucleotide sequence comprising a transcribable DNA
sequence encoding a non-coding RNA molecule, wherein the non-coding
RNA molecule comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% complementary to
at least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, or at least 27 consecutive nucleotides of a
mRNA molecule encoding an endogenous GA oxidase gene having at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 9, 12, 15, 30, and/or 33, and wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter, which can be heterologous with respect
to the transcribable DNA sequence and/or the corn plant.
[0369] As provided above, a corn plant or plant part can comprise a
first expression cassette comprising a first sequence encoding a
non-coding RNA molecule that targets one or more GA20 or GA3
oxidase gene(s) for suppression. In an aspect, the non-coding RNA
molecule can target one or more GA20 oxidase gene(s) for
suppression, such as a GA20 oxidase_3 gene, a GA20 oxidase_4 gene,
a GA20 oxidase_5 gene, or any combination thereof. According to
some embodiments, the first expression cassette comprises a first
transcribable DNA sequence encoding a non-coding RNA targeting a
GA20 oxidase_3 gene for suppression. According to other
embodiments, the first expression cassette comprises a first
transcribable DNA sequence encoding a non-coding RNA targeting a
GA20 oxidase_5 gene for suppression. According to yet further
embodiments, the first expression cassette comprises a first
transcribable DNA sequence encoding a non-coding RNA that targets
both the GA20 oxidase_3 gene and the GA20 oxidase_5 gene for
suppression. In addition to targeting a mature mRNA sequence
(including either or both of the untranslated or exonic sequences),
a non-coding RNA molecule can also target the intronic sequences of
a GA20 oxidase gene or transcript.
[0370] For suppression of a GA20 oxidase_3 gene, a first
transcribable DNA sequence comprises a sequence that is at least
80%, at least 81%, at least 82%, at least 83%, at least 84%, at
least 85%, at least 86%, at least 87%, at least 88%, at least 89%,
at least 90%, at least 91%, at least 92%, at least 93%, at least
94%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical or complementary to at
least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, at least 27, at least 28, at least 29, at
least 30, at least 31, at least 32, at least 33, at least 34, at
least 35, at least 36, at least 37, at least 38, at least 39, at
least 40, at least 41, at least 42, at least 43, at least 44, at
least 45, at least 46, at least 47, at least 48, at least 49, at
least 50, at least 51, at least 52, at least 53, at least 54, at
least 55, at least 56, at least 57, at least 58, at least 59, or at
least 60 consecutive nucleotides of a sequence selected from the
group consisting of SEQ ID NO: 7, SEQ ID NO: 8, and SEQ ID NO:
34.
[0371] For suppression of a GA20 oxidase_4 gene, a first
transcribable DNA sequence comprises a sequence that is at least
80%, at least 81%, at least 82%, at least 83%, at least 84%, at
least 85%, at least 86%, at least 87%, at least 88%, at least 89%,
at least 90%, at least 91%, at least 92%, at least 93%, at least
94%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical or complementary to at
least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, at least 27, at least 28, at least 29, at
least 30, at least 31, at least 32, at least 33, at least 34, at
least 35, at least 36, at least 37, at least 38, at least 39, at
least 40, at least 41, at least 42, at least 43, at least 44, at
least 45, at least 46, at least 47, at least 48, at least 49, at
least 50, at least 51, at least 52, at least 53, at least 54, at
least 55, at least 56, at least 57, at least 58, at least 59, or at
least 60 consecutive nucleotides of a sequence selected from the
group consisting of SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO:
38.
[0372] For suppression of a GA20 oxidase_5 gene, a first
transcribable DNA sequence comprises a sequence that is at least
80%, at least 81%, at least 82%, at least 83%, at least 84%, at
least 85%, at least 86%, at least 87%, at least 88%, at least 89%,
at least 90%, at least 91%, at least 92%, at least 93%, at least
94%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical or complementary to at
least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, at least 27, at least 28, at least 29, at
least 30, at least 31, at least 32, at least 33, at least 34, at
least 35, at least 36, at least 37, at least 38, at least 39, at
least 40, at least 41, at least 42, at least 43, at least 44, at
least 45, at least 46, at least 47, at least 48, at least 49, at
least 50, at least 51, at least 52, at least 53, at least 54, at
least 55, at least 56, at least 57, at least 58, at least 59, or at
least 60 consecutive nucleotides of a sequence selected from the
group consisting of SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO:
35.
[0373] For suppression of a GA20 oxidase_3 gene and a GA20
oxidase_5 gene, a transcribable DNA sequence comprises a sequence
that is at least 80%, at least 81%, at least 82%, at least 83%, at
least 84%, at least 85%, at least 86%, at least 87%, at least 88%,
at least 89%, at least 90%, at least 91%, at least 92%, at least
93%, at least 94%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical or
complementary to at least 15, at least 16, at least 17, at least
18, at least 19, at least 20, at least 21, at least 22, at least
23, at least 24, at least 25, at least 26, at least 27, at least
28, at least 29, at least 30, at least 31, at least 32, at least
33, at least 34, at least 35, at least 36, at least 37, at least
38, at least 39, at least 40, at least 41, at least 42, at least
43, at least 44, at least 45, at least 46, at least 47, at least
48, at least 49, at least 50, at least 51, at least 52, at least
53, at least 54, at least 55, at least 56, at least 57, at least
58, at least 59, or at least 60 consecutive nucleotides of a
sequence selected from the group consisting of SEQ ID NO: 7, SEQ ID
NO: 8, and SEQ ID NO: 34; and the transcribable DNA sequence
comprises a sequence that is at least 80%, at least 81%, at least
82%, at least 83%, at least 84%, at least 85%, at least 86%, at
least 87%, at least 88%, at least 89%, at least 90%, at least 91%,
at least 92%, at least 93%, at least 94%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical or complementary to at least 15, at least 16, at
least 17, at least 18, at least 19, at least 20, at least 21, at
least 22, at least 23, at least 24, at least 25, at least 26, at
least 27, at least 28, at least 29, at least 30, at least 31, at
least 32, at least 33, at least 34, at least 35, at least 36, at
least 37, at least 38, at least 39, at least 40, at least 41, at
least 42, at least 43, at least 44, at least 45, at least 46, at
least 47, at least 48, at least 49, at least 50, at least 51, at
least 52, at least 53, at least 54, at least 55, at least 56, at
least 57, at least 58, at least 59, or at least 60 consecutive
nucleotides of a sequence selected from the group consisting of SEQ
ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 35.
[0374] In another aspect, a first expression cassette comprises a
first transcribable DNA sequence encoding a non-coding RNA
targeting a GA3 oxidase gene(s) for suppression in corn, such as a
GA3 oxidase_1 gene or a GA3 oxidase_2 gene. In another aspect, a
first transcribable DNA sequence encoding a non-coding RNA targets
both the GA3 oxidase_1 gene and the GA3 oxidase_2 gene for
suppression. In addition to targeting a mature mRNA sequence
(including either or both of the untranslated or exonic sequences),
a non-coding RNA molecule can also target the intronic sequences of
a GA3 oxidase gene or transcript.
[0375] For suppression of a GA3 oxidase_1 gene, a first
transcribable DNA sequence comprises a sequence that is at least
80%, at least 81%, at least 82%, at least 83%, at least 84%, at
least 85%, at least 86%, at least 87%, at least 88%, at least 89%,
at least 90%, at least 91%, at least 92%, at least 93%, at least
94%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical or complementary to at
least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, at least 27, at least 28, at least 29, at
least 30, at least 31, at least 32, at least 33, at least 34, at
least 35, at least 36, at least 37, at least 38, at least 39, at
least 40, at least 41, at least 42, at least 43, at least 44, at
least 45, at least 46, at least 47, at least 48, at least 49, at
least 50, at least 51, at least 52, at least 53, at least 54, at
least 55, at least 56, at least 57, at least 58, at least 59, or at
least 60 consecutive nucleotides of a sequence selected from the
group consisting of SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO:
36.
[0376] For suppression of a GA3 oxidase_2 gene, a first
transcribable DNA sequence comprises a sequence that is at least
80%, at least 81%, at least 82%, at least 83%, at least 84%, at
least 85%, at least 86%, at least 87%, at least 88%, at least 89%,
at least 90%, at least 91%, at least 92%, at least 93%, at least
94%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical or complementary to at
least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, at least 27, at least 28, at least 29, at
least 30, at least 31, at least 32, at least 33, at least 34, at
least 35, at least 36, at least 37, at least 38, at least 39, at
least 40, at least 41, at least 42, at least 43, at least 44, at
least 45, at least 46, at least 47, at least 48, at least 49, at
least 50, at least 51, at least 52, at least 53, at least 54, at
least 55, at least 56, at least 57, at least 58, at least 59, or at
least 60 consecutive nucleotides of a sequence selected from the
group consisting of SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO:
37.
[0377] For suppression of a GA3 oxidase_1 gene and a GA3 oxidase_2
gene, a transcribable DNA sequence comprises a sequence that is at
least 80%, at least 81%, at least 82%, at least 83%, at least 84%,
at least 85%, at least 86%, at least 87%, at least 88%, at least
89%, at least 90%, at least 91%, at least 92%, at least 93%, at
least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical or complementary to
at least 15, at least 16, at least 17, at least 18, at least 19, at
least 20, at least 21, at least 22, at least 23, at least 24, at
least 25, at least 26, at least 27, at least 28, at least 29, at
least 30, at least 31, at least 32, at least 33, at least 34, at
least 35, at least 36, at least 37, at least 38, at least 39, at
least 40, at least 41, at least 42, at least 43, at least 44, at
least 45, at least 46, at least 47, at least 48, at least 49, at
least 50, at least 51, at least 52, at least 53, at least 54, at
least 55, at least 56, at least 57, at least 58, at least 59, or at
least 60 consecutive nucleotides of a sequence selected from the
group consisting of SEQ ID NO: 28, SEQ ID NO: 29, and SEQ ID NO:
36; and the transcribable DNA sequence comprises a sequence that is
at least 80%, at least 81%, at least 82%, at least 83%, at least
84%, at least 85%, at least 86%, at least 87%, at least 88%, at
least 89%, at least 90%, at least 91%, at least 92%, at least 93%,
at least 94%, at least 95%, at least 96%, at least 97%, at least
98%, at least 99%, at least 99.5%, or 100% identical or
complementary to at least 15, at least 16, at least 17, at least
18, at least 19, at least 20, at least 21, at least 22, at least
23, at least 24, at least 25, at least 26, at least 27, at least
28, at least 29, at least 30, at least 31, at least 32, at least
33, at least 34, at least 35, at least 36, at least 37, at least
38, at least 39, at least 40, at least 41, at least 42, at least
43, at least 44, at least 45, at least 46, at least 47, at least
48, at least 49, at least 50, at least 51, at least 52, at least
53, at least 54, at least 55, at least 56, at least 57, at least
58, at least 59, or at least 60 consecutive nucleotides of a
sequence selected from the group consisting of SEQ ID NO: 31, SEQ
ID NO: 32 and SEQ ID NO: 37.
[0378] In an aspect, a mutant allele of an endogenous GA20
oxidase_3 locus comprises a DNA segment inserted into the
endogenous GA20 oxidase_3 locus, where the DNA segment encodes an
antisense RNA sequence that is at least 70%, at least 80%, at least
90%, at least 91%, at least 92%, at least 93%, at least 94%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
or 100% complementary to at least 20, at least 30, at least 40, at
least 50, at least 100, at least 200, at least 300, at least 400,
at least 500, at least 600, at least 700, at least 800, at least
900, or at least 1000 consecutive nucleotides of one or more of SEQ
ID NOs: 182-184 and 186-188, and where the mutant allele of the
endogenous GA20 oxidase_3 locus produces an RNA transcript
comprising the antisense RNA sequence.
[0379] In an aspect, a mutant allele of an endogenous GA20
oxidase_5 locus comprises a DNA segment inserted into the
endogenous GA20 oxidase_5 locus, where the DNA segment encodes an
antisense RNA sequence that is at least 70%, at least 80%, at least
90%, at least 91%, at least 92%, at least 93%, at least 94%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
or 100% complementary to at least 20, at least 30, at least 40, at
least 50, at least 100, at least 200, at least 300, at least 400,
at least 500, at least 600, at least 700, at least 800, at least
900, or at least 1000 consecutive nucleotides of one or more of SEQ
ID NOs: 182-184 and 186-188, and where the mutant allele of the
endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence.
[0380] In an aspect, a mutant allele of the endogenous GA20
oxidase_3 locus suppresses the expression of a wild-type allele of
the endogenous GA20 oxidase_3 locus, a wild-type allele of the
endogenous GA20 oxidase_5 locus, or both. In an aspect, a mutant
allele of the endogenous GA20 oxidase_5 locus suppresses the
expression of a wild-type allele of the endogenous GA20 oxidase_3
locus, a wild-type allele of the endogenous GA20 oxidase_5 locus,
or both. In an aspect, a mutant allele comprises a deletion of at
least one portion of an endogenous GA20 oxidase_3 locus.
[0381] Without being bound by any scientific theory, if a genomic
region between the neighboring Zm.GA20ox5 and Zm.SAMT genes
(including possibly all or part of those genes) is deleted, then
the endogenous Zm.SAMT gene promoter can drive expression of an
antisense RNA transcript through all or part of the Zm.GA20ox5 gene
that can hybridize to a separate RNA transcript expressed from one
or both of the copies or alleles of the Zm.GA20ox5 and/or
Zm.GA20ox3 gene(s). Thus, a mutant allele having a deletion between
the Zm.GA20ox5 and Zm.SAMT genes can behave as a dominant negative
mutation or allele by causing suppression or silencing of one or
both (wild-type and/or mutant) copies or alleles of the endogenous
Zm.GA20ox5 gene, in addition to possible further suppression or
silencing of one or both copies or alleles of the endogenous
Zm.GA20ox3 gene.
[0382] According to aspects of the present disclosure, a mutant or
edited allele of the endogenous GA20 oxidase_5 (GA20ox5) gene or
locus is provided comprising a deletion between the neighboring
Zm.GA20ox5 and Zm.SAMT genes, such that an antisense RNA molecule
that is complementary to all or part of the coding sequence of the
GA20ox5 gene may be transcribed under the control of the endogenous
Zm.SAMT gene promoter. It is contemplated that the antisense RNA
molecule transcribed from the mutant or edited allele of the
endogenous GA20 oxidase_5 gene or locus may affect the expression
level(s) of the GA20 oxidase_5 and/or endogenous GA20 oxidase_3
gene(s) through different mechanisms, such as nonsense mediated
decay, non-stop decay, no-go decay, DNA or histone methylation or
other epigenetic changes, inhibition or decreased efficiency of
transcription and/or translation, ribosomal interference,
interference with mRNA processing or splicing, and/or
ubiquitin-mediated protein degradation via the proteasome. See,
e.g., Nickless, A. et al., "Control of gene expression through the
nonsense-mediated RNA decay pathway", Cell Biosci 7:26 (2017);
Karamyshev, A. et al., "Lost in Translation: Ribosome-Associated
mRNA and Protein Quality Controls", Frontiers in Genetics 9:431
(2018); Inada, T., "Quality controls induced by aberrant
translation", Nucleic Acids Res 48:3 (2020); and Szadeczky-Kardoss,
I. et al., "The nonstop decay and the RNA silencing systems operate
cooperatively in plants", Nucleic Acids Res 46:9 (2018), the entire
contents and disclosures of which are incorporated herein by
reference. Each of these different mechanisms may act alternatively
or in addition to RNA interference (RNAi), transcriptional gene
silencing (PGS) and/or post transcriptional gene silencing (PTGS)
mechanisms. See, e.g., Wilson, R. C. et al., "Molecular Mechanisms
of RNA Interference", Annu Rev Biophysics 42:217-39 (2013); and
Guo, Q. et al., "RNA Silencing in Plants: Mechanism, Technologies
and Applications in Horticulture Crops", Current Genomics
17:476-489 (2016), the entire contents and disclosures of which is
incorporated herein by reference. Some of the above mechanisms may
reduce expression of the edited allele itself, while others may
also reduce the expression of other copy/-ies or allele(s) of the
endogenous GA20 oxidase_5 and/or GA20 oxidase_3 locus/loci or
gene(s). Indeed, it is envisioned that the edited endogenous GA20
oxidase_5 locus, gene or allele may not only reduce or eliminate
its own expression and/or activity level, but may also have a
dominant or semi-dominant effect(s) on the other copy/-ies or
allele(s) of the endogenous GA20 oxidase_5 and/or GA20 oxidase_3
locus/loci or gene(s). Such dominant or semi-dominant effect(s) on
the GA20 oxidase_5 and/or GA20 oxidase_3 gene(s) may operate
through non-canonical suppression mechanisms that do not involve
RNAi and/or formation of targeted small RNAs at a significant or
detectable level.
[0383] As used herein, an "intergenic region" or "intergenic
sequence" refers to a genomic region or a polynucleotide sequence
located in between transcribed regions of two neighboring genes.
For example, the endogenous Zm.GA20ox5 gene and its neighboring
gene in the corn or maize genome, the s-adenosyl methyl transferase
(SAMT) or Zm.SAMT gene, contains an intergenic region between the
3' UTR of the Zm. GA20ox5 gene and the 3' UTR of the Zm.SAMT
gene.
[0384] In the corn genome, the Zm.GA20ox5 gene located next to the
Zm.SAMT gene. These two genes are separated by an intergenic region
of about 550 bp, with the Zm.SAMT gene positioned downstream and
oriented in the opposite orientation relative to the Zm.GA20ox5
gene. A reference genomic sequence of the region encompassing the
Zm.GA20ox5 and Zm.SAMT genes is provided in SEQ ID NOs. 226 and
227. SEQ ID NO. 226 represents the sequence of the sense strand of
the Zm.GA20ox5 gene encompassing both Zm.GA20ox5 and Zm.SAMT genes.
SEQ ID NO: 226 partially overlaps with SEQ ID NO: 222 and has a
shorter Zm.GA20ox5 upstream sequence and a longer Zm.GA20ox5
downstream sequence compared to the SEQ ID NO: 222. SEQ ID NO. 227
represents the sequence of the sense strand of the Zm.SAMT gene
(i.e., the antisense strand of the Zm.GA20ox5 gene) encompassing
both Zm.GA20ox5 and Zm.SAMT genes.
[0385] In an aspect, a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
transcription termination sequence of the endogenous Zm.SAMT gene,
and wherein the mutant allele produces a RNA molecule comprising an
antisense sequence complementary to all or part of the sense strand
of the endogenous GA20 oxidase_5 gene.
[0386] In an aspect, a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene.
[0387] In an aspect, a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of one or
more of the following: 5' UTR, 1.sup.st exon, 1.sup.st intron,
2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1.sup.st exon, 1.sup.st intron,
2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3.sup.rd intron,
4.sup.th exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th intron,
6.sup.th exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th intron,
8.sup.th exon, 3' UTR, and any portion thereof, of the endogenous
Zm.SAMT gene.
[0388] In an aspect, a mutant allele comprises a genome
modification which results in the transcription of an antisense
strand of at least an exon, an intron, or an untranslated region
(UTR) of an endogenous GA20 oxidase_5 gene, or any portion
thereof.
[0389] In an aspect, a mutant allele comprises the Zm.SAMT gene
promoter, or a functional part thereof, operably linked to at least
one transcribable antisense sequence of at least an exon, intron or
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof.
[0390] In an aspect, a mutant allele comprises a sequence selected
from the group consisting of SEQ ID NOs: 304-322.
[0391] In an aspect, a mutant allele comprises a first sequence and
a second sequence; wherein the first sequence comprises one or more
of the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon,
2.sup.nd intron, 3.sup.rd exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.GA20 oxidase_5 gene; and wherein the second sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3rd intron,
4.sup.th exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th intron,
6.sup.th exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th intron,
8.sup.th exon, 3' UTR, and any complementary sequence thereof, and
any portion of the foregoing, of the endogenous Zm.SAMT gene;
wherein the first sequence and the second sequence are contiguous
or separated only by an intervening sequence of fewer than 555
nucleotides. In an aspect, a mutant allele comprises a genomic
deletion relative to a wild type allele of the endogenous GA20
oxidase_5 locus, wherein the genomic deletion is flanked by a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene. In an aspect, a mutant allele comprises a
genomic sequence comprising a first sequence and a second sequence;
wherein the first sequence comprises at least 15, at least 20, at
least 25, at least 30, at least 35, at least 40, at least 45, at
least 50, at least 100, at least 200, at least 300, at least 400,
at least 500, at least 750, or at least 1000 consecutive
nucleotides of one or more of SEQ ID NOs: 228-235 and 276-283;
wherein the second sequence comprises at least 15, at least 20, at
least 25, at least 30, at least 35, at least 40, at least 45, at
least 50, at least 100, at least 200, at least 300, at least 400,
at least 500, at least 750, or at least 1000 consecutive
nucleotides of one or more of SEQ ID NOs: 235-276; and wherein the
genomic sequence is at least 50, at least 100, at least 250, at
least 500, or at least 1000 consecutive nucleotides in length,
and/or fewer than 9000 consecutive nucleotides in length. In an
aspect, a first sequence comprises one or more of SEQ ID NOs:
228-235 and 276-283, or any portion thereof, and a second sequence
comprises one or more of SEQ ID NOs: 235-276, or any portion
thereof. In an aspect, a first sequence comprises one or more of
SEQ ID NOs: 226-235 and 276-283, or any portion thereof, and a
second sequence comprises one or more of SEQ ID NOs: 226, 227, and
235-276, or any portion thereof. In an aspect, a first sequence
comprises at least 15, at least 20, at least 25, at least 30, at
least 35, at least 40, at least 45, at least 50, at least 100, at
least 200, at least 300, at least 400, at least 500, at least 750,
or at least 1000 consecutive nucleotides of one or more of SEQ ID
NOs: 226-235 and 276-283, and a second sequence comprises at least
15, at least 20, at least 25, at least 30, at least 35, at least
40, at least 45, at least 50, at least 100, at least 200, at least
300, at least 400, at least 500, at least 750, or at least 1000
consecutive nucleotides of one or more of SEQ ID NOs: 226, 227, and
235-276.
[0392] In an aspect, a genome modification further comprises the
deletion of at least a portion of the transcription termination
sequence of an endogenous GA20 oxidase_5 gene. In an aspect, a
genome modification comprises a deletion of one or both of the
transcription termination sequences of an endogenous GA20 oxidase_5
gene and an endogenous Zm.SAMT gene. In an aspect, a genome
modification comprises a deletion of at least 10, at least 15, at
least 20, at least 25, at least 30, at least 35, at least 40, at
least 45, at least 50, at least 100, at least 200, at least 300, at
least 400, at least 500, at least 750, or at least 1000 consecutive
nucleotides of the intergenic region between the endogenous GA20
oxidase_5 and SAMT genes. In an aspect, a genome modification
comprises a deletion of the entire intergenic region between the
endogenous GA20 oxidase_5 and SAMT genes. In an aspect, a genome
modification comprises a deletion of one or more sequence elements
selected from the group consisting of the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any portion of the foregoing, of an endogenous GA20
oxidase_5 gene. In an aspect, a genome modification comprises a
deletion of one or more sequence elements selected from the group
consisting of the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd
exon, 2.sup.nd intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th
exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th
exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th
exon, 3' UTR, and any portion of the foregoing, of the endogenous
Zm.SAMT locus. In an aspect, a genome modification results in the
production of an RNA molecule comprising an antisense sequence from
a genomic segment of selected from the group consisting of an exon,
a portion of an exon, an intron, a portion of an intron, a 5' or 3'
untranslated region (UTR), a portion of an UTR, and any combination
of the foregoing, of the endogenous GA20 oxidase_5 locus. In an
aspect, a genome modification comprises two or more, three or more,
four or more, five or more, or six or more non-contiguous
deletions.
[0393] In an aspect, a genomic deletion comprises a deletion of the
intergenic region between the endogenous Zm.GA20 oxidase_5 and
Zm.SAMT genes. In an aspect, a genomic deletion has a length of at
least 50, at least 100, at least 250, at least 500, at least 750,
at least 1000, at least 2500, or at least 5000 nucleotides. In an
aspect, a genomic deletion has a length of at most 7500, at most
7000, at most 6000, at most 5000, at most 4000, at most 3000, at
most 2500, at most 2000, at most 1000, or at most 500 nucleotides.
In an aspect, a genomic deletion corresponds to a deletion of one
or more genomic regions comprising a sequence selected from the
group consisting of SEQ ID NOs: 228-283. In an aspect a genome
deletion results in the production of an RNA transcript comprising
an antisense sequence from a genomic segment of the endogenous GA20
oxidase_5 locus selected from the group consisting of an exon,
portion of an exon, an intron, portion of an intron, an
untranslated region (UTR), portion of an UTR, and any combination
of the foregoing.
[0394] In an aspect, a mutant allele comprises the endogenous
Zm.SAMT gene promoter, or a portion thereof, operably linked to a
transcribable DNA sequence encoding a RNA molecule that causes
suppression of one or both of the endogenous GA20 oxidase_3 gene
and the endogenous GA20 oxidase_5 gene. In an aspect, a mutant
allele comprises the endogenous Zm.SAMT gene promoter, or a portion
thereof, operably linked to a transcribable DNA sequence encoding a
RNA molecule comprising an antisense sequence that is at least 80%,
at least 85%, at least 90%, at least 91%, at least 92%, at least
93%, at least 94%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, or 100% complementary to all or part of
the endogenous GA20 oxidase_3 or GA20 oxidase_5 gene. In an aspect,
a transcribable DNA sequence is at least 80%, at least 85%, at
least 90%, at least 91%, at least 92%, at least 93%, at least 94%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, or 100% complementary to an RNA transcript sequence, or a
portion thereof, encoded by an endogenous GA20 oxidase_3 or GA20
oxidase_5 gene. In an aspect, a transcribable DNA sequence is at
least 80% complementary to at least 15, at least 16, at least 17,
at least 18, at least 19, at least 20, at least 21, at least 22, at
least 23, at least 24, at least 25, at least 26, at least 27, at
least 28, at least 29, at least 30, at least 40, at least 50, at
least 60, at least 70, at least 80, at least 90, at least 100, or
at least 200 consecutive nucleotides of one or more of SEQ ID NOs:
218-220, 222-224, 226, and 228-255. In an aspect, a transcribable
DNA sequence is at least 80% complementary to at least 15, at least
16, at least 17, at least 18, at least 19, at least 20, at least
21, at least 22, at least 23, at least 24, at least 25, at least
26, at least 27, at least 28, at least 29, at least 30, at least
40, at least 50, at least 60, at least 70, at least 80, at least
90, at least 100, or at least 200 consecutive nucleotides of one or
more of SEQ ID NOs: 222-224 and 228-235.
[0395] In an aspect, a DNA segment comprises a nucleotide sequence
originating from the endogenous GA20 oxidase_3 locus. In an aspect,
a DNA segment corresponds to an inverted genomic fragment of the
endogenous GA20 oxidase_3 locus. In an aspect, a DNA segment
comprises a nucleotide sequence originating from an endogenous GA20
oxidase_5 locus. In an aspect, a DNA segment is inserted near or
adjacent to a corresponding endogenous DNA segment of the
endogenous GA20 oxidase_3 locus. In an aspect, the sense strand of
a DNA segment comprises a sequence at least 70%, at least 80%, at
least 90%, at least 91%, at least 92%, at least 93%, at least 94%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, or 100% complementary to an exon sequence of the endogenous
GA20 oxidase_3 or GA20 oxidase_5 locus. In an aspect, the sense
strand of a DNA segment comprises a sequence at least 70%, at least
80%, at least 90%, at least 91%, at least 92%, at least 93%, at
least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, or 100% complementary to an untranslated region (UTR)
sequence of the endogenous GA20 oxidase_3 or GA20 oxidase_5 locus.
In an aspect, the sense strand of a DNA segment comprises a
sequence at least 70%, at least 80%, at least 90%, at least 91%, at
least 92%, at least 93%, at least 94%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, or 100% complementary to
an exon sequence and an intron sequence of the endogenous GA20
oxidase_3 or GA20 oxidase_5 locus, the exon sequence and the intron
sequence being contiguous within the endogenous locus. In an
aspect, a DNA segment comprises a sequence having at least at least
70%, at least 80%, at least 90%, at least 91%, at least 92%, at
least 93%, at least 94%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, or 100% identity to one or more of SEQ
ID NOs: 194, 195, 207, 209, 211, 213, and 217.
[0396] In an aspect, a corresponding endogenous sequence of an RNA
transcript is at least 85%, at least 90%, at least 91%, at least
92%, at least 93%, at least 94%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, or 100% identical to at
least 20, at least 30, at least 40, at least 50, at least 100, at
least 200, at least 300, at least 400, at least 500, at least 600,
at least 700, at least 800, at least 900, or at least 1000
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188. In an aspect, an antisense RNA sequence forms a stem-loop
structure with the corresponding endogenous sequence of the RNA
transcript. In an aspect, an RNA transcript further comprises one
or more sequence elements of the endogenous GA20 oxidase_5 locus
selected from the group consisting of 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any portion thereof.
[0397] In an aspect, an RNA transcript sequence comprises a
sequence that is at least 85%, at least 90%, at least 91%, at least
92%, at least 93%, at least 94%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, or 100% identical to at
least 20, at least 30, at least 40, at least 50, at least 100, at
least 200, at least 300, at least 400, at least 500, at least 600,
at least 700, at least 800, at least 900, or at least 1000
consecutive nucleotides of one or more of SEQ ID NOs: 218-220,
222-224, 226, and 228-255. In an aspect, an RNA transcript sequence
comprises a sequence that is at least 85%, at least 90%, at least
91%, at least 92%, at least 93%, at least 94%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, or 100%
identical to at least 20, at least 30, at least 40, at least 50, at
least 100, at least 200, at least 300, at least 400, at least 500,
at least 600, at least 700, at least 800, at least 900, or at least
1000 consecutive nucleotides of one or more of SEQ ID NOs: 222-224
and 228-235.
[0398] In an aspect, an inserted DNA segment is located upstream
(e.g. on the 5' side) of a corresponding endogenous DNA segment. In
an aspect, an inserted DNA segment is located downstream (e.g. on
the 3' side) of a corresponding endogenous DNA segment. In an
aspect, a DNA segment comprises a length of at least 10, at least
15, at least 20, at least 25, at least 30, at least 40, at least
50, at least 75, at least 100, at least 200, at least 300, at least
400, at least 500, at least 600, at least 700, at least 800, or at
least 900 nucleotides. In an aspect, a DNA segment comprises a
length of at most 2000, at most 1500, at most 1000, at most 900, at
most 800, at most 700, at most 600, at most 500, at most 400, at
most 300, at most 200, at most 100, at most 75, at most 50, or at
most 25 nucleotides.
[0399] In an aspect, an inserted DNA segment and the corresponding
endogenous DNA segment of a mutant allele are separated by an
intervening DNA sequence. In an aspect, an intervening DNA sequence
comprises a length of at least 1 nucleotide. In an aspect, an
intervening DNA sequence comprises a length of at least 2, at least
3, at least 4, at least 5, at least 10, at least 20, at least 30,
at least 40, at least 50, at least 75, at least 100, at least 200,
at least 300, at least 400, at least 500, at least 600, at least
700, at least 800, at least 900, at least 1000, at least 1500, at
least 2000, at least 2500, at least 3000, or at least 40000
consecutive nucleotides. In an aspect, an intervening DNA sequence
comprises at most 5000, at most 4000, at most 3000, at most 2000,
at most 1500, at most 1000, at most 750, at most 500, at most 250,
at most 100, at most 75, at most 50, at most 25, at most 10, or at
most 5 consecutive nucleotides.
[0400] In an aspect, an intervening DNA sequence encodes an
intervening RNA sequence between an antisense RNA sequence and a
corresponding endogenous sequence of an RNA transcript. In an
aspect, an RNA transcript forms a stem-loop structure with an
intervening RNA sequence forming the loop portion of the stem-loop
structure. In an aspect, a stem-loop secondary structure comprises
a near-perfect-complement stem with mismatches. In an aspect, a
near-perfect-complement stem comprises fewer than 10, fewer than 9,
fewer than 8, fewer than 7, fewer than 6, fewer than 5, fewer than
4, fewer than 3, or fewer than 2 mismatches. In an aspect, a
stem-loop secondary structure comprises a perfect complement stem
with zero mismatches. In an aspect, an intervening DNA sequence
comprises an intron sequence. In an aspect, an intervening DNA
sequence does not comprise an intron sequence.
[0401] In an aspect, an intervening DNA sequence comprises a native
sequence of the endogenous GA20 oxidase_3 locus. In an aspect, an
intervening DNA sequence comprises an exogenous sequence inserted
into the endogenous GA20 oxidase_3 locus. In an aspect, an
intervening DNA sequence comprises a native sequence of the
endogenous GA20 oxidase_5 locus. In an aspect, an intervening DNA
sequence comprises an exogenous sequence inserted into the
endogenous GA20 oxidase_5 locus. In an aspect, a DNA segment is
inserted within a region selected from the group consisting of 5'
untranslated region (UTR), 1.sup.st exon, 1.sup.st intron, 2.sup.nd
exon, 2.sup.nd intron, 3.sup.rd exon and 3' UTR of the endogenous
GA20 oxidase_3 locus, and a combination thereof. In an aspect a DNA
segment is inserted at a genomic site recognized by a targeted
editing technique to create a double-stranded break (DSB). In an
aspect, a DNA segment comprises a nucleotide sequence originating
from an endogenous GA20 oxidase_3 locus. In an aspect, a DNA
segment corresponds to an inverted genomic fragment of the
endogenous GA20 oxidase_3 locus. In an aspect, a DNA segment
comprises a nucleotide sequence originating from the endogenous
GA20 oxidase_5 locus. In an aspect, a DNA segment corresponds to an
inverted genomic fragment of the endogenous GA20 oxidase_5 locus.
