U.S. patent application number 14/973174 was filed with the patent office on 2016-04-07 for wheat plants having increased resistance to imidazolinone herbicides.
This patent application is currently assigned to University of Saskatchewan. The applicant listed for this patent is University of Saskatchewan. Invention is credited to Pierre Hucl, Curtis J. POZNIAK.
Application Number | 20160097057 14/973174 |
Document ID | / |
Family ID | 23206205 |
Filed Date | 2016-04-07 |
United States Patent
Application |
20160097057 |
Kind Code |
A1 |
POZNIAK; Curtis J. ; et
al. |
April 7, 2016 |
WHEAT PLANTS HAVING INCREASED RESISTANCE TO IMIDAZOLINONE
HERBICIDES
Abstract
The present invention is directed to wheat plants having
increased resistance to an imidazolinone herbicide. More
particularly, the present invention includes wheat plants
containing one or more IMI nucleic acids such as a Teal IMI
cultivar. The nucleic acids are preferably located on or derived
from different genomes. The present invention also includes seeds
produced by these wheat plants and methods of controlling weeds in
the vicinity of these wheat plants.
Inventors: |
POZNIAK; Curtis J.;
(Saskatoon, CA) ; Hucl; Pierre; (Saskatoon,
CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
University of Saskatchewan |
Saskatoon |
|
CA |
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|
Assignee: |
University of Saskatchewan
|
Family ID: |
23206205 |
Appl. No.: |
14/973174 |
Filed: |
December 17, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13366907 |
Feb 6, 2012 |
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14973174 |
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12363087 |
Jan 30, 2009 |
8110727 |
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13366907 |
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10486605 |
Feb 9, 2004 |
7897845 |
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PCT/CA2002/001051 |
Jul 10, 2002 |
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12363087 |
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60311282 |
Aug 9, 2001 |
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Current U.S.
Class: |
800/300 ;
435/6.11; 504/253; 536/23.2 |
Current CPC
Class: |
C12N 9/88 20130101; C12N
15/8278 20130101; C12N 15/8274 20130101; C12Q 2600/156 20130101;
A01H 5/10 20130101; A01N 43/50 20130101; C12Q 2600/13 20130101;
C12Q 1/6895 20130101; C12Y 401/03 20130101 |
International
Class: |
C12N 15/82 20060101
C12N015/82; A01N 43/50 20060101 A01N043/50; C12N 9/88 20060101
C12N009/88 |
Claims
1. A wheat plant, wherein the wheat plant was obtained by a process
comprising crossing a plant of line Teal11A with a plant of another
wheat line, a representative sample of seed of the line Teal11A
having been deposited with American Type Culture Collection (ATCC)
under Patent Deposit Designation Number PTA-3953 and the wheat
plant comprises a Teal11A Imi2 nucleic acid, said Teal11A Imi2
nucleic acid comprising a polynucleotide sequence encoding an IMI
polypeptide having a serine to asparagine substitution in Domain E,
said Imi2 nucleic acid being that of the B genome, and said wheat
plant exhibiting increased tolerance to an imidazolinone herbicide
as compared to that of a wild-type wheat plant.
2. The plant of claim 1, wherein the imidazolinone herbicide
comprises at least one of: imazethapyr, imazapic, imazamox,
imazaquin, imazethabenz, imazapyr, a mixture of imazapyr and
imazamox, or a combination thereof.
3. The plant of claim 1, wherein the Teal11A Imi2 nucleic acid
comprises the polynucleotide sequence set forth in SEQ ID NO:3.
4. The plant of claim 1, wherein the Teal11A Imi2 nucleic acid
encodes an IMI polypeptide comprising the amino acid sequence set
forth in SEQ ID NO:4.
5. The plant of claim 1, wherein the plant is non-transgenic.
6. The plant of claim 1, wherein the plant is transgenic.
7. The plant of claim 1, wherein the plant comprises a non-mutated
AHAS gene at its Als1 or Als3 loci.
8. A wheat seed of a wheat plant wherein the wheat plant was
obtained by a process comprising crossing a plant of line Teal11A
with a plant of another wheat line, a representative sample of seed
of the line Teal11A having been deposited with American Type
Culture Collection (ATCC) under Patent Deposit Designation Number
PTA-3953 and the wheat seed comprises a Teal11A Imi2 nucleic acid,
said Teal11A Imi2 nucleic acid comprising a polynucleotide sequence
encoding an IMI polypeptide having a serine to asparagine
substitution in Domain E, said Imi2 nucleic acid being that of the
B genome, and a wheat plant grown from said seed exhibiting
increased tolerance to an imidazolinone herbicide as compared to
that of a wild-type wheat plant.
9. The seed of claim 8, wherein said seed further comprises a seed
treatment.
10. The seed of claim 9, wherein said seed treatment comprises an
herbicidal composition.
11. The seed of claim 10, wherein said herbicidal composition
comprises an imidazolinone herbicide.
12. The seed of claim 11, wherein said imidazolinone herbicide
comprises at least one of: imazethapyr, imazapic, imazamox,
imazaquin, imazethabenz, imazapyr, a mixture of imazapyr and
imazamox, or a combination thereof.
13. The seed of claim 11, wherein the imidazolinone herbicide
comprises imazethapyr.
14. The seed of claim 11, wherein the imidazolinone herbicide
comprises imazamox.
15. A method for treating a seed comprising: a. providing the seed
of claim 8; and b. treating said seed with an herbicidal
composition comprising an imidazolinone herbicide.
16. A method for controlling weeds in the vicinity of a wheat plant
comprising; a. providing the treated seed of claim 11; and b.
growing a wheat plant from said treated seed in a field thereby
controlling weeds in the vicinity of the wheat plant grown from
said treated seed.
17. A method for controlling weeds in the vicinity of a wheat plant
comprising: a. providing a wheat plant, wherein said plant was
obtained by a process comprising crossing a plant of line Teal11A,
a representative sample of seed of the line having been deposited
with American Type Culture Collection (ATCC) under Patent Deposit
Designation Number PTA-3953, with a plant of another wheat line and
the wheat plant comprises a Teal11A Imi2 nucleic acid, said Teal11A
Imi2 nucleic acid comprising a polynucleotide sequence encoding an
IMI polypeptide having a serine to asparagine substitution in
Domain E, said polynucleotide sequence is that of the B genome, and
said plant having increased tolerance to an imidazolinone herbicide
as compared to that of a wild-type wheat; and b. applying an
herbicidal composition comprising an imidazolinone herbicide to the
wheat plant and the vicinity thereof, thereby controlling
weeds.
18. The method of claim 15, wherein said imidazolinone herbicide
comprises at least one of: imazethapyr, imazapic, imazamox,
imazaquin, imazethabenz, imazapyr, a mixture of imazapyr and
imazamox, or a combination thereof.
19. The method of claim 15, wherein the imidazolinone herbicide
comprises imazethapyr.
20. The method of claim 15, wherein the imidazolinone herbicide
comprises imazamox.
21. The method of claim 16, wherein said imidazolinone herbicide
comprises at least one of: imazethapyr, imazapic, imazamox,
imazaquin, imazethabenz, imazapyr, a mixture of imazapyr and
imazamox, or a combination thereof.
22. The method of claim 16, wherein the imidazolinone herbicide
comprises imazethapyr.
23. The method of claim 16, wherein the imidazolinone herbicide
comprises imazamox.
24. The method of claim 17, wherein said imidazolinone herbicide
comprises at least one of: imazethapyr, imazapic, imazamox,
imazaquin, imazethabenz, imazapyr, a mixture of imazapyr and
imazamox, or a combination thereof.
25. The method of claim 17, wherein the imidazolinone herbicide
comprises imazethapyr.
26. The method of claim 17, wherein the imidazolinone herbicide
comprises imazamox.
27. An isolated Imi2 nucleic acid, wherein said Imi2 nucleic acid
is the Imi2 from a wheat plant, or from a seed thereof, the wheat
plant having been obtained by a process comprising crossing a plant
of line Teal11A, a representative sample of seed of the line having
been deposited with American Type Culture Collection (ATCC) under
Patent Deposit Designation Number PTA-3953, with a plant of another
wheat line, wherein the Imi2 nucleic acid of said wheat plant or
seed is a Teal11A Imi2 nucleic acid, said Teal11A Imi2 nucleic acid
comprising a polynucleotide sequence encoding an IMI polypeptide
having a serine to asparagine substitution in Domain E, said
polynucleotide sequence is that of the B genome, and said plant
having increased tolerance to an imidazolinone herbicide as
compared to that of a wild-type wheat.
28. A method for identifying the presence of the nucleic acid
described in claim 1 comprising: a. providing biological material
from a wheat plant or a seed thereof; b. assaying for the presence
of said nucleic acid using a PCR methodology or employing a labeled
probe; and c. identifying, based on the results of step (b), that
the wheat plant or wheat seed of step (a) comprises the nucleic
acid.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application
Ser. No. 12/363,087, filed Jan. 30, 2009, which is now U.S. Pat.
No. 7,521,599, issued on Apr. 21, 2009, which is a divisional of
U.S. patent application Ser. No. 10/486,605, filed Feb. 9, 2004,
which is now U.S. Pat. No. 7,897,845, issued on Mar. 1, 2011, which
is the U.S. National State of International Application
PCT/CA02/01051, filed Jul. 10, 2002, which published in English on
Feb. 20, 2003 and designates the U.S., which claims priority
benefit of U.S. Provisional Patent Application Ser. No. 60/311,282,
filed Aug. 9, 2001; all of which are hereby incorporated herein in
their entirety by reference.
FIELD OF THE INVENTION
[0002] The present invention relates in general to plants having an
increased resistance to imidazolinone herbicides. More
specifically, the present invention relates to wheat plants
obtained by mutagenesis and cross-breeding and transformation that
have an increased resistance to imidazolinone herbicides.
BACKGROUND OF THE INVENTION
[0003] Acetohydroxyacid synthase (AHAS; EC 4.1.3.18) is the first
enzyme that catalyzes the biochemical synthesis of the branched
chain amino acids valine, leucine and isoleucine (Singh B. K., 1999
Biosynthesis of valine, leucine and isoleucine in: Singh B. K. (Ed)
Plant amino acids. Marcel Dekker Inc. New York, N.Y. Pg 227-247).
AHAS is the site of action of four structurally diverse herbicide
families including the sulfonylureas (LaRossa R A and Falco S C,
1984 Trends Biotechnol 2:158-161), the imidazolinones (Shaner et
al., 1984 Plant Physiol 76:545-546), the triazolopyrimidines
(Subramanian and Gerwick, 1989 Inhibition of acetolactate synthase
by triazolopyrimidines in (ed) Whitaker J R, Sonnet P E
Biocatalysis in agricultural biotechnology. ACS Symposium Series,
American Chemical Society. Washington, D.C. Pg 277-288), and the
pyrimidyloxybenzoates (Subramanian et al., 1990 Plant Physiol 94:
239-244). Imidazolinone and sulfonylurea herbicides are widely used
in modem agriculture due to their effectiveness at very low
application rates and relative non-toxicity in animals. By
inhibiting AHAS activity, these families of herbicides prevent
further growth and development of susceptible plants including many
weed species. Several examples of commercially available
imidazolinone herbicides are PURSUIT.RTM. (imazethapyr),
SCEPTER.RTM. (imazaquin) and ARSENAL.RTM. (imazapyr). Examples of
sulfonylurea herbicides are chlorsulfuron, metsulfuron methyl,
sulfometuron methyl, chlorimuron ethyl, thifensulfuron methyl,
tribenuron methyl, bensulfuron methyl, nicosulfuron,
ethametsulfuron methyl, rimsulfuron, triflusulfuron methyl,
triasulfuron, primisulfuron methyl, cinosulfuron, amidosulfuron,
fluzasulfuron, imazosulfuron, pyrazosulfuron ethyl and
halosulfuron.
[0004] Due to their high effectiveness and low-toxicity,
imidazolinone herbicides are favored for application by spraying
over the top of a wide area of vegetation. The ability to spray an
herbicide over the top of a wide range of vegetation decreases the
costs associated with plantation establishment and maintenance and
decreases the need for site preparation prior to use of such
chemicals. Spraying over the top of a desired tolerant species also
results in the ability to achieve maximum yield potential of the
desired species due to the absence of competitive species. However,
the ability to use such spray-over techniques is dependent upon the
presence of imidazolinone resistant species of the desired
vegetation in the spray over area.
[0005] Among the major agricultural crops, some leguminous species
such as soybean are naturally resistant to imidazolinone herbicides
due to their ability to rapidly metabolize the herbicide compounds
(Shaner and Robinson, 1985 Weed Sci. 33:469-471). Other crops such
as corn (Newhouse et al., 1992 Plant Physiol. 100:882-886) and rice
(Barrette et al., 1989 Crop Safeners for Herbicides, Academic Press
New York, pp. 195-220) are somewhat susceptible to imidazolinone
herbicides. The differential sensitivity to the imidazolinone
herbicides is dependent on the chemical nature of the particular
herbicide and differential metabolism of the compound from a toxic
to a non-toxic form in each plant (Shaner et al., 1984 Plant
Physiol. 76:545-546; Brown et al., 1987 Pestic. Biochm. Physiol.
27:24-29). Other plant physiological differences such as absorption
and translocation also play an important role in sensitivity
(Shaner and Robinson, 1985 Weed Sci. 33:469-471).
[0006] Crop cultivars resistant to imidazolinones, sulfonylureas
and triazolopyrimidines have been successfully produced using seed,
microspore, pollen, and callus mutagenesis in Zea mays, Arabidopsis
thaliana, Brassica napus, Glycine max, and Nicotiana tabacum
(Sebastian et al., 1989 Crop Sci. 29:1403-1408; Swanson et al.,
1989 Theor. Appl. Genet. 78:525-530; Newhouse et al., 1991 Theor.
Appl. Genet. 83:65-70; Sathasivan et al., 1991 Plant Physiol.
97:1044-1050; Mourand et al., 1993 J. Heredity 84: 91-96). In all
cases, a single, partially dominant nuclear gene conferred
resistance. Four imidazolinone resistant wheat plants were also
previously isolated following seed mutagenesis of Triticum aestivum
L. cv Fidel (Newhouse et al., 1992 Plant Physiol. 100:882-886).
Inheritance studies confirmed that a single, partially dominant
gene conferred resistance. Based on allelic studies, the authors
concluded that the mutations in the four identified lines were
located at the same locus. One of the Fidel cultivar resistance
genes was designated FS-4 (Newhouse et al., 1992 Plant Physiol.
100:882-886).
[0007] Computer-based modeling of the three dimensional
conformation of the AHAS-inhibitor complex predicts several amino
acids in the proposed inhibitor binding pocket as sites where
induced mutations would likely confer selective resistance to
imidazolinones (Ott et al., 1996 J. Mol. Biol. 263:359-368) Wheat
plants produced with some of these rationally designed mutations in
the proposed binding sites of the AHAS enzyme have in fact
exhibited specific resistance to a single class of herbicides (Ott
et al., 1996 J. Mol. Biol. 263:359-368).
[0008] Plant resistance to imidazolinone herbicides has also been
reported in a number of patents. U.S. Pat. Nos. 4,761,373,
5,331,107, 5,304,732, 6,211,438, 6,211,439 and 6,222,100 generally
describe the use of an altered AHAS gene to elicit herbicide
resistance in plants, and specifically discloses certain
imidazolinone resistant corn lines. U.S. Pat. No. 5,013,659
discloses plants exhibiting herbicide resistance possessing
mutations in at least one amino acid in one or more conserved
regions. The mutations described therein encode either
cross-resistance for imidazolinones and sulfonylureas or
sulfonylurea-specific resistance, but imidazolinone-specific
resistance is not described. Additionally, U.S. Pat. No. 5,731,180
and U.S. Pat. No. 5,767,361 discuss an isolated gene having a
single amino acid substitution in a wild-type monocot AHAS amino
acid sequence that results in imidazolinone-specific
resistance.
[0009] To date, the prior art has not described imidazolinone
resistant wheat plants containing more than one altered AHAS gene.
Nor has the prior art described imidazolinone resistant wheat
plants containing mutations on genomes other than the genome from
which the FS-4 gene is derived. Therefore, what is needed in the
art is the identification of imidazolinone resistance genes from
additional genomes. What are also needed in the art are wheat
plants having increased resistance to herbicides such as
imidazolinone and containing more than one altered AHAS gene. Also
needed are methods for controlling weed growth in the vicinity of
such wheat plants. These compositions and methods would allow for
the use of spray over techniques when applying herbicides to areas
containing wheat plants.
SUMMARY OF THE INVENTION
[0010] The present invention provides wheat plants comprising IMI
nucleic acids, wherein the wheat plant has increased resistance to
an imidazolinone herbicide as compared to a wild-type variety of
the plant. The wheat plants can contain one, two, three or more IMI
nucleic acids. In one embodiment, the wheat plant comprises
multiple IMI nucleic acids located on different genomes.
Preferably, the IMI nucleic acids encode proteins comprising a
mutation in a conserved amino acid sequence selected from the group
consisting of a Domain A, a Domain B, a Domain C, a Domain D and a
Domain E. More preferably, the mutation is in a conserved Domain E
or a conserved Domain C. Also provided are plant parts and plant
seeds derived from the wheat plants described herein. In another
embodiment, the wheat plant comprises an IMI nucleic acid that is
not an Imi1 nucleic acid. The IMI nucleic acid can be an Imi2 or
Imi3 nucleic acid, for example.
[0011] The IMI nucleic acids of the present invention can comprise
a nucleotide sequence selected from the group consisting of: a
polynucleotide of SEQ ID NO:1; a polynucleotide of SEQ ID NO:3; a
polynucleotide sequence encoding a polypeptide of SEQ ID NO:2; a
polynucleotide sequence encoding a polypeptide of SEQ ID NO:4, a
polynucleotide comprising at least 60 consecutive nucleotides of
any of the aforementioned polynucleotides; and a polynucleotide
complementary to any of the aforementioned polynucleotides.
[0012] The plants of the present invention can be transgenic or
non-transgenic. Examples of non-transgenic wheat plants having
increased resistance to imidazolinone herbicides include a wheat
plant having an ATCC Patent Deposit Designation Number PTA-3953 or
PTA-3955; or a mutant, recombinant, or genetically engineered
derivative of the plant with ATCC Patent Deposit Designation Number
PTA-3953 or PTA-3955; or of any progeny of the plant with ATCC
Patent Deposit Designation Number PTA-3953 or PTA-3955; or a plant
that is a progeny of any of these plants.
[0013] In addition to the compositions of the present invention,
several methods are provided. Described herein are methods of
modifying a plant's tolerance to an imidazolinone herbicide
comprising modifying the expression of an IMI nucleic acid in the
plant. Also described are methods of producing a transgenic plant
having increased tolerance to an imidazolinone herbicide
comprising, transforming a plant cell with an expression vector
comprising one or more IMI nucleic acids and generating the plant
from the plant cell. The invention further includes a method of
controlling weeds within the vicinity of a wheat plant, comprising
applying an imidazolinone herbicide to the weeds and to the wheat
plant, wherein the wheat plant has increased resistance to the
imidazolinone herbicide as compared to a wild type variety of the
wheat plant and wherein the plant comprises one or more IMI nucleic
acids. In some preferred embodiments of these methods, the plants
comprise multiple IMI nucleic acids that are located on different
wheat genomes.
BRIEF DESCRIPTION OF THE DRAWINGS
[0014] FIG. 1 is a table showing the results of single plant
evaluation of imazamox resistance in parental and F.sub.1
populations resulting from reciprocal crosses between resistant
lines and CDC Teal. The numbers presented represent the number of
plants scored into each phenotypic class. Parental lines are
indicated in bold. The number of parental lines scored include
those scored with the F.sub.2 populations.
[0015] FIG. 2 is a table showing the reaction to imazamox in
F.sub.2 and BCF.sub.1 populations resulting from crosses between
resistant lines and CDC Teal and Chi-square tests of single locus
and two locus models (15A.times.Teal) for control of resistance.
The symbols used in FIG. 2 indicate the following: a--Chi-square P
value (1 df) represents the probability that deviations from the
tested ratio are due to chance alone. Chi-square P values greater
than 0.05 indicate that observed values were not significantly
different from expected values; b--Chi-square P value representing
the probability that deviations between F.sub.2 populations
resulting from reciprocal crosses between CDC Teal and resistant
lines are due to chance alone. Chi-square values greater than 0.05
indicate that reciprocal F.sub.2 populations were homogeneous, and
data from the two reciprocal populations was pooled; c--CDC Teal
was used as the recurrent parent; d--Ratios tested were based on
the results of the F.sub.2 generation; and e--Chi-square P value (1
df) for BCF.sub.1 ratio.
[0016] FIG. 3 is a table showing the results of an evaluation of
resistance to imazamox in F.sub.2:3 families resulting from crosses
between resistant lines and CDC Teal and Chi-square tests of
single-locus and two locus models (15A.times.Teal) for control of
resistance. The symbols used in FIG. 3 indicate the following:
a--Family segregation ratios tested were based on the results of
the F.sub.2 and BCF.sub.1 populations; b--Chi-square P value (2 df)
representing the probability that deviations from the tested ratio
are due to chance alone. Chi-square P values greater than 0.05
indicate that observed values were not significantly different from
expected values.
[0017] FIG. 4 is a table showing the results of a single plant
evaluation of imazamox resistance in F.sub.2 populations resulting
from inter-crosses between resistant lines. Chi-square ratios
tested were based on the results of the F.sub.2 and F.sub.2:3
family results obtained from crosses between resistant lines and
CDC Teal. The 15:1 ratio tested is for a two locus model and the
63:1 ratio tested is for a three locus model. The "a" symbol used
in FIG. 4 indicates the following: Chi-square P value (1 df)
representing the probability that deviations from the tested ratio
are due to chance alone. Chi-square P values greater than 0.05
indicate that observed values were not significantly different from
expected values.
[0018] FIG. 5 is a table showing the results of an evaluation of
imazamox resistance in F.sub.2:3 families resulting from
segregating inter-crosses between resistant lines. The symbols used
in FIG. 5 indicate the following: a--Family segregation ratios
tested were based on the results of the F.sub.2 populations
examined; b--Chi-square P value (2 df) representing the probability
that deviations from the tested ratio are due to chance alone.
Chi-square P values greater than 0.05 indicate that observed values
were not significantly different from expected values.