In an aspect, a DNA segment is inserted near or adjacent to a
corresponding endogenous DNA segment of the endogenous GA20
oxidase_5 locus.
[0402] In an aspect, this disclosure provides a corn plant where
the level of one or more active GAs in at least one internode
tissue of the stem or stalk of the modified corn plant is lower
than the same internode tissue of an unmodified control plant.
[0403] Any method known in the art for suppression of a target gene
can be used to suppress GA oxidase or brachytic gene(s) according
to aspects of the present disclosure including expression of
antisense RNAs, double stranded RNAs (dsRNAs) or inverted repeat
RNA sequences, or via co-suppression or RNA interference (RNAi)
through expression of small interfering RNAs (siRNAs), short
hairpin RNAs (shRNAs), trans-acting siRNAs (ta-siRNAs), or micro
RNAs (miRNAs). Collectively, antisense RNAs, dsRNAs, inverted
repeat RNA sequences, siRNAs, shRNAs, ta-siRNAs, and miRNAs are
referred to herein as "non-coding RNAs." Furthermore, sense and/or
antisense RNA molecules can be used that target the non-coding
genomic sequences or regions within or near a gene to cause
silencing of the gene. Accordingly, any of these methods can be
used for the targeted suppression of an endogenous GA oxidase
gene(s) or br2 in a tissue-specific or tissue-preferred manner.
See, e.g., U.S. Patent Application Publication Nos. 2009/0070898,
2011/0296555, and 2011/0035839, the contents and disclosures of
which are incorporated herein by reference.
[0404] In an aspect, an RNA molecule is an antisense RNA.
[0405] In an aspect, at least a portion of an antisense RNA
sequence is at least 70%, at least 80%, at least 90%, at least 91%,
at least 92%, at least 93%, at least 94%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, or 100%
complementary to a corresponding endogenous sequence of an RNA
transcript. In an aspect, an antisense RNA sequence hybridizes to a
corresponding endogenous sequence of an RNA transcript. In an
aspect, an antisense RNA sequence encoded by an inserted DNA
segment hybridizes to a corresponding endogenous sequence of an RNA
transcript encoded by a corresponding endogenous DNA segment. In an
aspect, an antisense RNA sequence forms a stem-loop structure with
a corresponding endogenous sequence of an RNA transcript.
[0406] In an aspect, a mutant allele produces a RNA molecule
comprising an antisense sequence that is at least 80%, at least
85%, at least 90%, at least 91%, at least 92%, at least 93%, at
least 94%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, or 100% complementary to a RNA transcript sequence,
or a portion thereof, encoded by the endogenous GA20 oxidase_5
gene. In an aspect, an anti sense sequence of an RNA molecule is at
least 80%, at least 85%, at least 90%, at least 91%, at least 92%,
at least 93%, at least 94%, at least 95%, at least 96%, at least
97%, at least 98%, at least 99%, or 100% complementary to at least
15, at least 16, at least 17, at least 18, at least 19, at least
20, at least 21, at least 22, at least 23, at least 24, at least
25, at least 26, at least 27, at least 28, at least 29, at least
30, at least 40, at least 50, at least 75, at least 100, or at
least 200 consecutive nucleotides of one or more of SEQ ID NOs:
218-220, 222-224, 226, and 228-255. In an aspect, an antisense
sequence of an RNA molecule is at least 80%, at least 85%, at least
90%, at least 91%, at least 92%, at least 93%, at least 94%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
or 100% complementary to at least 15, at least 16, at least 17, at
least 18, at least 19, at least 20, at least 21, at least 22, at
least 23, at least 24, at least 25, at least 26, at least 27, at
least 28, at least 29, at least 30, at least 40, at least 50, at
least 75, at least 100, or at least 200 consecutive nucleotides of
one or more of SEQ ID NOs: 222-224 and 228-235.
[0407] In an aspect, an antisense sequence can hybridize with an
RNA transcript encoded by a wild-type allele of one or both of the
endogenous GA20 oxidase_3 gene and the endogenous GA20 oxidase_5
gene. In an aspect, an antisense sequence can hybridize with a
sense RNA transcript encoded by an endogenous GA20 oxidase_5 gene.
In an aspect, an antisense sequence can hybridize with a sense RNA
transcript encoded by the mutant allele of the endogenous GA20
oxidase_5 gene. In an aspect, hybridization between an antisense
RNA and a sense RNA can cause suppression of a wild-type or mutant
allele of the endogenous GA20 oxidase_3 gene, a wild-type or mutant
allele of the endogenous GA20 oxidase_5 gene, or a wild-type or
mutant allele of both genes.
[0408] In an aspect, a sense RNA transcript encoded by the mutant
allele of the endogenous GA20 oxidase_5 gene is shortened or
truncated relative to a wild-type allele of the endogenous GA20
oxidase_5 gene.
[0409] In an aspect, a mutant allele can suppress the expression of
a wild-type allele of the endogenous GA20 oxidase_3 locus, a
wild-type allele of the endogenous GA20 oxidase_5 locus, or
both.
[0410] In an aspect, an expression level(s) of one or more
endogenous GA20 oxidase and/or GA3 oxidase gene(s) is/are reduced
or eliminated in the corn plant, thereby suppressing the endogenous
GA20 oxidase and/or GA3 oxidase gene(s).
[0411] According to an aspect, a corn plant is provided having the
expression level(s) of one or more GA20 oxidase gene(s) reduced in
at least one plant tissue by at least 5%, at least 10%, at least
20%, at least 25%, at least 30%, at least 40%, at least 50%, at
least 60%, at least 70%, at least 75%, at least 80%, at least 90%,
or 100%, as compared to a control corn plant.
[0412] According to an aspect, a corn plant is provided having the
expression level(s) of one or more GA3 oxidase gene(s) reduced in
at least one plant tissue by at least 5%, at least 10%, at least
20%, at least 25%, at least 30%, at least 40%, at least 50%, at
least 60%, at least 70%, at least 75%, at least 80%, at least 90%,
or 100%, as compared to a control corn plant.
[0413] According to an aspect, a corn plant is provided having the
expression level(s) of one or more GA20 oxidase gene(s) reduced in
at least one plant tissue by 5%-20%, 5%-25%, 5%-30%, 5%-40%,
5%-50%, 5%-60%, 5%-70%, 5%-75%, 5%-80%, 5%-90%, 5%-100%, 75%-100%,
50%-100%, 50%-90%, 50%-75%, 25%-75%, 30%-80%, or 10%-75%, as
compared to a control corn plant.
[0414] According to an aspect, a corn plant is provided having the
expression level(s) of one or more GA3 oxidase gene(s) reduced in
at least one plant tissue by 5%-20%, 5%-25%, 5%-30%, 5%-40%,
5%-50%, 5%-60%, 5%-70%, 5%-75%, 5%-80%, 5%-90%, 5%-100%, 75%-100%,
50%-100%, 50%-90%, 50%-75%, 25%-75%, 30%-80%, or 10%-75%, as
compared to a control corn plant.
[0415] According to an aspect, the at least one tissue of a corn
plant having a reduced expression level of a GA20 oxidase and/or
GA3 oxidase gene(s) includes one or more active GA producing
tissue(s) of the plant, such as the vascular and/or leaf tissue(s)
of the plant, during one or more vegetative stage(s) of
development.
[0416] In an aspect, suppression of an endogenous GA20 oxidase gene
or a GA3 oxidase gene is tissue-specific (e.g., only in leaf and/or
vascular tissue). Suppression of a GA20 oxidase gene can be
constitutive and/or vascular or leaf tissue specific or preferred.
In other aspects, suppression of a GA20 oxidase gene or a GA3
oxidase gene is constitutive and not tissue-specific. According to
an aspect, expression of an endogenous GA20 oxidase gene and/or a
GA3 oxidase gene is reduced in one or more tissue types (e.g., in
leaf and/or vascular tissue(s)) of a modified or transgenic plant
as compared to the same tissue(s) of a control plant.
[0417] In an aspect, at least 10% of the corn plants in a field
comprise a mutation in a GA20 oxidase locus as compared to a
wildtype GA20 oxidase locus. In an aspect, at least 20% of the corn
plants in a field comprise a mutation in a GA20 oxidase locus as
compared to a wildtype GA20 oxidase locus. In an aspect, at least
30% of the corn plants in a field comprise a mutation in a GA20
oxidase locus as compared to a wildtype GA20 oxidase locus. In an
aspect, at least 40% of the corn plants in a field comprise a
mutation in a GA20 oxidase locus as compared to a wildtype GA20
oxidase locus. In an aspect, at least 50% of the corn plants in a
field comprise a mutation in a GA20 oxidase locus as compared to a
wildtype GA20 oxidase locus. In an aspect, at least 60% of the corn
plants in a field comprise a mutation in a GA20 oxidase locus as
compared to a wildtype GA20 oxidase locus. In an aspect, at least
70% of the corn plants in a field comprise a mutation in a GA20
oxidase locus as compared to a wildtype GA20 oxidase locus. In an
aspect, at least 80% of the corn plants in a field comprise a
mutation in a GA20 oxidase locus as compared to a wildtype GA20
oxidase locus. In an aspect, at least 90% of the corn plants in a
field comprise a mutation in a GA20 oxidase locus as compared to a
wildtype GA20 oxidase locus. In an aspect, 100% of the corn plants
in a field comprise a mutation in a GA20 oxidase locus as compared
to a wildtype GA20 oxidase locus.
[0418] In an aspect, between 1% and 100% of the corn plants in a
field comprise a mutation in a GA20 oxidase locus as compared to a
wildtype GA20 oxidase locus. In an aspect, between 10% and 100% of
the corn plants in a field comprise a mutation in a GA20 oxidase
locus as compared to a wildtype GA20 oxidase locus. In an aspect,
between 20% and 100% of the corn plants in a field comprise a
mutation in a GA20 oxidase locus as compared to a wildtype GA20
oxidase locus. In an aspect, between 30% and 100% of the corn
plants in a field comprise a mutation in a GA20 oxidase locus as
compared to a wildtype GA20 oxidase locus. In an aspect, between
40% and 100% of the corn plants in a field comprise a mutation in a
GA20 oxidase locus as compared to a wildtype GA20 oxidase locus. In
an aspect, between 50% and 100% of the corn plants in a field
comprise a mutation in a GA20 oxidase locus as compared to a
wildtype GA20 oxidase locus. In an aspect, between 60% and 100% of
the corn plants in a field comprise a mutation in a GA20 oxidase
locus as compared to a wildtype GA20 oxidase locus. In an aspect,
between 70% and 100% of the corn plants in a field comprise a
mutation in a GA20 oxidase locus as compared to a wildtype GA20
oxidase locus. In an aspect, between 80% and 100% of the corn
plants in a field comprise a mutation in a GA20 oxidase locus as
compared to a wildtype GA20 oxidase locus. In an aspect, between
90% and 100% of the corn plants in a field comprise a mutation in a
GA20 oxidase locus as compared to a wildtype GA20 oxidase
locus.
[0419] In an aspect, at least 10% of the corn plants in a field
comprise a mutation in a GA3 oxidase locus as compared to a
wildtype GA3 oxidase locus. In an aspect, at least 20% of the corn
plants in a field comprise a mutation in a GA3 oxidase locus as
compared to a wildtype GA3 oxidase locus. In an aspect, at least
30% of the corn plants in a field comprise a mutation in a GA3
oxidase locus as compared to a wildtype GA3 oxidase locus. In an
aspect, at least 40% of the corn plants in a field comprise a
mutation in a GA3 oxidase locus as compared to a wildtype GA3
oxidase locus. In an aspect, at least 50% of the corn plants in a
field comprise a mutation in a GA3 oxidase locus as compared to a
wildtype GA3 oxidase locus. In an aspect, at least 60% of the corn
plants in a field comprise a mutation in a GA3 oxidase locus as
compared to a wildtype GA3 oxidase locus. In an aspect, at least
70% of the corn plants in a field comprise a mutation in a GA3
oxidase locus as compared to a wildtype GA3 oxidase locus. In an
aspect, at least 80% of the corn plants in a field comprise a
mutation in a GA3 oxidase locus as compared to a wildtype GA3
oxidase locus. In an aspect, at least 90% of the corn plants in a
field comprise a mutation in a GA3 oxidase locus as compared to a
wildtype GA3 oxidase locus. In an aspect, 100% of the corn plants
in a field comprise a mutation in a GA3 oxidase locus as compared
to a wildtype GA3 oxidase locus.
[0420] In an aspect, between 1% and 100% of the corn plants in a
field comprise a mutation in a GA3 oxidase locus as compared to a
wildtype GA3 oxidase locus. In an aspect, between 10% and 100% of
the corn plants in a field comprise a mutation in a GA3 oxidase
locus as compared to a wildtype GA3 oxidase locus. In an aspect,
between 20% and 100% of the corn plants in a field comprise a
mutation in a GA3 oxidase locus as compared to a wildtype GA3
oxidase locus. In an aspect, between 30% and 100% of the corn
plants in a field comprise a mutation in a GA3 oxidase locus as
compared to a wildtype GA3 oxidase locus. In an aspect, between 40%
and 100% of the corn plants in a field comprise a mutation in a GA3
oxidase locus as compared to a wildtype GA3 oxidase locus. In an
aspect, between 50% and 100% of the corn plants in a field comprise
a mutation in a GA3 oxidase locus as compared to a wildtype GA3
oxidase locus. In an aspect, between 60% and 100% of the corn
plants in a field comprise a mutation in a GA3 oxidase locus as
compared to a wildtype GA3 oxidase locus. In an aspect, between 70%
and 100% of the corn plants in a field comprise a mutation in a GA3
oxidase locus as compared to a wildtype GA3 oxidase locus. In an
aspect, between 80% and 100% of the corn plants in a field comprise
a mutation in a GA3 oxidase locus as compared to a wildtype GA3
oxidase locus. In an aspect, between 90% and 100% of the corn
plants in a field comprise a mutation in a GA3 oxidase locus as
compared to a wildtype GA3 oxidase locus.
[0421] In an aspect, at least 10% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, at least 20% of the corn plants in a field comprise a
heterologous polynucleotide capable of suppressing expression of a
GA20 oxidase gene or an mRNA transcribed therefrom. In an aspect,
at least 30% of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 40%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 50%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 60%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 70%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 80%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 90%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA20 oxidase
gene or an mRNA transcribed therefrom. In an aspect, 100% of the
corn plants in a field comprise a heterologous polynucleotide
capable of suppressing expression of a GA20 oxidase gene or an mRNA
transcribed therefrom.
[0422] In an aspect, between 1% and 100% of the corn plants in a
field comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 10% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 20% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 30% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 40% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 50% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 60% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 70% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 80% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 90% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA20 oxidase gene or an mRNA transcribed
therefrom.
[0423] In an aspect, at least 10% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, at least 20% of the corn plants in a field comprise a
heterologous polynucleotide capable of suppressing expression of a
GA3 oxidase gene or an mRNA transcribed therefrom. In an aspect, at
least 30% of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 40%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 50%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 60%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 70%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 80%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, at least 90%
of the corn plants in a field comprise a heterologous
polynucleotide capable of suppressing expression of a GA3 oxidase
gene or an mRNA transcribed therefrom. In an aspect, 100% of the
corn plants in a field comprise a heterologous polynucleotide
capable of suppressing expression of a GA3 oxidase gene or an mRNA
transcribed therefrom.
[0424] In an aspect, between 1% and 100% of the corn plants in a
field comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 10% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 20% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 30% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 40% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 50% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 60% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 70% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 80% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed therefrom.
In an aspect, between 90% and 100% of the corn plants in a field
comprise a heterologous polynucleotide capable of suppressing
expression of a GA3 oxidase gene or an mRNA transcribed
therefrom.
[0425] Without being bound by theory, it is proposed that ectopic
expression or overexpression of GA2 oxidase transgene(s) may be
effective in achieving a short stature, semi-dwarf phenotype with
increased resistance to lodging, but without reproductive off-types
in the ear. It is further proposed, without being limited by
theory, that restricting the expression of GA2 oxidase gene(s) to
certain active GA-producing tissues, such as the vascular and/or
leaf tissues of the plant, may be sufficient to produce a
short-stature plant with increased lodging resistance, but without
significant off-types in reproductive tissues. Expression of a GA2
oxidase transgene in a tissue-specific or tissue-preferred manner
may be sufficient and effective at producing plants with the short
stature phenotype, while avoiding potential off-types in
reproductive tissues that were previously observed with GA mutants
in corn (e.g., by avoiding or limiting the expression of the GA2
oxidase gene(s) in those reproductive tissues). For example, the
GA2 oxidase transgene(s) may be expressed using a vascular
promoter, such as a rice tungro bacilliform virus (RTBV) promoter,
that drives expression in vascular tissues of plants. The
expression pattern of the RTBV promoter is enriched in vascular
tissues of corn plants relative to non-vascular tissues, which is
sufficient to produce a semi-dwarf phenotype in corn plants when
operably linked to a transcribable DNA sequence encoding a GA2
oxidase gene(s). Lowering of active GA levels in tissue(s) of a
corn plant that produce active GAs may reduce plant height and
increase lodging resistance, and off-types may be avoided in those
plants if active GA levels are not also significantly impacted or
lowered in reproductive tissues, such as the developing female
organ or ear of the plant. If active GA levels could be reduced in
the stalk, stem, or internode(s) of corn plants without
significantly affecting GA levels in reproductive tissues (e.g.,
the female or male reproductive organs or inflorescences), then
corn plants having reduced plant height and increased lodging
resistance could be created without off-types in the reproductive
tissues of the plant.
[0426] Thus, recombinant DNA constructs and transgenic plants are
provided herein comprising a transcribable DNA sequence encoding a
GA2 oxidase mRNA and protein operably linked to a plant expressible
promoter, which may be a tissue-specific or tissue-preferred
promoter. Such a tissue-specific or tissue-preferred promoter may
drive expression of its associated GA2 oxidase coding sequence in
one or more active GA-producing tissue(s) of the plant to reduce
the level of active GAs produced in those tissue(s). Such a
tissue-specific or tissue-preferred promoter may drive expression
of its associated GA2 oxidase transgene or coding sequence during
one or more vegetative stage(s) of development. Such a
tissue-specific or tissue-preferred promoter may also have little
or no expression in one or more cell(s) or tissue(s) of the
developing female organ or ear of the plant to avoid the
possibility of off-types in those reproductive tissues. According
to some embodiments, the tissue-specific or tissue-preferred
promoter is a vascular promoter, such as the RTBV promoter. The
sequence of the RTBV promoter is provided herein as SEQ ID NO: 656,
and a truncated version of the RTBV promoter is further provided
herein as SEQ ID NO: 657.
[0427] Active or bioactive gibberellic acids (i.e., "active
gibberellins" or "active GAs") are known in the art for a given
plant species, as distinguished from inactive GAs. For example,
active GAs in corn and higher plants include the following: GA1,
GA3, GA4, and GA7. Thus, an "active GA-producing tissue" is a plant
tissue that produces one or more active GAs.
[0428] In addition to suppressing GA20 oxidase genes in active
GA-producing tissues of the plant with a vascular tissue promoter,
it is further proposed that GA2 oxidase transgenes may also be
expressed with various constitutive promoters to cause the short,
semi-dwarf stature phenotypes in corn, without any visible
off-types in the ear. Thus, it is further proposed that expression
of one or more GA2 oxidase transgenes could be carried out using a
constitutive promoter to create a short stature, lodging-resistant
corn plant without any significant or observable reproductive
off-types in the plant.
[0429] Without being limited by theory, it is proposed that short
stature, semi-dwarf phenotypes in corn plants may result from a
sufficient level of expression of a GA2 oxidase transgene(s) in
active GA-producing tissue(s) of the plant, and restricting the
pattern of expression to avoid reproductive ear tissues may not be
necessary to avoid reproductive off-types in the developing ear. It
is proposed that the semi-dwarf phenotype with GA2 oxidase
overexpression can be the result of shortening the stem internodes
of the plant. Without being bound by theory, it is proposed that
expression of GA2 oxidase transgene(s) in tissue(s) and/or cell(s)
of the plant where active GAs are produced, and not necessarily in
stem or internode tissue(s), may be sufficient to produce
semi-dwarf plants, even though the short stature trait is due to
shortening of the stem internodes. Given that GAs can migrate
through the vasculature of the plant, it is proposed that
manipulating GA oxidase genes in plant tissue(s) where active GAs
are produced may result in a short stature, semi-dwarf plant, even
though this may be largely achieved by reducing the level of active
GAs produced in non-stem tissues (i.e., away from the site of
action in the stem where reduced internode elongation leads to the
semi-dwarf phenotype). However, without being bound by theory,
expression of a GA2 oxidase transgene at low levels, and/or in a
limited number of plant tissues, may be insufficient to cause a
significant short stature, semi-dwarf phenotype.
[0430] The plant hormone gibberellin plays an important role in a
number of plant developmental processes including germination, cell
elongation, flowering, embryogenesis and seed development. Certain
biosynthetic enzymes (e.g., GA20 oxidase and GA3 oxidase) and
catabolic enzymes (e.g., GA2 oxidase) in the GA pathway are
critical to affecting active GA levels in plant tissues. While the
biosynthetic enzymes can increase the level of active GAs, the
catabolic enzymes can reduce the level(s) of active GAs in plants
or plant cells. Thus, it is proposed that overexpression or ectopic
expression of a GA2 oxidase transgene in a constitutive or
tissue-specific or tissue-preferred manner may produce corn plants
having a short stature phenotype and increased lodging resistance,
with possible increased yield, but without off-types in the ear.
Thus, according to some embodiments, constructs and transgenes are
provided comprising a transcribable DNA sequence encoding a GA2
oxidase protein operably linked to a constitutive or
tissue-specific or tissue-preferred promoter, such as a vascular or
leaf promoter. According to some embodiments, the tissue-specific
or tissue-preferred promoter is a vascular promoter, such as the
RTBV promoter. However, other types of tissue-specific or tissue
preferred promoters may potentially be used for GA2 oxidase
expression in active GA-producing tissues of a corn plant to
produce a semi-dwarf phenotype without significant off-types.
[0431] According to some embodiments, a modified or transgenic
plant is provided having a GA2 oxidase gene expression level that
is increased in at least one plant tissue by at least 5%, at least
10%, at least 20%, at least 25%, at least 30%, at least 40%, at
least 50%, at least 60%, at least 70%, at least 75%, at least 80%,
at least 90%, or 100%, as compared to a control plant. According to
some embodiments, a modified or transgenic plant is provided having
a GA2 oxidase gene expression level that is increased in at least
one plant tissue by 5%-20%, 5%-25%, 5%-30%, 5%-40%, 5%-50%, 5%-60%,
5%-70%, 5%-75%, 5%-80%, 5%-90%, 5%-100%, 75%-100%, 50%-100%,
50%-90%, 50%-75%, 25%-75%, 30%-80%, or 10%-75%, as compared to a
control plant. According to these embodiments, the at least one
tissue of a modified or transgenic plant having an increased
expression level of a GA2 oxidase gene(s) includes one or more
active GA producing tissue(s) of the plant, such as the vascular
and/or leaf tissue(s) of the plant, during one or more vegetative
stage(s) of development.
[0432] In some embodiments, transgenic expression of a GA2 oxidase
transgene is constitutive or tissue-specific (e.g., only in leaf
and/or vascular tissue). For example, expression of a GA2 oxidase
transgene may be vascular or leaf tissue specific or preferred. In
other embodiments, expression of a GA2 oxidase transgene is
constitutive and not tissue-specific. According to some
embodiments, expression of a GA2 oxidase transgene is increased in
one or more tissue types (e.g., in leaf and/or vascular tissue(s))
of a modified or transgenic plant as compared to the same tissue(s)
of a control plant.
[0433] According to embodiments of the present disclosure, a
recombinant DNA molecule, construct or vector is provided
comprising an expression cassette comprising a GA2 oxidase coding
sequence or transcribable DNA sequence that is operably linked to a
plant-expressible constitutive or tissue-specific or
tissue-preferred promoter. The expression cassette may comprise a
transcribable DNA sequence having a percent identity to all or part
of a GA2 oxidase gene or coding sequence. A transgene having a
coding sequence with a lower percent identity to all or part of a
GA2 oxidase gene may encode a protein having or retaining a GA
catabolic activity in a corn plant or plant cell similar to GA2
oxidase genes in general.
[0434] A single GA2 oxidase transgene or expression cassette may be
present in a construct, molecule or vector, or multiple GA2 oxidase
transgenes or expression cassettes may be arranged serially in
tandem or arranged in tandem segments or repeats, in a construct,
molecule or vector, which may also be interrupted by one or more
spacer sequence(s). The sequence of each transgene or expression
cassette may encode a GA2 oxidase mRNA and protein. A transcribable
DNA sequence or coding sequence of a GA2 oxidase transgene may
encode a protein having at least 70%, at least 75%, at least 80%,
at least 85%, at least 90%, at least 95%, at least 96%, at least
97%, at least 98%, at least 99%, at least 99.5% or 100% identity to
all or part of a GA2 oxidase gene sequence.
[0435] According to embodiments of the present disclosure, a
recombinant DNA molecule, construct or vector is provided
comprising a transcribable DNA sequence encoding a GA2 oxidase.
According to some embodiments, a recombinant DNA molecule, vector
or construct is provided comprising a transcribable DNA sequence
encoding a GA2 oxidase mRNA and protein in a plant cell, and
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter, such as a constitutive or
tissue-specific or tissue-preferred promoter. According to
embodiments of the present disclosure, suitable tissue-specific or
tissue preferred promoters for expression of a GA2 oxidase may
include those promoters that drive or cause expression of its
associated suppression element or sequence at least in the vascular
and/or leaf tissue(s) of a corn plant. Expression of the GA2
oxidase with a tissue-specific or tissue-preferred promoter may
also occur in other tissues of the corn plant outside of the
vascular and leaf tissues, but active GA levels in the developing
reproductive tissues of the plant (particularly in the female
reproductive organ or ear) are preferably not significantly reduced
or impacted (relative to wild type or control plants), such that
development of the female organ or ear may proceed normally in the
transgenic plant without off-types in the ear and a loss in yield
potential. According to many embodiments, the plant-expressible
promoter may preferably drive expression constitutively or in at
least a portion of the vascular and/or leaf tissues of the plant.
However, some tissue-specific and tissue-preferred promoters
driving expression of a GA2 oxidase transgene in a plant may not
produce a significant short stature or anti-lodging phenotypes due
to the spatial-temporal pattern of expression of the promoter
during plant development, and/or the amount or strength of
expression of the promoter being too low or weak. A sufficient
level of expression of a transcribable DNA sequence encoding a GA2
oxidase may be necessary to produce a short stature, semi-dwarf
phenotype that resists lodging, since lower levels of expression
may be insufficient to lower active GA levels in the plant to a
sufficient extent to cause a significant phenotype. Thus,
tissue-specific and tissue-preferred promoters that drive, etc., a
moderate or strong level of expression of their associated
transcribable DNA sequence in active GA-producing tissue(s) of a
plant may be preferred. Furthermore, such tissue-specific and
tissue-preferred should drive, etc., expression of their associated
transcribable DNA sequence during one or more vegetative stage(s)
of plant development when the plant is growing and/or elongating
including one or more of the following vegetative stage(s):
V.sub.E, V1, V2, V3, V4, V5, V6, V7, V8, V9, V10, V11, V12, V13,
V14, Vn, V.sub.T, such as expression at least during V3-V12,
V4-V12, V5-V12, V6-V12, V7-V12, V8-V12, V3-V14, V5-V14, V6-V14,
V7-V14, V8-V14, V9-V14, V10-V14, etc., or during any other range of
vegetative stages when growth and/or elongation of the plant is
occurring.
[0436] Any vascular promoters known in the art may potentially be
used as the tissue-specific or tissue-preferred promoter. Examples
of vascular promoters include the RTBV promoter (see, e.g., SEQ ID
NO: 656), a known sucrose synthase gene promoter, such as a corn
sucrose synthase-1 (Sus1 or Sh1) promoter (see, e.g., SEQ ID NO:
658), a corn Sh1 gene paralog promoter, a barley sucrose synthase
promoter (Ss1) promoter, a rice sucrose synthase-1 (RSs1) promoter
(see, e.g., SEQ ID NO: 659), or a rice sucrose synthase-2 (RSs2)
promoter (see, e.g., SEQ ID NO: 660), a known sucrose transporter
gene promoter, such as a rice sucrose transporter promoter (SUT1)
(see, e.g., SEQ ID NO: 661), or various known viral promoters, such
as a Commelina yellow mottle virus (CoYMV) promoter, a wheat dwarf
geminivirus (WDV) large intergenic region (LIR) promoter, a maize
streak geminivirus (MSV) coat protein (CP) promoter, or a rice
yellow stripe 1 (YS1)-like or OsYSL2 promoter (SEQ ID NO: 662), and
any functional sequence portion or truncation of any of the
foregoing promoters with a similar pattern of expression, such as a
truncated RTBV promoter (see, e.g., SEQ ID NO: 657). Any other
vascular promoters known in the art may also be used, including
promoter sequences from related genes (e.g., sucrose synthase,
sucrose transporter, and viral gene promoter sequences) from the
same or different plant species, microbe or virus that have a
similar pattern of expression. Further provided are promoter
sequences with a high degree of homology to any of the foregoing.
For example, a vascular promoter may comprise a DNA sequence that
is at least at least 70%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5% or 100% identical to one or more of SEQ ID NOs:
656, 657, 658, 659, 660, 661, and 662, any functional sequence
portion or truncation thereof, and/or any sequence complementary to
any of the foregoing sequences. Examples of vascular promoters may
further include other known, engineered and/or later-identified
promoter sequences shown to have a pattern of expression in
vascular tissue(s) of a corn plant.
[0437] Any leaf promoters known in the art may potentially be used
as the tissue-specific or tissue-preferred promoter. Examples of
leaf promoters include a corn pyruvate phosphate dikinase or PPDK
promoter (see, e.g., SEQ ID NO: 663), a corn fructose 1,6
bisphosphate aldolase or FDA promoter (see, e.g., SEQ ID NO: 664),
and a rice Nadh-Gogat promoter (see, e.g., SEQ ID NO: 665), and any
functional sequence portion or truncation of any of the foregoing
promoters with a similar pattern of expression. Other examples of
leaf promoters from monocot plant genes include a ribulose
biphosphate carboxylase (RuBisCO) or RuBisCO small subunit (RBCS)
promoter, a chlorophyll a/b binding protein gene promoter, a
phosphoenolpyruvate carboxylase (PEPC) promoter, and a Myb gene
promoter, and any functional sequence portion or truncation of any
of these promoters with a similar pattern of expression. Any other
leaf promoters known in the art may also be used, including
promoter sequences from related genes from the same or different
plant species, microbe or virus that have a similar pattern of
expression. Further provided are promoter sequences with a high
degree of homology to any of the foregoing. For example, a leaf
promoter may comprise a DNA sequence that is at least at least 70%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5% or
100% identical to one or more of SEQ ID NOs: 663, 664, and 665, any
functional sequence portion or truncation thereof, and/or any
sequence complementary to any of the foregoing sequences. Examples
of leaf promoters may further include other known, engineered
and/or later-identified promoter sequences shown to have a pattern
of expression in leaf tissue(s) of a corn plant.
[0438] Any constitutive promoters known in the art may potentially
be used. Examples of constitutive promoters that may be used in
corn plants include, for example, various actin gene promoters,
such as a rice Actin 1 promoter (see, e.g., U.S. Pat. No.
5,641,876; see also SEQ ID NO: 666 or SEQ ID NO: 667) and a rice
Actin 2 promoter (see, e.g., U.S. Pat. No. 6,429,357; see also,
e.g., SEQ ID NO: 668 or SEQ ID NO: 669), a CaMV 35S or 19S promoter
(see, e.g., U.S. Pat. No. 5,352,605; see also, e.g., SEQ ID NO: 670
for CaMV 35S), a maize ubiquitin promoter (see, e.g., U.S. Pat. No.
5,510,474), a Coix lacryma-jobi polyubiquitin promoter (see, e.g.,
SEQ ID NO: 671), a rice or maize Gos2 promoter (see, e.g., Pater et
al., The Plant Journal, 2(6): 837-44 1992; see also, e.g., SEQ ID
NO: 672 for the rice Gos2 promoter), a FMV 35S promoter (see, e.g.,
U.S. Pat. No. 6,372,211), a dual enhanced CMV promoter (see, e.g.,
U.S. Pat. No. 5,322,938), a MMV promoter (see, e.g., U.S. Pat. No.
6,420,547; see also, e.g., SEQ ID NO: 673), a PCLSV promoter (see,
e.g., U.S. Pat. No. 5,850,019; see also, e.g., SEQ ID NO: 674), an
Emu promoter (see, e.g., Last et al., Theor. Appl. Genet. 81:581
(1991); and Mcelroy et al., Mol. Gen. Genet. 231:150 (1991)), a
tubulin promoter from maize, rice or other species, a nopaline
synthase (nos) promoter, an octopine synthase (ocs) promoter, a
mannopine synthase (mas) promoter, or a plant alcohol dehydrogenase
(e.g., maize Adhl) promoter, any other promoters including viral
promoters known or later-identified in the art to provide
constitutive expression in a corn plant, any other constitutive
promoters known in the art that may be used in corn plants, and any
functional sequence portion or truncation of any of the foregoing
promoters.