[0019] FIG. 6 is a table comparing the percent uninhibited in vitro
AHAS activity in four wheat lines in the presence of increasing
concentrations of the imidazolinone herbicide imazamox. Teal is a
wild type line with no tolerance to imidazolinone herbicides while
BW755 contains the FS4 mutant gene.
[0020] FIG. 7 is a table comparing injury sustained by three wheat
genotypes when treated with either a 10.times. or 30.times. rate of
imazamox. The 1.times. rate is 20 g/ha. BW755 contains the FS4
mutant gene. 15A/11A is a bulk of selfed progeny from the cross of
Teal11A and Teal15A. The population was not yet homozygous at all
three non-allelic loci.
[0021] FIG. 8 shows a DNA sequence alignment of partial Als1 and
Imi1 wheat genes amplified from genomic DNA: CDC Teal (row 2; SEQ
ID NO:15 and SEQ ID NO:16), BW755 (row 3; SEQ ID NO:17 and SEQ ID
NO:18), TealIMI 10A (row 4; SEQ ID NO:19 and SEQ ID NO:20), TealIMI
11A (row 5; SEQ ID NO:21 and SEQ ID NO:22), and TealIMI 15A (row 6;
SEQ ID NO:23 and SEQ ID NO:24). Partial sequences were aligned with
a complete rice ALS gene sequence (row 1; SEQ ID NO:13 and SEQ ID
NO:14) derived from Genbank (Accession no. AB049822) and translated
to protein sequences (presented on top of the DNA sequences). The
five highly conserved amino acid domains known to house mutations
that confer resistance to AHAS inhibitors are indicated in bold.
Note the guanine to adenine substitutions in BW755, TealIMI 10A,
and TealIMI 15A result in a serine to asparagine substitution
(serine-627 in rice) in the IPSGG domain (Domain E) of the Als1
gene. Accordingly, the resistance genes present in the BW755,
TealIMI 10A, and TealIMI 15A plants have been designated as part of
the Imi1 class. These Teal resistance genes are referred to herein
as TealIMI 10A and TealIMI 15A.
[0022] FIG. 9 shows a DNA sequence alignment of partial Als2 and
Imi2 wheat genes amplified from genomic DNA: CDC Teal (row 2; SEQ
ID NO:25 and SEQ ID NO:26), BW755 (row 3; SEQ ID NO:27 and SEQ ID
NO:28), TealIMI 10A (row 4; SEQ ID NO:29 and SEQ ID NO:30), TealIMI
11A (row 5; SEQ ID NO:31 and SEQ ID NO:32) and TealIMI 15A (row 6;
SEQ ID NO:33 and SEQ ID NO:34). Partial. AHAS sequences were
aligned with a complete rice AHAS sequence (row 1; SEQ ID NO:13 and
SEQ ID NO:14) derived from GenBank (Accession no. AB049822) and
translated into protein sequences (presented above the DNA
sequences). The five highly conserved domains known to house
mutations that confer resistance to AHAS inhibitors are indicated
in bold. Note the guanine to adenine substitution in TealIMI 11A,
resulting in a serine to asparagine substitution (serine-627 in
rice) in the IPSGG domain of the Als2 gene. Accordingly, the
resistance gene present in TealIMI 11A plant has been designated as
part of the Imi2 class of nucleic acids. This Teal resistance gene
is referred to herein as TealIMI2 11A.
[0023] FIG. 10 shows the partial DNA sequence of TealIMI 15A (SEQ
ID NO:1) and the deduced amino acid sequence of the same (SEQ ID
NO:2).
[0024] FIG. 11 shows the partial DNA sequence of TealIMI2 11A (SEQ
ID NO:3) and the deduced amino acid sequence of the same (SEQ ID
NO:4).
[0025] FIG. 12 shows the wild type nucleic acid sequence of the
Teal ALS1 ORF (SEQ ID NO:5), the Teal ALS2 ORF (SEQ ID NO:6) the
Teal ALS3 ORF (SEQ ID NO:7).
[0026] FIG. 13 is a schematic representation of the conserved amino
acid sequences in the AHAS genes implicated in resistance to
various AHAS inhibitors. The specific amino acid site responsible
for resistance is indicated by an underline. (Modified from Devine,
M. D. and Eberlein, C. V., 1997 Physiological, biochemical and
molecular aspects of herbicide resistance based on altered target
sites in Herbicide Activity Toxicity, Biochemistry, and Molecular
Biology, IOS Press Amsterdam, p. 159-185).
DETAILED DESCRIPTION
[0027] The present invention is directed to wheat plants, wheat
plant parts and wheat plant cells having increased resistance to
imidazolinone herbicides. The present invention also includes seeds
produced by the wheat plants described herein and methods for
controlling weeds in the vicinity of the wheat plants described
herein. It is to be understood that as used in the specification
and in the claims, "a" or "an" can mean one or more, depending upon
the context in which it is used. Thus, for example, reference to "a
cell" can mean that at least one cell can be utilized.
[0028] As used herein, the term "wheat plant" refers to a plant
that is a member of the Triticum genus. The wheat plants of the
present invention can be members of a Triticum genus including, but
not limited to, T. aestivum, T. turgidum, T. timopheevii, T.
monococcum, T. zhukovskyi and T. urartu and hybrids thereof.
Examples of T. aestivum subspecies included within the present
invention are aestivum (common wheat), compactum (club wheat),
macha (macha wheat), vavilovi (vavilovi wheat), spelta and
sphaecrococcum (shot wheat). Examples of T. turgidum subspecies
included within the present invention are turgidum, carthlicum,
dicoccon, durum, paleocolchicum, polonicum, turanicun and
dicoccoides. Examples of T. monococcum subspecies included within
the present invention are monococcum (einkorn) and aegilopoides. In
one embodiment of the present invention, the wheat plant is a
member of the Triticum aestivum species, and more particularly, the
CDC Teal cultivar.
[0029] The term "wheat plant" is intended to encompass wheat plants
at any stage of maturity or development as well as any tissues or
organs (plant parts) taken or derived from any such plant unless
otherwise clearly indicated by context. Plant parts include, but
are not limited to, stems, roots, flowers, ovules, stamens, leaves,
embryos, meristematic regions, callus tissue, anther cultures,
gametophytes, sporophytes, pollen, microspores, protoplasts and the
like. The present invention also includes seeds produced by the
wheat plants of the present invention. In one embodiment, the seeds
are true breeding for an increased resistance to an imidazolinone
herbicide as compared to a wild type variety of the wheat plant
seed.
[0030] The present invention describes a wheat plant comprising one
or more IMI nucleic acids, wherein the wheat plant has increased
resistance to an imidazolinone herbicide as compared to a wild-type
variety of the plant. As used herein, the term "IMI nucleic acid"
refers to a nucleic acid that is mutated from an AHAS nucleic acid
in a wild type wheat plant that confers increased imidazolinone
resistance to a plant in which it is transcribed. Wild type Teal
AHAS nucleic acids are shown in SEQ ID NO:5 (Teal ALS1 ORF), SEQ ID
NO:6 (Teal ALS2 ORF) and SEQ ID NO:7 (Teal ALS3 ORF). In one
embodiment, the wheat plant comprises multiple IMI nucleic acids.
As used when describing the IMI nucleic acids, the term "multiple"
refers to IMI nucleic acids that have different nucleotide
sequences and does not refer to a mere increase in number of the
same IMI nucleic acid. For example, the IMI nucleic acids can be
different due to the fact that they are derived from or located on
different wheat genomes.
[0031] It is possible for the wheat plants of the present invention
to have multiple IMI nucleic acids from different genomes since
these plants can contain more than one genome. For example, a
Triticum aestivum wheat plant contains three genomes sometimes
referred to as the A, B and D genomes. Because AHAS is a required
metabolic enzyme, it is assumed that each genome has at least one
gene coding for the AHAS enzyme, commonly seen with other metabolic
enzymes in hexaploid wheat that have been mapped. The AHAS nucleic
acid on each genome can, and usually does, differ in its nucleotide
sequence from an AHAS nucleic acid on another genome. One of skill
in the art can determine the genome of origin of each AHAS nucleic
acid through genetic crossing and/or either sequencing methods or
exonuclease digestion methods known to those of skill in the art
and as also described in Example 2 below. For the purposes of this
invention, IMI nucleic acids derived from one of the A, B or D
genomes are distinguished and designated as Imi1, Imi2 or Imi3
nucleic acids.
[0032] It is not stated herein that any particular Imi nucleic acid
class correlates with any particular A, B or D genome. For example,
it is not stated herein that the Imi1 nucleic acids correlate to A
genome nucleic acids, that Imi2 nucleic acids correlate to B genome
nucleic acids, etc. The Imi1, Imi2 and Imi3 designations merely
indicate that the IMI nucleic acids within each such class do not
segregate independently, whereas two IMI nucleic acids from
different classes do segregate independently and may therefore be
derived from different wheat genomes. The Imi1 class of nucleic
acids includes the FS-4 gene as described by Newhouse et al. (1992
Plant Physiol. 100:882-886) and the TealIMI 15A gene described in
more detail below. The Imi2 class of nucleic acids includes the
TealIMI2 11A gene described below. Each Imi class can include
members from different wheat species. Therefore, each imi class
includes IMI nucleic acids that differ in their nucleotide sequence
but that are nevertheless designated as originating from, or being
located on, the same wheat genome using inheritance studies as
described in the Examples below and known to those of ordinary
skill in the art.
[0033] Accordingly, the present invention includes a wheat plant
comprising one or more IMI nucleic acids, wherein the wheat plant
has increased resistance to an imidazolinone herbicide as compared
to a wild-type variety of the plant and wherein the one or more IMI
nucleic acids are selected from a group consisting of an Imi1, Imi2
and Imi3 nucleic acid. In one embodiment, the plant comprises an
Imi1 nucleic and an Imi3 nucleic acid. In a preferred embodiment,
the Imi1 nucleic acid comprises the polynucleotide sequence shown
in SEQ ID NO:1. In another embodiment, the plant comprises an Imi2
nucleic acid. In a preferred embodiment, the Imi2 nucleic acid
comprises the polynucleotide sequence shown in SEQ ID NO:3.
[0034] As used herein with regard to nucleic acids, the term "from"
refers to a nucleic acid "located on" or "derived from" a
particular genome. The term "located on" refers to a nucleic acid
contained within that particular genome. As also used herein with
regard to a genome, the term "derived from" refers to a nucleic
acid that has been removed or isolated from that genome. The term
"isolated" is defined in more detail below.
[0035] In another embodiment, the wheat plant comprises an IMI
nucleic acid, wherein the nucleic acid is a non-Imi1 nucleic acid.
The term "non-Imi1", refers to an IMI nucleic acid that is not a
member of the Imi1 class as described above. Examples of nucleic
acids from the Imi1 class are shown in rows 3, 4 and 5 of FIG. 8.
One example of non-Imi1 nucleic acid is shown in row 5 of FIG. 8.
Accordingly, in a preferred embodiment, the wheat plant comprises
an IMI nucleic acid comprising the polynucleotide sequence encoding
the polypeptide of SEQ ID NO:4. The polynucleotide sequence can
comprise the sequence shown in SEQ ID NO:3.
[0036] The present invention includes wheat plants comprising one,
two, three or more IMI nucleic acids, wherein the wheat plant has
increased resistance to an imidazolinone herbicide as compared to a
wild-type variety of the plant. The IMI nucleic acids can comprise
a nucleotide sequence selected from the group consisting of a
polynucleotide of SEQ ID NO:1; a polynucleotide of SEQ ID NO:3; a
polynucleotide sequence encoding a polypeptide of SEQ ID NO:2; a
polynucleotide sequence encoding a polypeptide of SEQ ID NO:4, a
polynucleotide comprising at least 60 consecutive nucleotides of
any of the aforementioned polynucleotides; and a polynucleotide
complementary to any of the aforementioned polynucleotides.
[0037] The imidazolinone herbicide can be selected from, but is not
limited to, PURSUIT.RTM. (imazethapyr), CADRE.RTM. (imazapic),
RAPTOR.RTM. (imazamox), SCEPTER.RTM. (imazaquin), ASSERT.RTM.
(imazethabenz), ARSENAL.RTM. (imazapyr), a derivative of any of the
aforementioned herbicides, or a mixture of two or more of the
aforementioned herbicides, for example, imazapyr/imazamox
(ODYSSEY.RTM.). More specifically, the imidazolinone herbicide can
be selected from, but is not limited to,
2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-nicotinic acid,
2-(4-isopropyl)-4-methyl-5-oxo-2-imidazolin-2-yl)-3-quinolinecarboxylic
acid,
5-ethyl-2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-nicotinic
acid,
2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-5-(methoxymethyl)--
nicotinic acid,
2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-5-methylnicotinic
acid, and a mixture of methyl
6-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-m-toluate and
methyl 2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-p-toluate.
The use of
5-ethyl-2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-nicotinic
acid and
2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-5-(methoxymethyl)-ni-
cotinic acid is preferred. The use of
2-(4-isopropyl-4-methyl-5-oxo-2-imidazolin-2-yl)-5-(methoxymethyl)-nicoti-
nic acid is particularly preferred.
[0038] In one embodiment, the wheat plant comprises two IMI nucleic
acids, wherein the nucleic acids are derived from or located on
different wheat genomes. Preferably, one of the two nucleic acids
is an Imi1 nucleic acid, and more preferably comprises the
polynucleotide sequence of SEQ ID NO: 1. In another embodiment, the
wheat plant comprises one IMI nucleic acid, wherein the nucleic
acid comprises the polynucleotide sequence of SEQ ID NO:1 or SEQ ID
NO:3. In yet another embodiment, the wheat plant comprises three or
more IMI nucleic acids wherein each nucleic acid is from a
different genome. Preferably, at least one of the three IMI nucleic
acids comprises a polynucleotide sequence selected from the group
consisting of SEQ ID NO:1 and SEQ ID NO:3.
[0039] In a preferred embodiment of the present invention, the one
or more IMI nucleic acids contained within the plant encode an
amino acid sequence comprising a mutation in a domain that is
conserved among several AHAS proteins. These conserved domains are
referred to herein as Domain A, Domain B, Domain C, Domain D and
Domain E. FIG. 13 shows the general location of each domain in an
AHAS protein. As used herein, Domain A contains the amino acid
sequence AITGQVPRRMIGT (SEQ ID NO:8); Domain B contains the amino
acid sequence QWED (SEQ ID NO:9); Domain C contains the amino acid
sequence VFAYPGGASMEIHQALTRS (SEQ ID NO:10); Domain D contains the
amino acid sequence AFQETP (SEQ ID NO:11); Domain E contains the
amino acid sequence IPSGG (SEQ ID NO:12). The present invention
also contemplates that there may be slight variations in the
conserved domains, for example, in cockleberry plants, the serine
residue in Domain E is replaced by an alanine residue.
[0040] Accordingly, the present invention includes a wheat plant
comprising an IMI nucleic acid that encodes an amino acid sequence
having a mutation in a conserved domain selected from the group
consisting of a Domain A, a Domain B, a Domain C, a Domain D and a
Domain E. In one embodiment, the wheat plant comprises an IMI
nucleic acid that encodes an amino acid sequence having a mutation
in a Domain E. In further preferred embodiments, the mutations in
the conserved domains occur at the locations indicated by the
following underlining: AITGQVPRRMIGT (SEQ ID NO:8); QWED (SEQ ID
NO:9); VFAYPGGASMEIHQALTRS (SEQ ID NO:10); AFQETP (SEQ ID NO:11)
and IPSGG (SEQ ID NO:12). One preferred substitution is asparagine
for serine in Domain E (SEQ ID NO:12).
[0041] The wheat plants described herein can be either transgenic
wheat plants or non-transgenic wheat plants. As used herein, the
term "transgenic" refers to any plant, plant cell, callus, plant
tissue or plant part, that contains all or part of at least one
recombinant polynucleotide. In many cases, all or part of the
recombinant polynucleotide is stably integrated into a chromosome
or stable extra-chromosomal element, so that it is passed on to
successive generations. For the purposes of the invention, the term
"recombinant polynucleotide" refers to a polynucleotide that has
been altered, rearranged or modified by genetic engineering.
Examples include any cloned polynucleotide, or polynucleotides,
that are linked or joined to heterologous sequences. The term
"recombinant" does not refer to alterations of polynucleotides that
result from naturally occurring events, such as spontaneous
mutations, or from non-spontaneous mutagenesis followed by
selective breeding. Plants containing mutations arising due to
non-spontaneous mutagenesis and selective breeding are referred to
herein as non-transgenic plants and are included in the present
invention. In embodiments wherein the wheat plant is transgenic and
comprises multiple IMI nucleic acids, the nucleic acids can be
derived from different genomes or the same genome. Alternatively,
in embodiments wherein the wheat plant is non-transgenic and
comprises multiple IMI nucleic acids, the nucleic acids are located
on different genomes.
[0042] An example of a non-transgenic wheat plant cultivar
comprising one IMI nucleic acid is the plant cultivar deposited
with the ATCC under Patent Deposit Designation Number PTA-3953 and
designated herein as the TealIMI 11A wheat cultivar. The TealIMI
11A wheat cultivar contains an Imi2 nucleic acid. The partial
nucleotide and deduced amino acid sequences corresponding to the
TealIMI2 11A gene are shown in SEQ ID NO:3 and SEQ ID NO:4,
respectively. The only portion of the sequences not included in SEQ
ID NO:3 and SEQ ID NO:4 are those sequences encoding and
corresponding to a signal sequence that is cleaved from the mature
TealIMI2 11A protein. Accordingly, SEQ ID NO:4 represents the full
deduced sequence of the mature TealIMI2 11A protein.
[0043] An example of a wheat plant cultivar comprising two IMI
nucleic acids on different genomes is the plant cultivar deposited
with the ATCC under Patent Deposit Designation Number PTA-3955 and
designated herein as the TealIMI 15A wheat cultivar. The TealIMI
15A wheat cultivar contains Imi1 and Imi3 nucleic acids. The Imi1
nucleic acid comprises a mutation that results in a serine to
asparagine change in the IMI protein encoded thereby. The mutated
AHAS genes are designated herein as TealIMI 15A and TealIMI3 15A.
The partial nucleotide and deduced amino acid sequences
corresponding to the TealIMI 15A gene are shown in SEQ ID NO:1 and
SEQ ID NO:2, respectively. The only portion of the sequences not
included in SEQ ID NO:1 and SEQ ID NO:2 are those sequences
encoding and corresponding to approximately 100-150 base pairs at
the 5' end and approximately 5 base pairs at the 3' end of the
coding region.
[0044] Separate deposits of 2500 seeds of the TealIMI 11A and
TealIMI 15A wheat cultivars were made with the American Type
Culture Collection, Manassas, Va. on Jan. 3, 2002. These deposits
were made in accordance with the terms and provisions of the
Budapest Treaty relating to the deposit of microorganisms. The
deposits were made for a term of at least thirty years and at least
five years after the most recent request for the furnishing of a
sample of the deposit is received by the ATCC. The deposited seeds
were accorded Patent Deposit Designation Numbers PTA-3953 (TealIMI
11A) and PTA-3955 (TealIMI 15A).
[0045] The present invention includes the wheat plant having a
Patent Deposit Designation Number PTA-3953 or PTA-3955; a mutant,
recombinant, or genetically engineered derivative of the plant with
Patent Deposit Designation Number PTA-3953 or PTA-3955; any progeny
of the plant with Patent Deposit Designation Number PTA-3953 or
PTA-3955; and a plant that is the progeny of any of these plants.
In a preferred embodiment, the wheat plant of the present invention
additionally has the herbicide resistance characteristics of the
plant with Patent Deposit Designation Number PTA-3953 or
PTA-3955.
[0046] Also included in the present invention are hybrids of the
TealIMI 11A and TealIMI 15A wheat cultivars described herein.
Example 5 demonstrates TealIMI11 .ANG./TealIMI15A hybrids having
increased resistance to an imidazolinone herbicide. The present
invention further includes hybrids of the TealIMI 11 .ANG. or
TealIMI 15A wheat cultivars and another wheat cultivar. The other
wheat cultivar includes, but is not limited to, T. aestivum L. cv
Fidel and any wheat cultivar harboring a mutant gene FS-1, FS-2,
FS-3 or FS-4. (See U.S. Pat. No. 6,339,184 and U.S. patent
application Ser. No. 08/474,832). In a preferred embodiment, the
wheat plant is a hybrid between a TealIMI 11A cultivar and a Fidel
FS-4 cultivar. The TealIMI 11A/FS-4 hybrids comprise an Imi1
nucleic acid and an Imi2 nucleic acid. A hybrid of TealIMI 11A and
a Fidel cultivar harboring the FS-4 gene is included in the present
invention and was deposited with the American Type Culture
Collection, Manassas, Va. on Jan. 3, 2002. This deposit was made in
accordance with the terms and provisions of the Budapest Treaty
relating to the deposit of microorganisms. The deposit was made for
a term of at least thirty years and at least five years after the
most recent request for the furnishing of a sample of the deposit
is received by the ATCC. The deposited seeds were accorded Patent
Deposit Designation Number PTA-3954.
[0047] The terms "cultivar" and "variety" refer to a group of
plants within a species defined by the sharing of a common set of
characteristics or traits accepted by those skilled in the art as
sufficient to distinguish one cultivar or variety from another
cultivar or variety. There is no implication in either term that
all plants of any given cultivar or variety will be genetically
identical at either the whole gene or molecular level or that any
given plant will be homozygous at all loci. A cultivar or variety
is considered "true breeding" for a particular trait if, when the
true-breeding cultivar or variety is self-pollinated, all of the
progeny contain the trait. In the present invention, the trait
arises from a mutation in an AHAS gene of the wheat plant or
seed.
[0048] It is to be understood that the wheat plant of the present
invention can comprise a wild type or non-mutated AHAS gene in
addition to an IMI gene. As described in Example 4, it is
contemplated that wheat cultivar TealIMI 11A contains a mutation in
only one of multiple AHAS isoenzymes and that wheat cultivar
TealIMI 15A contains a mutation in only two of multiple AHAS
isoenzymes. Therefore, the present invention includes a wheat plant
comprising one or more IMI nucleic acids in addition to one or more
wild type or non-mutated AHAS nucleic acids.