[0439] Any other constitutive promoters known in the art may also
be used, including promoter sequences from related genes from the
same or different plant species, microbe or virus that have a
similar pattern of expression. Further provided are promoter
sequences with a high degree of homology to any of the foregoing.
For example, a constitutive promoter may comprise a DNA sequence
that is at least at least 70%, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5% or 100% identical to one or more of SEQ
ID NOs: 666, 667, 668, 669, 670, 671, 672, 673, and 674, any
functional sequence portion or truncation thereof, and/or any
sequence complementary to any of the foregoing sequences. Examples
of constitutive promoters may further include other known,
engineered and/or later-identified promoter sequences shown to have
a constitutive pattern of expression in a corn plant. Furthermore,
any known or later-identified constitutive promoter may also be
used.
[0440] According to embodiments of the present disclosure, a
recombinant DNA molecule, construct or vector is provided
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein from a monocot or cereal plant, such as a corn plant, or
encoding a GA2 oxidase protein having at least a certain percent
homology to a GA2 oxidase protein from a monocot or cereal plant,
such as a corn plant. A family of at least thirteen GA2 oxidase
genes have been identified in corn (Zea mays) including Zm.GA2
oxidase_1, Zm.GA2 oxidase_2, Zm.GA2 oxidase_3, Zm.GA2 oxidase_4,
Zm.GA2 oxidase_5, Zm.GA2 oxidase_6, Zm.GA2 oxidase_7, Zm.GA2
oxidase_8, Zm.GA2 oxidase_9, Zm.GA2 oxidase_10, Zm.GA2 oxidase_11,
Zm.GA2 oxidase_12, and Zm.GA2 oxidase_13. The DNA and protein
sequences by SEQ ID NO for each of these GA2 oxidase genes are
provided in Table 3.
TABLE-US-00003 TABLE 3 DNA and protein sequences for GA2 oxidase
genes in corn. Coding Sequence GA2 oxidase Gene (CDS) Protein
Zm.GA2 oxidase_1 SEQ ID NO: 324 SEQ ID NO: 325 Zm.GA2 oxidase_2 SEQ
ID NO: 326 SEQ ID NO: 327 Zm.GA2 oxidase_3 SEQ ID NO: 328 SEQ ID
NO: 329 Zm.GA2 oxidase_4 SEQ ID NO: 330 SEQ ID NO: 331 Zm.GA2
oxidase_5 SEQ ID NO: 332 SEQ ID NO: 333 Zm.GA2 oxidase_6 SEQ ID NO:
334 SEQ ID NO: 335 Zm.GA2 oxidase_7 SEQ ID NO: 336 SEQ ID NO: 337
Zm.GA2 oxidase_8 SEQ ID NO: 338 SEQ ID NO: 339 Zm.GA2 oxidase_9 SEQ
ID NO: 340 SEQ ID NO: 341 Zm.GA2 oxidase_10 SEQ ID NO: 342 SEQ ID
NO: 343 Zm.GA2 oxidase_11 SEQ ID NO: 344 SEQ ID NO: 345 Zm.GA2
oxidase_12 SEQ ID NO: 346 SEQ ID NO: 347 Zm.GA2 oxidase_13 SEQ ID
NO: 348 SEQ ID NO: 349
[0441] GA2 oxidase genes from other monocot or cereal plant species
may also be used, such as rice, barley, wheat and sorghum.
According to embodiments of the present disclosure, a recombinant
DNA molecule, construct or vector is provided comprising a
transcribable DNA sequence encoding a GA2 oxidase protein from a
monocot or cereal plant other than corn, or encoding a GA2 oxidase
protein having at least a certain percent homology to a GA2 oxidase
protein from a monocot or cereal plant other than corn. A family of
at least ten GA2 oxidase genes have been identified in rice (Oryza
sativa) plants including Os.GA2 oxidase_1, Os.GA2 oxidase_2, Os.GA2
oxidase_3, Os.GA2 oxidase_4, Os.GA2 oxidase_5, Os.GA2 oxidase_6,
Os.GA2 oxidase_7, Os.GA2 oxidase_8, Os.GA2 oxidase_9, and Os.GA2
oxidase_10. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes are provided in Table 4.
TABLE-US-00004 TABLE 4 DNA and protein sequences for GA2 oxidase
genes in rice. Coding Sequence GA2 oxidase Gene (CDS) Protein
Os.GA2 oxidase_1 SEQ ID NO: 350 SEQ ID NO: 351 Os.GA2 oxidase_2 SEQ
ID NO: 352 SEQ ID NO: 353 Os.GA2 oxidase_3 SEQ ID NO: 354 SEQ ID
NO: 355 Os.GA2 oxidase_4 SEQ ID NO: 356 SEQ ID NO: 357 Os.GA2
oxidase_5 SEQ ID NO: 358 SEQ ID NO: 359 Os.GA2 oxidase_6 SEQ ID NO:
360 SEQ ID NO: 361 Os.GA2 oxidase_7 SEQ ID NO: 362 SEQ ID NO: 363
Os.GA2 oxidase_8 SEQ ID NO: 364 SEQ ID NO: 365 Os.GA2 oxidase_9 SEQ
ID NO: 366 SEQ ID NO: 367 Os.GA2 oxidase_10 SEQ ID NO: 368 SEQ ID
NO: 369
[0442] A family of at least eight GA2 oxidase genes have been
identified in barley (Hordeum vulgare) plants including Hv.GA2
oxidase_1, Hv.GA2 oxidase_2, Hv.GA2 oxidase_3, Hv.GA2 oxidase_4,
Hv.GA2 oxidase_5, Hv.GA2 oxidase_6, Hv.GA2 oxidase_7, and Hv.GA2
oxidase_8. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes are provided in Table 5.
TABLE-US-00005 TABLE 5 DNA and protein sequences for GA2 oxidase
genes in barley. Coding Sequence GA2 oxidase Gene (CDS) Protein
Hv.GA2 oxidase_1 SEQ ID NO: 370 SEQ ID NO: 371 Hv.GA2 oxidase_2 SEQ
ID NO: 372 SEQ ID NO: 373 Hv.GA2 oxidase_3 SEQ ID NO: 374 SEQ ID
NO: 375 Hv.GA2 oxidase_4 SEQ ID NO: 376 SEQ ID NO: 377 Hv.GA2
oxidase_5 SEQ ID NO: 378 SEQ ID NO: 379 Hv.GA2 oxidase_6 SEQ ID NO:
380 SEQ ID NO: 381 Hv.GA2 oxidase_7 SEQ ID NO: 382 SEQ ID NO: 383
Hv.GA2 oxidase_8 SEQ ID NO: 384 SEQ ID NO: 385
[0443] A family of at least sixteen GA2 oxidase genes have been
identified in sorghum (Sorghum bicolor) plants including Hv.GA2
oxidase_1, Sb.GA2 oxidase_2, Sb.GA2 oxidase_3, Sb.GA2 oxidase_4,
Sb.GA2 oxidase_5, Sb.GA2 oxidase_6, Sb.GA2 oxidase_7, Sb.GA2
oxidase_8, Sb.GA2 oxidase_9, Sb.GA2 oxidase_10, Sb.GA2 oxidase_11,
Sb.GA2 oxidase_12, Sb.GA2 oxidase_13, Sb.GA2 oxidase_14, Sb.GA2
oxidase_15, and Sb.GA2 oxidase_16. The DNA and protein sequences by
SEQ ID NO for each of these GA2 oxidase genes are provided in Table
6.
TABLE-US-00006 TABLE 6 DNA and protein sequences for GA2 oxidase
genes in sorghum. Coding Sequence GA2 oxidase Gene (CDS) Protein
Sb.GA2 oxidase_1 SEQ ID NO: 386 SEQ ID NO: 387 Sb.GA2 oxidase_2 SEQ
ID NO: 388 SEQ ID NO: 389 Sb.GA2 oxidase_3 SEQ ID NO: 390 SEQ ID
NO: 391 Sb.GA2 oxidase_4 SEQ ID NO: 392 SEQ ID NO: 393 Sb.GA2
oxidase_5 SEQ ID NO: 394 SEQ ID NO: 395 Sb.GA2 oxidase_6 SEQ ID NO:
396 SEQ ID NO: 397 Sb.GA2 oxidase_7 SEQ ID NO: 398 SEQ ID NO: 399
Sb.GA2 oxidase_8 SEQ ID NO: 400 SEQ ID NO: 401 Sb.GA2 oxidase_9 SEQ
ID NO: 402 SEQ ID NO: 403 Sb.GA2 oxidase_10 SEQ ID NO: 404 SEQ ID
NO: 405 Sb.GA2 oxidase_11 SEQ ID NO: 406 SEQ ID NO: 407 Sb.GA2
oxidase_12 SEQ ID NO: 408 SEQ ID NO: 409 Sb.GA2 oxidase_13 SEQ ID
NO: 410 SEQ ID NO: 411 Sb.GA2 oxidase_14 SEQ ID NO: 412 SEQ ID NO:
413 Sb.GA2 oxidase_15 SEQ ID NO: 414 SEQ ID NO: 415 Sb.GA2
oxidase_16 SEQ ID NO: 416 SEQ ID NO: 417
[0444] A family of at least fifteen GA2 oxidase genes have been
identified in wheat (Triticum aestivum) including Ta.GA2 oxidase_1,
Ta.GA2 oxidase_2, Ta.GA2 oxidase_3, Ta.GA2 oxidase_4, Ta.GA2
oxidase_5, Ta.GA2 oxidase_6, Ta.GA2 oxidase_7, Ta.GA2 oxidase_8,
Ta.GA2 oxidase_9, Ta.GA2 oxidase_10, Ta.GA2 oxidase_11, Ta.GA2
oxidase_12, Ta.GA2 oxidase_13, Ta.GA2 oxidase_14, and Ta.GA2
oxidase_15. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes from wheat are provided in Table 7.
TABLE-US-00007 TABLE 7 DNA and protein sequences for GA2 oxidase
genes in wheat. Coding Sequence GA2 oxidase Gene (CDS) Protein
Ta.GA2 oxidase_1 SEQ ID NO: 418 SEQ ID NO: 419 Ta.GA2 oxidase_2 SEQ
ID NO: 420 SEQ ID NO: 421 Ta.GA2 oxidase_3 SEQ ID NO: 422 SEQ ID
NO: 423 Ta.GA2 oxidase_4 SEQ ID NO: 424 SEQ ID NO: 425 Ta.GA2
oxidase_5 SEQ ID NO: 426 SEQ ID NO: 427 Ta.GA2 oxidase_6 SEQ ID NO:
428 SEQ ID NO: 429 Ta.GA2 oxidase_7 SEQ ID NO: 430 SEQ ID NO: 431
Ta.GA2 oxidase_8 SEQ ID NO: 432 SEQ ID NO: 433 Ta.GA2 oxidase_9 SEQ
ID NO: 434 SEQ ID NO: 435 Ta.GA2 oxidase_10 SEQ ID NO: 436 SEQ ID
NO: 437 Ta.GA2 oxidase_11 SEQ ID NO: 438 SEQ ID NO: 439 Ta.GA2
oxidase_12 SEQ ID NO: 440 SEQ ID NO: 441 Ta.GA2 oxidase_13 SEQ ID
NO: 442 SEQ ID NO: 443 Ta.GA2 oxidase_14 SEQ ID NO: 444 SEQ ID NO:
445 Ta.GA2 oxidase_15 SEQ ID NO: 446 SEQ ID NO: 447
[0445] In addition to the corn sequences listed in Table 3, a
family of at least eleven GA2 oxidase genes have been identified in
another corn (Zea mays) germplasm line including Zm2.GA2 oxidase_1,
Zm2.GA2 oxidase_2, Zm2.GA2 oxidase_3, Zm2.GA2 oxidase_4, Zm2.GA2
oxidase_5, Zm2.GA2 oxidase_6, Zm2.GA2 oxidase_7, Zm2.GA2 oxidase_8,
Zm2.GA2 oxidase_9, Zm2.GA2 oxidase_10, and Zm2.GA2 oxidase_11. The
DNA and protein sequences by SEQ ID NO for each of these GA2
oxidase genes are provided in Table 8.
TABLE-US-00008 TABLE 8 Additional DNA and protein sequences for GA2
oxidase genes in corn. Coding Sequence GA2 oxidase Gene (CDS)
Protein Zm2.GA2 oxidase_1 SEQ ID NO: 448 SEQ ID NO: 449 Zm2.GA2
oxidase_2 SEQ ID NO: 450 SEQ ID NO: 451 Zm2.GA2 oxidase_3 SEQ ID
NO: 452 SEQ ID NO: 453 Zm2.GA2 oxidase_4 SEQ ID NO: 454 SEQ ID NO:
455 Zm2.GA2 oxidase_5 SEQ ID NO: 456 SEQ ID NO: 457 Zm2.GA2
oxidase_6 SEQ ID NO: 458 SEQ ID NO: 459 Zm2.GA2 oxidase_7 SEQ ID
NO: 460 SEQ ID NO: 461 Zm2.GA2 oxidase_8 SEQ ID NO: 462 SEQ ID NO:
463 Zm2.GA2 oxidase_9 SEQ ID NO: 464 SEQ ID NO: 465 Zm2.GA2
oxidase_10 SEQ ID NO: 466 SEQ ID NO: 467 Zm2.GA2 oxidase_11 SEQ ID
NO: 468 SEQ ID NO: 469
[0446] According to embodiments of the present disclosure, a
recombinant DNA molecule, construct or vector is provided
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein from a dicot plant, such as a soybean, cotton, canola,
Arabidopsis, moss (Physcomitrella patens), common bean (Phaseolus
vulgaris), cottonwood (Populus trichocarpa), barrel clover
(Medicago truncatula), pea (Pisum sativum), spinach (Spinacia
oleracea) or whorled honey flower (Paris polyphylla) plant, or
encoding a GA2 oxidase protein having at least a certain percent
homology to a GA2 oxidase protein from a monocot or cereal plant,
such as a corn plant.
[0447] A family of at least sixteen GA2 oxidase genes have been
identified in soybean (Glycine max) including Gm.GA2 oxidase_1,
Gm.GA2 oxidase_2, Gm.GA2 oxidase_3, Gm.GA2 oxidase_4, Gm.GA2
oxidase_5, Gm.GA2 oxidase_6, Gm.GA2 oxidase_7, Gm.GA2 oxidase_8,
Gm.GA2 oxidase_9, Gm.GA2 oxidase_10, Gm.GA2 oxidase_11, Gm.GA2
oxidase_12, Gm.GA2 oxidase_13, Gm.GA2 oxidase_14, Gm.GA2
oxidase_15, and Gm.GA2 oxidase_16. The DNA and protein sequences by
SEQ ID NO for each of these GA2 oxidase genes from soybean are
provided in Table 9.
TABLE-US-00009 TABLE 9 Additional DNA and protein sequences for GA2
oxidase genes in soybean. Coding Sequence GA2 oxidase Gene (CDS)
Protein Gm.GA2 oxidase_1 SEQ ID NO: 470 SEQ ID NO: 471 Gm.GA2
oxidase_2 SEQ ID NO: 472 SEQ ID NO: 473 Gm.GA2 oxidase_3 SEQ ID NO:
474 SEQ ID NO: 475 Gm.GA2 oxidase_4 SEQ ID NO: 476 SEQ ID NO: 477
Gm.GA2 oxidase_5 SEQ ID NO: 478 SEQ ID NO: 479 Gm.GA2 oxidase_6 SEQ
ID NO: 480 SEQ ID NO: 481 Gm.GA2 oxidase_7 SEQ ID NO: 482 SEQ ID
NO: 483 Gm.GA2 oxidase_8 SEQ ID NO: 484 SEQ ID NO: 485 Gm.GA2
oxidase_9 SEQ ID NO: 486 SEQ ID NO: 487 Gm.GA2 oxidase_10 SEQ ID
NO: 488 SEQ ID NO: 489 Gm.GA2 oxidase_11 SEQ ID NO: 490 SEQ ID NO:
491 Gm.GA2 oxidase_12 SEQ ID NO: 492 SEQ ID NO: 493 Gm.GA2
oxidase_13 SEQ ID NO: 494 SEQ ID NO: 495 Gm.GA2 oxidase_14 SEQ ID
NO: 496 SEQ ID NO: 497 Gm.GA2 oxidase_15 SEQ ID NO: 498 SEQ ID NO:
499 Gm.GA2 oxidase_16 SEQ ID NO: 500 SEQ ID NO: 501
[0448] A family of at least fifteen related GA2 oxidase genes have
been identified in cotton (Gossypium hirsutum) including Gh.GA2
oxidase_1, Gh.GA2 oxidase_2, Gh.GA2 oxidase_3, Gh.GA2 oxidase_4,
Gh.GA2 oxidase_5, Gh.GA2 oxidase_6, Gh.GA2 oxidase_7, Gh.GA2
oxidase_8, Gh.GA2 oxidase_9, Gh.GA2 oxidase_10, Gh.GA2 oxidase_11,
Gh.GA2 oxidase_12, Gh.GA2 oxidase_13, Gh.GA2 oxidase_14, and Gh.GA2
oxidase_15. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes from cotton are provided in Table 10.
TABLE-US-00010 TABLE 10 DNA and protein sequences for GA2 oxidase
genes in cotton. Coding Sequence GA2 oxidase Gene (CDS) Protein
Gh.GA2 oxidase_1 SEQ ID NO: 502 SEQ ID NO: 503 Gh.GA2 oxidase_2 SEQ
ID NO: 504 SEQ ID NO: 505 Gh.GA2 oxidase_3 SEQ ID NO: 506 SEQ ID
NO: 507 Gh.GA2 oxidase_4 SEQ ID NO: 508 SEQ ID NO: 509 Gh.GA2
oxidase_5 SEQ ID NO: 510 SEQ ID NO: 511 Gh.GA2 oxidase_6 SEQ ID NO:
512 SEQ ID NO: 513 Gh.GA2 oxidase_7 SEQ ID NO: 514 SEQ ID NO: 515
Gh.GA2 oxidase_8 SEQ ID NO: 516 SEQ ID NO: 517 Gh.GA2 oxidase_9 SEQ
ID NO: 518 SEQ ID NO: 519 Gh.GA2 oxidase_10 SEQ ID NO: 520 SEQ ID
NO: 521 Gh.GA2 oxidase_11 SEQ ID NO: 522 SEQ ID NO: 523 Gh.GA2
oxidase_12 SEQ ID NO: 524 SEQ ID NO: 525 Gh.GA2 oxidase_13 SEQ ID
NO: 526 SEQ ID NO: 527 Gh.GA2 oxidase_14 SEQ ID NO: 528 SEQ ID NO:
529 Gh.GA2 oxidase_15 SEQ ID NO: 530 SEQ ID NO: 531
[0449] A family of at least fifteen GA2 oxidase genes have been
identified in canola (Brassica napus) including Bn.GA2 oxidase_1,
Bn.GA2 oxidase_2, Bn.GA2 oxidase_3, Bn.GA2 oxidase_4, Bn.GA2
oxidase_5, Bn.GA2 oxidase_6, Bn.GA2 oxidase_7, Bn.GA2 oxidase_8,
Bn.GA2 oxidase_9, Bn.GA2 oxidase_10, Bn.GA2 oxidase_11, Bn.GA2
oxidase_12, Bn.GA2 oxidase_13, Bn.GA2 oxidase_14, and Bn.GA2
oxidase_15. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes from canola are provided in Table 11.
TABLE-US-00011 TABLE 11 DNA and protein sequences for GA2 oxidase
genes in canola. Coding Sequence GA2 oxidase Gene (CDS) Protein
Bn.GA2 oxidase_1 SEQ ID NO: 532 SEQ ID NO: 533 Bn.GA2 oxidase_2 SEQ
ID NO: 534 SEQ ID NO: 535 Bn.GA2 oxidase_3 SEQ ID NO: 536 SEQ ID
NO: 537 Bn.GA2 oxidase_4 SEQ ID NO: 538 SEQ ID NO: 539 Bn.GA2
oxidase_5 SEQ ID NO: 540 SEQ ID NO: 541 Bn.GA2 oxidase_6 SEQ ID NO:
542 SEQ ID NO: 543 Bn.GA2 oxidase_7 SEQ ID NO: 544 SEQ ID NO: 545
Bn.GA2 oxidase_8 SEQ ID NO: 546 SEQ ID NO: 547 Bn.GA2 oxidase_9 SEQ
ID NO: 548 SEQ ID NO: 549 Bn.GA2 oxidase_10 SEQ ID NO: 550 SEQ ID
NO: 551 Bn.GA2 oxidase_11 SEQ ID NO: 552 SEQ ID NO: 553 Bn.GA2
oxidase_12 SEQ ID NO: 554 SEQ ID NO: 555 Bn.GA2 oxidase_13 SEQ ID
NO: 556 SEQ ID NO: 557 Bn.GA2 oxidase_14 SEQ ID NO: 558 SEQ ID NO:
559 Bn.GA2 oxidase_15 SEQ ID NO: 560 SEQ ID NO: 561
[0450] A family of at least seven GA2 oxidase genes have been
identified in thale cress (Arabidopsis thaliana) including At.GA2
oxidase_1, At.GA2 oxidase_2, At.GA2 oxidase_3, At.GA2 oxidase_4,
At.GA2 oxidase_6, and At.GA2 oxidase_7, and At.GA2 oxidase_8. The
DNA and protein sequences by SEQ ID NO for each of these GA2
oxidase genes from Arabidopsis are provided in Table 12.
TABLE-US-00012 TABLE 12 DNA and protein sequences for GA2 oxidase
genes in thale cress. Coding Sequence GA2 oxidase Gene (CDS)
Protein At.GA2 oxidase_1 SEQ ID NO: 562 SEQ ID NO: 563 At.GA2
oxidase_2 SEQ ID NO: 564 SEQ ID NO: 565 At.GA2 oxidase_3 SEQ ID NO:
566 SEQ ID NO: 567 At.GA2 oxidase_4 SEQ ID NO: 568 SEQ ID NO: 569
At.GA2 oxidase_6 SEQ ID NO: 570 SEQ ID NO: 571 At.GA2 oxidase_7 SEQ
ID NO: 572 SEQ ID NO: 573 At.GA2 oxidase_8 SEQ ID NO: 574 SEQ ID
NO: 575
[0451] A family of at least seven GA2 oxidase genes have been
identified in moss (Physcomitrella patens) including Pp.GA2
oxidase_1, Pp.GA2 oxidase_2, Pp.GA2 oxidase_3, Pp.GA2 oxidase_4,
Pp.GA2 oxidase_5, Pp.GA2 oxidase_6, and Pp.GA2 oxidase_7. The DNA
and protein sequences by SEQ ID NO for each of these GA2 oxidase
genes from moss are provided in Table 13.
TABLE-US-00013 TABLE 13 DNA and protein sequences for GA2 oxidase
genes in moss. Coding Sequence GA2 oxidase Gene (CDS) Protein
Pp.GA2 oxidase_1 SEQ ID NO: 576 SEQ ID NO: 577 Pp.GA2 oxidase_2 SEQ
ID NO: 578 SEQ ID NO: 579 Pp.GA2 oxidase_3 SEQ ID NO: 580 SEQ ID
NO: 581 Pp.GA2 oxidase_4 SEQ ID NO: 582 SEQ ID NO: 583 Pp.GA2
oxidase_5 SEQ ID NO: 584 SEQ ID NO: 585 Pp.GA2 oxidase_6 SEQ ID NO:
586 SEQ ID NO: 587 Pp.GA2 oxidase_7 SEQ ID NO: 588 SEQ ID NO:
589
[0452] A family of at least nine GA2 oxidase genes have been
identified in barrel clover (Medicago truncatula) including Mt.GA2
oxidase_1, Mt.GA2 oxidase_2, Mt.GA2 oxidase_3, Mt.GA2 oxidase_4,
Mt.GA2 oxidase_5, Mt.GA2 oxidase_6, Mt.GA2 oxidase_7, Mt.GA2
oxidase_8, and Mt.GA2 oxidase_9. The DNA and protein sequences by
SEQ ID NO for each of these GA2 oxidase genes from Medicago are
provided in Table 14.
TABLE-US-00014 TABLE 14 DNA and protein sequences for GA2 oxidase
genes in M. truncatula. Coding Sequence GA2 oxidase Gene (CDS)
Protein Mt.GA2 oxidase_1 SEQ ID NO: 590 SEQ ID NO: 591 Mt.GA2
oxidase_2 SEQ ID NO: 592 SEQ ID NO: 593 Mt.GA2 oxidase_3 SEQ ID NO:
594 SEQ ID NO: 595 Mt.GA2 oxidase_4 SEQ ID NO: 596 SEQ ID NO: 597
Mt.GA2 oxidase_5 SEQ ID NO: 598 SEQ ID NO: 599 Mt.GA2 oxidase_6 SEQ
ID NO: 600 SEQ ID NO: 601 Mt.GA2 oxidase_7 SEQ ID NO: 602 SEQ ID
NO: 603 Mt.GA2 oxidase_8 SEQ ID NO: 604 SEQ ID NO: 605 Mt.GA2
oxidase_9 SEQ ID NO: 606 SEQ ID NO: 607
[0453] A family of at least four related GA2 oxidase genes have
been identified in whorled honey flower (Paris polyphylla)
including Ppo.GA2 oxidase_1, Ppo.GA2 oxidase_2, Ppo.GA2 oxidase_3,
and Ppo.GA2 oxidase_4. The DNA and protein sequences by SEQ ID NO
for each of these GA2 oxidase genes from honey flower are provided
in Table 15.
TABLE-US-00015 TABLE 15 DNA and protein sequences for GA2 oxidase
genes in P. polyphylla. Coding Sequence GA2 oxidase Gene (CDS)
Protein Ppo.GA2 oxidase_1 SEQ ID NO: 608 SEQ ID NO: 609 Ppo.GA2
oxidase_2 SEQ ID NO: 610 SEQ ID NO: 611 Ppo.GA2 oxidase_3 SEQ ID
NO: 612 SEQ ID NO: 613 Ppo.GA2 oxidase_4 SEQ ID NO: 614 SEQ ID NO:
615
[0454] A family of at least eight GA2 oxidase genes have been
identified in common bean (Phaseolus vulgaris) including Pv.GA2
oxidase_1, Pv.GA2 oxidase_2, Pv.GA2 oxidase_3, Pv.GA2 oxidase_4,
Pv.GA2 oxidase_5, Pv.GA2 oxidase_6, Pv.GA2 oxidase_7, and Pv.GA2
oxidase_8. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes from common bean are provided in Table
16.
TABLE-US-00016 TABLE 16 DNA and protein sequences for GA2 oxidase
genes in common bean. Coding Sequence GA2 oxidase Gene (CDS)
Protein Pv.GA2 oxidase_1 SEQ ID NO: 616 SEQ ID NO: 617 Pv.GA2
oxidase_2 SEQ ID NO: 618 SEQ ID NO: 619 Pv.GA2 oxidase_3 SEQ ID NO:
620 SEQ ID NO: 621 Pv.GA2 oxidase_4 SEQ ID NO: 622 SEQ ID NO: 623
Pv.GA2 oxidase_5 SEQ ID NO: 624 SEQ ID NO: 625 Pv.GA2 oxidase_6 SEQ
ID NO: 626 SEQ ID NO: 627 Pv.GA2 oxidase_7 SEQ ID NO: 628 SEQ ID
NO: 629 Pv.GA2 oxidase_8 SEQ ID NO: 630 SEQ ID NO: 631
[0455] A family of at least seven related GA2 oxidase genes have
been identified in cottonwood (Populus trichocarpa) including
Pt.GA2 oxidase_1, Pt.GA2 oxidase_2, Pt.GA2 oxidase_3, Pt.GA2
oxidase_4, Pt.GA2 oxidase_5, Pt.GA2 oxidase_6, and Pt.GA2
oxidase_7. The DNA and protein sequences by SEQ ID NO for each of
these GA2 oxidase genes from cottonwood are provided in Table
17.
TABLE-US-00017 TABLE 17 DNA and protein sequences for GA2 oxidase
genes in cottonwood. Coding Sequence GA2 oxidase Gene (CDS) Protein
Pt.GA2 oxidase_1 SEQ ID NO: 632 SEQ ID NO: 633 Pt.GA2 oxidase_2 SEQ
ID NO: 634 SEQ ID NO: 635 Pt.GA2 oxidase_3 SEQ ID NO: 636 SEQ ID
NO: 637 Pt.GA2 oxidase_4 SEQ ID NO: 638 SEQ ID NO: 639 Pt.GA2
oxidase_5 SEQ ID NO: 640 SEQ ID NO: 641 Pt.GA2 oxidase_6 SEQ ID NO:
642 SEQ ID NO: 643 Pt.GA2 oxidase_7 SEQ ID NO: 644 SEQ ID NO:
645
[0456] A family of at least two GA2 oxidase genes have been
identified in pea (Pisum sativum) including Ps.GA2 oxidase_1 and
Ps.GA2 oxidase_2. The DNA and protein sequences by SEQ ID NO for
these GA2 oxidase genes from pea are provided in Table 18.
TABLE-US-00018 TABLE 18 DNA and protein sequences for GA2 oxidase
genes in pea. Coding Sequence GA2 oxidase Gene (CDS) Protein Ps.GA2
oxidase_1 SEQ ID NO: 646 SEQ ID NO: 647 Ps.GA2 oxidase_2 SEQ ID NO:
648 SEQ ID NO: 649
[0457] A family of at least three related GA2 oxidase genes have
been identified in spinach (Spinacia oleracea) including So.GA2
oxidase_1, So.GA2 oxidase_2, and So.GA2 oxidase_3. The DNA and
protein sequences by SEQ ID NO for each of these GA2 oxidase genes
from spinach are provided in Table 19.
TABLE-US-00019 TABLE 19 DNA and protein sequences for GA2 oxidase
genes in spinach. Coding Sequence GA2 oxidase Gene (CDS) Protein
So.GA2 oxidase_1 SEQ ID NO: 650 SEQ ID NO: 651 So.GA2 oxidase_2 SEQ
ID NO: 652 SEQ ID NO: 653 So.GA2 oxidase_3 SEQ ID NO: 654 SEQ ID
NO: 655
[0458] According to some embodiments, a GA2 oxidase protein encoded
by a transcribable DNA sequence of a recombinant DNA construct is,
or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 325, 327, 329, 331, 333, 335, 337, 339, 341, 343, 345,
347, and/or 349. According to some embodiments, a transcribable DNA
sequence of a recombinant DNA molecule, vector or construct is or
comprises a sequence that is at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346,
and/or 348.
[0459] According to some embodiments, a GA2 oxidase protein encoded
by a transcribable DNA sequence of a recombinant DNA molecule,
vector or construct is or comprises a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 351, 353, 355, 357, 359,
361, 363, 365, 367 and/or 397. According to some embodiments, a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 350, 352, 354, 356, 358, 360, 362, 364, 366
and/or 368. According to some embodiments, a GA2 oxidase protein
encoded by a transcribable DNA sequence of a recombinant DNA
molecule, vector or construct is or comprises a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 371, 373, 375, 377, 379,
381, 383 and/or 385. According to some embodiments, a transcribable
DNA sequence of a recombinant DNA molecule, vector or construct is
or comprises a sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 370, 372, 374, 376, 378, 380, 382 and/or 384. According
to some embodiments, a GA2 oxidase protein encoded by a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 387, 389, 391, 393, 395, 397, 399, 401, 403,
405, 407, 409, 411, 413, 415 and/or 417. According to some
embodiments, a transcribable DNA sequence of a recombinant DNA
molecule, vector or construct is or comprises a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 386, 388, 390, 392, 394,
396, 398, 400, 402, 404, 406, 408, 410, 412, 414 and/or 416.
According to some embodiments, a GA2 oxidase protein encoded by a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 419, 421, 423, 425, 427, 429, 431, 433, 435,
437, 439, 441, 443, 445 and/or 447. According to some embodiments,
a transcribable DNA sequence of a recombinant DNA molecule, vector
or construct is or comprises a sequence that is at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 418, 420, 422, 424, 426, 428, 430, 432,
434, 436, 438, 440, 442, 444 and/or 446. According to some
embodiments, a GA2 oxidase protein encoded by a transcribable DNA
sequence of a recombinant DNA molecule, vector or construct is or
comprises a sequence that is at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 449, 451, 453, 455, 457, 459, 461, 463, 465, 467 and/or
469. According to some embodiments, a transcribable DNA sequence of
a recombinant DNA molecule, vector or construct is or comprises a
sequence that is at least 80%, at least 85%, at least 90%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99%, at
least 99.5%, or 100% identical to one or more of SEQ ID NOs: 448,
450, 452, 454, 456, 458, 460, 462, 464, 466 and/or 468.
[0460] According to some embodiments, a GA2 oxidase protein encoded
by a transcribable DNA sequence of a recombinant DNA molecule,
vector or construct is or comprises a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to a protein sequence from a dicot or leguminous plant.
According to some embodiments, a GA2 oxidase protein encoded by a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 471, 473, 475, 477, 479, 481, 483, 485, 487,
489, 491, 493, 495, 497, 499 and/or 501. According to some
embodiments, a transcribable DNA sequence of a recombinant DNA
molecule, vector or construct is or comprises a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 470, 472, 474, 476, 478,
480, 482, 484, 486, 488, 490, 492, 494, 496, 498 and/or 500.
According to some embodiments, a GA2 oxidase protein encoded by a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 503, 505, 507, 509, 511, 513, 515, 517, 519,
521, 523, 525, 527, 529 and/or 531. According to some embodiments,
a transcribable DNA sequence of a recombinant DNA molecule, vector
or construct is or comprises a sequence that is at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 502, 504, 506, 508, 510, 512, 514, 516,
518, 520, 522, 524, 526, 528 and/or 530. According to some
embodiments, a GA2 oxidase protein encoded by a transcribable DNA
sequence of a recombinant DNA molecule, vector or construct is or
comprises a sequence that is at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555,
557, 559 and/or 561. According to some embodiments, a transcribable
DNA sequence of a recombinant DNA molecule, vector or construct is
or comprises a sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552,
554, 556, 558 and/or 560. According to some embodiments, a GA2
oxidase protein encoded by a transcribable DNA sequence of a
recombinant DNA molecule, vector or construct is or comprises a
sequence that is at least 80%, at least 85%, at least 90%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99%, at
least 99.5%, or 100% identical to one or more of SEQ ID NOs: 563,
565, 567, 569, 571, 573 and/or 575. According to some embodiments,
a transcribable DNA sequence of a recombinant DNA molecule, vector
or construct is or comprises a sequence that is at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 562, 564, 566, 568, 570, 572 and/or 574.
According to some embodiments, a GA2 oxidase protein encoded by a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 577, 579, 581, 583, 585, 587 and/or 589.
According to some embodiments, a transcribable DNA sequence of a
recombinant DNA molecule, vector or construct is or comprises a
sequence that is at least 80%, at least 85%, at least 90%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99%, at
least 99.5%, or 100% identical to one or more of SEQ ID NOs: 576,
578, 580, 582, 584, 586 and/or 588. According to some embodiments,
a GA2 oxidase protein encoded by a transcribable DNA sequence of a
recombinant DNA molecule, vector or construct is or comprises a
sequence that is at least 80%, at least 85%, at least 90%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99%, at
least 99.5%, or 100% identical to one or more of SEQ ID NOs: 591,
593, 595, 597, 599, 601, 603, 605 and/or 607. According to some
embodiments, a transcribable DNA sequence of a recombinant DNA
molecule, vector or construct is or comprises a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 590, 592, 594, 596, 598,
600, 602, 604 and/or 606. According to some embodiments, a GA2
oxidase protein encoded by a transcribable DNA sequence of a
recombinant DNA molecule, vector or construct is or comprises a
sequence that is at least 80%, at least 85%, at least 90%, at least
95%, at least 96%, at least 97%, at least 98%, at least 99%, at
least 99.5%, or 100% identical to one or more of SEQ ID NOs: 609,
611, 613 and/or 615. According to some embodiments, a transcribable
DNA sequence of a recombinant DNA molecule, vector or construct is
or comprises a sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 608, 610, 612 and/or 614. According to some
embodiments, a GA2 oxidase protein encoded by a transcribable DNA
sequence of a recombinant DNA molecule, vector or construct is or
comprises a sequence that is at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 617, 619, 621, 623, 625, 627, 629 and/or 631. According to
some embodiments, a transcribable DNA sequence of a recombinant DNA
molecule, vector or construct is or comprises a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 616, 618, 620, 622, 624,
626, 628 and/or 630. According to some embodiments, a GA2 oxidase
protein encoded by a transcribable DNA sequence of a recombinant
DNA molecule, vector or construct is or comprises a sequence that
is at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 633, 635, 637, 639,
641, 643 and/or 645. According to some embodiments, a transcribable
DNA sequence of a recombinant DNA molecule, vector or construct is
or comprises a sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 632, 634, 636, 638, 640, 642 and/or 644. According to
some embodiments, a GA2 oxidase protein encoded by a transcribable
DNA sequence of a recombinant DNA molecule, vector or construct is
or comprises a sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 647 and/or 649. According to some embodiments, a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 646 and/or 648. According to some
embodiments, a GA2 oxidase protein encoded by a transcribable DNA
sequence of a recombinant DNA molecule, vector or construct is or
comprises a sequence that is at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 651, 653 and/or 655. According to some embodiments, a
transcribable DNA sequence of a recombinant DNA molecule, vector or
construct is or comprises a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 650, 652 and/or 654.
[0461] According to some embodiments, a GA2 oxidase protein encoded
by a transcribable DNA sequence of a recombinant DNA molecule,
vector or construct is or comprises a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 325, 327, 329, 331, 333,
335, 337, 339, 341, 343, 345, 347, 349, 351, 353, 355, 357, 359,
361, 363, 365, 367, 369, 371, 373, 375, 377, 379, 381, 383, 385,
387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411,
413, 415, 417, 419, 421, 423, 425, 427, 429, 431, 433, 435, 437,
439, 441, 443, 445, 447, 449, 451, 453, 455, 457, 459, 461, 463,
465, 467 and/or 469. According to some embodiments, a transcribable
DNA sequence of a recombinant DNA molecule, vector or construct is
or comprises a sequence that is at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344,
346, 348, 350, 352, 354, 356, 358, 360, 362, 364, 366, 368, 370,
372, 374, 376, 378, 380, 382, 384, 386, 388, 390, 392, 394, 396,
398, 400, 402, 404, 406, 408, 410, 412, 414, 416, 418, 420, 422,
424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, 446, 448,
450, 452, 454, 456, 458, 460, 462, 464, 466 and/or 468.
[0462] According to some embodiments, a GA2 oxidase protein encoded
by a transcribable DNA sequence of a recombinant DNA molecule,
vector or construct is or comprises a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 471, 473, 475, 477, 479,
481, 483, 485, 487, 489, 491, 493, 495, 497, 499, 501, 503, 505,
507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527, 529, 531,
533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557,
559, 561, 563, 565, 567, 569, 571, 573, 575, 577, 579, 581, 583,
585, 587, 589, 591, 593, 595, 597, 599, 601, 603, 605, 607, 609,
611, 613, 615, 617, 619, 621, 623, 625, 627, 629, 631, 633, 635,
637, 639, 641, 643, 645, 647, 649, 651, 653 and/or 655. According
to some embodiments, a transcribable DNA sequence of a recombinant
DNA molecule, vector or construct is or comprises a sequence that
is at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 470, 472, 474, 476,
478, 480, 482, 484, 486, 488, 490, 492, 494, 496, 498, 500, 502,
504, 506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528,
530, 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552, 554,
556, 558, 560, 562, 564, 566, 568, 570, 572, 574, 576, 578, 580,
582, 584, 586, 588, 590, 592, 594, 596, 598, 600, 602, 604, 606,
608, 610, 612, 614, 616, 618, 620, 622, 624, 626, 628, 630, 632,
634, 636, 638, 640, 642, 644, 646, 648, 650, 652 and/or 654.
[0463] According to embodiments of the present invention, the
level(s) of one or more active GAs may be reduced in the stalk or
stem of a corn plant by ectopically expressing a catabolic GA2
oxidase gene to produce the short stature phenotype and resistance
to lodging in transgenic plants, but without off-types in the
reproductive or ear tissues of the plant.
[0464] According to embodiments of the present invention,
expression of a GA2 oxidase transgene may be driven by a variety of
different plant-expressible promoter types including constitutive
and tissue-specific or tissue-preferred promoters, such as a
vascular or leaf promoter. According to present embodiments, a
recombinant DNA molecule, vector or construct for expression of a
GA2 oxidase transgene in a plant is provided comprising a
transcribable DNA sequence encoding a protein that is at least 80%,
at least 85%, at least 90%, at least 95%, at least 96%, at least
97%, at least 98%, at least 99%, at least 99.5%, or 100% identical
to a GA2 oxidase protein sequence provided herein, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter, such as a constitutive, vascular or
leaf promoter. According to some embodiments, a recombinant DNA
molecule, vector or construct is provided comprising a
transcribable DNA sequence encoding a GA2 oxidase protein that
comprises or consists of a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 325, 327, 329, 331, 333, 335, 337, 339, 341,
343, 345, 347 and/or 349, wherein the transcribable DNA sequence is
operably linked to a plant-expressible promoter. According to some
of these embodiments, the transcribable DNA sequence of the
recombinant DNA molecule, vector or construct comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
324, 326, 328, 330, 332, 334, 336, 338, 340, 342, 344, 346, and/or
348. According to some embodiments, the plant expressible promoter
is a vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0465] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 324, 326, 328, 330, 332,
334, 336, 338, 340, 342, 344, 346, and/or 348, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least 75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0466] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
351, 353, 355, 357, 359, 361, 363, 365, 367 and/or 369, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some of these embodiments,
the transcribable DNA sequence of the recombinant DNA molecule,
vector or construct comprises or consists of a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 350, 352, 354, 356, 358,
360, 362, 364, 366 and/or 368. According to some embodiments, the
plant expressible promoter is a vascular promoter. According to
some embodiments, the plant expressible promoter is a vascular
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 656, 657, 658, 659,
660, 661, and/or 662, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a RTBV promoter (e.g., a promoter comprising the RTBV
(SEQ ID NO: 656) or truncated RTBV (SEQ ID NO: 657) sequence) or a
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0467] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 350, 352, 354, 356, 358,
360, 362, 364, 366 and/or 368, wherein the transcribable DNA
sequence is operably linked to a plant-expressible promoter.
According to some embodiments, the transcribable DNA sequence
encodes a GA2 oxidase protein. According to some embodiments, the
plant expressible promoter is a vascular promoter. According to
some embodiments, the plant expressible promoter is a vascular
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 656, 657, 658, 659,
660, 661, and/or 662, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a RTBV promoter (e.g., a promoter comprising the RTBV
(SEQ ID NO: 656) or truncated RTBV (SEQ ID NO: 657) sequence) or a
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0468] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
371, 373, 375, 377, 379, 381, 383, and/or 385, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some of these embodiments,
the transcribable DNA sequence of the recombinant DNA molecule,
vector or construct comprises or consists of a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 370, 372, 374, 376, 378,
380, 382, and/or 384. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0469] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 370, 372, 374, 376, 378,
380, 382, and/or 384, wherein the transcribable DNA sequence is
operably linked to a plant-expressible promoter. According to some
embodiments, the transcribable DNA sequence encodes a GA2 oxidase
protein. According to some embodiments, the plant expressible
promoter is a vascular promoter. According to some embodiments, the
plant expressible promoter is a vascular promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or
a functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0470] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
387, 389, 391, 393, 395, 397, 399, 401, 403, 405, 407, 409, 411,
413, 415, and/or 417, wherein the transcribable DNA sequence is
operably linked to a plant-expressible promoter. According to some
of these embodiments, the transcribable DNA sequence of the
recombinant DNA molecule, vector or construct comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
386, 388, 390, 392, 394, 396, 398, 400, 402, 404, 406, 408, 410,
412, 414, and/or 416. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0471] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 386, 388, 390, 392, 394,
396, 398, 400, 402, 404, 406, 408, 410, 412, 414, and/or 416,
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least 75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0472] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
419, 421, 423, 425, 427, 429, 431, 433, 435, 437, 439, 441, 443,
445, and/or 447, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some of these
embodiments, the transcribable DNA sequence of the recombinant DNA
molecule, vector or construct comprises or consists of a sequence
that is at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 418, 420,
421, 424, 426, 428, 430, 432, 434, 436, 438, 440, 442, 444, and/or
446. According to some embodiments, the plant expressible promoter
is a vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0473] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 418, 420, 421, 424, 426,
428, 430, 432, 434, 436, 438, 440, 442, 444, and/or 446, wherein
the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least 75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0474] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
449, 451, 453, 455, 457, 459, 461, 463, 465, 467, and/or 469,
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some of these embodiments,
the transcribable DNA sequence of the recombinant DNA molecule,
vector or construct comprises or consists of a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 448, 450, 452, 454, 456,
458, 460, 462, 464, 466, and/or 468. According to some embodiments,
the plant expressible promoter is a vascular promoter. According to
some embodiments, the plant expressible promoter is a vascular
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 656, 657, 658, 659,
660, 661, and/or 662, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a RTBV promoter (e.g., a promoter comprising the RTBV
(SEQ ID NO: 656) or truncated RTBV (SEQ ID NO: 657) sequence) or a
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0475] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 448, 450, 452, 454, 456,
458, 460, 462, 464, 466, and/or 468, wherein the transcribable DNA
sequence is operably linked to a plant-expressible promoter.
According to some embodiments, the transcribable DNA sequence
encodes a GA2 oxidase protein. According to some embodiments, the
plant expressible promoter is a vascular promoter. According to
some embodiments, the plant expressible promoter is a vascular
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 656, 657, 658, 659,
660, 661, and/or 662, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a RTBV promoter (e.g., a promoter comprising the RTBV
(SEQ ID NO: 656) or truncated RTBV (SEQ ID NO: 657) sequence) or a
promoter comprising a sequence that is at least 70%, at least 75%,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0476] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493, 495,
497, 499, and/or 501, wherein the transcribable DNA sequence is
operably linked to a plant-expressible promoter. According to some
of these embodiments, the transcribable DNA sequence of the
recombinant DNA molecule, vector or construct comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490, 492, 494,
496, 498, and/or 500. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0477] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 470, 472, 474, 476, 478,
480, 482, 484, 486, 488, 490, 492, 494, 496, 498, and/or 500,
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least 75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0478] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525, 527,
529, and/or 531, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some of these
embodiments, the transcribable DNA sequence of the recombinant DNA
molecule, vector or construct comprises or consists of a sequence
that is at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 502, 504,
506, 508, 510, 512, 514, 516, 518, 520, 522, 524, 526, 528, and/or
530. According to some embodiments, the plant expressible promoter
is a vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0479] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 502, 504, 506, 508, 510,
512, 514, 516, 518, 520, 522, 524, 526, 528, and/or 530, wherein
the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least 75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0480] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555, 557,
559, and/or 561, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some of these
embodiments, the transcribable DNA sequence of the recombinant DNA
molecule, vector or construct comprises or consists of a sequence
that is at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 532, 534,
536, 538, 540, 542, 544, 546, 548, 550, 552, 554, 556, 558, and/or
560. According to some embodiments, the plant expressible promoter
is a vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0481] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 532, 534, 536, 538, 540,
542, 544, 546, 548, 550, 552, 554, 556, 558, and/or 560, wherein
the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least 75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0482] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
563, 565, 567, 569, 571, 573, and/or 575, wherein the transcribable
DNA sequence is operably linked to a plant-expressible promoter.
According to some of these embodiments, the transcribable DNA
sequence of the recombinant DNA molecule, vector or construct
comprises or consists of a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 562, 564, 566, 568, 570, 572, and/or 574.
According to some embodiments, the plant expressible promoter is a
vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0483] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 562, 564, 566, 568, 570,
572, and/or 574, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some
embodiments, the transcribable DNA sequence encodes a GA2 oxidase
protein. According to some embodiments, the plant expressible
promoter is a vascular promoter. According to some embodiments, the
plant expressible promoter is a vascular promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or
a functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0484] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
577, 579, 581, 583, 585, 587, and/or 589, wherein the transcribable
DNA sequence is operably linked to a plant-expressible promoter.
According to some of these embodiments, the transcribable DNA
sequence of the recombinant DNA molecule, vector or construct
comprises or consists of a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 576, 578, 580, 582, 584, 586, and/or 588.
According to some embodiments, the plant expressible promoter is a
vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0485] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 576, 578, 580, 582, 584,
586, and/or 588, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some
embodiments, the transcribable DNA sequence encodes a GA2 oxidase
protein. According to some embodiments, the plant expressible
promoter is a vascular promoter. According to some embodiments, the
plant expressible promoter is a vascular promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or
a functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0486] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
591, 593, 595, 597, 599, 601, 603, 605, and/or 607, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some of these embodiments,
the transcribable DNA sequence of the recombinant DNA molecule,
vector or construct comprises or consists of a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 590, 592, 594, 596, 598,
600, 602, 604, and/or 608. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0487] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 590, 592, 594, 596, 598,
600, 602, 604, and/or 608, wherein the transcribable DNA sequence
is operably linked to a plant-expressible promoter. According to
some embodiments, the transcribable DNA sequence encodes a GA2
oxidase protein. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0488] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
609, 611, 613, and/or 615, wherein the transcribable DNA sequence
is operably linked to a plant-expressible promoter. According to
some of these embodiments, the transcribable DNA sequence of the
recombinant DNA molecule, vector or construct comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
608, 610, 612, and/or 614. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0489] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 608, 610, 612, and/or 614,
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least \75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0490] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
617, 619, 621, 623, 625, 627, 629, and/or 631, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some of these embodiments,
the transcribable DNA sequence of the recombinant DNA molecule,
vector or construct comprises or consists of a sequence that is at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 616, 618, 620, 622, 624,
626, 628, and/or 630. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0491] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 616, 618, 620, 622, 624,
626, 628, and/or 630, wherein the transcribable DNA sequence is
operably linked to a plant-expressible promoter. According to some
embodiments, the transcribable DNA sequence encodes a GA2 oxidase
protein. According to some embodiments, the plant expressible
promoter is a vascular promoter. According to some embodiments, the
plant expressible promoter is a vascular promoter comprising a
sequence that is at least 70%, at least \75%, at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or
662, or a functional portion of any of the foregoing. According to
some embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0492] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
633, 635, 637, 639, 641, 643, and/or 645, wherein the transcribable
DNA sequence is operably linked to a plant-expressible promoter.
According to some of these embodiments, the transcribable DNA
sequence of the recombinant DNA molecule, vector or construct
comprises or consists of a sequence that is at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 632, 634, 636, 638, 640, 642, and/or 644.
According to some embodiments, the plant expressible promoter is a
vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0493] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 632, 634, 636, 638, 640,
642, and/or 644, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some
embodiments, the transcribable DNA sequence encodes a GA2 oxidase
protein. According to some embodiments, the plant expressible
promoter is a vascular promoter. According to some embodiments, the
plant expressible promoter is a vascular promoter comprising a
sequence that is at least 70%, at least \75%, at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or
662, or a functional portion of any of the foregoing. According to
some embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0494] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs: 647
and/or 649, wherein the transcribable DNA sequence is operably
linked to a plant-expressible promoter. According to some of these
embodiments, the transcribable DNA sequence of the recombinant DNA
molecule, vector or construct comprises or consists of a sequence
that is at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 646 and/or
648. According to some embodiments, the plant expressible promoter
is a vascular promoter. According to some embodiments, the plant
expressible promoter is a vascular promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a
functional portion of any of the foregoing. According to some
embodiments, the plant expressible promoter is a RTBV promoter
(e.g., a promoter comprising the RTBV (SEQ ID NO: 656) or truncated
RTBV (SEQ ID NO: 657) sequence) or a promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to SEQ ID NO: 656
and/or 657. According to some embodiments, the plant expressible
promoter is a leaf promoter. According to some embodiments, the
plant expressible promoter is a leaf promoter comprising a sequence
that is at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 663, 664, and/or 665, or a functional portion of any of
the foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0495] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 646 and/or 648, wherein the
transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least \75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0496] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence encoding a GA2 oxidase protein that comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
651, 653, and/or 655, wherein the transcribable DNA sequence is
operably linked to a plant-expressible promoter. According to some
of these embodiments, the transcribable DNA sequence of the
recombinant DNA molecule, vector or construct comprises or consists
of a sequence that is at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
650, 652, and/or 654. According to some embodiments, the plant
expressible promoter is a vascular promoter. According to some
embodiments, the plant expressible promoter is a vascular promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 656, 657, 658, 659, 660,
661, and/or 662, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
RTBV promoter (e.g., a promoter comprising the RTBV (SEQ ID NO:
656) or truncated RTBV (SEQ ID NO: 657) sequence) or a promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to SEQ ID NO: 656 and/or 657. According to some
embodiments, the plant expressible promoter is a leaf promoter.
According to some embodiments, the plant expressible promoter is a
leaf promoter comprising a sequence that is at least 70%, at least
75%, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 663, 664,
and/or 665, or a functional portion of any of the foregoing.
According to some embodiments, the plant expressible promoter is a
constitutive promoter. According to some embodiments, the plant
expressible promoter is a constitutive promoter comprising a
sequence that is at least 70%, at least 75%, at least 80%, at least
85%, at least 90%, at least 95%, at least 96%, at least 97%, at
least 98%, at least 99%, at least 99.5%, or 100% identical to one
or more of SEQ ID NOs: 666, 667, 668, 669, 670, 671, 672, 673,
and/or 674, or a functional portion of any of the foregoing.
[0497] According to some embodiments, a recombinant DNA molecule,
vector or construct is provided comprising a transcribable DNA
sequence that comprises or consists of a sequence that is at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 650, 652, and/or 654,
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter. According to some embodiments, the
transcribable DNA sequence encodes a GA2 oxidase protein. According
to some embodiments, the plant expressible promoter is a vascular
promoter. According to some embodiments, the plant expressible
promoter is a vascular promoter comprising a sequence that is at
least 70%, at least \75%, at least 80%, at least 85%, at least 90%,
at least 95%, at least 96%, at least 97%, at least 98%, at least
99%, at least 99.5%, or 100% identical to one or more of SEQ ID
NOs: 656, 657, 658, 659, 660, 661, and/or 662, or a functional
portion of any of the foregoing. According to some embodiments, the
plant expressible promoter is a RTBV promoter (e.g., a promoter
comprising the RTBV (SEQ ID NO: 656) or truncated RTBV (SEQ ID NO:
657) sequence) or a promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to SEQ ID NO: 656 and/or 657.
According to some embodiments, the plant expressible promoter is a
leaf promoter. According to some embodiments, the plant expressible
promoter is a leaf promoter comprising a sequence that is at least
70%, at least 75%, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
663, 664, and/or 665, or a functional portion of any of the
foregoing. According to some embodiments, the plant expressible
promoter is a constitutive promoter. According to some embodiments,
the plant expressible promoter is a constitutive promoter
comprising a sequence that is at least 70%, at least 75%, at least
80%, at least 85%, at least 90%, at least 95%, at least 96%, at
least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 666, 667, 668, 669, 670,
671, 672, 673, and/or 674, or a functional portion of any of the
foregoing.
[0498] According to many embodiments, a modified or transgenic corn
plant is provided comprising and/or transformed with a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein as provided herein. According to some
embodiments, a modified or transgenic corn plant is provided that
is transformed with a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase mRNA and protein,
wherein the transcribable DNA sequence is operably linked to a
plant-expressible promoter, wherein the GA2 oxidase mRNA and/or
protein is identical to an endogenous GA2 oxidase protein, and
wherein the expression level of the GA2 oxidase mRNA and/or protein
is increased in one or more plant tissue(s) of the modified or
transgenic plant as compared to a wild type or control plant, such
as increased in one or more vascular and/or leaf tissue(s) of the
modified or transgenic plant, such as by at least 5%, at least 10%,
at least 20%, at least 25%, at least 30%, at least 40%, at least
50%, at least 60%, at least 70%, at least 75%, at least 80%, at
least 90%, or 100%, as compared to a wild type or control
plant.
[0499] According to present embodiments, a modified or transgenic
corn plant is provided comprising a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein as provided herein, wherein the level of one or more active
GAs, such as GA1, GA3, GA4, and/or GA7, is reduced or lowered in
one or more plant tissue(s), such as one or more stem, internode,
vascular and/or leaf tissue(s) or one or more stem and/or internode
tissue(s), of the modified or transgenic plant, such as by at least
5%, at least 10%, at least 20%, at least 25%, at least 30%, at
least 40%, at least 50%, at least 60%, at least 70%, at least 75%,
at least 80%, at least 90%, or 100%, as compared to a wild type or
control plant.
[0500] According to many embodiments, a modified or transgenic
plant is provided that is transformed with a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein as provided herein, wherein the transcribable DNA
sequence is operably linked to a constitutive promoter or a
tissue-specific or tissue-preferred promoter, such as a vascular
promoter or a leaf promoter, and wherein the modified or transgenic
plant has one or more of the following traits: a semi-dwarf or
reduced plant height or stature, decreased stem internode length,
increased lodging resistance, and/or increased stem or stalk
diameter. Such a modified or transgenic plant may not have any
significant reproductive off-types. A modified or transgenic plant
may have one or more of the following additional traits: reduced
green snap, deeper roots, increased leaf area, earlier canopy
closure, higher stomatal conductance, lower ear height, increased
foliar water content, improved drought tolerance, increased
nitrogen use efficiency, increased water use efficiency, reduced
anthocyanin content and anthocyanin area in leaves under normal
and/or nitrogen or water limiting stress conditions, increased ear
weight, increased kernel number, increased kernel weight, increased
yield, and/or increased harvest index. According to many
embodiments, the level of one or more active GAs, such as GA1, GA3,
GA4, and/or GA7, is/are reduced or lowered in one or more plant
tissue(s), such as one or more stem, internode, vascular and/or
leaf tissue(s), or one or more stem and/or internode tissue(s), of
the modified or transgenic plant, such as by at least 5%, at least
10%, at least 20%, at least 25%, at least 30%, at least 40%, at
least 50%, at least 60%, at least 70%, at least 75%, at least 80%,
at least 90%, or 100%, as compared to a wild type or control
plant.
[0501] A recombinant DNA molecule, construct or vector of the
present disclosure may comprise a transcribable DNA sequence
encoding a GA2 oxidase as provided herein, wherein the
transcribable DNA sequence is operatively linked to a
plant-expressible promoter, such as a constitutive or vascular
and/or leaf promoter. In addition to its associated promoter, a
transcribable DNA sequence encoding a GA2 oxidase may also be
operatively linked to one or more additional regulatory element(s),
such as an enhancer(s), leader, transcription start site (TSS),
linker, 5' and 3' untranslated region(s) (UTRs), intron(s),
polyadenylation signal, termination region or sequence, etc., that
are suitable, necessary or preferred for strengthening, regulating
or allowing expression of the transcribable DNA sequence in a corn
plant cell. Such additional regulatory element(s) may be optional
and/or used to enhance or optimize expression of the transgene or
transcribable DNA sequence. As provided herein, an "enhancer" may
be distinguished from a "promoter" in that an enhancer typically
lacks a transcription start site, TATA box, or equivalent sequence
and is thus insufficient alone to drive transcription. As used
herein, a "leader" may be defined generally as the DNA sequence of
the 5'-UTR of a gene (or transgene) between the transcription start
site (TSS) and 5' end of the transcribable DNA sequence or protein
coding sequence start site of the transgene.
[0502] Transgenic plants expressing a GA2 oxidase transgene may
have an earlier canopy closure (e.g., approximately one day
earlier, or 12-48 hours, 12-36 hours, 18-36 hours, or about 24
hours earlier canopy closure) than a wild type or control plant.
Although transgenic plants expressing a GA2 oxidase transgene may
have a lower ear height than a wild type or control plant, the
height of the ear may generally be at least 18 inches above the
ground. Transgenic plants expressing a GA2 oxidase may have greater
biomass and/or leaf area during one or more late vegetative stages
(e.g., V8-V12) than a wild type or control plant. Transgenic plants
expressing a GA2 oxidase may have deeper roots during later
vegetative stages when grown in the field, than a wild type or
control plant, which may be due to an increased root front
velocity. These transgenic plants may reach a depth 90 cm below
ground sooner (e.g., 5-25 days sooner, 5-20 days sooner, 5-15 days
sooner, 10-25 days sooner, or 15-25 days sooner, or about 5, 10,
15, 20 Or 25 days sooner) than a wild type or control plant, which
may occur by or prior to the vegetative to reproductive transition
of the plant (e.g., by V16/R1 at about 50 days after planting as
opposed to about 70 days after planting for control plants).
[0503] According to some embodiments, a recombinant DNA construct
or vector may comprise two or more expression elements or cassettes
that may be stacked together in a construct or vector either in
tandem in a single expression cassette or separately in two or more
expression cassettes. A recombinant DNA construct or vector may
comprise either a single expression cassette comprising a
transcribable DNA sequence that encodes a GA2 oxidase mRNA and
protein or two or more expression cassettes comprising two or more
transcribable DNA sequences that encode two or more GA2 oxidase
mRNAs and proteins, including at least a first GA2 oxidase mRNA and
protein and a second GA2 oxidase mRNA and protein, wherein the two
or more transcribable DNA sequences, GA2 oxidase mRNAs and/or GA2
oxidase proteins are the same or different, and wherein each
transcribable DNA sequence is operably linked to a
plant-expressible promoter. The plant-expressible promoter may be a
constitutive promoter, or a tissue-specific or tissue-preferred
promoter, as provided herein. If two or more transcribable DNA
sequences are present in a recombinant DNA construct or vector or a
modified or transgenic plant, plant part, cell, or explant, each
transcribable DNA sequence may be operably linked to the same or
different plant-expressible promoters.
[0504] According to other embodiments, a recombinant DNA construct
or vector may comprise two or more expression cassettes including a
first expression cassette and a second expression cassette, wherein
the first expression cassette comprises a first transcribable DNA
sequence operably linked to a first plant-expressible promoter, and
the second expression cassette comprises a second transcribable DNA
sequence operably linked to a second plant-expressible promoter,
wherein the first transcribable DNA sequence encodes a first GA2
oxidase and the second transcribable DNA sequence encodes a second
GA2 oxidase. The first and second plant-expressible promoters may
each be a constitutive promoter, or a tissue-specific or
tissue-preferred promoter, as provided herein, and the first and
second plant-expressible promoters may be the same or different
promoters.
[0505] According to other embodiments, two or more constructs,
expression cassettes or transgenes encoding one or more GA2 oxidase
proteins may be combined in a modified plant by crossing two or
more plants together in one or more generations to produce a
modified plant having a desired combination of the constructs,
expression cassettes or transgenes. According to these embodiments,
a first modified plant comprising a first construct, expression
cassette or transgene encoding a first GA2 oxidase protein may be
crossed to a second modified plant comprising a second construct,
expression cassette or transgene encoding a second GA2 oxidase
protein, such that a modified progeny plant may be made comprising
the first construct, expression cassette or transgene and the
second construct, expression cassette or transgene. Alternatively,
a modified plant comprising two or more constructs, expression
cassettes or transgenes encoding two or more GA2 oxidase proteins
may be made by (i) co-transforming a first construct, expression
cassette or transgene and a second construct, expression cassette
or transgene (each encoding a GA2 oxidase protein) in the same or
different transformation molecules or vectors, (ii) transforming a
modified plant with a second construct, expression cassette or
transgene in a transformation molecule or vector, wherein the
modified plant already comprises a first construct, expression
cassette or transgene, or (iii) transforming a plant with a first
construct, expression cassette or transgene in a first
transformation molecule or vector, and then transforming the plant
with a second construct, expression cassette or transgene in a
second transformation molecule or vector.
[0506] According to embodiments of the present disclosure, modified
plants are provided comprising two or more constructs comprising
GA2 oxidase transgene(s) including a first recombinant DNA
construct and a second recombinant DNA construct, wherein the first
recombinant DNA construct comprises a first transcribable DNA
sequence encoding a first GA2 oxidase mRNA and protein, and the
second recombinant DNA construct comprises a second transcribable
DNA sequence encoding a second GA2 oxidase mRNA and protein. The
first and second recombinant DNA constructs may be stacked in a
single vector and transformed into a plant as a single event, or
present in separate vectors or constructs that may be transformed
as separate events. According to some embodiments, the first and
second GA2 oxidase transgenes may be the same or different GA
oxidase gene(s).
[0507] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein fewer than or equal to 50%
of said corn plants have lodged at the time of harvest, and wherein
at least one corn plant of said plurality of corn plants comprises
a recombinant DNA construct comprising a transcribable DNA sequence
encoding a GA2 oxidase protein and a plant-expressible promoter,
wherein the transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0508] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0509] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter.
[0510] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0511] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average yield of said
field is at least 170 bushels per acre, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a recombinant DNA construct comprising a transcribable
DNA sequence encoding a GA2 oxidase protein and a plant-expressible
promoter, wherein the transcribable DNA sequence is operably linked
to the plant-expressible promoter.
[0512] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter.
[0513] In some embodiments, this disclosure provides a method
comprising harvesting a plurality of corn plants from a field at
least 1 day after the average kernel moisture content of at least
50% of said plurality of corn plants is between 10% and 30%,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible
promoter.
[0514] In an aspect, at least 10% of the corn plants in a field
comprise a recombinant DNA construct comprising a transcribable DNA
sequence encoding a GA2 oxidase protein and a plant-expressible
promoter, wherein the transcribable DNA sequence is operably linked
to the plant-expressible promoter. In an aspect, at least 20% of
the corn plants in a field comprise a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible promoter.
In an aspect, at least 30% of the corn plants in a field comprise a
recombinant DNA construct comprising a transcribable DNA sequence
encoding a GA2 oxidase protein and a plant-expressible promoter,
wherein the transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, at least 40% of the corn
plants in a field comprise a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. In an aspect,
at least 50% of the corn plants in a field comprise a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, at least 60% of the corn
plants in a field comprise a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. In an aspect,
at least 70% of the corn plants in a field comprise a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, at least 80% of the corn
plants in a field comprise a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. In an aspect,
at least 90% of the corn plants in a field comprise a recombinant
DNA construct comprising a transcribable DNA sequence encoding a
GA2 oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, 100% of the corn plants
in a field comprise a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter.
[0515] In an aspect, between 1% and 100% of the corn plants in a
field comprise a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. In an aspect,
between 10% and 100% of the corn plants in a field comprise a
recombinant DNA construct comprising a transcribable DNA sequence
encoding a GA2 oxidase protein and a plant-expressible promoter,
wherein the transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, between 20% and 100% of
the corn plants in a field comprise a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible promoter.
In an aspect, between 30% and 100% of the corn plants in a field
comprise a recombinant DNA construct comprising a transcribable DNA
sequence encoding a GA2 oxidase protein and a plant-expressible
promoter, wherein the transcribable DNA sequence is operably linked
to the plant-expressible promoter. In an aspect, between 40% and
100% of the corn plants in a field comprise a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, between 50% and 100% of
the corn plants in a field comprise a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible promoter.