[0049] In addition to wheat plants, the present invention
encompasses isolated IMI proteins and nucleic acids. The nucleic
acids comprise a polynucleotide selected from the group consisting
of a polynucleotide of SEQ ID NO:1; a polynucleotide of SEQ ID
NO:3; a polynucleotide sequence encoding a polypeptide of SEQ ID
NO:2; a polynucleotide sequence encoding a polypeptide of SEQ ID
NO:4, a polynucleotide comprising at least 60 consecutive
nucleotides of any of the aforementioned polynucleotides; and a
polynucleotide complementary to any of the aforementioned
polynucleotides. In a preferred embodiment, the IMI nucleic acid
comprises a polynucleotide sequence encoding a polypeptide of SEQ
ID NO:2 or SEQ ID NO:4. In a further preferred embodiment, the IMI
nucleic acid comprises a polynucleotide sequence of SEQ ID NO:1 or
SEQ ID NO:3.
[0050] The term "AHAS protein" refers to an acetohydroxyacid
synthase protein and the term "IMI protein" refers to any AHAS
protein that is mutated from a wild type AHAS protein and that
confers increased imidazolinone resistance to a plant, plant cell,
plant part, plant seed or plant tissue when it is expressed
therein. In a preferred embodiment, the IMI protein comprises a
polypeptide of SEQ ID NO:2 or SEQ ID NO:4. As also used herein, the
terms "nucleic acid" and "polynucleotide" refer to RNA or DNA that
is linear or branched, single or double stranded, or a hybrid
thereof. The term also encompasses RNA/DNA hybrids. These terms
also encompass untranslated sequence located at both the 3' and 5'
ends of the coding region of the gene: at least about 1000
nucleotides of sequence upstream from the 5' end of the coding
region and at least about 200 nucleotides of sequence downstream
from the 3' end of the coding region of the gene. Less common
bases, such as inosine, 5-methylcytosine, 6-methyladenine,
hypoxanthine and others can also be used for antisense, dsRNA and
ribozyme pairing. For example, polynucleotides that contain C-5
propyne analogues of uridine and cytidine have been shown to bind
RNA with high affinity and to be potent antisense inhibitors of
gene expression. Other modifications, such as modification to the
phosphodiester backbone, or the 2'-hydroxy in the ribose sugar
group of the RNA can also be made. The antisense polynucleotides
and ribozymes can consist entirely of ribonucleotides, or can
contain mixed ribonucleotides and deoxyribonucleotides. The
polynucleotides of the invention may be produced by any means,
including genomic preparations, cDNA preparations, in vitro
synthesis, RT-PCR and in vitro or in vivo transcription.
[0051] An "isolated" nucleic acid molecule is one that is
substantially separated from other nucleic acid molecules, which
are present in the natural source of the nucleic acid (i.e.,
sequences encoding other polypeptides). Preferably, an "isolated"
nucleic acid is free of some of the sequences that naturally flank
the nucleic acid (i.e., sequences located at the 5' and 3' ends of
the nucleic acid) in its naturally occurring replicon. For example,
a cloned nucleic acid is considered isolated. In various
embodiments, the isolated IMI nucleic acid molecule can contain
less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of
nucleotide sequences which naturally flank the nucleic acid
molecule in genomic DNA of the cell from which the nucleic acid is
derived (e.g., a Triticum aestivum cell). A nucleic acid is also
considered isolated if it has been altered by human intervention,
or placed in a locus or location that is not its natural site, or
if it is introduced into a cell by agroinfection or biolistics.
Moreover, an "isolated" nucleic acid molecule, such as a cDNA
molecule, can be free from some of the other cellular material with
which it is naturally associated, or culture medium when produced
by recombinant techniques, or chemical precursors or other
chemicals when chemically synthesized.
[0052] Specifically excluded from the definition of "isolated
nucleic acids" are: naturally-occurring chromosomes (such as
chromosome spreads), artificial chromosome libraries, genomic
libraries, and cDNA libraries that exist either as an in vitro
nucleic acid preparation or as a transfected/transformed host cell
preparation, wherein the host cells are either an in vitro
heterogeneous preparation or plated as a heterogeneous population
of single colonies. Also specifically excluded are the above
libraries wherein a specified nucleic acid makes up less than 5% of
the number of nucleic acid inserts in the vector molecules. Further
specifically excluded are whole cell genomic DNA or whole cell RNA
preparations (including whole cell preparations that are
mechanically sheared or enzymatically digested). Even further
specifically excluded are the whole cell preparations found as
either an in vitro preparation or as a heterogeneous mixture
separated by electrophoresis wherein the nucleic acid of the
invention has not further been separated from the heterologous
nucleic acids in the electrophoresis medium (e.g., further
separating by excising a single band from a heterogeneous band
population in an agarose gel or nylon blot).
[0053] A nucleic acid molecule of the present invention, e.g., a
nucleic acid molecule containing a nucleotide sequence of SEQ ID
NO:1, SEQ ID NO:3 or a portion thereof, can be isolated using
standard molecular biology techniques and the sequence information
provided herein. For example, a T. aestivum IMI cDNA can be
isolated from a T. aestivum library using all or a portion of the
sequence of SEQ ID NO:1 or SEQ ID NO:3. Moreover, a nucleic acid
molecule encompassing all or a portion of SEQ ID NO:1 or SEQ ID
NO:3 can be isolated by the polymerase chain reaction using
oligonucleotide primers designed based upon this sequence. For
example, mRNA can be isolated from plant cells (e.g., by the
guanidinium-thiocyanate extraction procedure of Chirgwin et al.,
1979 Biochemistry 18:5294-5299) and cDNA can be prepared using
reverse transcriptase (e.g., Moloney MLV reverse transcriptase,
available from Gibco/BRL, Bethesda, Md.; or AMV reverse
transcriptase, available from Seikagaku America, Inc., St.
Petersburg, Fla.). Synthetic oligonucleotide primers for polymerase
chain reaction amplification can be designed based upon the
nucleotide sequence shown in SEQ ID NO:1 or SEQ ID NO:3. A nucleic
acid molecule of the invention can be amplified using cDNA or,
alternatively, genomic DNA, as a template and appropriate
oligonucleotide primers according to standard PCR amplification
techniques. The nucleic acid molecule so amplified can be cloned
into an appropriate vector and characterized by DNA sequence
analysis. Furthermore, oligonucleotides corresponding to an IMI
nucleotide sequence can be prepared by standard synthetic
techniques, e.g., using an automated DNA synthesizer.
[0054] The IMI nucleic acids of the present invention can comprise
sequences encoding an IMI protein (i.e., "coding regions"), as well
as 5' untranslated sequences and 3' untranslated sequences.
Alternatively, the nucleic acid molecules of the present invention
can comprise only the coding regions of an IMI gene, or can contain
whole genomic fragments isolated from genomic DNA. A coding region
of these sequences is indicated as an "ORF position". Moreover, the
nucleic acid molecule of the invention can comprise a portion of a
coding region of an IMI gene, for example, a fragment that can be
used as a probe or primer. The nucleotide sequences determined from
the cloning of the IMI genes from T. aestivum allow for the
generation of probes and primers designed for use in identifying
and/or cloning IMI homologs in other cell types and organisms, as
well as IMI homologs from other wheat plants and related species.
The portion of the coding region can also encode a biologically
active fragment of an IMI protein.
[0055] As used herein, the term "biologically active portion of" an
IMI protein is intended to include a portion, e.g., a domain/motif,
of an IMI protein that, when produced in a plant increases the
plant's resistance to an imidazolinone herbicide as compared to a
wild-type variety of the plant. Methods for quantitating increased
resistance to imidazolinone herbicides are provided in the Examples
provided below. Biologically active portions of an IMI protein
include peptides encoded by polynucleotide sequences comprising SEQ
ID NO:1 or SEQ ID NO:3 which include fewer amino acids than a full
length IMI protein and impart increased resistance to an
imidazolinone herbicide upon expression in a plant. Typically,
biologically active portions (e.g., peptides which are, for
example, 5, 10, 15, 20, 30, 35, 36, 37, 38, 39, 40, 50, 100 or more
amino acids in length) comprise a domain or motif with at least one
activity of an IMI protein. Moreover, other biologically active
portions in which other regions of the polypeptide are deleted, can
be prepared by recombinant techniques and evaluated for one or more
of the activities described herein. Preferably, the biologically
active portions of an IMI protein include one or more conserved
domains selected from the group consisting of a Domain A, a Domain
B, a Domain C, a Domain D and a Domain E, wherein the conserved
domain contains a mutation.
[0056] The invention also provides IMI chimeric or fusion
polypeptides. As used herein, an IMI "chimeric polypeptide" or
"fusion polypeptide" comprises an IMI polypeptide operatively
linked to a non-IMI polypeptide. A "non-IMI polypeptide" refers to
a polypeptide having an amino acid sequence that is not
substantially identical to an IMI polypeptide, e.g., a polypeptide
that is not an IMI isoenzyme, which peptide performs a different
function than an IMI polypeptide. Within the fusion polypeptide,
the term "operatively linked" is intended to indicate that the IMI
polypeptide and the non-IMI polypeptide are fused to each other so
that both sequences fulfill the proposed function attributed to the
sequence used. The non-IMI polyp eptide can be fused to the
N-terminus or C-terminus of the IMI polypeptide. For example, in
one embodiment, the fusion polypeptide is a GST-IMI fusion
polypeptide in which the IMI sequence is fused to the C-terminus of
the GST sequence. Such fusion polypeptides can facilitate the
purification of recombinant IMI polypeptides. In another
embodiment, the fusion polypeptide is an IMI polypeptide containing
a heterologous signal sequence at its N-terminus. In certain host
cells (e.g., mammalian host cells), expression and/or secretion of
an IMI polypeptide can be increased through use of a heterologous
signal sequence.
[0057] An isolated nucleic acid molecule encoding an IMI
polypeptide having sequence identity to a polypeptide encoded by a
polynucleotide sequence of SEQ ID NO:1 or SEQ ID NO:3 can be
created by introducing one or more nucleotide substitutions,
additions or deletions into a nucleotide sequence of SEQ ID NO:1 or
SEQ ID NO:3 such that one or more amino acid substitutions,
additions or deletions are introduced into the encoded polypeptide.
Mutations can be introduced into a sequence of SEQ ID NO:1 or SEQ
ID NO:3 by standard techniques, such as site-directed mutagenesis
and PCR-mediated mutagenesis. Preferably, conservative amino acid
substitutions are made at one or more predicted non-essential amino
acid residues.
[0058] A "conservative amino acid substitution" is one in which the
amino acid residue is replaced with an amino acid residue having a
similar side chain. Families of amino acid residues having similar
side chains have been defined in the art. These families include
amino acids with basic side chains (e.g., lysine, arginine,
histidine), acidic side chains (e.g., aspartic acid, glutamic
acid), uncharged polar side chains (e.g., glycine, asparagine,
glutamine, serine, threonine, tyrosine, cysteine), nonpolar side
chains (e.g., alanine, valine, leucine, isoleucine, proline,
phenylalanine, methionine, tryptophan), beta-branched side chains
(e.g., threonine, valine, isoleucine) and aromatic side chains
(e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a
predicted nonessential amino acid residue in an IMI polypeptide is
preferably replaced with another amino acid residue from the same
side chain family. Alternatively, in another embodiment, mutations
can be introduced randomly along all or part of an IMI coding
sequence, such as by saturation mutagenesis, and the resultant
mutants can be screened for an IMI activity described herein to
identify mutants that retain IMI activity. Following mutagenesis of
the sequence of SEQ ID NO:1 or SEQ ID NO:3, the encoded polypeptide
can be expressed recombinantly and the activity of the polypeptide
can be determined by analyzing the imidazolinone resistance of a
plant expressing the polypeptide as described in the Examples
below.
[0059] To determine the percent sequence identity of two amino acid
sequences (e.g., SEQ ID NO:2 or SEQ ID NO:4 and a mutant form
thereof), the sequences are aligned for optimal comparison purposes
(e.g., gaps can be introduced in the sequence of one polypeptide
for optimal alignment with the other polypeptide). The amino acid
residues at corresponding amino acid positions are then compared.
When a position in one sequence (e.g., SEQ ID NO:2 or SEQ ID NO:4)
is occupied by the same amino acid residue as the corresponding
position in the other sequence (e.g., a mutant form of SEQ ID NO:2
or SEQ ID NO:4), then the molecules are identical at that position.
The same type of comparison can be made between two nucleic acid
sequences. The percent sequence identity between the two sequences
is a function of the number of identical positions shared by the
sequences (i.e., percent sequence identity=numbers of identical
positions/total numbers of positions.times.100). For the purposes
of the invention, the percent sequence identity between two nucleic
acid or polypeptide sequences is determined using the Vector NTI
6.0 (PC) software package (InforMax, 7600 Wisconsin Ave., Bethesda,
Md. 20814). A gap opening penalty of 15 and a gap extension penalty
of 6.66 are used for determining the percent identity of two
nucleic acids. A gap opening penalty of 10 and a gap extension
penalty of 0.1 are used for determining the percent identity of two
polypeptides. All other parameters are set at the default settings.
It is to be understood that for the purposes of determining
sequence identity, when comparing a DNA sequence to an RNA
sequence, a thymidine nucleotide is equivalent to a uracil
nucleotide. Preferably, the isolated IMI polypeptides included in
the present invention are at least about 50-60%, preferably at
least about 60-70%, and more preferably at least about 70-75%,
75-80%, 80-85%, 85-90% or 90-95%, and most preferably at least
about 96%, 97%, 98%, 99% or more identical to an entire amino acid
sequence encoded by a polynucleotide sequence shown in SEQ ID NO:1
or SEQ ID NO:3. In another embodiment, the isolated IMI
polypeptides included in the present invention are at least about
50-60%, preferably at least about 60-70%, and more preferably at
least about 70-75%, 75-80%, 80-85%, 85-90% or 90-95%, and most
preferably at least about 96%, 97%, 98%, 99% or more identical to
an entire amino acid sequence shown in SEQ ID NO:2 or SEQ ID
NO:4.
[0060] Additionally, optimized IMI nucleic acids can be created.
Preferably, an optimized IMI nucleic acid encodes an IMI
polypeptide that modulates a plant's tolerance to imidazolinone
herbicides, and more preferably increases a plant's tolerance to an
imidazolinone herbicide upon its over-expression in the plant. As
used herein, "optimized" refers to a nucleic acid that is
genetically engineered to increase its expression in a given plant
or animal. To provide plant optimized IMI nucleic acids, the DNA
sequence of the gene can be modified to 1) comprise codons
preferred by highly expressed plant genes; 2) comprise an A+T
content in nucleotide base composition to that substantially found
in plants; 3) form a plant initiation sequence, 4) eliminate
sequences that cause destabilization, inappropriate
polyadenylation, degradation and termination of RNA, or that form
secondary structure hairpins or RNA splice sites. Increased
expression of IMI nucleic acids in plants can be achieved by
utilizing the distribution frequency of codon usage in plants in
general or a particular plant. Methods for optimizing nucleic acid
expression in plants can be found in EPA 0359472; EPA 0385962; PCT
Application No. WO 91/16432; U.S. Pat. No. 5,380,831; U.S. Pat. No.
5,436,391; Perlack et al., 1991 Proc. Natl. Acad. Sci. USA
88:3324-3328; and Murray et al., 1989 Nucleic Acids Res.
17:477-498.
[0061] As used herein, "frequency of preferred codon usage" refers
to the preference exhibited by a specific host cell in usage of
nucleotide codons to specify a given amino acid. To determine the
frequency of usage of a particular codon in a gene, the number of
occurrences of that codon in the gene is divided by the total
number of occurrences of all codons specifying the same amino acid
in the gene. Similarly, the frequency of preferred codon usage
exhibited by a host cell can be calculated by averaging frequency
of preferred codon usage in a large number of genes expressed by
the host cell. It is preferable that this analysis be limited to
genes that are highly expressed by the host cell. The percent
deviation of the frequency of preferred codon usage for a synthetic
gene from that employed by a host cell is calculated first by
determining the percent deviation of the frequency of usage of a
single codon from that of the host cell followed by obtaining the
average deviation over all codons. As defined herein, this
calculation includes unique codons (i.e., ATG and TGG). In general
terms, the overall average deviation of the codon usage of an
optimized gene from that of a host cell is calculated using the
equation 1A=n 1 Z X.sub.n-Y.sub.nX.sub.n times 100 Z where
X.sub.n=frequency of usage for codon n in the host cell;
Y.sub.n=frequency of usage for codon n in the synthetic gene, n
represents an individual codon that specifies an amino acid and the
total number of codons is Z. The overall deviation of the frequency
of codon usage, A, for all amino acids should preferably be less
than about 25%, and more preferably less than about 10%.
[0062] Hence, an IMI nucleic acid can be optimized such that its
distribution frequency of codon usage deviates, preferably, no more
than 25% from that of highly expressed plant genes and, more
preferably, no more than about 10%. In addition, consideration is
given to the percentage G+C content of the degenerate third base
(monocotyledons appear to favor G+C in this position, whereas
dicotyledons do not). It is also recognized that the XCG (where X
is A, T, C, or G) nucleotide is the least preferred codon in dicots
whereas the XTA codon is avoided in both monocots and dicots.
Optimized IMI nucleic acids of this invention also preferably have
CG and TA doublet avoidance indices closely approximating those of
the chosen host plant (i.e., Triticum aestivum). More preferably
these indices deviate from that of the host by no more than about
10-15%.
[0063] In addition to the nucleic acid molecules encoding the IMI
polypeptides described above, another aspect of the invention
pertains to isolated nucleic acid molecules that are antisense
thereto. Antisense polynucleotides are thought to inhibit gene
expression of a target polynucleotide by specifically binding the
target polynucleotide and interfering with transcription, splicing,
transport, translation and/or stability of the target
polynucleotide. Methods are described in the prior art for
targeting the antisense polynucleotide to the chromosomal DNA, to a
primary RNA transcript or to a processed mRNA. Preferably, the
target regions include splice sites, translation initiation codons,
translation termination codons, and other sequences within the open
reading frame.
[0064] The term "antisense", for the purposes of the invention,
refers to a nucleic acid comprising a polynucleotide that is
sufficiently complementary to all or a portion of a gene, primary
transcript or processed mRNA, so as to interfere with expression of
the endogenous gene. "Complementary" polynucleotides are those that
are capable of base pairing according to the standard Watson-Crick
complementarity rules. Specifically, purines will base pair with
pyrimidines to form a combination of guanine paired with cytosine
(G:C) and adenine paired with either thymine (A:T) in the case of
DNA, or adenine paired with uracil (A:U) in the case of RNA. It is
understood that two polynucleotides may hybridize to each other
even if they are not completely complementary to each other,
provided that each has at least one region that is substantially
complementary to the other. The term "antisense nucleic acid"
includes single stranded RNA as well as double-stranded DNA
expression cassettes that can be transcribed to produce an
antisense RNA. "Active" antisense nucleic acids are antisense RNA
molecules that are capable of selectively hybridizing with a
primary transcript or mRNA encoding a polypeptide having at least
80% sequence identity with the polypeptide encoded by the
polynucleotide sequence of SEQ ID NO:1 or SEQ ID NO:3.
[0065] In addition to the IMI nucleic acids and polypeptides
described above, the present invention encompasses these nucleic
acids and polypeptides attached to a moiety. These moieties
include, but are not limited to, detection moieties, hybridization
moieties, purification moieties, delivery moieties, reaction
moieties, binding moieties, and the like. A typical group of
nucleic acids having moieties attached are probes and primers.
Probes and primers typically comprise a substantially isolated
oligonucleotide. The oligonucleotide typically comprises a region
of nucleotide sequence that hybridizes under stringent conditions
to at least about 12, preferably about 25, more preferably about
40, 50 or 75 consecutive nucleotides of a sense strand of the
sequence set forth in SEQ ID NO:1 or SEQ ID NO:3, an anti-sense
sequence of the sequence set forth in SEQ ID NO:1 or SEQ ID NO:3,
or naturally occurring mutants thereof. Primers based on a
nucleotide sequence of SEQ ID NO:1 or SEQ ID NO:3 can be used in
PCR reactions to clone IMI homologs. Probes based on the IMI
nucleotide sequences can be used to detect transcripts or genomic
sequences encoding the same or homologous polypeptides. In
preferred embodiments, the probe further comprises a label group
attached thereto, e.g. the label group can be a radioisotope, a
fluorescent compound, an enzyme, or an enzyme co-factor. Such
probes can be used as a part of a genomic marker test kit for
identifying cells which express an IMI polypeptide, such as by
measuring a level of an IMI-encoding nucleic acid, in a sample of
cells, e.g., detecting IMI mRNA levels or determining whether a
genomic IMI gene has been mutated or deleted.
[0066] The invention further provides an isolated recombinant
expression vector comprising an IMI nucleic acid as described
above, wherein expression of the vector in a host cell results in
increased resistance to an imidazolinone herbicide as compared to a
wild type variety of the host cell. As used herein, the term
"vector" refers to a nucleic acid molecule capable of transporting
another nucleic acid to which it has been linked. One type of
vector is a "plasmid", which refers to a circular double stranded
DNA loop into which additional DNA segments can be ligated. Another
type of vector is a viral vector, wherein additional DNA segments
can be ligated into the viral genome. Certain vectors are capable
of autonomous replication in a host cell into which they are
introduced (e.g., bacterial vectors having a bacterial origin of
replication and episomal mammalian vectors). Other vectors (e.g.,
non-episomal mammalian vectors) are integrated into the genome of a
host cell upon introduction into the host cell, and thereby are
replicated along with the host genome. Moreover, certain vectors
are capable of directing the expression of genes to which they are
operatively linked. Such vectors are referred to herein as
"expression vectors". In general, expression vectors of utility in
recombinant DNA techniques are often in the form of plasmids. In
the present specification, "plasmid" and "vector" can be used
interchangeably as the plasmid is the most commonly used form of
vector. However, the invention is intended to include such other
forms of expression vectors, such as viral vectors (e.g.,
replication defective retroviruses, adenoviruses and
adeno-associated viruses), which serve equivalent functions.