In an aspect, between 60% and 100% of the corn plants in a field
comprise a recombinant DNA construct comprising a transcribable DNA
sequence encoding a GA2 oxidase protein and a plant-expressible
promoter, wherein the transcribable DNA sequence is operably linked
to the plant-expressible promoter. In an aspect, between 70% and
100% of the corn plants in a field comprise a recombinant DNA
construct comprising a transcribable DNA sequence encoding a GA2
oxidase protein and a plant-expressible promoter, wherein the
transcribable DNA sequence is operably linked to the
plant-expressible promoter. In an aspect, between 80% and 100% of
the corn plants in a field comprise a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible promoter.
In an aspect, between 90% and 100% of the corn plants in a field
comprise a recombinant DNA construct comprising a transcribable DNA
sequence encoding a GA2 oxidase protein and a plant-expressible
promoter, wherein the transcribable DNA sequence is operably linked
to the plant-expressible promoter.
[0516] A corn field is considered to be "harvested" when at least
one ear has been removed from most, all, or a majority of the corn
plants in the field. A corn plant(s) is/are considered to be
"harvested" when at least one ear has been removed from the corn
plant(s), or at least from most or a majority of the corn
plants.
[0517] As used herein, a "field" refers to an outdoor location that
is suitable for growing corn, and a "corn field" refers to a field
that has been planted with a plurality or population of corn
plants. The field or location can be irrigated or non-irrigated. A
corn field can comprise a land area planted with corn seed and/or
at least one corn plant or a plurality of corn plants, which can be
at one or more stages of development. According to some aspects, a
plurality of corn plants in a field can be at a homogeneous or the
same (or nearly homogeneous or nearly the same) stage of
development, such that the plurality of corn plants have
approximately the same height. In an aspect, a corn plant provided
herein is planted in a field.
[0518] In another aspect, a corn plant provided herein is not
planted in the field, but is planted indoors, such as in a
greenhouse, and/or in a container holding a growth medium or
soil.
[0519] A corn field can comprise one or more rows of corn plants of
the same or different lengths. As used herein, a "row" comprises a
plurality of corn plants in a linear or near linear arrangement. In
an aspect, a row comprises at least two corn plants. Without being
limiting, a row of corn plants is planted in a line, and if a corn
field comprises two or more rows, they are typically planted
parallel to each other. A corn field can comprise one or more rows
of corn plants where the rows are of the same or different lengths.
Without being limiting, a corn field comprises at least 1 row of
corn plants. In another aspect, a corn field comprises at least 10
rows of corn plants. In another aspect, a corn field comprises at
least 50 rows of corn plants. In another aspect, a corn field
comprises at least 500 rows of corn plants. In another aspect, a
corn field comprises at least 1,000 rows of corn plants. In another
aspect, a corn field comprises at least 5,000 rows of corn plants.
In another aspect, a corn field comprises at least 10,000 rows of
corn plants.
[0520] In an aspect, a corn field comprises rows that are spaced at
least 5 inches apart. In another aspect, a corn field comprises
rows that are spaced at least 10 inches apart. In a further aspect,
a corn field comprises rows that are spaced at least 15 inches
apart. In an aspect, a corn field comprises rows of corn plants
that are spaced at least 20 inches apart. In another aspect, a corn
field comprises rows of corn that are spaced at least 25 inches
apart. In another aspect, a corn field comprises rows of corn that
are spaced at least 30 inches apart. According to some aspects, a
corn field can comprise two or more pluralities of corn plants with
the pluralities of corn plants being planted with different corn
varieties, at different times, at different densities, in different
arrangements (e.g., in rows or scattered or random placement),
and/or at different row spacings and/or row lengths, such that the
pluralities of corn plants have different heights, spacings, etc.,
at different time points during the growing season, although each
plurality of corn plants can be relatively uniform with respect to
plant height and other growth metrics.
[0521] In an aspect, a field comprises a single plot. In another
aspect, a field comprises multiple plots. In another aspect, one or
more edges of a field are bordered by a fence. In another aspect,
one or more edges of a field are unfenced. In another aspect, one
or more edges of a field are bordered by hedges. In an aspect, a
field comprises a physically contiguous space. In another aspect,
the field comprises a physically non-contiguous space. In still
another aspect, the field comprises a biologically contiguous
space. As used herein, a "biologically contiguous space" refers to
a space where the pollen can move from one section of a field to
another. In an aspect, a biologically contiguous field is
physically contiguous. In another aspect, a biologically contiguous
field is physically non-contiguous (e.g., plots within the field or
a single plot within the field can be separated by a structure,
without being limiting, such as a road, creek, irrigation ditch,
trail, hedgerow, fence, irrigation pipes, fallow field, empty
field, or non-corn plants).
[0522] In an aspect, a field comprises at least 0.5 acres. In an
aspect, a field comprises at least 1 acre. In another aspect, a
field comprises at least 5 acres. In another aspect, a field
comprises at least 10 acres. In another aspect, a field comprises
at least 15 acres. In another aspect, a field comprises at least 20
acres. In another aspect, a field comprises at least 25 acres. In
another aspect, a field comprises at least 30 acres. In another
aspect, a field comprises at least 35 acres. In another aspect, a
field comprises at least 40 acres. In another aspect, a field
comprises at least 45 acres. In another aspect, a field comprises
at least 50 acres. In another aspect, a field comprises at least 75
acres. In another aspect, a field comprises at least 100 acres. In
another aspect, a field comprises at least 150 acres. In another
aspect, a field comprises at least 200 acres. In another aspect, a
field comprises at least 250 acres. In another aspect, a field
comprises at least 300 acres. In another aspect, a field comprises
at least 350 acres. In another aspect, a field comprises at least
400 acres. In another aspect, a field comprises at least 450 acres.
In another aspect, a field comprises at least 500 acres. In another
aspect, a field comprises at least 750 acres. In another aspect, a
field comprises at least 1000 acres. In another aspect, a field
comprises at least 1500 acres. In another aspect, a field comprises
at least 2000 acres. In another aspect, a field comprises at least
2500 acres. In another aspect, a field comprises at least 3000
acres. In another aspect, a field comprises at least 4000 acres. In
another aspect, a field comprises at least 5000 acres. In another
aspect, a field comprises at least 10,000 acres.
[0523] In an aspect, a field comprises between 0.5 acres and 10,000
acres. In another aspect, a field comprises between 1 acre and
10,000 acres. In another aspect, a field comprises between 5 acres
and 10,000 acres. In another aspect, a field comprises between 10
acres and 10,000 acres. In another aspect, a field comprises
between 15 acres and 10,000 acres. In another aspect, a field
comprises between 20 acres and 10,000 acres. In another aspect, a
field comprises between 25 acres and 10,000 acres. In another
aspect, a field comprises between 30 acres and 10,000 acres. In
another aspect, a field comprises between 35 acres and 10,000
acres. In another aspect, a field comprises between 40 acres and
10,000 acres. In another aspect, a field comprises between 45 acres
and 10,000 acres. In another aspect, a field comprises between 50
acres and 10,000 acres. In another aspect, a field comprises
between 75 acres and 10,000 acres. In another aspect, a field
comprises between 100 acres and 10,000 acres. In another aspect, a
field comprises between 150 acres and 10,000 acres. In another
aspect, a field comprises between 200 acres and 10,000 acres. In
another aspect, a field comprises between 250 acres and 10,000
acres. In another aspect, a field comprises between 300 acres and
10,000 acres. In another aspect, a field comprises between 350
acres and 10,000 acres. In another aspect, a field comprises
between 400 acres and 10,000 acres. In another aspect, a field
comprises between 450 acres and 10,000 acres. In another aspect, a
field comprises between 500 acres and 10,000 acres. In another
aspect, a field comprises between 750 acres and 10,000 acres. In
another aspect, a field comprises between 1000 acres and 10,000
acres. In another aspect, a field comprises between 1500 acres and
10,000 acres. In another aspect, a field comprises between 2000
acres and 10,000 acres. In another aspect, a field comprises
between 2500 acres and 10,000 acres. In another aspect, a field
comprises between 3000 acres and 10,000 acres. In another aspect, a
field comprises between 4000 acres and 10,000 acres. In another
aspect, a field comprises between 5000 acres and 10,000 acres. In
another aspect, a field comprises between 1 acre and 5000 acres. In
another aspect, a field comprises between 1 acre and 2500 acres. In
another aspect, a field comprises between 1 acre and 1000 acres. In
another aspect, a field comprises between 1 acre and 500 acres. In
another aspect, a field comprises between 1 acre and 250 acres. In
another aspect, a field comprises between 1 acre and 100 acres. In
another aspect, a field comprises between 1 acre and 75 acres. In
another aspect, a field comprises between 1 acre and 50 acres. In
another aspect, a field comprises between 1 acre and 25 acres. In
another aspect, a field comprises between 1 acre and 10 acres.
[0524] In an aspect, a corn field can further comprise plants other
than corn plants including, without being limiting, cotton,
alfalfa, sunflowers, sorghum, wheat, barley, oat, rice, rye,
soybean, vegetables (e.g., potato, tomato, carrot), grass (e.g.,
bluegrass, Triticale), and weeds.
[0525] In an aspect, a corn field comprises a density of at least
10,000 corn plants per acre. In another aspect, a corn field
comprises a density of at least 15,000 corn plants per acre. In
another aspect, a corn field comprises a density of at least 20,000
corn plants per acre. In another aspect, a corn field comprises a
density of at least 22,000 corn plants per acre. In another aspect,
a corn field comprises a density of at least 24,000 corn plants per
acre. In another aspect, a corn field comprises a density of at
least 26,000 corn plants per acre. In another aspect, a corn field
comprises a density of at least 28,000 corn plants per acre. In
another aspect, a corn field comprises a density of at least 30,000
corn plants per acre. In another aspect, a corn field comprises a
density of at least 32,000 corn plants per acre. In another aspect,
a corn field comprises a density of at least 34,000 corn plants per
acre. In another aspect, a corn field comprises a density of at
least 36,000 corn plants per acre. In another aspect, a corn field
comprises a density of at least 38,000 corn plants per acre. In
another aspect, a corn field comprises a density of at least 40,000
corn plants per acre. In another aspect, a corn field comprises a
density of at least 42,000 corn plants per acre. In another aspect,
a corn field comprises a density of at least 44,000 corn plants per
acre. In another aspect, a corn field comprises a density of at
least 46,000 corn plants per acre. In another aspect, a corn field
comprises a density of at least 48,000 corn plants per acre. In
another aspect, a corn field comprises a density of at least 50,000
corn plants per acre. In another aspect, a corn field comprises a
density of at least 52,000 corn plants per acre. In another aspect,
a corn field comprises a density of at least 54,000 corn plants per
acre. In another aspect, a corn field comprises a density of at
least 56,000 corn plants per acre. In another aspect, a corn field
comprises a density of at least 58,000 corn plants per acre. In
another aspect, a corn field comprises a density of at least 60,000
corn plants per acre.
[0526] In an aspect, a corn field comprises a density of between
10,000 and 50,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 10,000 and 40,000 corn plants per
acre. In an aspect, a corn field comprises a density of between
10,000 and 30,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 10,000 and 25,000 corn plants per
acre. In an aspect, a corn field comprises a density of between
10,000 and 20,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 20,000 corn plants and 60,000 corn
plants per acre. In an aspect, a corn field comprises a density of
between 20,000 corn plants and 58,000 corn plants per acre. In an
aspect, a corn field comprises a density of between 20,000 corn
plants and 55,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 20,000 corn plants and 50,000 corn
plants per acre. In an aspect, a corn field comprises a density of
between 20,000 corn plants and 45,000 corn plants per acre. In an
aspect, a corn field comprises a density of between 20,000 corn
plants and 42,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 20,000 corn plants and 40,000 corn
plants per acre. In an aspect, a corn field comprises a density of
between 20,000 corn plants and 38,000 corn plants per acre. In an
aspect, a corn field comprises a density of between 20,000 corn
plants and 36,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 20,000 corn plants and 34,000 corn
plants per acre. In an aspect, a corn field comprises a density of
between 20,000 corn plants and 32,000 corn plants per acre. In an
aspect, a corn field comprises a density of between 20,000 corn
plants and 30,000 corn plants per acre. In an aspect, a corn field
comprises a density of between 24,000 corn plants and 58,000 corn
plants per acre. In an aspect, a corn field comprises a density of
between 38,000 corn plants and 60,000 corn plants per acre. In an
aspect, a corn field comprises a density of between 38,000 corn
plants and 50,000 corn plants per acre.
[0527] In an aspect, a corn field comprises at least 10 corn
plants. In another aspect, a corn field comprises at least 10 corn
plants per acre. In an aspect, a corn field comprises at least 100
corn plants. In another aspect, a corn field comprises at least 100
corn plants per acre. In an aspect, a corn field comprises at least
500 corn plants. In another aspect, a corn field comprises at least
500 corn plants per acre. In an aspect, a corn field comprises at
least 1000 corn plants. In another aspect, a corn field comprises
at least 1000 corn plants per acre. In an aspect, a corn field
comprises at least 5000 corn plants. In another aspect, a corn
field comprises at least 5000 corn plants per acre. In an aspect, a
corn field comprises at least 10,000 corn plants. In an aspect, a
corn field comprises at least 10,000 corn plants per acre. In an
aspect, a corn field comprises at least 12,000 corn plants. In an
aspect, a corn field comprises at least 12,000 corn plants per
acre. In an aspect, a corn field comprises at least 15,000 corn
plants. In an aspect, a corn field comprises at least 15,000 corn
plants per acre. In an aspect, a corn field comprises at least
18,000 corn plants. In an aspect, a corn field comprises at least
18,000 corn plants per acre. In an aspect, a corn field comprises
at least 20,000 corn plants. In an aspect, a corn field comprises
at least 20,000 corn plants per acre. In an aspect, a corn field
comprises at least 22,000 corn plants. In an aspect, a corn field
comprises at least 22,000 corn plants per acre. In an aspect, a
corn field comprises at least 24,000 corn plants. In an aspect, a
corn field comprises at least 24,000 corn plants per acre. In an
aspect, a corn field comprises at least 26,000 corn plants. In an
aspect, a corn field comprises at least 26,000 corn plants per
acre. In an aspect, a corn field comprises at least 28,000 corn
plants. In an aspect, a corn field comprises at least 28,000 corn
plants per acre. In an aspect, a corn field comprises at least
30,000 corn plants. In an aspect, a corn field comprises at least
30,000 corn plants per acre. In an aspect, a corn field comprises
at least 32,000 corn plants. In an aspect, a corn field comprises
at least 32,000 corn plants per acre. In an aspect, a corn field
comprises at least 34,000 corn plants. In an aspect, a corn field
comprises at least 34,000 corn plants per acre. In an aspect, a
corn field comprises at least 36,000 corn plants. In an aspect, a
corn field comprises at least 36,000 corn plants per acre. In an
aspect, a corn field comprises at least 38,000 corn plants. In an
aspect, a corn field comprises at least 38,000 corn plants per
acre. In an aspect, a corn field comprises at least 40,000 corn
plants. In an aspect, a corn field comprises at least 40,000 corn
plants per acre. In an aspect, a corn field comprises at least
42,000 corn plants. In an aspect, a corn field comprises at least
42,000 corn plants per acre. In an aspect, a corn field comprises
at least 44,000 corn plants. In an aspect, a corn field comprises
at least 44,000 corn plants per acre. In an aspect, a corn field
comprises at least 46,000 corn plants. In an aspect, a corn field
comprises at least 46,000 corn plants per acre. In an aspect, a
corn field comprises at least 48,000 corn plants. In an aspect, a
corn field comprises at least 48,000 corn plants per acre. In an
aspect, a corn field comprises at least 50,000 corn plants. In an
aspect, a corn field comprises at least 50,000 corn plants per
acre. In an aspect, a corn field comprises at least 52,000 corn
plants. In an aspect, a corn field comprises at least 52,000 corn
plants per acre. In an aspect, a corn field comprises at least
54,000 corn plants. In an aspect, a corn field comprises at least
54,000 corn plants per acre. In an aspect, a corn field comprises
at least 56,000 corn plants. In an aspect, a corn field comprises
at least 56,000 corn plants per acre. In an aspect, a corn field
comprises at least 58,000 corn plants. In an aspect, a corn field
comprises at least 58,000 corn plants per acre. In an aspect, a
corn field comprises at least 60,000 corn plants. In an aspect, a
corn field comprises at least 60,000 corn plants per acre.
[0528] In an aspect, a corn field comprises between 10,000 corn
plants per acre and 50,000 corn plants per acre. In an aspect, a
corn field comprises between 10,000 corn plants per acre and 40,000
corn plants per acre. In an aspect, a corn field comprises between
10,000 corn plants per acre and 30,000 corn plants per acre. In an
aspect, a corn field comprises between 10,000 corn plants per acre
and 25,000 corn plants per acre. In an aspect, a corn field
comprises between 10,000 corn plants per acre and 20,000 corn
plants per acre. In an aspect, a corn field comprises between
20,000 corn plants per acre and 60,000 corn plants per acre. In an
aspect, a corn field comprises between 20,000 corn plants per acre
and 58,000 corn plants per acre. In an aspect, a corn field
comprises between 20,000 corn plants per acre and 55,000 corn
plants per acre. In an aspect, a corn field comprises between
20,000 corn plants per acre and 50,000 corn plants per acre. In an
aspect, a corn field comprises between 20,000 corn plants per acre
and 45,000 corn plants per acre. In an aspect, a corn field
comprises between 20,000 corn plants per acre and 42,000 corn
plants per acre. In an aspect, a corn field comprises between
20,000 corn plants per acre and 40,000 corn plants per acre. In an
aspect, a corn field comprises between 20,000 corn plants per acre
and 38,000 corn plants per acre. In an aspect, a corn field
comprises between 20,000 corn plants per acre and 36,000 corn
plants per acre. In an aspect, a corn field comprises between
20,000 corn plants per acre and 34,000 corn plants per acre. In an
aspect, a corn field comprises between 20,000 corn plants per acre
and 32,000 corn plants per acre. In an aspect, a corn field
comprises between 20,000 corn plants per acre and 30,000 corn
plants per acre. In an aspect, a corn field comprises between
24,000 corn plants per acre and 58,000 corn plants per acre. In an
aspect, a corn field comprises between 38,000 corn plants per acre
and 60,000 corn plants per acre. In an aspect, a corn field
comprises between 38,000 corn plants per acre and 50,000 corn
plants per acre.
[0529] In an aspect, a corn field comprises at least 0.5 acres. In
another aspect, a corn field comprises at least 1 acre. In another
aspect, a corn field comprises at least 3 acres. In another aspect,
a corn field comprises at least 5 acres. In another aspect, a corn
field comprises at least 10 acres. In another aspect, a corn field
comprises at least 15 acres. In another aspect, a corn field
comprises at least 20 acres. In another aspect, a corn field
comprises at least 25 acres. In another aspect, a corn field
comprises at least 50 acres. In another aspect, a corn field
comprises at least 75 acres. In another aspect, a corn field
comprises at least 100 acres. In another aspect, a corn field
comprises at least 150 acres. In another aspect, a corn field
comprises at least 200 acres. In another aspect, a corn field
comprises at least 250 acres. In another aspect, a corn field
comprises at least 300 acres. In another aspect, a corn field
comprises at least 350 acres. In another aspect, a corn field
comprises at least 400 acres. In another aspect, a corn field
comprises at least 500 acres. In another aspect, a corn field
comprises at least 750 acres. In another aspect, a corn field
comprises at least 1000 acres. In another aspect, a corn field
comprises at least 2500 acres. In another aspect, a corn field
comprises at least 5 acres. In another aspect, a corn field
comprises at least 5000 acres.
[0530] In an aspect, a corn field comprises between 0.5 acres and
5000 acres. In another aspect, a corn field comprises between 1
acre and 5000 acres. In another aspect, a corn field comprises
between 5 acres and 5000 acres. In another aspect, a corn field
comprises between 10 acres and 5000 acres. In another aspect, a
corn field comprises between 25 acres and 5000 acres. In another
aspect, a corn field comprises between 50 acres and 5000 acres. In
another aspect, a corn field comprises between 100 acres and 5000
acres. In another aspect, a corn field comprises between 200 acres
and 5000 acres. In another aspect, a corn field comprises between
500 acres and 5000 acres. In another aspect, a corn field comprises
between 1000 acres and 5000 acres. In another aspect, a corn field
comprises between 1 acre and 500 acres. In another aspect, a corn
field comprises between 1 acre and 400 acres. In another aspect, a
corn field comprises between 1 acre and 300 acres. In another
aspect, a corn field comprises between 1 acre and 250 acres. In
another aspect, a corn field comprises between 1 acre and 200
acres. In another aspect, a corn field comprises between 1 acre and
150 acres. In another aspect, a corn field comprises between 1 acre
and 100 acres. In another aspect, a corn field comprises between 1
acre and 75 acres. In another aspect, a corn field comprises
between 1 acre and 50 acres. In another aspect, a corn field
comprises between 1 acre and 25 acres. In another aspect, a corn
field comprises between 10 acres and 25 acres. In another aspect, a
corn field comprises between 10 acres and 50 acres. In another
aspect, a corn field comprises between 10 acres and 100 acres. In
another aspect, a corn field comprises between 10 acres and 250
acres. In another aspect, a corn field comprises between 10 acres
and 500 acres. In another aspect, a corn field comprises between
100 acres and 250 acres. In another aspect, a corn field comprises
between 100 acres and 500 acres.
[0531] As used herein, the term "yield" refers to the amount of
harvested plant material or grain, such as kernels or seeds, but
may also or instead include the amount of biomass harvested
(including for example, stalk, leaves, and/or kernels), from the
plant(s). Harvested grain can be used in a variety of applications
including food processing, animal feed, etc., and biomass may be
used for a variety of applications including sileage, biofuel,
etc., as known in the art. In an aspect, yield is measured as the
amount of biomass or sileage harvested from the plant(s). In
another aspect, yield is measured in bushels per acre. In another
aspect, yield is measured in average number of kernels per ear. In
still another aspect, yield is measured in grams per dry kernel. In
still another aspect, yield is measured in terms of average kernel
weight and the average number of kernels per ear. In still another
aspect, yield is measured in Standard Seed Units (SSU) per acre.
One SSU for corn is equivalent to 80,000 corn seed kernels. The
number of Standard Seed Units (SSUs) is appropriate for seed
production since it quantifies the number of plants that can
potentially be grown from the quantity of seeds, whereas yield
takes into account both seed number and seed size.
[0532] In an aspect, the average yield of a corn field comprises at
least 100 bushels per acre. In an aspect, the average yield of a
corn field comprises at least 120 bushels per acre. In an aspect,
the average yield of a corn field comprises at least 130 bushels
per acre. In an aspect, the average yield of a corn field comprises
at least 140 bushels per acre. In an aspect, the average yield of a
corn field comprises at least 150 bushels per acre. In an aspect,
the average yield of a corn field comprises at least 160 bushels
per acre. In an aspect, the average yield of a corn field comprises
at least 170 bushels per acre. In an aspect, the average yield of a
corn field comprises at least 180 bushels per acre. In an aspect,
the average yield of a corn field comprises at least 190 bushels
per acre. In an aspect, the average yield of a corn field comprises
at least 200 bushels per acre. In an aspect, the average yield of a
corn field comprises at least 210 bushels per acre. In an aspect,
the average yield of a corn field comprises at least 220 bushels
per acre. In an aspect, the average yield of a corn field comprises
at least 230 bushels per acre. In an aspect, the average yield of a
corn field comprises at least 240 bushels per acre. In an aspect,
the average yield of a corn field comprises at least 250 bushels
per acre. In an aspect, the average yield of a corn field comprises
at least 260 bushels per acre.
[0533] In an aspect, the average yield of a corn field comprises
between 100 bushels per acre and 260 bushels per acre. In an
aspect, the average yield of a corn field comprises between 120
bushels per acre and 260 bushels per acre. In an aspect, the
average yield of a corn field comprises between 140 bushels per
acre and 260 bushels per acre. In an aspect, the average yield of a
corn field comprises between 160 bushels per acre and 260 bushels
per acre. In an aspect, the average yield of a corn field comprises
between 180 bushels per acre and 260 bushels per acre. In an
aspect, the average yield of a corn field comprises between 200
bushels per acre and 260 bushels per acre. In an aspect, the
average yield of a corn field comprises between 220 bushels per
acre and 260 bushels per acre. In an aspect, the average yield of a
corn field comprises between 240 bushels per acre and 260 bushels
per acre. In an aspect, the average yield of a corn field comprises
between 100 bushels per acre and 200 bushels per acre. In an
aspect, the average yield of a corn field comprises between 150
bushels per acre and 250 bushels per acre. In an aspect, the
average yield of a corn field comprises between 150 bushels per
acre and 200 bushels per acre.
[0534] In an aspect, the average yield of a corn field harvested at
least 30 days, at least 40 days, at least 50 days, at least 60
days, at least 70 days, at least 80 days at, least 90 days, at
least 100 days, or at least 110 days after fertilization or silking
is within 5% of the average yield of a corn field comprising plants
of the same genetic background harvested between 20 and 30 days
after fertilization or silking. In an aspect, the average yield of
a corn field harvested at least 30 days, at least 40 days, at least
50 days, at least 60 days, at least 70 days, at least 80 days at,
least 90 days, at least 100 days, or at least 110 days after
fertilization or silking is within 10% of the average yield of a
corn field comprising plants of the same genetic background
harvested between 20 and 30 days after fertilization or silking. In
an aspect, the average yield of a corn field harvested at least 30
days, at least 40 days, at least 50 days, at least 60 days, at
least 70 days, at least 80 days, at least 90 days, at least 100
days, or at least 110 days after fertilization or silking is within
15% of the average yield of a corn field comprising plants of the
same genetic background harvested between 20 and 30 days after
fertilization or silking. In an aspect, the average yield of a corn
field harvested at least 30 days, at least 40 days, at least 50
days, at least 60 days, at least 70 days, at least 80 days, at
least 90 days, at least 100 days, or at least 110 days after
fertilization or silking is within 20% of the average yield of a
corn field comprising plants of the same genetic background
harvested between 20 and 30 days after fertilization or silking. In
an aspect, the average yield of a corn field comprising plants of
the same genetic background harvested at least 30 days, at least 40
days, at least 50 days, at least 60 days, at least 70 days, at
least 80 days, at least 90 days, at least 100 days, or at least 110
days after fertilization or silking is within 25% of the average
yield of a corn field comprising plants of the same genetic
background harvested between 20 and 30 days after fertilization or
silking. In an aspect, the average yield of a corn field comprising
plants of the same genetic background harvested at least 30 days,
at least 40 days, at least 50 days, at least 60 days, at least 70
days, at least 80 days, at least 90 days, at least 100 days, or at
least 110 days after fertilization or silking is within 30% of the
average yield of a corn field comprising plants of the same genetic
background harvested between 20 and 30 days after fertilization or
silking.
[0535] Kernel moisture content can be measured by any means
typically used in the art. Non-limiting examples for measuring
kernel moisture content include the use of an electronic grain
moisture tester (e.g., infrared monitors); direct measurement of
water content via a chemical reaction (e.g., the Karl Fischer
method); and drying whole kernel samples and measuring weight loss
during drying.
[0536] In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of less
than or equal to 30%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of less than or equal to 29%. In an aspect, a
method provided herein comprises harvesting corn plants comprising
an average kernel moisture content of less than or equal to 28%. In
an aspect, a method provided herein comprises harvesting corn
plants comprising an average kernel moisture content of less than
or equal to 27%. In an aspect, a method provided herein comprises
harvesting corn plants comprising an average kernel moisture
content of less than or equal to 26%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of less than or equal to 25%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of less than or equal
to 24%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of less
than or equal to 23%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of less than or equal to 22%. In an aspect, a
method provided herein comprises harvesting corn plants comprising
an average kernel moisture content of less than or equal to 21%. In
an aspect, a method provided herein comprises harvesting corn
plants comprising an average kernel moisture content of less than
or equal to 20%. In an aspect, a method provided herein comprises
harvesting corn plants comprising an average kernel moisture
content of less than or equal to 19%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of less than or equal to 18%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of less than or equal
to 17%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of less
than or equal to 16%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of less than or equal to 15%. In an aspect, a
method provided herein comprises harvesting corn plants comprising
an average kernel moisture content of less than or equal to 14%. In
an aspect, a method provided herein comprises harvesting corn
plants comprising an average kernel moisture content of less than
or equal to 13%. In an aspect, a method provided herein comprises
harvesting corn plants comprising an average kernel moisture
content of less than or equal to 12%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of less than or equal to 10%. Each
of the above average kernel moisture content ranges may also apply
to the kernel moisture content of a corn plant, such as the kernel
moisture content of a corn plant of a plurality of corn plants.
[0537] In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 10% and 30%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 11% and 30%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 12% and 30%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 13% and
30%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 14% and 30%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 15% and 30%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 16% and 30%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 17% and
30%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 18% and 30%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 19% and 30%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 20% and 30%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 21% and
30%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 22% and 30%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 23% and 30%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 24% and 30%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 25% and
30%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 26% and 30%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 27% and 30%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 28% and 30%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 29% and
30%. Each of the above average kernel moisture content ranges may
also apply to the kernel moisture content of a corn plant of the
plurality of corn plants.
[0538] In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 10% and 25%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 10% and 20%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 10% and 15%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 10% and
13%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 13% and 25%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 13% and 20%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average kernel moisture content of between 13% and 15%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average kernel moisture content of between 15% and
25%. In an aspect, a method provided herein comprises harvesting
corn plants comprising an average kernel moisture content of
between 15% and 20%. In an aspect, a method provided herein
comprises harvesting corn plants comprising an average kernel
moisture content of between 20% and 25%. Each of the above average
kernel moisture content ranges may also apply to the kernel
moisture content of a corn plant of the plurality of corn
plants.
[0539] In an aspect, methods are provided comprising harvesting a
plurality of corn plants in a field at least 1 day, at least 2
days, at least 3 days, at least 4 days, at least 5 days, at least 6
days, at least 7 days, at least 8 days, at least 9 days, at least
10 days, at least 11 days, at least 12 days, at least 13 days, at
least 14 days, at least 15 days, at least 20 days, at least 25
days, at least 30 days, at least 35 days, at least 40 days, at
least 45 days, at least 50 days, at least 55 days, at least 60
days, at least 65 days, at least 70 days, at least 75 days, at
least 80 days, at least 90 days, at least 95 days, at least 100
days, or at least 1 week, at least 2 weeks, at least 3 weeks, at
least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7
weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at
least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14
weeks, or at least 15 weeks after a desired or acceptable kernel
moisture content or average kernel moisture content is reached
(without being limiting, for example, between 10% and 30%, between
10% and 25%, between 13% and 25%, between 15% and 25%, between 10%
and 20%, or between 15% and 30% or any other specific moisture
content percentage within any of such moisture content ranges or as
provided herein), which may comprise measuring the moisture content
of one or more ears or kernels of a corn plant of the plurality of
corn plants in the field (or an average kernel moisture content for
two or more corn plants of the plurality of corn plants), and
harvesting the plurality of corn plants if the desired or
acceptable kernel moisture content or desired or acceptable average
kernel moisture content is reached.
[0540] As used herein, an "acceptable" or "desired" kernel moisture
content can be any specific kernel moisture content percentage, or
any kernel moisture content within a range of kernel moisture
content percentages, provided herein. An "average kernel moisture
content" for a plurality of corn plants is the average moisture
content of kernels from two or more corn plants, such as from two
or more corn plants of a plurality of corn plants. An "average
kernel moisture content" for a single corn plant is the average
moisture content of two or more kernels from a corn plant.
[0541] Leaf moisture content or stalk moisture content can also be
measured using techniques standard in the art.
[0542] In an aspect, a method provided herein comprises harvesting
corn plants comprising an average stalk moisture content and/or an
average leaf moisture content of equal to or less than 30%. In an
aspect, a method provided herein comprises harvesting corn plants
comprising an average stalk moisture content and/or an average leaf
moisture content of equal to or less than 25%. In an aspect, a
method provided herein comprises harvesting corn plants comprising
an average stalk moisture content and/or an average leaf moisture
content of equal to or less than 20%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average stalk moisture content and/or an average leaf moisture
content of equal to or less than 15%. In an aspect, a method
provided herein comprises harvesting corn plants comprising an
average stalk moisture content and/or an average leaf moisture
content of equal to or less than 10%. Each of the above average
stalk and/or leaf moisture content ranges may also apply to the
stalk or leaf moisture content of a corn plant of the plurality of
corn plants.
[0543] In another aspect, a method provided herein comprises
harvesting corn plants comprising an average stalk moisture content
and/or an average leaf moisture content of between 10% and 30%. In
another aspect, a method provided herein comprises harvesting corn
plants comprising an average stalk moisture content and/or an
average leaf moisture content of between 15% and 30%. In another
aspect, a method provided herein comprises harvesting corn plants
comprising an average stalk moisture content and/or an average leaf
moisture content of between 15% and 25% or a stalk or leaf moisture
content of a corn plant, such as a corn plant of a plurality of
corn plants, of between 15% and 25%.
[0544] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field where fewer than or equal
to 95% of the corn plants have lodged at the time of harvest. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a field where fewer than or equal to 90% of the
corn plants have lodged at the time of harvest. In an aspect, a
method provided herein comprises harvesting a plurality of corn
plants from a field where fewer than or equal to 80% of the corn
plants have lodged at the time of harvest. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a field where fewer than or equal to 70% of the corn plants
have lodged at the time of harvest. In an aspect, a method provided
herein comprises harvesting a plurality of corn plants from a field
where fewer than or equal to 60% of the corn plants have lodged at
the time of harvest. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a field where
fewer than or equal to 50% of the corn plants have lodged at the
time of harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 45% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 40% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 35% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 30% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 25% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 20% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 15% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 10% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 5% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where fewer than
or equal to 1% of the corn plants have lodged at the time of
harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where 0% of the
corn plants have lodged at the time of harvest.