[0067] The recombinant expression vectors of the invention comprise
a nucleic acid of the invention in a form suitable for expression
of the nucleic acid in a host cell, which means that the
recombinant expression vectors include one or more regulatory
sequences, selected on the basis of the host cells to be used for
expression, which is operably linked to the nucleic acid sequence
to be expressed. Within a recombinant expression vector, "operably
linked" is intended to mean that the nucleotide sequence of
interest is linked to the regulatory sequence(s) in a manner which
allows for expression of the nucleotide sequence (e.g., in an in
vitro transcription/translation system or in a host cell when the
vector is introduced into the host cell). The term "regulatory
sequence" is intended to include promoters, enhancers and other
expression control elements (e.g., polyadenylation signals). Such
regulatory sequences are described, for example, in Goeddel, Gene
Expression Technology: Methods in Enzymology 185, Academic Press,
San Diego, Calif. (1990) or see: Gruber and Crosby, in: Methods in
Plant Molecular Biology and Biotechnology, eds. Glick and Thompson,
Chapter 7, 89-108, CRC Press: Boca Raton, Fla., including the
references therein. Regulatory sequences include those that direct
constitutive expression of a nucleotide sequence in many types of
host cells and those that direct expression of the nucleotide
sequence only in certain host cells or under certain conditions. It
will be appreciated by those skilled in the art that the design of
the expression vector can depend on such factors as the choice of
the host cell to be transformed, the level of expression of
polypeptide desired, etc. The expression vectors of the invention
can be introduced into host cells to thereby produce polypeptides
or peptides, including fusion polypeptides or peptides, encoded by
nucleic acids as described herein (e.g., IMI polypeptides, fusion
polypeptides, etc.).
[0068] In a preferred embodiment of the present invention, the IMI
polypeptides are expressed in plants and plants cells such as
unicellular plant cells (such as algae) (see Falciatore et al.,
1999 Marine Biotechnology 1(3):239-251 and references therein) and
plant cells from higher plants (e.g., the spermatophytes, such as
crop plants). An IMI polynucleotide may be "introduced" into a
plant cell by any means, including transfection, transformation or
transduction, electroporation, particle bombardment, biolistics,
agroinfection and the like. One transformation method known to
those of skill in the art is the dipping of a flowering plant into
an Agrobacteria solution, wherein the Agrobacteria contains the IMI
nucleic acid, followed by breeding of the transformed gametes.
[0069] Other suitable methods for transforming or transfecting host
cells including plant cells can be found in Sambrook, et al.
(Molecular Cloning: A Laboratory Manual. 2.sup.nd, ed., Cold Spring
Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring
Harbor, N. Y., 1989) and other laboratory manuals such as Methods
in Molecular Biology, 1995, Vol. 44, Agrobacterium protocols, ed:
Gartland and Davey, Humana Press, Totowa, N.J. As increased
resistance to imidazolinone herbicides is a general trait wished to
be inherited into a wide variety of plants like maize, wheat, rye,
oat, triticale, rice, barley, soybean, peanut, cotton, rapeseed and
canola, manihot, pepper, sunflower and tagetes, solanaceous plants
like potato, tobacco, eggplant, and tomato, Vicia species, pea,
alfalfa, bushy plants (coffee, cacao, tea), Salix species, trees
(oil palm, coconut), perennial grasses and forage crops, these crop
plants are also preferred target plants for a genetic engineering
as one further embodiment of the present invention. Forage crops
include, but are not limited to, Wheatgrass, Canarygrass,
Bromegrass, Wildrye Grass, Bluegrass, Orchardgrass, Alfalfa,
Salfoin, Birdsfoot Trefoil, Alsike Clover, Red Clover and Sweet
Clover.
[0070] In one embodiment of the present invention, transfection of
an IMI polynucleotide into a plant is achieved by Agrobacterium
mediated gene transfer. Agrobacterium mediated plant transformation
can be performed using for example the GV3101 (pMP90) (Koncz and
Schell, 1986 Mol. Gen. Genet. 204:383-396) or LBA4404 (Clontech)
Agrobacterium tumefaciens strain. Transformation can be performed
by standard transformation and regeneration techniques (Deblaere et
al., 1994 Nucl. Acids. Res. 13:4777-4788; Gelvin, Stanton B. and
Schilperoort, Robert A, Plant Molecular Biology Manual, 2.sup.nd
Ed.-Dordrecht: Kluwer Academic Publ., 1995.--in Sect., Ringbuc
Zentrale Signatur: BT11-P ISBN 0-7923-27314; Glick, Bernard R. and
Thompson, John E., Methods in Plant Molecular Biology and
Biotechnology, Boca Raton: CRC Press, 1993 360 S., ISBN
0-8493-5164-2). For example, rapeseed can be transformed via
cotyledon or hypocotyl transformation (Moloney et al., 1989 Plant
cell Report 8:238-242; De Block et al., 1989 Plant Physiol.
91:694-701). Use of antibiotica for Agrobacterium and plant
selection depends on the binary vector and the Agrobacterium strain
used for transformation. Rapeseed selection is normally performed
using kanamycin as selectable plant marker. Agrobacterium mediated
gene transfer to flax can be performed using, for example, a
technique described by Mlynarova et al., 1994 Plant Cell Report
13:282-285. Additionally, transformation of soybean can be
performed using, for example, a technique described in European
Patent No. 0424 047, U.S. Pat. No. 5,322,783, European Patent No.
0397 687, U.S. Pat. No. 5,376,543 or U.S. Pat. No. 5,169,770.
Transformation of maize can be achieved by particle bombardment,
polyethylene glycol mediated DNA uptake or via the silicon carbide
fiber technique. (See, for example, Freeling and Walbot "The maize
handbook" Springer Verlag: New York (1993) ISBN 3-540-97826-7). A
specific example of maize transformation is found in U.S. Pat. No.
5,990,387 and a specific example of wheat transformation can be
found in PCT Application No. WO 93/07256.
[0071] According to the present invention, the introduced IMI
polynucleotide may be maintained in the plant cell stably if it is
incorporated into a non-chromosomal autonomous replicon or
integrated into the plant chromosomes. Alternatively, the
introduced IMI polynucleotide may be present on an
extra-chromosomal non-replicating vector and be transiently
expressed or transiently active. In one embodiment, a homologous
recombinant microorganism can be created wherein the IMI
polynucleotide is integrated into a chromosome, a vector is
prepared which contains at least a portion of an AHAS gene into
which a deletion, addition or substitution has been introduced to
thereby alter, e.g., functionally disrupt, the endogenous AHAS gene
and to create an IMI gene. To create a point mutation via
homologous recombination, DNA-RNA hybrids can be used in a
technique known as chimeraplasty (Cole-Strauss et al., 1999 Nucleic
Acids Research 27(5):1323-1330 and Kmiec, 1999 Gene therapy
American Scientist 87(3):240-247). Other homologous recombination
procedures in Triticum species are also well known in the art and
are contemplated for use herein.
[0072] In the homologous recombination vector, the IMI gene can be
flanked at its 5' and 3' ends by an additional nucleic acid
molecule of the AHAS gene to allow for homologous recombination to
occur between the exogenous IMI gene carried by the vector and an
endogenous AHAS gene, in a microorganism or plant. The additional
flanking AHAS nucleic acid molecule is of sufficient length for
successful homologous recombination with the endogenous gene.
Typically, several hundreds of base pairs up to kilobases of
flanking DNA (both at the 5' and 3' ends) are included in the
vector (see e.g., Thomas, K. R., and Capecchi, M. R., 1987 Cell
51:503 for a description of homologous recombination vectors or
Strepp et al., 1998 PNAS, 95(8):4368-4373 for cDNA based
recombination in Physcomitrella patens). However, since the IMI
gene normally differs from the AHAS gene at very few amino acids, a
flanking sequence is not always necessary. The homologous
recombination vector is introduced into a microorganism or plant
cell (e.g., via polyethylene glycol mediated DNA), and cells in
which the introduced IMI gene has homologously recombined with the
endogenous AHAS gene are selected using art-known techniques.
[0073] In another embodiment, recombinant microorganisms can be
produced that contain selected systems that allow for regulated
expression of the introduced gene. For example, inclusion of an IMI
gene on a vector placing it under control of the lac operon permits
expression of the IMI gene only in the presence of IPTG. Such
regulatory systems are well known in the art.
[0074] Whether present in an extra-chromosomal non-replicating
vector or a vector that is integrated into a chromosome, the IMI
polynucleotide preferably resides in a plant expression cassette. A
plant expression cassette preferably contains regulatory sequences
capable of driving gene expression in plant cells that are operably
linked so that each sequence can fulfill its function, for example,
termination of transcription by polyadenylation signals. Preferred
polyadenylation signals are those originating from Agrobacterium
tumefaciens t-DNA such as the gene 3 known as octopine synthase of
the Ti-plasmid pTiACH5 (Gielen et al., 1984 EMBO J. 3:835) or
functional equivalents thereof, but also all other terminators
functionally active in plants are suitable. As plant gene
expression is very often not limited on transcriptional levels, a
plant expression cassette preferably contains other operably linked
sequences like translational enhancers such as the
overdrive-sequence containing the 5'-untranslated leader sequence
from tobacco mosaic virus enhancing the polypeptide per RNA ratio
(Gallie et al., 1987 Nucl. Acids Research 15:8693-8711). Examples
of plant expression vectors include those detailed in: Becker, D.
et al., 1992 New plant binary vectors with selectable markers
located proximal to the left border, Plant Mol. Biol. 20:1195-1197;
Bevan, M. W., 1984 Binary Agrobacterium vectors for plant
transformation, Nucl. Acid. Res. 12:8711-8721; and Vectors for Gene
Transfer in Higher Plants; in: Transgenic Plants, Vol. 1,
Engineering and Utilization, eds.: Kung and R. Wu, Academic Press,
1993, S. 15-38.
[0075] Plant gene expression should be operably linked to an
appropriate promoter conferring gene expression in a timely, cell
or tissue specific manner. Promoters useful in the expression
cassettes of the invention include any promoter that is capable of
initiating transcription in a plant cell. Such promoters include,
but are not limited to those that can be obtained from plants,
plant viruses and bacteria that contain genes that are expressed in
plants, such as Agrobacterium and Rhizobium.
[0076] The promoter may be constitutive, inducible, developmental
stage-preferred, cell type-preferred, tissue-preferred or
organ-preferred. Constitutive promoters are active under most
conditions. Examples of constitutive promoters include the CaMV 19S
and 35 S promoters (Odell et al. 1985 Nature 313:810-812), the sX
CaMV 35S, promoter (Kay et al. 1987 Science 236:1299-1302) the Sep1
promoter, the rice actin promoter (McElroy et al. 1990 Plant Cell
2:163-171), the Arabidopsis actin promoter, the ubiquitan promoter
(Christensen et al. 1989 Plant Molec Biol. 18:675-689); pEmu (Last
et al. 1991 Theor Appl Genet. 81:581-588), the figwort mosaic virus
35S promoter, the Smas promoter (Velten et al. 1984 EMBO J.
3:2723-2730), the GRP1-8 promoter, the cinnamyl alcohol
dehydrogenase promoter (U.S. Pat. No. 5,683,439), promoters from
the T-DNA of Agrobacterium, such as mannopine synthase, nopaline
synthase, and octopine synthase, the small subunit of ribulose
biphosphate carboxylase (ssuRUBISCO) promoter, and the like.
[0077] Inducible promoters are active under certain environmental
conditions, such as the presence or absence of a nutrient or
metabolite, heat or cold, light, pathogen attack, anaerobic
conditions, and the like. For example, the hsp80 promoter from
Brassica is induced by heat shock, the PPDK promoter is induced by
light, the PR-1 promoter from tobacco, Arabidopsis and maize are
inducible by infection with a pathogen, and the Adhl promoter is
induced by hypoxia and cold stress. Plant gene expression can also
be facilitated via an inducible promoter (for review see Gatz, 1997
Annu. Rev. Plant Physiol. Plant Mol. Biol. 48:89-108). Chemically
inducible promoters are especially suitable if gene expression is
wanted to occur in a time specific manner. Examples of such
promoters are a salicylic acid inducible promoter (PCT Application
No. WO 95/19443), a tetracycline inducible promoter (Gatz et al.,
1992 Plant J. 2:397-404) and an ethanol inducible promoter (PCT
Application No. WO 93/21334).