[0545] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a field where between 0% and 100%
of the corn plants have lodged at the time of harvest. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a field where between 0% and 90% of the corn
plants have lodged at the time of harvest. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a field where between 0% and 80% of the corn plants have
lodged at the time of harvest. In an aspect, a method provided
herein comprises harvesting a plurality of corn plants from a field
where between 0% and 70% of the corn plants have lodged at the time
of harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where between 0%
and 60% of the corn plants have lodged at the time of harvest. In
an aspect, a method provided herein comprises harvesting a
plurality of corn plants from a field where between 0% and 50% of
the corn plants have lodged at the time of harvest. In an aspect, a
method provided herein comprises harvesting a plurality of corn
plants from a field where between 0% and 40% of the corn plants
have lodged at the time of harvest. In an aspect, a method provided
herein comprises harvesting a plurality of corn plants from a field
where between 0% and 30% of the corn plants have lodged at the time
of harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where between 0%
and 25% of the corn plants have lodged at the time of harvest. In
an aspect, a method provided herein comprises harvesting a
plurality of corn plants from a field where between 0% and 20% of
the corn plants have lodged at the time of harvest. In an aspect, a
method provided herein comprises harvesting a plurality of corn
plants from a field where between 0% and 15% of the corn plants
have lodged at the time of harvest. In an aspect, a method provided
herein comprises harvesting a plurality of corn plants from a field
where between 0% and 10% of the corn plants have lodged at the time
of harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where between
10% and 50% of the corn plants have lodged at the time of harvest.
In an aspect, a method provided herein comprises harvesting a
plurality of corn plants from a field where between 10% and 20% of
the corn plants have lodged at the time of harvest. In an aspect, a
method provided herein comprises harvesting a plurality of corn
plants from a field where between 10% and 30% of the corn plants
have lodged at the time of harvest. In an aspect, a method provided
herein comprises harvesting a plurality of corn plants from a field
where between 20% and 50% of the corn plants have lodged at the
time of harvest. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a field where between
30% and 50% of the corn plants have lodged at the time of harvest.
In an aspect, a method provided herein comprises harvesting a
plurality of corn plants from a field where between 40% and 50% of
the corn plants have lodged at the time of harvest.
[0546] The height of a corn plant can be determined based on a
variety of anatomical locations on a corn plant. In an aspect, the
height of a corn plant is measured as the distance between the top
of the soil or ground and the ligule or collar of the uppermost
fully-expanded leaf of a corn plant. As used herein, a
"fully-expanded leaf" is a leaf where the leaf blade is exposed,
and both the ligule and auricle are visible at the blade/sheath
boundary. In another aspect, the height of a corn plant is measured
as the distance between the top of the soil or ground and the upper
leaf surface of the leaf farthest from the soil or ground. In a
further aspect, the height of a corn plant is measured as the
distance between the top of the soil or ground and the arch of the
highest corn leaf that is at least 50% developed. In still a
further aspect, the height of a corn plant is measured as the
distance between the top of the soil or ground and the anatomical
part of the corn plant that is farthest from the top of the soil or
ground. Exemplary, non-limiting methods of measuring plant height
include comparing photographs of corn plants to a height reference,
or physically measuring individual corn plants with a suitable
ruler. If not otherwise stated, the height of a corn plant for the
present disclosure is measured as the distance between the top of
the soil or ground and the collar of the uppermost fully-expanded
leaf of a corn plant. If not otherwise stated, all descriptions
herein with regard to the plant height of a population of plants
can refer to either the average plant height among the population
of plants or, if stated, the percentage(s) of plants among the
population of plants.
[0547] Short stature corn plants typically have improved
standability and reduced lodging as compared to taller corn plants.
In an aspect, the average height of corn plants in a corn field
provided herein is less than or equal to 1.9 meters at the time of
harvest. In an aspect, the average height of corn plants in a corn
field provided herein is less than or equal to 1.8 meters at the
time of harvest. In an aspect, the average height of corn plants in
a corn field provided herein is less than or equal to 1.7 meters at
the time of harvest. In an aspect, the average height of corn
plants in a corn field provided herein is less than or equal to 1.6
meters at the time of harvest. In an aspect, the average height of
corn plants in a corn field provided herein is less than or equal
to 1.5 meters at the time of harvest. In an aspect, the average
height of corn plants in a corn field provided herein is less than
or equal to 1.4 meters at the time of harvest. In an aspect, the
average height of corn plants in a corn field provided herein is
less than or equal to 1.3 meters at the time of harvest.
[0548] In an aspect, the average height of corn plants in a corn
field provided herein is less than or equal to 1.9 meters at R1
stage, R2 stage, R3 stage, R4 stage, or R5 stage or later. In an
aspect, the average height of corn plants in a corn field provided
herein is less than or equal to 1.8 meters at R1 stage, R2 stage,
R3 stage, R4 stage, or R5 stage or later. In an aspect, the average
height of corn plants in a corn field provided herein is less than
or equal to 1.7 meters at R1 stage, R2 stage, R3 stage, R4 stage,
or R5 stage or later. In an aspect, the average height of corn
plants in a corn field provided herein is less than or equal to 1.6
meters at R1 stage, R2 stage, R3 stage, R4 stage, or R5 stage or
later. In an aspect, the average height of corn plants in a corn
field provided herein is less than or equal to 1.5 meters at R1
stage, R2 stage, R3 stage, R4 stage, or R5 stage or later. In an
aspect, the average height of corn plants in a corn field provided
herein is less than or equal to 1.4 meters at R1 stage, R2 stage,
R3 stage, R4 stage, or R5 stage or later. In an aspect, the average
height of corn plants in a corn field provided herein is less than
or equal to 1.3 meters at R1 stage, R2 stage, R3 stage, R4 stage,
or R5 stage or later. In an aspect, the average height of corn
plants in a corn field provided herein is less than or equal to 1.2
meters at R1 stage, R2 stage, R3 stage, R4 stage, or R5 stage or
later. In an aspect, the average height of corn plants in a corn
field provided herein is less than or equal to 1.1 meters at R1
stage, R2 stage, R3 stage, R4 stage, or R5 stage or later.
[0549] In an aspect, the average height of corn plants in a corn
field provided herein is between 1.1 meters and 1.9 meters at the
time of harvest. In an aspect, the average height of corn plants in
a corn field provided herein is between 1.3 meters and 1.8 meters
at the time of harvest. In an aspect, the average height of corn
plants in a corn field provided herein is between 1.3 meters and
1.7 meters at the time of harvest. In an aspect, the average height
of corn plants in a corn field provided herein is between 1.4
meters and 1.7 meters at the time of harvest. In an aspect, the
average height of corn plants in a corn field provided herein is
between 1.5 meters and 1.7 meters at the time of harvest. In an
aspect, the average height of corn plants in a corn field provided
herein is between 1.6 meters and 1.7 meters at the time of
harvest.
[0550] Corn leaves consist of four main anatomical parts: a
proximal sheath, a ligule, an auricle, and a distal blade. The
sheath wraps around the stem and younger leaves, while the blade is
flattened in the mediolateral axis (midrib to margin). The ligule
and auricle are found at the blade/sheath boundary; the ligule is
an adaxial (upper) membranous structure that acts as a collar
around the stem, and the auricle is a projection on the lower
surface of the blade base that connects the blade to the sheath.
Stages of corn plant growth are divided into vegetative (V) stages
and reproductive (R) stages. Upon germination, a corn plant is said
to be in VE stage (emergence). Once the first leaf collar (e.g.,
the ligule) is visible, the corn plant is in the V1 stage. The
emergence of the second leaf collar signifies V2 stage; the
emergence of the third leaf collar signifies the V3 stage; and so
on until the tassel emerges. For example, if twelve leaf collars
are visible, the plant is a V12 stage plant. Once the bottom-most
branch of the tassel emerges the plant is in VT stage, which is the
final vegetative stage. The reproductive stage of growth occurs
after the vegetative stage. The number of vegetative stages prior
to VT stage can vary by environment and corn line. The first
reproductive stage (R1; silking stage; "silking") occurs when silk
is visible outside the husk leaves surrounding an ear of corn. R2
(blistering stage) occurs when corn kernels are white on the
outside and are filled with a clear liquid inside. R3 (milk stage)
occurs when the kernels are yellow on the outside and are filled
with a milky white fluid inside. R4 (dough stage) occurs when the
kernels are filled with a thick, or pasty, fluid. In some corn
lines the cob will also turn pink or red at this stage. R5 (dent
stage) occurs when a majority of the kernels are at least partially
dented. The final reproductive stage, R6 (physiological maturity),
occurs when the kernels have attained their maximum dry weight.
[0551] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 20 days after
at least 50% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days after
at least 50% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 40 days after
at least 50% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days, at
least 40 days, at least 50 days, at least 60 days, at least 70
days, at least 80 days, at least 90 days, at least 100 days, or at
least 110 days after at least 50%, at least 60%, at least 70%, at
least 80%, at least 90% or 100% of corn plants in the corn field
have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 55 days after at least 50% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 60 days after at least 50% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 75 days after at least 50% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 90 days after at least 50% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 120 days after at least 50% of corn plants in the corn
field have reached R3 stage.
[0552] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 20 days and
120 days after at least 50% of corn plants in the corn field have
reached R3 stage. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a corn field between 30
days and 120 days after at least 50% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 40 days and 120 days after at least 50% of corn plants in
the corn field have reached R3 stage. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 50 days and 120 days after at least 50%
of corn plants in the corn field have reached R3 stage. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field between 50 days and 90 days after
at least 50% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and 80
days after at least 50% of corn plants in the corn field have
reached R3 stage. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a corn field between 50
days and 70 days after at least 50% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 60 days and 120 days after at least 50% of corn plants in
the corn field have reached R3 stage. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 60 days and 90 days after at least 50% of
corn plants in the corn field have reached R3 stage.
[0553] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 20 days after
at least 75% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days after
at least 75% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 40 days after
at least 75% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days, at
least 40 days, at least 50 days, at least 60 days, at least 70
days, at least 80 days, at least 90 days, at least 100 days, or at
least 110 days after at least 75% of corn plants in the corn field
have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 55 days after at least 75% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 60 days after at least 75% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 75 days after at least 75% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 90 days after at least 75% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 120 days after at least 75% of corn plants in the corn
field have reached R3 stage.
[0554] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 20 days and
120 days after at least 75% of corn plants in the corn field have
reached R3 stage. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a corn field between 30
days and 120 days after at least 75% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 40 days and 120 days after at least 75% of corn plants in
the corn field have reached R3 stage. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 50 days and 120 days after at least 75%
of corn plants in the corn field have reached R3 stage. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field between 50 days and 90 days after
at least 75% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and 80
days after at least 75% of corn plants in the corn field have
reached R3 stage. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a corn field between 50
days and 70 days after at least 75% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 60 days and 120 days after at least 75% of corn plants in
the corn field have reached R3 stage. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 60 days and 90 days after at least 75% of
corn plants in the corn field have reached R3 stage.
[0555] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 20 days after
at least 90% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days after
at least 90% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 40 days after
at least 90% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days, at
least 40 days, at least 50 days, at least 60 days, at least 70
days, at least 80 days, at least 90 days, at least 100 days, or at
least 110 days after at least 90% of corn plants in the corn field
have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 55 days after at least 90% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 60 days after at least 90% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 75 days after at least 90% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 90 days after at least 90% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
at least 120 days after at least 90% of corn plants in the corn
field have reached R3 stage.
[0556] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 20 days and
120 days after at least 90% of corn plants in the corn field have
reached R3 stage. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a corn field between 30
days and 120 days after at least 90% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 40 days and 120 days after at least 90% of corn plants in
the corn field have reached R3 stage. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 50 days and 120 days after at least 90%
of corn plants in the corn field have reached R3 stage. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field between 50 days and 90 days after
at least 90% of corn plants in the corn field have reached R3
stage. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and 80
days after at least 90% of corn plants in the corn field have
reached R3 stage. In an aspect, a method provided herein comprises
harvesting a plurality of corn plants from a corn field between 50
days and 70 days after at least 90% of corn plants in the corn
field have reached R3 stage. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 60 days and 120 days after at least 90% of corn plants in
the corn field have reached R3 stage. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 60 days and 90 days after at least 90% of
corn plants in the corn field have reached R3 stage.
[0557] As used herein, the term "fertilization" refers to the union
of a male gamete and a female gamete to produce a kernel, or
fertilized egg, following pollination. In an aspect, fertilization
is performed by wind. In another aspect, fertilization is performed
by human intervention. In another aspect, fertilization is
performed by an animal or insect.
[0558] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 35 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 40 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 45 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 50 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 55 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 60 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 65 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 70 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 75 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 80 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 85 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 90 days after
fertilization or silking of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field at least 120 days after
fertilization or silking of the plurality of corn plants.
[0559] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and
120 days after fertilization or silking of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and 90
days after fertilization or silking of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and 80
days after fertilization or silking of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and 70
days after fertilization or silking of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 60 days and
120 days after fertilization or silking of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 60 days and 90
days after fertilization or silking of the plurality of corn
plants.
[0560] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 30 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 35 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 40 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 45 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 50 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 55 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 60 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 65 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 70 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 75 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 80 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 85 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 90 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field at least 120 days
after fertilization or silking of at least 50%, at least 60%, at
least 70%, at least 80%, at least 90%, or 100% of the plurality of
corn plants.
[0561] In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 50 days and
120 days after fertilization or silking of at least 50%, at least
60%, at least 70%, at least 80%, at least 90%, or 100% of the
plurality of corn plants. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 50 days and 90 days after fertilization or silking of at
least 50%, at least 60%, at least 70%, at least 80%, at least 90%,
or 100% of the plurality of corn plants. In an aspect, a method
provided herein comprises harvesting a plurality of corn plants
from a corn field between 50 days and 80 days after fertilization
or silking of at least 50%, at least 60%, at least 70%, at least
80%, at least 90%, or 100% of the plurality of corn plants. In an
aspect, a method provided herein comprises harvesting a plurality
of corn plants from a corn field between 50 days and 70 days after
fertilization or silking of at least 50%, at least 60%, at least
70%, at least 80%, at least 90%, or 100% of the plurality of corn
plants. In an aspect, a method provided herein comprises harvesting
a plurality of corn plants from a corn field between 60 days and
120 days after fertilization or silking of at least 50%, at least
60%, at least 70%, at least 80%, at least 90%, or 100% of the
plurality of corn plants. In an aspect, a method provided herein
comprises harvesting a plurality of corn plants from a corn field
between 60 days and 90 days after fertilization or silking of at
least 50%, at least 60%, at least 70%, at least 80%, at least 90%,
or 100% of the plurality of corn plants.
[0562] The following non-limiting embodiments are specifically
envisioned: [0563] 1. A method comprising harvesting a plurality of
corn plants from a field at least 50 days after fertilization or
silking of at least 50% of said plurality of corn plants, wherein
fewer than or equal to 50% of said corn plants have lodged at the
time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0564] 2. A method comprising harvesting a plurality of corn plants
from a field at least 50 days after fertilization or silking of at
least 50% of said plurality of corn plants, wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_5 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence. [0565] 3. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein fewer than or equal to 50%
of said corn plants have lodged at the time of harvest, and wherein
at least one corn plant of said plurality of corn plants comprises
a mutant allele of an endogenous Brachytic2 (br2) locus, wherein
the mutant allele comprises a DNA segment inserted into the
endogenous br2 locus, wherein the DNA segment encodes an antisense
RNA that is at least 70% complementary to at least 20 consecutive
nucleotides of SEQ ID NO: 132 or 180, and wherein the mutant allele
of the endogenous br2 locus produces an RNA transcript comprising
the antisense RNA sequence. [0566] 4. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a deletion of at least one nucleotide from an endogenous
br2 locus as compared to SEQ ID NO: 132. [0567] 5. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein fewer than or equal to 50%
of said corn plants have lodged at the time of harvest, and wherein
at least one corn plant of said plurality of corn plants comprises
a dominant or semi-dominant transgene or mutant allele of a gene,
and wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant. [0568] 6. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein fewer than or equal to 50%
of said corn plants have lodged at the time of harvest, and wherein
at least one corn plant of said plurality of corn plants comprises
a premature stop codon within a nucleic acid sequence encoding a
Brachytic2 protein as compared to a control corn plant. [0569] 7. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification
comprising a deletion of at least a portion of the transcription
termination sequence of the endogenous Zm.SAMT gene, and wherein
the mutant allele produces a RNA molecule comprising an antisense
sequence complementary to all or part of the sense strand of the
endogenous GA20 oxidase_5 gene. [0570] 8. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after fertilization or silking of at least 50% of said plurality of
corn plants, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene. [0571] 9. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after fertilization or silking
of at least 50% of said plurality of corn plants, wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of one or
more of the following: 5' UTR, 1.sup.st exon, 1.sup.st intron,
2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1.sup.st exon, 1.sup.st intron,
2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3.sup.rd intron,
4.sup.th exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th intron,
6.sup.th exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th intron,
8.sup.th exon, 3' UTR, and any portion thereof, of the endogenous
Zm.SAMT gene. [0572] 10. A method comprising harvesting a plurality
of corn plants from a field at least 50 days after fertilization or
silking of at least 50% of said plurality of corn plants, wherein
fewer than or equal to 50% of said corn plants have lodged at the
time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof [0573] 11. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after
fertilization or silking of at least 50% of said plurality of corn
plants, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof [0574] 12. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a sequence selected from the
group consisting of SEQ ID NOs: 87-105. [0575] 13. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein fewer than or equal to 50%
of said corn plants have lodged at the time of harvest, and wherein
at least one corn plant of said plurality of corn plants comprises
a mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR,
1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon, 3' UTR, and any complementary sequence thereof, and
any portion of the foregoing, of the endogenous Zm.GA20 oxidase_5
gene; and wherein the second sequence comprises one or more of the
5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th
intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th
intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene; wherein the first
sequence and the second sequence are contiguous or separated only
by an intervening sequence of fewer than 555 nucleotides. [0576]
14. A method comprising harvesting a plurality of corn plants from
a field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genomic deletion relative to
a wild type allele of the endogenous GA20 oxidase_5 locus, wherein
the genomic deletion is flanked by a first sequence and a second
sequence; wherein the first sequence comprises one or more of the
5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3' UTR, and any complementary sequence
thereof, and any portion of the foregoing, of the endogenous
Zm.GA20 oxidase_5 gene; and wherein the second sequence comprises
one or more of the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd
exon, 2.sup.nd intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th
exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th
exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene. [0577]
15. A method comprising harvesting a plurality of corn plants from
a field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genomic sequence comprising a
first sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length. [0578] 16. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0579] 17. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein fewer than or equal to
50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a DNA segment inserted into the
endogenous GA20 oxidase_5 locus, wherein the DNA segment encodes an
antisense RNA sequence that is at least 70% complementary to at
least 20 consecutive nucleotides of one or more of SEQ ID NOs:
182-184 and 186-188, and wherein the mutant allele of the
endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence. [0580] 18. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
an endogenous Brachytic2 (br2) locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous br2 locus,
wherein the DNA segment encodes an antisense RNA that is at least
70% complementary to at least 20 consecutive nucleotides of SEQ ID
NO: 132 or 180, and wherein the mutant allele of the endogenous br2
locus produces an RNA transcript comprising the antisense RNA
sequence. [0581] 19. A method comprising harvesting a plurality of
corn plants from a field at least 50 days after at least 50% of
said corn plants have reached R3 stage, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of an endogenous Brachytic2 (br2) locus,
wherein the mutant allele comprises a deletion of at least one
nucleotide from an endogenous br2 locus as compared to SEQ ID NO:
132. [0582] 20. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein fewer than or equal to
50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a dominant or semi-dominant transgene or mutant allele of
a gene, and wherein the transgene or mutant allele causes a short
stature phenotype in the at least one corn plant.
[0583] 21. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein fewer than or equal to
50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a premature stop codon within a nucleic acid sequence
encoding a Brachytic2 protein as compared to a control corn plant.
[0584] 22. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein fewer than or equal to
50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification
comprising a deletion of at least a portion of the transcription
termination sequence of the endogenous Zm.SAMT gene, and wherein
the mutant allele produces a RNA molecule comprising an antisense
sequence complementary to all or part of the sense strand of the
endogenous GA20 oxidase_5 gene. [0585] 23. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the intergenic region between the endogenous GA20
oxidase_5 and Zm.SAMT genes, and wherein the mutant allele produces
a RNA molecule comprising an antisense sequence complementary to
all or part of the sense strand of the endogenous GA20 oxidase_5
gene. [0586] 24. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein fewer than or equal to
50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification
comprising a deletion of at least a portion of one or more of the
following: 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon,
2.sup.nd intron, 3.sup.rd exon, 3' UTR, and any portion thereof,
and the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon,
2.sup.nd intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon,
4.sup.th intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th exon,
6.sup.th intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3'
UTR, and any portion thereof, of the endogenous Zm.SAMT gene.
[0587] 25. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein fewer than or equal to
50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification which
results in the transcription of an antisense strand of at least an
exon, an intron, or an untranslated region (UTR) of the endogenous
GA20 oxidase_5 gene, or any portion thereof [0588] 26. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof. [0589] 27.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a sequence selected from the group consisting of
SEQ ID NOs: 87-105. [0590] 28. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
least 50% of said corn plants have reached R3 stage, wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides. [0591] 29. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein fewer than or equal to 50% of said corn plants
have lodged at the time of harvest, and wherein at least one corn
plant of said plurality of corn plants comprises a mutant allele of
the endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd i.sub.nt.sub.ron,
3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron,
5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron,
7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any
complementary sequence thereof, and any portion of the foregoing,
of the endogenous Zm.SAMT gene. [0592] 30. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic sequence comprising a first sequence and a
second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length. [0593] 31. A method comprising harvesting a plurality of
corn plants from a field at least 50 days after fertilization or
silking of at least 50% of said plurality of corn plants, wherein
the average kernel moisture content is less than or equal to 30%,
and wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0594] 32. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after fertilization or silking
of at least 50% of said plurality of corn plants, wherein the
average kernel moisture content is less than or equal to 30%, and
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0595] 33. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after fertilization or silking
of at least 50% of said plurality of corn plants, wherein the
average kernel moisture content is less than or equal to 30%, and
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a DNA
segment inserted into the endogenous br2 locus, wherein the DNA
segment encodes an antisense RNA that is at least 70% complementary
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180,
and wherein the mutant allele of the endogenous br2 locus produces
an RNA transcript comprising the antisense RNA sequence. [0596] 34.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein the average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a deletion of at
least one nucleotide from an endogenous br2 locus as compared to
SEQ ID NO: 132. [0597] 35. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant. [0598] 36. A method comprising harvesting
a plurality of corn plants from a field at least 50 days after
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a premature
stop codon within a nucleic acid sequence encoding a Brachytic2
protein as compared to a control corn plant. [0599] 37. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
transcription termination sequence of the endogenous Zm.SAMT gene,
and wherein the mutant allele produces a RNA molecule comprising an
antisense sequence complementary to all or part of the sense strand
of the endogenous GA20 oxidase_5 gene. [0600] 38. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene. [0601] 39.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein the average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of one or
more of the following: 5' UTR, 1.sup.st exon, 1.sup.st intron,
2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and any
portion thereof, and the 5' UTR, 1
.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron,
5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron,
7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any
portion thereof, of the endogenous Zm.SAMT gene. [0602] 40. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein the average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification which results in the transcription of an antisense
strand of at least an exon, an intron, or an untranslated region
(UTR) of the endogenous GA20 oxidase_5 gene, or any portion thereof
[0603] 41. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after fertilization or silking
of at least 50% of said plurality of corn plants, wherein the
average kernel moisture content is less than or equal to 30%, and
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof. [0604] 42.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein the average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a sequence
selected from the group consisting of SEQ ID NOs: 87-105. [0605]
43. A method comprising harvesting a plurality of corn plants from
a field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein the average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides. [0606] 44. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
deletion relative to a wild type allele of the endogenous GA20
oxidase_5 locus, wherein the genomic deletion is flanked by a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene. [0607] 45. A method comprising harvesting
a plurality of corn plants from a field at least 50 days after
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genomic sequence comprising a first sequence and
a second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length. [0608] 46. A method comprising harvesting a plurality of
corn plants from a field at least 50 days after at least 50% of
said corn plants have reached R3 stage, wherein average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_3 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_3 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_3 locus produces a RNA transcript
comprising the antisense RNA sequence. [0609] 47. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a DNA segment inserted into the endogenous GA20
oxidase_5 locus, wherein the DNA segment encodes an antisense RNA
sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_5 locus produces a RNA transcript comprising the antisense
RNA sequence. [0610] 48. A method comprising harvesting a plurality
of corn plants from a field at least 50 days after at least 50% of
said corn plants have reached R3 stage, wherein average kernel
moisture content is less than or equal to 30%, and wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous br2 locus, wherein the DNA segment
encodes an antisense RNA that is at least 70% complementary to at
least 20 consecutive nucleotides of SEQ ID NO: 132 or 180, and
wherein the mutant allele of the endogenous br2 locus produces an
RNA transcript comprising the antisense RNA sequence. [0611] 49. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a deletion of at least one nucleotide from
an endogenous br2 locus as compared to SEQ ID NO: 132. [0612] 50. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant. [0613] 51. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a premature
stop codon within a nucleic acid sequence encoding a Brachytic2
protein as compared to a control corn plant. [0614] 52. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genome modification comprising a deletion of at
least a portion of the transcription termination sequence of the
endogenous Zm.SAMT gene, and wherein the mutant allele produces a
RNA molecule comprising an antisense sequence complementary to all
or part of the sense strand of the endogenous GA20 oxidase_5 gene.
[0615] 53. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene. [0616] 54.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of one or more of the following: 5' UTR,
1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon, 3' UTR, and any portion thereof, and the 5' UTR,
1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron,
5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron,
7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any
portion thereof, of the endogenous Zm.SAMT gene. [0617] 55. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein average kernel moisture content is less
than or equal to 30%, and wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification which results in the
transcription of an antisense strand of at least an exon, an
intron, or an untranslated region (UTR) of the endogenous GA20
oxidase_5 gene, or any portion thereof. [0618] 56. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises the Zm.SAMT gene promoter, or a functional part
thereof, operably linked to at least one transcribable antisense
sequence of at least an exon, intron or untranslated region (UTR)
of the endogenous GA20 oxidase_5 gene, or any portion thereof
[0619] 57. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a sequence
selected from the group consisting of SEQ ID NOs: 87-105.
[0620] 58. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides. [0621] 59. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genomic deletion relative to a wild type allele
of the endogenous GA20 oxidase_5 locus, wherein the genomic
deletion is flanked by a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR,
1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon, 3' UTR, and any complementary sequence thereof, and
any portion of the foregoing, of the endogenous Zm.GA20 oxidase_5
gene; and wherein the second sequence comprises one or more of the
5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th
intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th
intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene. [0622] 60. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein average kernel moisture content is less than or
equal to 30%, and wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genomic sequence comprising a first sequence and
a second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length. [0623] 61. A method comprising harvesting a plurality of
corn plants from a field at least 50 days after at fertilization or
silking of at least 50% of said plurality of corn plants, wherein
the average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0624] 62. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at fertilization or
silking of at least 50% of said plurality of corn plants, wherein
the average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_5
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0625] 63. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at fertilization or
silking of at least 50% of said plurality of corn plants, wherein
the average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a DNA
segment inserted into the endogenous br2 locus, wherein the DNA
segment encodes an antisense RNA that is at least 70% complementary
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180,
and wherein the mutant allele of the endogenous br2 locus produces
an RNA transcript comprising the antisense RNA sequence. [0626] 64.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after at fertilization or silking of at
least 50% of said plurality of corn plants, wherein the average
yield of said field is at least 170 bushels per acre, wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of an endogenous Brachytic2
(br2) locus, wherein the mutant allele comprises a deletion of at
least one nucleotide from an endogenous br2 locus as compared to
SEQ ID NO: 132. [0627] 65. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant. [0628] 66. A method comprising harvesting
a plurality of corn plants from a field at least 50 days after at
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a premature
stop codon within a nucleic acid sequence encoding a Brachytic2
protein as compared to a control corn plant. [0629] 67. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average yield of said
field is at least 170 bushels per acre, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification
comprising a deletion of at least a portion of the transcription
termination sequence of the endogenous Zm.SAMT gene, and wherein
the mutant allele produces a RNA molecule comprising an antisense
sequence complementary to all or part of the sense strand of the
endogenous GA20 oxidase_5 gene. [0630] 68. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at fertilization or silking of at least 50% of said plurality
of corn plants, wherein the average yield of said field is at least
170 bushels per acre, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the intergenic region between the
endogenous GA20 oxidase_5 and Zm.SAMT genes, and wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
complementary to all or part of the sense strand of the endogenous
GA20 oxidase_5 gene. [0631] 69. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genome modification comprising a deletion of at
least a portion of one or more of the following: 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3' UTR, and any portion thereof, and the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any portion
thereof, of the endogenous Zm.SAMT gene. [0632] 70. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average yield of said
field is at least 170 bushels per acre, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification which
results in the transcription of an antisense strand of at least an
exon, an intron, or an untranslated region (UTR) of the endogenous
GA20 oxidase_5 gene, or any portion thereof. [0633] 71. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at fertilization or silking of at least 50% of
said plurality of corn plants, wherein the average yield of said
field is at least 170 bushels per acre, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises the Zm.SAMT gene promoter, or a
functional part thereof, operably linked to at least one
transcribable antisense sequence of at least an exon, intron or
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof. [0634] 72. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a sequence selected from the group consisting of
SEQ ID NOs: 87-105. [0635] 73. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
fertilization or silking of at least 50% of said plurality of corn
plants, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a first sequence and a second sequence; wherein
the first sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.GA20 oxidase_5 gene;
and wherein the second sequence comprises one or more of the 5'
UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th
intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th
intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene; wherein the first
sequence and the second sequence are contiguous or separated only
by an intervening sequence of fewer than 555 nucleotides. [0636]
74. A method comprising harvesting a plurality of corn plants from
a field at least 50 days after at fertilization or silking of at
least 50% of said plurality of corn plants, wherein the average
yield of said field is at least 170 bushels per acre, wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
deletion relative to a wild type allele of the endogenous GA20
oxidase_5 locus, wherein the genomic deletion is flanked by a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5
' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3' UTR, and any complementary sequence
thereof, and any portion of the foregoing, of the endogenous
Zm.GA20 oxidase_5 gene; and wherein the second sequence comprises
one or more of the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd
exon, 2.sup.nd intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th
exon, 4.sup.th intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th
exon, 6.sup.th intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th
exon, 3' UTR, and any complementary sequence thereof, and any
portion of the foregoing, of the endogenous Zm.SAMT gene. [0637]
75. A method comprising harvesting a plurality of corn plants from
a field at least 50 days after at fertilization or silking of at
least 50% of said plurality of corn plants, wherein the average
yield of said field is at least 170 bushels per acre, wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genomic
sequence comprising a first sequence and a second sequence; wherein
the first sequence comprises at least 15 consecutive nucleotides of
one or more of SEQ ID NOs: 228-235 and 276-283; wherein the second
sequence comprises at least 15 consecutive nucleotides of one or
more of SEQ ID NOs: 235-276; and wherein the genomic sequence is at
least 50 consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length. [0638] 76. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_3 locus, wherein the mutant allele
comprises a DNA segment inserted into the endogenous GA20 oxidase_3
locus, wherein the DNA segment encodes an antisense RNA sequence
that is at least 70% complementary to at least 20 consecutive
nucleotides of one or more of SEQ ID NOs: 182-184 and 186-188, and
wherein the mutant allele of the endogenous GA20 oxidase_3 locus
produces a RNA transcript comprising the antisense RNA sequence.
[0639] 77. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein the average yield of
said field is at least 170 bushels per acre, wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a DNA segment
inserted into the endogenous GA20 oxidase_5 locus, wherein the DNA
segment encodes an antisense RNA sequence that is at least 70%
complementary to at least 20 consecutive nucleotides of one or more
of SEQ ID NOs: 182-184 and 186-188, and wherein the mutant allele
of the endogenous GA20 oxidase_5 locus produces a RNA transcript
comprising the antisense RNA sequence. [0640] 78. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of an endogenous Brachytic2 (br2) locus, wherein the mutant
allele comprises a DNA segment inserted into the endogenous br2
locus, wherein the DNA segment encodes an antisense RNA that is at
least 70% complementary to at least 20 consecutive nucleotides of
SEQ ID NO: 132 or 180, and wherein the mutant allele of the
endogenous br2 locus produces an RNA transcript comprising the
antisense RNA sequence. [0641] 79. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
least 50% of said corn plants have reached R3 stage, wherein the
average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of an endogenous
Brachytic2 (br2) locus, wherein the mutant allele comprises a
deletion of at least one nucleotide from an endogenous br2 locus as
compared to SEQ ID NO: 132. [0642] 80. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a dominant or
semi-dominant transgene or mutant allele of a gene, and wherein the
transgene or mutant allele causes a short stature phenotype in the
at least one corn plant. [0643] 81. A method comprising harvesting
a plurality of corn plants from a field at least 50 days after at
least 50% of said corn plants have reached R3 stage, wherein the
average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a premature stop codon within a
nucleic acid sequence encoding a Brachytic2 protein as compared to
a control corn plant. [0644] 82. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
least 50% of said corn plants have reached R3 stage, wherein the
average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of the transcription termination sequence of the endogenous
Zm.SAMT gene, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene. [0645] 83.
A method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein the average yield of said field is at
least 170 bushels per acre, wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a genome modification comprising a deletion
of at least a portion of the intergenic region between the
endogenous GA20 oxidase_5 and Zm.SAMT genes, and wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
complementary to all or part of the sense strand of the endogenous
GA20 oxidase_5 gene. [0646] 84. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
least 50% of said corn plants have reached R3 stage, wherein the
average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any portion thereof, and the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon,
3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th exon,
5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th exon,
7.sup.th intron, 8.sup.th exon, 3' UTR, and any portion thereof, of
the endogenous Zm.SAMT gene. [0647] 85. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification which results in the transcription
of an antisense strand of at least an exon, an intron, or an
untranslated region (UTR) of the endogenous GA20 oxidase_5 gene, or
any portion thereof. [0648] 86. A method comprising harvesting a
plurality of corn plants from a field at least 50 days after at
least 50% of said corn plants have reached R3 stage, wherein the
average yield of said field is at least 170 bushels per acre,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises the Zm.SAMT gene promoter, or a functional part thereof,
operably linked to at least one transcribable antisense sequence of
at least an exon, intron or untranslated region (UTR) of the
endogenous GA20 oxidase_5 gene, or any portion thereof [0649] 87. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein the average yield of said field is at
least 170 bushels per acre, wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a sequence selected from the group
consisting of SEQ ID NOs: 87-105. [0650] 88. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein the average yield of said field is at least 170 bushels per
acre, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a first sequence and a second sequence; wherein the first
sequence comprises one or more of the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides. [0651] 89. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genomic deletion relative to a wild type allele
of the endogenous GA20 oxidase_5 locus, wherein the genomic
deletion is flanked by a first sequence and a second sequence;
wherein the first sequence comprises one or more of the 5' UTR,
1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon, 3' UTR, and any complementary sequence thereof, and
any portion of the foregoing, of the endogenous Zm.GA20 oxidase_5
gene; and wherein the second sequence comprises one or more of the
5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th
intron, 5.sup.th exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th
intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.SAMT gene. [0652] 90. A method
comprising harvesting a plurality of corn plants from a field at
least 50 days after at least 50% of said corn plants have reached
R3 stage, wherein the average yield of said field is at least 170
bushels per acre, wherein fewer than or equal to 50% of said corn
plants have lodged at the time of harvest, and wherein at least one
corn plant of said plurality of corn plants comprises a mutant
allele of the endogenous GA20 oxidase_5 locus, wherein the mutant
allele comprises a genomic sequence comprising a first sequence and
a second sequence; wherein the first sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 228-235 and
276-283; wherein the second sequence comprises at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 235-276; and
wherein the genomic sequence is at least 50 consecutive nucleotides
in length, and/or fewer than 9000 consecutive nucleotides in
length. [0653] 91. A method comprising harvesting a plurality of
corn plants from a field at least 1 day after the average kernel
moisture content of at least 50% of said plurality of corn plants
is between 10% and 30%, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_3 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_3 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_3 locus produces a RNA transcript comprising the antisense
RNA sequence.
[0654] 92. A method comprising harvesting a plurality of corn
plants from a field at least 1 day after the average kernel
moisture content of at least 50% of said plurality of corn plants
is between 10% and 30%, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
GA20 oxidase_5 locus, wherein the DNA segment encodes an antisense
RNA sequence that is at least 70% complementary to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188, and wherein the mutant allele of the endogenous GA20
oxidase_5 locus produces a RNA transcript comprising the antisense
RNA sequence. [0655] 93. A method comprising harvesting a plurality
of corn plants from a field at least 1 day after the average kernel
moisture content of at least 50% of said plurality of corn plants
is between 10% and 30%, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of an endogenous Brachytic2 (br2) locus, wherein the
mutant allele comprises a DNA segment inserted into the endogenous
br2 locus, wherein the DNA segment encodes an antisense RNA that is
at least 70% complementary to at least 20 consecutive nucleotides
of SEQ ID NO: 132 or 180, and wherein the mutant allele of the
endogenous br2 locus produces an RNA transcript comprising the
antisense RNA sequence. [0656] 94. A method comprising harvesting a
plurality of corn plants from a field at least 1 day after the
average kernel moisture content of at least 50% of said plurality
of corn plants is between 10% and 30%, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of an endogenous Brachytic2 (br2) locus,
wherein the mutant allele comprises a deletion of at least one
nucleotide from an endogenous br2 locus as compared to SEQ ID NO:
132. [0657] 95. A method comprising harvesting a plurality of corn
plants from a field at least 1 day after the average kernel
moisture content of at least 50% of said plurality of corn plants
is between 10% and 30%, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
dominant or semi-dominant transgene or mutant allele of a gene, and
wherein the transgene or mutant allele causes a short stature
phenotype in the at least one corn plant. [0658] 96. A method
comprising harvesting a plurality of corn plants from a field at
least 1 day after the average kernel moisture content of at least
50% of said plurality of corn plants is between 10% and 30%,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a premature stop codon within a
nucleic acid sequence encoding a Brachytic2 protein as compared to
a control corn plant. [0659] 97. A method comprising harvesting a
plurality of corn plants from a field at least 1 day after the
average kernel moisture content of at least 50% of said plurality
of corn plants is between 10% and 30%, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genome modification
comprising a deletion of at least a portion of the transcription
termination sequence of the endogenous Zm.SAMT gene, and wherein
the mutant allele produces a RNA molecule comprising an antisense
sequence complementary to all or part of the sense strand of the
endogenous GA20 oxidase_5 gene. [0660] 98. A method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification comprising a deletion of at least a portion of the
intergenic region between the endogenous GA20 oxidase_5 and Zm.SAMT
genes, and wherein the mutant allele produces a RNA molecule
comprising an antisense sequence complementary to all or part of
the sense strand of the endogenous GA20 oxidase_5 gene. [0661] 99.
A method comprising harvesting a plurality of corn plants from a
field at least 1 day after the average kernel moisture content of
at least 50% of said plurality of corn plants is between 10% and
30%, wherein fewer than or equal to 50% of said corn plants have
lodged at the time of harvest, and wherein at least one corn plant
of said plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genome modification comprising a deletion of at least a
portion of one or more of the following: 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any portion thereof, and the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon,
3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th exon,
5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th exon,
7.sup.th intron, 8.sup.th exon, 3' UTR, and any portion thereof, of
the endogenous Zm.SAMT gene. [0662] 100. A method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a genome
modification which results in the transcription of an antisense
strand of at least an exon, an intron, or an untranslated region
(UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof. [0663] 101. A method comprising harvesting a plurality of
corn plants from a field at least 1 day after the average kernel
moisture content of at least 50% of said plurality of corn plants
is between 10% and 30%, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises the Zm.SAMT gene promoter, or a functional
part thereof, operably linked to at least one transcribable
antisense sequence of at least an exon, intron or untranslated
region (UTR) of the endogenous GA20 oxidase_5 gene, or any portion
thereof. [0664] 102. A method comprising harvesting a plurality of
corn plants from a field at least 1 day after the average kernel
moisture content of at least 50% of said plurality of corn plants
is between 10% and 30%, wherein fewer than or equal to 50% of said
corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
mutant allele of the endogenous GA20 oxidase_5 locus, wherein the
mutant allele comprises a sequence selected from the group
consisting of SEQ ID NOs: 87-105. [0665] 103. A method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a mutant allele of the endogenous GA20
oxidase_5 locus, wherein the mutant allele comprises a first
sequence and a second sequence; wherein the first sequence
comprises one or more of the 5' UTR, 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3' UTR, and
any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene; wherein the first sequence and the second
sequence are contiguous or separated only by an intervening
sequence of fewer than 555 nucleotides. [0666] 104. A method
comprising harvesting a plurality of corn plants from a field at
least 1 day after the average kernel moisture content of at least
50% of said plurality of corn plants is between 10% and 30%,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a mutant allele of the
endogenous GA20 oxidase_5 locus, wherein the mutant allele
comprises a genomic deletion relative to a wild type allele of the
endogenous GA20 oxidase_5 locus, wherein the genomic deletion is
flanked by a first sequence and a second sequence; wherein the
first sequence comprises one or more of the 5' UTR, 1.sup.st exon,
1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon, 3'
UTR, and any complementary sequence thereof, and any portion of the
foregoing, of the endogenous Zm.GA20 oxidase_5 gene; and wherein
the second sequence comprises one or more of the 5' UTR, 1.sup.st
exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd
exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th intron, 5.sup.th
exon, 5.sup.th intron, 6.sup.th exon, 6.sup.th intron, 7.sup.th
exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and any complementary
sequence thereof, and any portion of the foregoing, of the
endogenous Zm.SAMT gene. [0667] 105. A method comprising harvesting
a plurality of corn plants from a field at least 1 day after the
average kernel moisture content of at least 50% of said plurality
of corn plants is between 10% and 30%, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a mutant allele of the endogenous GA20 oxidase_5 locus,
wherein the mutant allele comprises a genomic sequence comprising a
first sequence and a second sequence; wherein the first sequence
comprises at least 15 consecutive nucleotides of one or more of SEQ
ID NOs: 228-235 and 276-283; wherein the second sequence comprises
at least 15 consecutive nucleotides of one or more of SEQ ID NOs:
235-276; and wherein the genomic sequence is at least 50
consecutive nucleotides in length, and/or fewer than 9000
consecutive nucleotides in length. [0668] 106. The method of any
one of embodiments 1-1(n), 3-Error! Reference source not found., or
4-Error! Reference source not found., wherein said harvesting is
performed at least 55 days, at least 60 days, at least 75 days, at
least 90 days, or at least 120 days after said fertilization or
silking. [0669] 107. The method of any one of embodiments 2-2(n),
4-Error! Reference source not found., or 5-6, wherein said
harvesting is performed at least 55 days, at least 60 days, at
least 75 days, at least 90 days, or at least 120 days after at
least 50% of said corn plants have reached R3 stage. [0670] 108.
The method of any one of embodiments 1-7(n), wherein fewer than or
equal to 40%, fewer than or equal to 30%, fewer than or equal to
20%, or fewer than or equal to 10% of said corn plants have lodged
at the time of harvest. [0671] 109. The method of any one of
embodiments 1-105, wherein the average height of said corn plants
is less than or equal to 1.7 meters, less than or equal to 1.6
meters, or less than or equal to 1.5 meters at the time of harvest.
[0672] 110. The method of embodiment 109, wherein said height is
measured as the distance between the soil and the ligule of the
uppermost fully-expanded leaf. [0673] 111. The method of any one of
embodiments 1-105, wherein at least 50% of said corn plants are
inbred corn plants. [0674] 112. The method of any one of
embodiments 1-105, wherein at least 50% of said corn plants are
hybrid corn plants. [0675] 113. The method of any one of
embodiments 1-105, wherein at least 50% of said corn plants are
semi-dwarf corn plants. [0676] 114. The method of any one of
embodiments 1-105, wherein at least 50% of said corn plants are
dwarf corn plants. [0677] 115. The method of any one of embodiments
1-105, wherein at least 50% of said corn plants are brachytic corn
plants. [0678] 116. The method of any one of embodiments 1-105,
wherein said field comprises a planting density of at least 10,000
corn plants per acre. [0679] 117. The method of any one of
embodiments 1-105, wherein the average yield of said field is at
least 180 bushels per acre, at least 190 bushels per acre, at least
200 bushels per acre, at least 210 bushels per acre, at least 220
bushels per acre, or at least 250 bushels per acre. [0680] 118. The
method of any one of embodiments 1-90, wherein the average kernel
moisture content is equal to or less than 29%, equal to or less
than 28%, equal to or less than 27%, equal to or less than 26%,
equal to or less than 25%, equal to or less than 24%, equal to or
less than 23%, equal to or less than 22%, equal to or less than
21%, equal to or less than 20%, equal to or less than 19%, equal to
or less than 15%, equal to or less than 10%. [0681] 119. The method
of any one of embodiments 1-105, wherein the average kernel
moisture content is between 10% and 20%, between 13% and 20%,
between 15% and 20%, between 10% and 25%, between 13% and 25%,
between 15% and 25%, between 20% and 25%, between 13% and 30%,
between 15% and 30%, between 20% and 30%, or between 25% and 30%.
[0682] 120. The method of any one of embodiments 91-105, wherein
said harvesting occurs at least 2 days, at least 3 days, at least 4
days, at least 5 days, at least 6 days, at least 7 days, at least 8
days, at least 9 days, at least 10 days, at least 15 days, at least
20 days, at least 25 days, at least 30 days, at least 35 days, at
least 40 days at least 45 days, at least 50 days, at least 55 days,
at least 60 days, at least 65 days, or at least 70 days after an
average kernel moisture content of said plurality of corn plants is
between 10% and 30%, or the kernel moisture content of a corn plant
of the plurality of corn plants is between 10% and 30%.
[0683] 121. The method of any one of embodiments 1-105, wherein
said method comprises growing said plurality of said corn plants in
said corn field prior to said harvesting. [0684] 122. The method of
any one of embodiments 1, 16, 31, 46, 61, 76, and 91, wherein the
mutant allele of the endogenous GA20 oxidase_3 locus suppresses the
expression of a wild-type allele of the endogenous GA20 oxidase_3
locus, a wild-type allele of the endogenous GA20 oxidase_5 locus,
or both. [0685] 123. The method of any one of embodiments 1, 16,
31, 46, 61, 76, and 91, wherein the DNA segment comprises a
nucleotide sequence originating from the endogenous GA20 oxidase_3
locus. [0686] 124. The method of any one of embodiments 1, 16, 31,
46, 61, 76, and 91, wherein the DNA segment corresponds to an
inverted genomic fragment of the endogenous GA20 oxidase_3 locus.
[0687] 125. The method of any one of embodiments 1, 16, 31, 46, 61,
76, and 91, wherein the DNA segment comprises a nucleotide sequence
originating from the endogenous GA20 oxidase_5 locus. [0688] 126.
The method of any one of embodiments 1, 16, 31, 46, 61, 76, and 91,
wherein at least a portion of the antisense RNA sequence is at
least 70% complementary to a corresponding endogenous sequence of
the RNA transcript. [0689] 127. The method of embodiment 126,
wherein the corresponding endogenous sequence of the RNA transcript
is at least 85% identical to at least 20 consecutive nucleotides of
one or more of SEQ ID NOs: 182-184 and 186-188. [0690] 128. The
method of any one of embodiments 1, 16, 31, 46, 61, 76, and 91,
wherein the DNA segment is inserted near or adjacent to a
corresponding endogenous DNA segment of the endogenous GA20
oxidase_3 locus. [0691] 129. The method of embodiment 128, wherein
the antisense RNA sequence forms a stem-loop structure with the
corresponding endogenous sequence of the RNA transcript. [0692]
130. The method of embodiment 128, wherein the inserted DNA segment
and the corresponding endogenous DNA segment of the mutant allele
are separated by an intervening DNA sequence. [0693] 131. The
method of embodiment 130, wherein the intervening DNA sequence
comprises a native sequence of the endogenous GA20 oxidase_3 locus.
[0694] 132. The method of embodiment 130, wherein the intervening
DNA sequence comprises an exogenous sequence inserted into the
endogenous GA20 oxidase_3 locus. [0695] 133. The method of
embodiment 126, wherein the DNA segment is inserted within a region
selected from the group consisting of 5' untranslated region (UTR),
1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd intron,
3.sup.rd exon and 3' UTR of the endogenous GA20 oxidase_3 locus,
and a combination thereof. [0696] 134. The method of embodiment
126, wherein the DNA segment is inserted at a genomic site
recognized by a targeted editing technique to create a
double-stranded break (DSB). [0697] 135. The method of embodiment
126, wherein the mutant allele further comprises a deletion of at
least one portion of the endogenous GA20 oxidase_3 locus. [0698]
136. The method of embodiment 126, wherein the sense strand of the
DNA segment comprises a sequence at least 70% complementary to an
exon sequence of the endogenous GA20 oxidase_3 or GA20 oxidase_5
locus. [0699] 137. The method of embodiment 126, wherein the sense
strand of the DNA segment comprises a sequence at least 70%
complementary to an untranslated region (UTR) sequence of the
endogenous GA20 oxidase_3 or GA20 oxidase_5 locus. [0700] 138. The
method of embodiment 126, wherein the sense strand of the DNA
segment comprises a sequence at least 70% complementary to an exon
sequence and an intron sequence of the endogenous GA20 oxidase_3 or
GA20 oxidase_5 locus, the exon sequence and the intron sequence
being contiguous within the endogenous locus. [0701] 139. The
method of embodiment 126, wherein the DNA segment comprises a
sequence having at least at least 70% identity to one or more of
SEQ ID Nos: 194, 195, 207, 209, 211, 213, and 217. [0702] 140. The
method of any one of embodiments 2, 17, 32, 47, 62, 77, or 92,
wherein the mutant allele of the endogenous GA20 oxidase_5 locus
suppresses the expression of a wild-type allele of the endogenous
GA20 oxidase_3 locus, a wild-type allele of the endogenous GA20
oxidase_5 locus, or both. [0703] 141. The method of any one of
embodiments 2, 17, 32, 47, 62, 77, or 92, wherein the RNA
transcript further comprises one or more sequence elements of the
endogenous GA20 oxidase_5 locus selected from the group consisting
of 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3' UTR, and any portion thereof [0704] 142.
The method of any one of embodiments 2, 17, 32, 47, 62, 77, or 92,
wherein the DNA segment comprises a nucleotide sequence originating
from the endogenous GA20 oxidase_3 locus. [0705] 143. The method of
embodiment 142, wherein the DNA segment corresponds to an inverted
genomic fragment of the endogenous GA20 oxidase_3 locus. [0706]
144. The method of any one of embodiments 2, 17, 32, 47, 62, 77, or
92, wherein the DNA segment comprises a nucleotide sequence
originating from the endogenous GA20 oxidase_5 locus. [0707] 145.
The method of embodiment 142, wherein the DNA segment corresponds
to an inverted genomic fragment of the endogenous GA20 oxidase_5
locus. [0708] 146. The method of any one of embodiments 2, 17, 32,
47, 62, 77, or 92, wherein at least a portion of the antisense RNA
sequence is at least 70% complementary to a corresponding
endogenous sequence of the RNA transcript. [0709] 147. The method
of embodiment 146, wherein the corresponding endogenous sequence of
the RNA transcript is at least 85% identical to at least 20
consecutive nucleotides of one or more of SEQ ID NOs: 182-184 and
186-188. [0710] 148. The method of embodiment 146, wherein the DNA
segment is inserted near or adjacent to a corresponding endogenous
DNA segment of the endogenous GA20 oxidase_5 locus. [0711] 149. The
method of embodiment 148, wherein the antisense RNA sequence forms
a stem-loop structure with the corresponding endogenous sequence of
the RNA transcript. [0712] 150. The method of embodiment 148,
wherein the inserted DNA segment and the corresponding endogenous
DNA segment of the mutant allele are separated by an intervening
DNA sequence. [0713] 151. The method of embodiment 150, wherein the
intervening DNA sequence comprises a native sequence of the
endogenous GA20 oxidase_5 locus. [0714] 152. The method of
embodiment 150, wherein the intervening DNA sequence comprises an
exogenous sequence inserted into the endogenous GA20 oxidase_5
locus. [0715] 153. The method of embodiment 146, wherein the DNA
segment is inserted within a region selected from the group
consisting of 5' untranslated region (UTR), 1.sup.st exon, 1.sup.st
intron, 2.sup.nd exon, 2.sup.nd intron, 3.sup.rd exon and 3' UTR of
the endogenous GA20 oxidase_5 locus, and a combination thereof.
[0716] 154. The method of embodiment 146, wherein the DNA segment
is inserted at a genomic site recognized by a targeted editing
technique to create a double-stranded break (DSB). [0717] 155. The
method of embodiment 146, wherein the mutant allele further
comprises a deletion of at least one portion of the endogenous GA20
oxidase_5 locus. [0718] 156. The method of embodiment 146, wherein
the sense strand of the DNA segment comprises a sequence at least
70% complementary to an exon sequence of the endogenous GA20
oxidase_3 or GA20 oxidase_5 locus. [0719] 157. The method of
embodiment 146, wherein the sense strand of the DNA segment
comprises a sequence at least 70% complementary to an untranslated
region (UTR) sequence of the endogenous GA20 oxidase_3 or GA20
oxidase_5 locus. [0720] 158. The method of embodiment 146, wherein
the sense strand of the DNA segment comprises a sequence at least
70% complementary to an exon sequence and an intron sequence of the
endogenous GA20 oxidase_3 or GA20 oxidase_5 locus, the exon
sequence and the intron sequence being contiguous within the
endogenous locus. [0721] 159. The method of embodiment 146, wherein
the DNA segment comprises a sequence having at least at least 70%
identity to one or more of SEQ ID Nos: 194, 195, 207, 209, 211,
213, and 217. [0722] 160. The method of any one of embodiments 1,
2, 16, 17, 31, 32, 46, 47, 61, 62, 76, 77, 91, or 92, wherein the
level of one or more active GAs in at least one internode tissue of
the stem or stalk of the modified corn plant is lower than the same
internode tissue of an unmodified control plant. [0723] 161. The
method of any one of embodiments 7-9, 22-24, 37-39, 52-54, 67-69,
82-84, or 97-99, wherein the mutant allele comprises the endogenous
Zm.SAMT gene promoter, or a portion thereof, operably linked to a
transcribable DNA sequence encoding a RNA molecule that causes
suppression of one or both of the endogenous GA20 oxidase_3 gene
and the endogenous GA20 oxidase_5 gene. [0724] 162. The method of
any one of embodiments 7-9, 22-24, 37-39, 52-54, 67-69, 82-84, or
97-99, wherein the mutant allele comprises the endogenous Zm.SAMT
gene promoter, or a portion thereof, operably linked to a
transcribable DNA sequence encoding a RNA molecule comprising an
antisense sequence that is at least 80% complementary to all or
part of the endogenous GA20 oxidase_3 or GA20 oxidase_5 gene.
[0725] 163. The method of any one of embodiments 11, 26, 41, 56,
71, 86, or 101, wherein the transcribable DNA sequence is at least
80% complementary to a RNA transcript sequence, or a portion
thereof, encoded by the endogenous GA20 oxidase_3 or GA20 oxidase_5
gene. [0726] 164. The method of any one of embodiments 11, 26, 41,
56, 71, 86, or 101, wherein the transcribable DNA sequence is at
least 80% complementary to at least 15 consecutive nucleotides of
one or more of SEQ ID NOs: 218-220, 222-224, 226, and 228-255.
[0727] 165. The method of any one of embodiments 11, 26, 41, 56,
71, 86, or 101, wherein the transcribable DNA sequence is at least
80% complementary to at least 15 consecutive nucleotides of one or
more of SEQ ID NOs: 222-224 and 228-235. [0728] 166. The method of
any one of embodiments 7-14, 22-29, 37-44, 52-59, 67-74, 82-89, or
97-104, wherein the genome modification further deletes at least a
portion of the transcription termination sequence of the endogenous
GA20 oxidase_5 gene. [0729] 167. The method of any one of
embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90, or 97-105,
wherein the genome modification comprises a deletion of one or both
of the transcription termination sequences of the endogenous GA20
oxidase_5 and SAMT genes. [0730] 168. The method of any one of
embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90, 97-105 or 167,
wherein the genome modification comprises a deletion of at least 25
consecutive nucleotides of the intergenic region between the
endogenous GA20 oxidase_5 and SAMT genes. [0731] 169. The method of
any one of embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90,
97-105, 167, or 168, wherein the genome modification comprises a
deletion of the entire intergenic region between the endogenous
GA20 oxidase_5 and SAMT genes. [0732] 170. The method of any one of
embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90, 97-105, or
167-169, wherein the genome modification comprises a deletion of
one or more sequence elements selected from the group consisting of
the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3' UTR, and any portion of the foregoing, of
the endogenous GA20 oxidase_5 gene. [0733] 171. The method of any
one of embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90, 97-105,
or 167-170, wherein the genome modification comprises a deletion of
one or more sequence elements selected from the group consisting of
the 5' UTR, 1.sup.st exon, 1.sup.st intron, 2.sup.nd exon, 2.sup.nd
intron, 3.sup.rd exon, 3.sup.rd intron, 4.sup.th exon, 4.sup.th
intron, 5.sup.th exon, 5.sup.thintron, 6.sup.th exon, 6.sup.th
intron, 7.sup.th exon, 7.sup.th intron, 8.sup.th exon, 3' UTR, and
any portion of the foregoing, of the endogenous Zm.SAMT locus.
[0734] 172. The method of any one of embodiments 7-15, 22-30,
37-45, 52-60, 67-75, 82-90, 97-105, or 167-171, wherein the mutant
allele produces a RNA molecule comprising an antisense sequence
that is at least 80% complementary to a RNA transcript sequence, or
a portion thereof, encoded by the endogenous GA20 oxidase_5 gene.
[0735] 173. The method of any one of embodiments 7-15, 22-30,
37-45, 52-60, 67-75, 82-90, 97-105, or 167-172, wherein the RNA
transcript sequence comprises a sequence that is at least 90%
identical to at least 15 consecutive nucleotides of one or more of
SEQ ID NOs: 218-220, 222-224, 226, and 228-255. [0736] 174. The
method of any one of embodiments 7-15, 22-30, 37-45, 52-60, 67-75,
82-90, 97-105, or 167-173, wherein the RNA transcript sequence
comprises a sequence that is at least 90 identical to at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 222-224 and
228-235. [0737] 175. The method of any one of embodiments 7-15,
22-30, 37-45, 52-60, 67-75, 82-90, 97-105, or 167-174, wherein the
antisense sequence of the RNA molecule is at least 80%
complementary to at least 15 consecutive nucleotides of one or more
of SEQ ID NOs: 218-220, 222-224, 226, and 228-255. [0738] 176. The
method of any one of embodiments 7-15, 22-30, 37-45, 52-60, 67-75,
82-90, 97-105, or 167-175, wherein the antisense sequence of the
RNA molecule is at least 80% complementary to at least 15
consecutive nucleotides of one or more of SEQ ID NOs: 222-224 and
228-235. [0739] 177. The method of any one of embodiments 7-15,
22-30, 37-45, 52-60, 67-75, 82-90, 97-105, or 167-176, wherein the
genome modification results in the production of an RNA molecule
comprising an antisense sequence from a genomic segment of selected
from the group consisting of an exon, a portion of an exon, an
intron, a portion of an intron, a 5' or 3' untranslated region
(UTR), a portion of an UTR, and any combination of the foregoing,
of the endogenous GA20 oxidase_5 locus. [0740] 178. The method of
any one of embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90,
97-105, or 167-177, wherein the antisense sequence can hybridize
with an RNA transcript encoded by a wild-type allele of one or both
of the endogenous GA20 oxidase_3 gene and the endogenous GA20
oxidase_5 gene. [0741] 179. The method of any one of embodiments
7-15, 22-30, 37-45, 52-60, 67-75, 82-90, 97-105, or 167-178,
wherein the antisense sequence can hybridize with a sense RNA
transcript encoded by an endogenous GA20 oxidase_5 gene. [0742]
180. The method of any one of embodiments 7-15, 22-30, 37-45,
52-60, 67-75, 82-90, 97-105, or 167-179, wherein the antisense
sequence can hybridize with a sense RNA transcript encoded by the
mutant allele of the endogenous GA20 oxidase_5 gene. [0743] 181.
The method of embodiment 179 or 180, wherein the sense RNA
transcript encoded by the mutant allele of the endogenous GA20
oxidase_5 gene is shortened or truncated relative to a wild-type
allele of the endogenous GA20 oxidase_5 gene.
[0744] 182. The method of any one of embodiments 178-181, wherein
the hybridization can cause suppression of a wild-type or mutant
allele of the endogenous GA20 oxidase_3 gene, a wild-type or mutant
allele of the endogenous GA20 oxidase_5 gene, or a wild-type or
mutant allele of both genes. [0745] 183. The method of any one of
embodiments 7-15, 22-30, 37-45, 52-60, 67-75, 82-90, 97-105, or
167-182, wherein the genome modification comprises two or more,
three or more, four or more, five or more, or six or more
non-contiguous deletions. [0746] 184. The method of any one of
embodiments 3, 18, 33, 48, 63, 78, or 93, wherein the mutant allele
of the endogenous br2 locus suppresses the expression of a
wild-type allele of the endogenous br2 locus. [0747] 185. The
method of any one of embodiments 3, 18, 33, 48, 63, 78, or 93,
wherein the mutant allele product of the endogenous br2 locus
disrupts the function of a wild-type allele product of the
endogenous br2 locus. [0748] 186. The method of any one of
embodiments 3, 18, 33, 48, 63, 78, or 93, wherein the RNA
transcript further comprises one or more sequence elements of the
endogenous br2 locus selected from the group consisting of 5'UTR,
first exon, first intron, second exon, second intron, third exon,
third intron, fourth exon, fourth intron, fifth exon, 3' UTR, and
any portion thereof. [0749] 187. The method of any one of
embodiments 3, 18, 33, 48, 63, 78, or 93, wherein the DNA segment
comprises a nucleotide sequence originating from the endogenous br2
locus. [0750] 188. The method of embodiment 187, wherein the DNA
segment corresponds to an inverted genomic fragment of the
endogenous br2 locus. [0751] 189. The method of any one of
embodiments 3, 18, 33, 48, 63, 78, or 93, wherein at least a
portion of the antisense RNA sequence is at least 70% complementary
to a corresponding endogenous sequence of the RNA transcript.
[0752] 190. The method of embodiment 189, wherein the corresponding
endogenous sequence of the RNA transcript is at least 85% identical
to at least 20 consecutive nucleotides of SEQ ID NO: 132 or 180.
[0753] 191. The method of embodiment 189 or 190, wherein the
antisense RNA sequence hybridizes to the corresponding endogenous
sequence of the RNA transcript. [0754] 192. The method of any one
of embodiments 189-191, wherein the DNA segment is inserted near or
adjacent to a corresponding endogenous DNA segment of the
endogenous br2 locus. [0755] 193. The method of embodiment 192,
wherein the antisense RNA sequence encoded by the inserted DNA
segment hybridizes to a corresponding endogenous sequence of the
RNA transcript encoded by the corresponding endogenous DNA segment.
[0756] 194. The method of embodiment 192 or 193, wherein the
antisense RNA sequence forms a stem-loop structure with the
corresponding endogenous sequence of the RNA transcript. [0757]
195. The method of embodiment 192, wherein the inserted DNA segment
and the corresponding endogenous DNA segment of the mutant allele
are separated by an intervening DNA sequence. [0758] 196. The
method of embodiment 195, wherein the intervening DNA sequence has
a length of at least 2 consecutive nucleotides. [0759] 197. The
method of embodiment 195, wherein the DNA segment and the
corresponding endogenous DNA segment are separated by an
intervening sequence of at most 4000 consecutive nucleotides.
[0760] 198. The method of any one of embodiments 195-197, wherein
the intervening DNA sequence encodes an intervening RNA sequence
between the antisense RNA sequence and the corresponding endogenous
sequence of the RNA transcript. [0761] 199. The method of
embodiment 198, wherein the RNA transcript forms a stem-loop
secondary structure with the intervening RNA sequence forming the
loop portion of the stem-loop secondary structure. [0762] 200. The
method of embodiment 199, wherein the stem-loop secondary structure
comprises a near-perfect-complement stem with mismatches. [0763]
201. The method of embodiment 199, wherein the stem-loop secondary
structure comprises a perfect-complement stem with no mismatch.
[0764] 202. The method of any one of embodiments 195-201, wherein
the intervening DNA sequence comprises a native sequence of the
endogenous br2 locus. [0765] 203. The method of any one of
embodiments 195-201, wherein the intervening DNA sequence comprises
an exogenous sequence inserted into the endogenous br2 locus.
[0766] 204. The method of any one of embodiments 195-201, wherein
the intervening DNA sequence comprises an intron sequence. [0767]
205. The method of any one of embodiments 195-201, wherein the
intervening DNA sequence does not comprise an intron sequence.
[0768] 206. The method of claim 192, wherein the inserted DNA
segment is located upstream of the corresponding endogenous DNA
segment. [0769] 207. The method of embodiment 192, wherein the
inserted DNA segment is located downstream of the corresponding
endogenous DNA segment. [0770] 208. The method of embodiment 189,
wherein the DNA segment is inserted within a region selected from
the group consisting of 5' untranslated region (UTR), first exon,
first intron, second exon, second intron, third exon, third intron,
fourth exon, fourth intron, fifth exon, and 3' UTR of the
endogenous br2 locus, and a combination thereof. [0771] 209. The
method of embodiment 189, wherein the mutant allele further
comprises a deletion of at least one portion of the endogenous br2
locus. [0772] 210. The method of embodiment 189, wherein the sense
strand of the DNA segment comprises a sequence at least 70%
complementary to an exon sequence of the endogenous br2 locus.
[0773] 211. The method of embodiment 189, wherein the sense strand
of the DNA segment comprises a sequence at least 70% complementary
to an untranslated region (UTR) sequence of the endogenous br2
locus. [0774] 212. The method of embodiment 189, wherein the sense
strand of the DNA segment comprises a sequence at least 70%
complementary to an exon sequence and an intron sequence of the
endogenous br2 locus, the exon sequence and the intron sequence
being contiguous within the endogenous locus. [0775] 213. The
method of embodiment 189, wherein the DNA segment comprises a
sequence having at least at least 70% identity to one or more of
SEQ ID Nos: 132 and 180. [0776] 214. The method of any one of
embodiments 3, 18, 33, 48, 63, 78, or 93, wherein the DNA segment
has a length of at least 15 nucleotides. [0777] 215. The method of
any one of embodiments 3, 18, 33, 48, 63, 78, or 93, wherein the
DNA segment has a length of at most 1000 nucleotides. [0778] 216.
The method of any one of embodiments 4, 19, 34, 49, 64, 79, or 94,
wherein the deletion further comprises the deletion of the at least
one exon of the endogenous br2 locus as compared to SEQ ID NO: 132.
[0779] 217. The method of any one of embodiments 4, 19, 34, 49, 64,
79, or 94, wherein the deletion further comprises the deletion of
the endogenous br2 locus. [0780] 218. The method of embodiment 216,
wherein the at least one exon is the first exon of the endogenous
br2 locus. [0781] 219. The method of embodiment 216, wherein the at
least one exon is the second exon of the endogenous br2 locus.
[0782] 220. The method of embodiment 216, wherein the at least one
exon is the third exon of the endogenous br2 locus. [0783] 221. The
method of embodiment 216, wherein the at least one exon is the
fourth exon of the endogenous br2 locus. [0784] 222. The method of
embodiment 216, wherein the at least one exon is the fifth exon of
the endogenous br2 locus. [0785] 223. The method of any one of
embodiments 4, 19, 34, 49, 64, 79, 94, or 216-222, wherein the
deletion further comprises a deletion of at least one nucleotide
from at least one intron of the endogenous br2 locus. [0786] 224.
The method of embodiment 222, wherein the deletion comprises the
deletion of the at least one intron. [0787] 225. The method of any
one of embodiments 4, 19, 34, 49, 64, 79, 94, or 216-224, wherein
the deletion further comprises the deletion of at least one
nucleotide of the 5'-untranslated region of the endogenous br2
locus. [0788] 226. The method of any one of embodiments 4, 19, 34,
49, 64, 79, or 94 or 216-225, wherein the deletion further
comprises the deletion of at least one nucleotide of the
3'-untranslated region of the endogenous br2 locus. [0789] 227. The
method of embodiment 216, wherein the deletion further comprises
the deletion of a second exon of the endogenous br2 locus. [0790]
228. The method of embodiment 227, wherein the two exons are
contiguous exons. [0791] 229. The method of embodiment 227, wherein
the two exons are not contiguous exons. [0792] 230. The method of
claim any one of embodiments 4, 19, 34, 49, 64, 79, or 94, wherein
the mutant allele encodes a truncated protein as compared to SEQ ID
NO: 181. [0793] 231. The method of any one of embodiments 4, 19,
34, 49, 64, 79, or 94 or 218-229, wherein the deletion comprises
between 10 nucleotides and 8000 nucleotides. [0794] 232. The method
of any one of embodiments 4, 19, 34, 49, 64, 79, or 94, wherein the
mutant allele encodes an mRNA transcript comprising a premature
stop codon as compared to SEQ ID NO: 180. [0795] 233. The method of
any one of embodiments 6, 21, 36, 51, 66, 81, or 96, wherein a
protein encoded by the nucleic acid sequence is truncated as
compared to SEQ ID NO: 181. [0796] 234. The method of any one of
embodiments 6, 21, 36, 51, 66, 81, or 96, wherein a protein encoded
by the nucleic acid sequence comprises 1378 or fewer amino acids.
[0797] 235. The method of any one of embodiments 6, 21, 36, 51, 66,
81, or 96, wherein the premature stop codon is present in a region
of the nucleic acid sequence selected from the group consisting of
the first exon, the second exon, the third exon, the fourth exon,
and the fifth exon. [0798] 236. The method of any one of
embodiments 6, 21, 36, 51, 66, 81, 96, or 233-235, wherein the
premature stop codon results from a nonsense mutation. [0799] 237.
The method of any one of embodiments 6, 21, 36, 51, 66, 81, 96, or
233-235, wherein the premature stop codon results from a missense
mutation. [0800] 238. The method of any one of embodiments 5, 20,
35, 50, 65, 80, or 95, wherein the gene is a GA20 oxidase gene.
[0801] 239. The method of embodiment 238, wherein the GA20 oxidase
gene is a GA20 oxidase_3 gene. [0802] 240. The method of embodiment
238, wherein the GA20 oxidase is a GA20 oxidase_5 gene. [0803] 241.
The method of any one of embodiments 5, 20, 35, 50, 65, 80, or 95,
wherein the gene is a GA3 oxidase gene. [0804] 242. The method of
any one of embodiments 5, 20, 35, 50, 65, 80, or 95, wherein the
gene is a brachytic2 gene. [0805] 243. The method of any one of
embodiments 5, 20, 35, 50, 65, 80, or 95, wherein the mutant allele
is a dominant negative mutant allele. [0806] 244. The method of
embodiment 243, wherein the dominant negative mutant allele
generates an antisense RNA transcript capable of triggering
suppression of an unmodified or wildtype allele of the gene. [0807]
245. The method of embodiment 243, wherein the dominant negative
mutant allele encodes a truncated protein as compared to an
unmodified allele of the gene. [0808] 246. The method of embodiment
243, wherein the dominant negative mutant allele generates at least
one RNA transcript capable of forming a hairpin-loop secondary
structure. [0809] 247. The method of embodiment 243, wherein the
coding sequence of the dominant negative mutant allele is operably
linked to a promoter of the native copy of the gene. [0810] 248.
The method of embodiment 243, wherein the dominant negative mutant
allele comprises an inverted copy of the gene, or a portion
thereof, adjacent to a wildtype copy of the gene at the endogenous
locus of the gene. [0811] 249. The method of embodiment 243,
wherein the dominant negative mutant allele comprises a deletion of
a portion of a chromosome between a first region of the gene and a
second region of the gene, wherein an antisense RNA transcript of
the first region of the gene is generated following the deletion of
the portion of the chromosome. [0812] 250. The method of embodiment
243, wherein the dominant negative mutant allele comprises a first
promoter and a second promoter separated by an intervening region,
wherein the first promoter and the second promoter are positioned
in opposite orientations, wherein the second promoter generates at
least one antisense RNA transcript, and wherein expression of the
gene is reduced as compared to a control corn plant that lacks the
dominant negative mutant allele. [0813] 251. The method of
embodiment 243, wherein the dominant negative mutant allele
comprises a tissue-specific or tissue-preferred promoter inserted
into the gene in reverse orientation as compared to the native
promoter of the gene, wherein the tissue-specific or
tissue-preferred promoter generates at least one antisense RNA
transcript, and wherein expression of the gene is reduced as
compared to a control corn plant that lacks the dominant negative
mutant allele. [0814] 252. The method of any one of embodiments 5,
20, 35, 50, 65, 80, or 95, wherein the mutant allele comprises an
insertion, an inversion, or a deletion as compared to a wildtype
allele of the gene. [0815] 253. The method of any one of
embodiments 1-252, wherein the at least one corn plant is
homozygous or biallelic for the mutant allele or transgene. [0816]
254. The method of any one of embodiments 1-252, wherein the at
least one corn plant is heterozygous for the mutant allele or
transgene. [0817] 255. The method of any one of embodiments 1-252,
wherein the at least one corn plant has improved lodging resistance
relative to an unmodified control plant. [0818] 256. The method of
any one of embodiments 1-252, wherein the mutant allele is a
dominant allele. [0819] 257. The method of any one of embodiments
1-252, wherein the mutant allele is a semi-dominant allele. [0820]
258. A method comprising harvesting a plurality of corn plants from
a field at least 50 days after fertilization or silking of at least
50% of said plurality of corn plants, wherein fewer than or equal
to 50% of said corn plants have lodged at the time of harvest, and
wherein at least one corn plant of said plurality of corn plants
comprises a recombinant DNA construct comprising a transcribable
DNA sequence encoding a GA2 oxidase protein and a plant-expressible
promoter, wherein the transcribable DNA sequence is operably linked
to the plant-expressible promoter. [0821] 259. A method comprising
harvesting a plurality of corn plants from a field at least 50 days
after at least 50% of said corn plants have reached R3 stage,
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible
promoter.
[0822] 260. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after fertilization or silking
of at least 50% of said plurality of corn plants, wherein the
average kernel moisture content is less than or equal to 30%, and
wherein fewer than or equal to 50% of said corn plants have lodged
at the time of harvest, and wherein at least one corn plant of said
plurality of corn plants comprises a recombinant DNA construct
comprising a transcribable DNA sequence encoding a GA2 oxidase
protein and a plant-expressible promoter, wherein the transcribable
DNA sequence is operably linked to the plant-expressible promoter.
[0823] 261. A method comprising harvesting a plurality of corn
plants from a field at least 50 days after at least 50% of said
corn plants have reached R3 stage, wherein average kernel moisture
content is less than or equal to 30%, and wherein fewer than or
equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. [0824] 262. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after at fertilization or silking of at
least 50% of said plurality of corn plants, wherein the average
yield of said field is at least 170 bushels per acre, wherein fewer
than or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. [0825] 263. A
method comprising harvesting a plurality of corn plants from a
field at least 50 days after at least 50% of said corn plants have
reached R3 stage, wherein the average yield of said field is at
least 170 bushels per acre, wherein fewer than or equal to 50% of
said corn plants have lodged at the time of harvest, and wherein at
least one corn plant of said plurality of corn plants comprises a
recombinant DNA construct comprising a transcribable DNA sequence
encoding a GA2 oxidase protein and a plant-expressible promoter,
wherein the transcribable DNA sequence is operably linked to the
plant-expressible promoter. [0826] 264. A method comprising
harvesting a plurality of corn plants from a field at least 1 day
after the average kernel moisture content of at least 50% of said
plurality of corn plants is between 10% and 30%, wherein fewer than
or equal to 50% of said corn plants have lodged at the time of
harvest, and wherein at least one corn plant of said plurality of
corn plants comprises a recombinant DNA construct comprising a
transcribable DNA sequence encoding a GA2 oxidase protein and a
plant-expressible promoter, wherein the transcribable DNA sequence
is operably linked to the plant-expressible promoter. [0827] 265.
The method of any one of embodiments 258-264, wherein the GA2
oxidase protein is, or comprises a sequence that is, at least 80%
identical to one or more of SEQ ID NOs: 325, 327, 329, 331, 333,
335, 337, 339, 341, 343, 345, 347, and 349. [0828] 266. The method
of any one of embodiments 258-265, wherein the transcribable DNA
sequence is, or comprises a sequence that is, at least 80%
identical to one or more of SEQ ID NOs: 324, 326, 328, 330, 332,
334, 336, 338, 340, 342, 344, 346, and 348. [0829] 267. The method
of any one of embodiments 258-264, wherein the GA2 oxidase protein
is, or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 351, 353, 355, 357, 359, 361, 363, 365, 367, and/or
369. [0830] 268. The method of any one of embodiments 258-264 or
267, wherein the transcribable DNA sequence is, or comprises a
sequence that is, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
350, 352, 354, 356, 358, 360, 362, 364, 366, and/or 368. [0831]
269. The method of any one of embodiments 258-264, wherein the GA2
oxidase protein is, or comprises a sequence that is, at least 80%,
at least 85%, at least 90%, at least 95%, at least 96%, at least
97%, at least 98%, at least 99%, at least 99.5%, or 100% identical
to one or more of SEQ ID NOs: 371, 373, 375, 377, 379, 381, 383,
and/or 385. [0832] 270. The method of any one of embodiments
258-264 or 269, wherein the transcribable DNA sequence is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 370, 372, 374, 376, 378, 380, 382, and/or 384. [0833] 271.
The method of any one of embodiments 258-264, wherein the GA2
oxidase protein is, or comprises a sequence that is, at least 80%,
at least 85%, at least 90%, at least 95%, at least 96%, at least
97%, at least 98%, at least 99%, at least 99.5%, or 100% identical
to one or more of SEQ ID NOs: 387, 389, 391, 393, 395, 397, 399,
401, 403, 405, 407, 409, 411, 413, 415, and/or 417. [0834] 272. The
method of any one of embodiments 258-264 or 271, wherein the
transcribable DNA sequence is, or comprises a sequence that is, at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 386, 388, 390, 392, 394,
396, 398, 400, 402, 404, 406, 408, 410, 412, 414, and/or 416.
[0835] 273. The method of any one of embodiments 258-264, wherein
the GA2 oxidase protein is, or comprises a sequence that is, at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 419, 421, 423, 425, 427,
429, 431, 433, 435, 437, 439, 441, 443, 445, and/or 447. [0836]
274. The method of any one of embodiments 258-264 or 273, wherein
the transcribable DNA sequence is, or comprises a sequence that is,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 418, 420, 421, 424,
426, 428, 430, 432, 434, 436, 438, 440, 442, 444, and/or 446.
[0837] 275. The method of any one of embodiments 258-264, wherein
the GA2 oxidase protein is, or comprises a sequence that is, at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 449, 451, 453, 455, 457,
459, 461, 463, 465, 467, and/or 469. [0838] 276. The method of any
one of embodiments 258-264 or 275, wherein the transcribable DNA
sequence is, or comprises a sequence that is, at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 448, 450, 452, 454, 456, 458, 460, 462,
464, 466, and/or 468. [0839] 277. The method of any one of
embodiments 258-264, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 471, 473, 475, 477, 479, 481, 483, 485, 487, 489, 491, 493,
495, 497, 499, and/or 501. [0840] 278. The method of any one of
embodiments 258-264 or 277, wherein the transcribable DNA sequence
is, or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 470, 472, 474, 476, 478, 480, 482, 484, 486, 488, 490,
492, 494, 496, 498, and/or 500. [0841] 279. The method of any one
of embodiments 258-264, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 503, 505, 507, 509, 511, 513, 515, 517, 519, 521, 523, 525,
527, 529, and/or 531. [0842] 280. The method of any one of
embodiments 258-264 or 279, wherein the transcribable DNA sequence
is, or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 502, 504, 506, 508, 510, 512, 514, 516, 518, 520, 522,
524, 526, 528, and/or 530. [0843] 281. The method of any one of
embodiments 258-264, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 533, 535, 537, 539, 541, 543, 545, 547, 549, 551, 553, 555,
557, 559, and/or 561. [0844] 282. The method of any one of
embodiments 258-264 or 281, wherein the transcribable DNA sequence
is, or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 532, 534, 536, 538, 540, 542, 544, 546, 548, 550, 552,
554, 556, 558, and/or 560. [0845] 283. The method of any one of
embodiments 258-264, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 563, 565, 567, 569, 571, 573, and/or 575. [0846] 284. The
method of any one of embodiments 258-264 or 283, wherein the
transcribable DNA sequence is, or comprises a sequence that is, at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 562, 564, 566, 568, 570,
572, and/or 574. [0847] 285. The method of any one of embodiments
258-264, wherein the GA2 oxidase protein is, or comprises a
sequence that is, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
577, 579, 581, 583, 585, 587, and/or 589. [0848] 286. The method of
any one of embodiments 258-264 or 285, wherein the transcribable
DNA sequence is, or comprises a sequence that is, at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 576, 578, 580, 582, 584, 586, and/or
588. [0849] 287. The method of any one of embodiments 258-264,
wherein the GA2 oxidase protein is, or comprises a sequence that
is, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 591, 593,
595, 597, 599, 601, 603, 605, and/or 607. [0850] 288. The method of
any one of embodiments 258-264 or 287, wherein the transcribable
DNA sequence is, or comprises a sequence that is, at least 80%, at
least 85%, at least 90%, at least 95%, at least 96%, at least 97%,
at least 98%, at least 99%, at least 99.5%, or 100% identical to
one or more of SEQ ID NOs: 590, 592, 594, 596, 598, 600, 602, 604,
and/or 606. [0851] 289. The method of any one of embodiments
258-264, wherein the GA2 oxidase protein is, or comprises a
sequence that is, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs:
609, 611, 613, and/or 615. [0852] 290. The method of any one of
embodiments 258-264 or 289, wherein the transcribable DNA sequence
is, or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 608, 610, 612, and/or 614. [0853] 291. The method of
any one of embodiments 258-264, wherein the GA2 oxidase protein is,
or comprises a sequence that is, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%, at least 99.5%, or 100% identical to one or more of
SEQ ID NOs: 617, 619, 621, 623, 625, 627, 629, and/or 631. [0854]
292. The method of any one of embodiments 258-264 or 291, wherein
the transcribable DNA sequence is, or comprises a sequence that is,
at least 80%, at least 85%, at least 90%, at least 95%, at least
96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or
100% identical to one or more of SEQ ID NOs: 616, 618, 620, 622,
624, 626, 628, and/or 630. [0855] 293. The method of any one of
embodiments 258-264, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 633, 635, 637, 639, 641, 643, and/or 645. [0856] 294. The
method of any one of embodiments 258-264 or 293, wherein the
transcribable DNA sequence is, or comprises a sequence that is, at
least 80%, at least 85%, at least 90%, at least 95%, at least 96%,
at least 97%, at least 98%, at least 99%, at least 99.5%, or 100%
identical to one or more of SEQ ID NOs: 632, 634, 636, 638, 640,
642, and/or 644. [0857] 295. The method of any one of embodiments
258-264, wherein the GA2 oxidase protein is, or comprises a
sequence that is, at least 80%, at least 85%, at least 90%, at
least 95%, at least 96%, at least 97%, at least 98%, at least 99%,
at least 99.5%, or 100% identical to one or more of SEQ ID NOs: 647
and/or 649. [0858] 296. The method of any one of embodiments
258-264 or 295, wherein the transcribable DNA sequence is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 646 and/or 648. [0859] 297. The method of any one of
embodiments 258-264, wherein the GA2 oxidase protein is, or
comprises a sequence that is, at least 80%, at least 85%, at least
90%, at least 95%, at least 96%, at least 97%, at least 98%, at
least 99%, at least 99.5%, or 100% identical to one or more of SEQ
ID NOs: 651, 653, and/or 655.
[0860] 298. The method of any one of embodiments 258-264 or 297,
wherein the transcribable DNA sequence is, or comprises a sequence
that is, at least 80%, at least 85%, at least 90%, at least 95%, at
least 96%, at least 97%, at least 98%, at least 99%, at least
99.5%, or 100% identical to one or more of SEQ ID NOs: 650, 652,
and/or 654. [0861] 299. The method of any one of embodiments
258-298, wherein the plant-expressible promoter is a vascular
promoter. [0862] 300. The method of embodiment 299, wherein the
vascular promoter comprises one of the following: a sucrose
synthase promoter, a sucrose transporter promoter, a Sh1 promoter,
Commelina yellow mottle virus (CoYMV) promoter, a wheat dwarf
geminivirus (WDV) large intergenic region (LIR) promoter, a maize
streak geminivirus (MSV) coat protein (CP) promoter, a rice yellow
stripe 1 (YS1)-like promoter, or a rice yellow stripe 2 (OsYSL2)
promoter. [0863] 301. The method of embodiment 299, wherein the
vascular promoter comprises a DNA sequence that is at least 80%
identical to one or more of SEQ ID NO: 658, SEQ ID NO: 659, SEQ ID
NO: 660, SEQ ID NO: 661, or SEQ ID NO: 662, or a functional portion
thereof. [0864] 302. The method of any one of embodiments 258-298,
wherein the plant-expressible promoter is a rice tungro bacilliform
virus (RTBV) promoter. [0865] 303. The method of embodiment 302,
wherein the RTBV promoter comprises a DNA sequence that is at least
80% identical to one or more of SEQ ID NOs: 656 and 657, or a
functional portion thereof. [0866] 304. The method of any one of
embodiments 258-298, wherein the plant-expressible promoter is a
leaf promoter. [0867] 305. The method of embodiment 304, wherein
the leaf promoter comprises one or more of the following: a RuBisCO
promoter, a PPDK promoter, a FDA promoter, a Nadh-Gogat promoter, a
chlorophyll a/b binding protein gene promoter, a
phosphoenolpyruvate carboxylase (PEPC) promoter, or a Myb gene
promoter. [0868] 306. The method of embodiment 304, wherein the
leaf promoter comprises a DNA sequence that is at least 80%
identical to one or more of SEQ ID NO: 663, SEQ ID NO: 664, or SEQ
ID NO: 665, or a functional portion thereof. [0869] 307. The method
of any one of embodiments 258-298, wherein the plant expressible
promoter is a constitutive promoter. [0870] 308. The method of
embodiment 307, wherein the constitutive promoter is selected from
the group consisting of: an actin promoter, a CaMV 35S or 19S
promoter, a plant ubiquitin promoter, a plant Gos2 promoter, a FMV
promoter, a CMV promoter, a MMV promoter, a PCLSV promoter, an Emu
promoter, a tubulin promoter, a nopaline synthase promoter, an
octopine synthase promoter, a mannopine synthase promoter, or a
maize alcohol dehydrogenase, or a functional portion thereof [0871]
309. The method of embodiment 307, wherein the constitutive
promoter comprises a DNA sequence that is at least 80% identical to
one or more of SEQ ID NOs: 666, SEQ ID NO: 667, SEQ ID NO: 668, SEQ
ID NO: 669, SEQ ID NO: 670, SEQ ID NO: 671, SEQ ID NO: 672, SEQ ID
NO: 673 or SEQ ID NO: 674, or a functional portion thereof. [0872]
310. The method of any one of embodiments 258, 260, or 262, wherein
said harvesting is performed at least 55 days, at least 60 days, at
least 75 days, at least 90 days, or at least 120 days after said
fertilization or silking. [0873] 311. The method of any one of
embodiments 259, 261, or 263, wherein said harvesting is performed
at least 55 days, at least 60 days, at least 75 days, at least 90
days, or at least 120 days after at least 50% of said corn plants
have reached R3 stage. [0874] 312. The method of any one of
embodiments 258-311, wherein fewer than or equal to 40%, fewer than
or equal to 30%, fewer than or equal to 20%, or fewer than or equal
to 10% of said corn plants have lodged at the time of harvest.
[0875] 313. The method of any one of embodiments 258-311, wherein
the average height of said corn plants is less than or equal to 1.7
meters, less than or equal to 1.6 meters, or less than or equal to
1.5 meters at the time of harvest. [0876] 314. The method of claim
313, wherein said height is measured as the distance between the
soil and the ligule of the uppermost fully-expanded leaf. [0877]
315. The method of any one of embodiments 258-314, wherein at least
50% of said corn plants are inbred corn plants. [0878] 316. The
method of any one of embodiments 258-314, wherein at least 50% of
said corn plants are hybrid corn plants. [0879] 317. The method of
any one of embodiments 258-314, wherein at least 50% of said corn
plants are semi-dwarf corn plants. [0880] 318. The method of any
one of embodiments 258-314, wherein at least 50% of said corn
plants are dwarf corn plants. [0881] 319. The method of any one of
embodiments 258-314, wherein at least 50% of said corn plants are
brachytic corn plants. [0882] 320. The method of any one of
embodiments 258-319, wherein said field comprises a planting
density of at least 10,000 corn plants per acre. [0883] 321. The
method of any one of embodiments 258-320, wherein the average yield
of said field is at least 180 bushels per acre, at least 190
bushels per acre, at least 200 bushels per acre, at least 210
bushels per acre, at least 220 bushels per acre, or at least 250
bushels per acre. [0884] 322. The method of any one of embodiments
258-263, wherein the average kernel moisture content is equal to or
less than 29%, equal to or less than 28%, equal to or less than
27%, equal to or less than 26%, equal to or less than 25%, equal to
or less than 24%, equal to or less than 23%, equal to or less than
22%, equal to or less than 21%, equal to or less than 20%, equal to
or less than 19%, equal to or less than 15%, equal to or less than
10%. [0885] 323. The method of any one of embodiments 258-321,
wherein the average kernel moisture content is between 10% and 20%,
between 13% and 20%, between 15% and 20%, between 10% and 25%,
between 13% and 25%, between 15% and 25%, between 20% and 25%,
between 13% and 30%, between 15% and 30%, between 20% and 30%, or
between 25% and 30%. [0886] 324. The method of any one of
embodiments 264, wherein said harvesting occurs at least 2 days, at
least 3 days, at least 4 days, at least 5 days, at least 6 days, at
least 7 days, at least 8 days, at least 9 days, at least 10 days,
at least 15 days, at least 20 days, at least 25 days, at least 30
days, at least 35 days, at least 40 days at least 45 days, at least
50 days, at least 55 days, at least 60 days, at least 65 days, or
at least 70 days after an average kernel moisture content of said
plurality of corn plants is between 10% and 30%, or the kernel
moisture content of a corn plant of the plurality of corn plants is
between 10% and 30%. [0887] 325. The method of any one of
embodiments 258-324, wherein said method comprises growing said
plurality of said corn plants in said corn field prior to said
harvesting.
[0888] Having described the present disclosure in detail, it will
be apparent that modifications, variations, and equivalent aspects
are possible without departing from the spirit and scope of the
present disclosure as described herein and in the appended claims.
Furthermore, it should be appreciated that all examples in the
present disclosure are provided as non-limiting examples.
EXAMPLES
Example 1: Semi-Dwarf Corn Improves Stalk Health for Late Season
Harvest
[0889] Three semi-dwarf (SD) and three wild-type control hybrids
were planted in 30 inch rows with a density of approximately 42,000
plants per acre in two separate locations. When hybrids reached
full maturity, Stalk Lodging Percent (STLP) and Stalk Health Good
Pith Percentage (SHGPP) were collected at two separate locations at
normal (October 10.sup.th) and late (November 12.sup.th) harvest
dates. Hybrids were replicated twelve times for each harvest date.
Hybrids harvested at normal harvest date had a moisture content of
approximately 25%. At late harvest date, hybrids had a moisture
content of approximately 15%. STLP was collected by counting the
number of lodged plants by the total number of plants per plot at
normal harvest and at late harvest prior to combine harvest. After
combine harvest, SHGPP was collected cutting the residual stalks
between first and second node then observing the intactness of the
pith. Percentage of intactness was designated for each stalk. As
shown in Table 20, SD hybrids have improved stalk health as
compared to WT hybrids.
TABLE-US-00020 TABLE 20 Stalk Health of SD versus WT Control
Hybrids at Normal and Delayed Harvest SHGPP STLP Normal Late Normal
Late Stature Line Harvest Harvest Harvest Harvest SD Line #1 59.3
62.1 0.6 1.1 Line #2 70.3 52.6 2.1 3.5 Line #3 62.0 53.5 1.9 2.2
Semi-Dwarf Total 63.8 56.1 1.5 2.2 WT Line #4 43.5 29.4 1.5 3.4
Line #5 23.9 18.1 2.6 5.5 Line #6 51.8 49.1 0.5 8.6 WT Total 39.7
32.3 1.5 5.9
[0890] In this example, SD hybrids overall improved stalk health
good pith percent as compared to WT at normal harvest, 63.8 versus
39.7 percent, respectively. This improved stalk health good pith
percent was observed at late harvest, 56.1 for SD hybrids compared
to 32.3 for WT hybrids (FIG. 1). At late harvest, the SD hybrids
significantly reduced stalk lodging as compared to WT, 2.2 versus
5.9, respectively. This experiment demonstrates that SD hybrids
allow growers flexibility in harvest by extending the window of
time to harvest due to improved stalk health and standability.
Example 2: Planting Date and Corn Maturity Impacts Harvest
Window
[0891] Each growing season farmers must balance planting the corn
crop after the threat of freezing temperatures has passed, yet with
enough time to allow maximum growing degree days (GDDs) for crop
development to maturity and dry down before the first frost. Corn
GDDs are calculated by subtracting the plant's lower base or
threshold temperature of 50.degree. F. (10.degree. C.) from the
average daily air temperature in .degree. F. or .degree. C. Average
daily air temperature is calculated by averaging the daily maximum
and minimum air temperatures measured in any 24-hour period. To
fully mature corn, a certain amount of accumulated GDDs are
required in relation to its relative maturity (RM) and geographical
location. In the northern hemisphere, including the North American
continent, the corn planting window is typically within the months
of April, May and June (see, e.g., FIG. 2). Emergence, growth, and
pollination of the crop follows through stages of silking, dough,
dent, maturity (black layer), and grain harvest (see, e.g., FIG.
2). When corn reaches physiological maturity (black layer), it is
around 30% moisture. Additionally, late-planted and full-season
corn products tend to dry more slowly. In general, it takes about
30 GDDs per point of moisture to dry corn from maturity to 25%
moisture content (see, e.g., Table 21). After reaching maturity,
typical drying rates may range from 0.4% to 0.8% loss of moisture
content per day (see, e.g., Table 21).
[0892] The optimum harvest moisture content for corn is
approximately 23% to 25%. At this moisture level, kernels shell
easily and stalks generally stand better, which can make harvesting
more efficient. A normal harvest loss level of a timely and
efficient harvest can be 1% to 2%. Knowing the grain moisture
content at maturity can help predict grain moisture at different
potential harvest dates. A year with wet weather and delays in
planting may result in slower field drying of corn. However, if
enough GDDs accumulate, the drying process may be hastened. Other
factors may also come into play if harvest is delayed. For example,
corn could have developed a shallow root system because of the
early-season moisture. In addition, conditions may have been
conducive for the development of stalk rots and stalk
cannibalization in corn. These factors could lead to higher than
normal harvest losses because of an increased risk for stalk
lodging in corn at maturity. Delaying harvest until corn dries down
to 17% to 19% moisture content can save on artificial drying costs.
However, as corn dries down in the field there is greater potential
for excess harvest losses from stalk lodging, in part due to normal
senescence of the plant through the maturation process. Most
harvest losses are mechanical, caused by corn grain never getting
into the combine harvester because it cannot be collected from
lodged plants on the ground. Allowing corn to dry down in the field
could lead to excess harvest losses, as much as 2% to 10% or more
above the normal level from a timely and efficient harvest.
[0893] Corn plants that are less susceptible to lodging that have
improved stalk health for late season harvest reduce the risk of
harvest losses (see Example 1). Farmers make planting decisions of
corn hybrids based on their geographical region and historical
weather data for spring and fall frosts. In one example, a farmer
in Minnesota, USA (MN) planting a 100 RM corn hybrid can expect
between 144 days and 149 days to accumulate enough GDDs for crop
maturity depending on the planting date based on a 30 year average
(1981-2010). The same farmer would need an additional 9 days of GDD
accumulation to reach 25% grain moisture for harvest, or 18 to 19
days of GDD accumulation to reach 20% grain moisture for harvest
(assuming optimal weather conditions). In another example, a farmer
in Illinois, USA (IL) planting a 110 RM corn hybrid can expect
between 128 days and 138 days to accumulate enough GDDs for crop
maturity depending on the planting date based on a 30 year average
(1981-2010). The same farmer would need an additional 7 to 8 days
of GDD accumulation to reach 25% grain moisture for harvest, or 15
to 16 days of GDD accumulation to reach 20% grain moisture for
harvest (assuming optimal weather conditions). As described herein,
at minimum an additional 30 GDDs per point of grain moisture
content to reach 25% or less would be required beyond the maturity
date.
[0894] In the northern hemisphere, or on the North America
continent, the corn harvest window is typically within the months
of September and November (see, e.g., FIG. 2). The longer the
amount of time from crop maturity to harvest the greater the risk
of increased harvest losses unless the corn variety can provide a
benefit of improved standability (less susceptible to lodging),
such as the semi-dwarf (SD) corn in Example 1. Stated another way,
semi-dwarf or shorter stature corn varieties or lines with improved
standability that are less susceptible to lodging (such as the
disclosed SD corn) provide a benefit to farmers or crop growers by
enabling grain dry down in the field for a duration of time beyond
standard expectations from conventional corn varieties. For
example, a farmer would have an increased probability to reach 20%
grain moisture prior to harvest, ranging from an extra 8 to 10 days
at minimum (beyond 25% grain moisture) for the crop to remain in
the field in the example provided (see, e.g., Table 21).
TABLE-US-00021 TABLE 2 Total Days to Seed Days Harvest Days to
Harvest GDD Maturity reach Average Moisture to reach Date reach
Date to Planting (Black Total GDD at 25% 25% 20% 20% Location
Year(s) RM Maturity Date Layer) GDD per day Maturity Moisture
Moisture Moisture Moisture MN 1981-2010 100 2401 Apr 20th Sep 16th
149 16 30% 9 Sep 25th 19 Oct 4th MN 1981-2010 100 2401 May 1st Sep
22nd 144 17 30% 9 Sep 30th 18 Oct 9th MN 1981-2010 100 2401 May
20th Oct 15th 148 16 30% 9 Oct 24th 18 Nov 2nd IL 1981-2010 110
2642 Apr 15th Aug 31st 138 19 30% 8 Sep 7th 16 Sep 15th IL
1981-2010 110 2642 May 1st Sep 7th 129 20 30% 7 Sep 14th 15 Sep
21st IL 1981-2010 110 2642 Jun 1st Oct 7th 128 21 30% 7 Oct 14th 15
Oct 21st
Sequence CWU 0 SQTB SEQUENCE LISTING The patent application
contains a lengthy "Sequence Listing" section. A copy of the
"Sequence Listing" is available in electronic form from the USPTO
web site
(https://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20220162632A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
0 SQTB SEQUENCE LISTING The patent application contains a lengthy
"Sequence Listing" section. A copy of the "Sequence Listing" is
available in electronic form from the USPTO web site
(https://seqdata.uspto.gov/?pageRequest=docDetail&DocID=US20220162632A1).
An electronic copy of the "Sequence Listing" will also be available
from the USPTO upon request and payment of the fee set forth in 37
CFR 1.19(b)(3).
* * * * *
References