[0078] Developmental stage-preferred promoters are preferentially
expressed at certain stages of development. Tissue and organ
preferred promoters include those that are preferentially expressed
in certain tissues or organs, such as leaves, roots, seeds, or
xylem. Examples of tissue preferred and organ preferred promoters
include, but are not limited to fruit-preferred, ovule-preferred,
male tissue-preferred, seed-preferred, integument-preferred,
tuber-preferred, stalk-preferred, pericarp-preferred, and
leaf-preferred, stigma-preferred, pollen-preferred,
anther-preferred, a petal-preferred, sepal-preferred,
pedicel-preferred, silique-preferred, stem-preferred,
root-preferred promoters and the like. Seed preferred promoters are
preferentially expressed during seed development and/or
germination. For example, seed preferred promoters can be
embryo-preferred, endosperm preferred and seed coat-preferred. See
Thompson et al. 1989 BioEssays 10:108. Examples of seed preferred
promoters include, but are not limited to cellulose synthase
(celA), Cim1, gamma-zein, globulin-1, maize 19 kD zein (cZ19B1) and
the like.
[0079] Other suitable tissue-preferred or organ-preferred promoters
include the napin-gene promoter from rapeseed (U.S. Pat. No.
5,608,152), the USP-promoter from Vicia faba (Baeumlein et al.,
1991 Mol Gen Genet. 225(3):459-67), the oleosin-promoter from
Arabidopsis (PCT Application No. WO 98/45461), the
phaseolin-promoter from Phaseolus vulgaris (U.S. Pat. No.
5,504,200), the Bce4-promoter from Brassica (PCT Application No. WO
91/13980) or the legumin B4 promoter (LeB4; Baeumlein et al., 1992
Plant Journal, 2(2):233-9) as well as promoters conferring seed
specific expression in monocot plants like maize, barley, wheat,
rye, rice, etc. Suitable promoters to note are the lpt2 or
lpt1-gene promoter from barley (PCT Application No. WO 95/15389 and
PCT Application No. WO 95/23230) or those described in PCT
Application No. WO 99/16890 (promoters from the barley
hordein-gene, rice glutelin gene, rice oryzin gene, rice prolamin
gene, wheat gliadin gene, wheat glutelin gene, oat glutelin gene,
Sorghum kasirin-gene and rye secalin gene).
[0080] Other promoters useful in the expression cassettes of the
invention include, but are not limited to, the major chlorophyll
a/b binding protein promoter, histone promoters, the Ap3 promoter,
the .beta.-conglycin promoter, the napin promoter, the soy bean
lectin promoter, the maize 15 kD zein promoter, the 22 kD zein
promoter, the 27 kD zein promoter, the g-zein promoter, the waxy,
shrunken 1, shrunken 2 and bronze promoters, the Zm13 promoter
(U.S. Pat. No. 5,086,169), the maize polygalacturonase promoters
(PG) (U.S. Pat. Nos. 5,412,085 and 5,545,546) and the SGB6 promoter
(U.S. Pat. No. 5,470,359), as well as synthetic or other natural
promoters.
[0081] Additional flexibility in controlling heterologous gene
expression in plants may be obtained by using DNA binding domains
and response elements from heterologous sources (i.e., DNA binding
domains from non-plant sources). An example of such a heterologous
DNA binding domain is the LexA DNA binding domain (Brent and
Ptashne, Cell 43:729-736 (1985)).
[0082] Another aspect of the invention pertains to host cells into
which a recombinant expression vector of the invention has been
introduced. The terms "host cell" and "recombinant host cell" are
used interchangeably herein. It is understood that such terms refer
not only to the particular subject cell but they also apply to the
progeny or potential progeny of such a cell. Because certain
modifications may occur in succeeding generations due to either
mutation or environmental influences, such progeny may not, in
fact, be identical to the parent cell, but are still included
within the scope of the term as used herein. A host cell can be any
prokaryotic or eukaryotic cell. For example, an IMI polynucleotide
can be expressed in bacterial cells such as C. glutamicum, insect
cells, fungal cells or mammalian cells (such as Chinese hamster
ovary cells (CHO) or COS cells), algae, ciliates, plant cells,
fungi or other microorganisms like C. glutamicum. Other suitable
host cells are known to those skilled in the art.
[0083] A host cell of the invention, such as a prokaryotic or
eukaryotic host cell in culture, can be used to produce (i.e.,
express) an IMI polynucleotide. Accordingly, the invention further
provides methods for producing IMI polypeptides using the host
cells of the invention. In one embodiment, the method comprises
culturing the host cell of invention (into which a recombinant
expression vector encoding an IMI polypeptide has been introduced,
or into which genome has been introduced a gene encoding a
wild-type or IMI polypeptide) in a suitable medium until IMI
polypeptide is produced. In another embodiment, the method further
comprises isolating IMI polypeptides from the medium or the host
cell. Another aspect of the invention pertains to isolated IMI
polypeptides, and biologically active portions thereof. An
"isolated" or "purified" polypeptide or biologically active portion
thereof is free of some of the cellular material when produced by
recombinant DNA techniques, or chemical precursors or other
chemicals when chemically synthesized. The language "substantially
free of cellular material" includes preparations of IMI polypeptide
in which the polypeptide is separated from some of the cellular
components of the cells in which it is naturally or recombinantly
produced. In one embodiment, the language "substantially free of
cellular material" includes preparations of an IMI polypeptide
having less than about 30% (by dry weight) of non-IMI material
(also referred to herein as a "contaminating polypeptide"), more
preferably less than about 20% of non-IMI material, still more
preferably less than about 10% of non-IMI material, and most
preferably less than about 5% non-IMI material.
[0084] When the IMI polypeptide, or biologically active portion
thereof, is recombinantly produced, it is also preferably
substantially free of culture medium, i.e., culture medium
represents less than about 20%, more preferably less than about
10%, and most preferably less than about 5% of the volume of the
polypeptide preparation. The language "substantially free of
chemical precursors or other chemicals" includes preparations of
IMI polypeptide in which the polypeptide is separated from chemical
precursors or other chemicals that are involved in the synthesis of
the polypeptide. In one embodiment, the language "substantially
free of chemical precursors or other chemicals" includes
preparations of an IMI polypeptide having less than about 30% (by
dry weight) of chemical precursors or non-IMI chemicals, more
preferably less than about 20% chemical precursors or non-IMI
chemicals, still more preferably less than about 10% chemical
precursors or non-IMI chemicals, and most preferably less than
about 5% chemical precursors or non-IMI chemicals. In preferred
embodiments, isolated polyp eptides, or biologically active
portions thereof, lack contaminating polypeptides from the same
organism from which the IMI polypeptide is derived. Typically, such
polypeptides are produced by recombinant expression of, for
example, a Triticum aestivum IMI polypeptide in plants other than
Triticum aestivum or microorganisms such as C. glutamicum,
ciliates, algae or fungi.
[0085] The IMI polynucleotide and polypeptide sequences of the
invention have a variety of uses. The nucleic acid and amino acid
sequences of the present invention can be used to transform plants,
thereby modulating the plant's resistance to imidazolinone
herbicides. Accordingly, the invention provides a method of
producing a transgenic plant having increased tolerance to an
imidazolinone herbicide comprising, (a) transforming a plant cell
with one or more expression vectors comprising one or more IMI
nucleic acids, and (b) generating from the plant cell a transgenic
plant with an increased resistance to an imidazolinone herbicide as
compared to a wild type variety of the plant. In one embodiment,
the multiple IMI nucleic acids are derived from different genomes.
Also included in the present invention are methods of producing a
transgenic plant having increased tolerance to an imidazolinone
herbicide comprising, (a) transforming a plant cell with an
expression vector comprising an IMI nucleic acid, wherein the
nucleic acid is a non-Imi1 nucleic acid and (b) generating from the
plant cell a transgenic plant with an increased resistance to an
imidazolinone herbicide as compared to a wild type variety of the
plant.
[0086] The present invention includes methods of modifying a
plant's tolerance to an imidazolinone herbicide comprising
modifying the expression of one or more IMI nucleic acids.
Preferably, the nucleic acids are located on or derived from
different genomes. The plant's resistance to the imidazolinone
herbicide can be increased or decreased as achieved by increasing
or decreasing the expression of an IMI polynucleotide,
respectively. Preferably, the plant's resistance to the
imidazolinone herbicide is increased by increasing expression of an
IMI polynucleotide. Expression of an IMI polynucleotide can be
modified by any method known to those of skill in the art. The
methods of increasing expression of IMI polynucleotides can be used
wherein the plant is either transgenic or not transgenic. In cases
when the plant is transgenic, the plant can be transformed with a
vector containing any of the above described IMI coding nucleic
acids, or the plant can be transformed with a promoter that directs
expression of endogenous IMI polynucleotides in the plant, for
example. The invention provides that such a promoter can be tissue
specific or developmentally regulated. Alternatively,
non-transgenic plants can have endogenous IMI polynucleotide
expression modified by inducing a native promoter. The expression
of polynucleotides comprising SEQ ID NO:1 or SEQ ID NO:3 in target
plants can be accomplished by, but is not limited to, one of the
following examples: (a) constitutive promoter, (b) chemical-induced
promoter, and (c) engineered promoter over-expression with for
example zinc-finger derived transcription factors (Greisman and
Pabo, 1997 Science 275:657).
[0087] In a preferred embodiment, transcription of the IMI
polynucleotide is modulated using zinc-finger derived transcription
factors (ZFPs) as described in Greisman and Pabo, 1997 Science
275:657 and manufactured by Sangamo Biosciences, Inc. These ZFPs
comprise both a DNA recognition domain and a functional domain that
causes activation or repression of a target nucleic acid such as an
IMI nucleic acid. Therefore, activating and repressing ZFPs can be
created that specifically recognize the IMI polynucleotide
promoters described above and used to increase or decrease IMI
polynucleotide expression in a plant, thereby modulating the
herbicide resistance of the plant.
[0088] As described in more detail above, the plants produced by
the methods of the present invention can be monocots or dicots. The
plants can be selected from maize, wheat, rye, oat, triticale,
rice, barley, soybean, peanut, cotton, rapeseed, canola, manihot,
pepper, sunflower, tagetes, solanaceous plants, potato, tobacco,
eggplant, tomato, Vicia species, pea, alfalfa, coffee, cacao, tea,
Salix species, oil palm, coconut, perennial grass and forage crops,
for example. Forage crops include, but are not limited to,
Wheatgrass, Canarygrass, Bromegrass, Wildrye Grass, Bluegrass,
Orchardgrass, Alfalfa, Salfoin, Birdsfoot Trefoil, Alsike Clover,
Red Clover and Sweet Clover. In a preferred embodiment, the plant
is a wheat plant. In each of the methods described above, the plant
cell includes, but is not limited to, a protoplast, gamete
producing cell, and a cell that regenerates into a whole plant. As
used herein, the term "transgenic" refers to any plant, plant cell,
callus, plant tissue or plant part, that contains all or part of at
least one recombinant polynucleotide. In many cases, all or part of
the recombinant polynucleotide is stably integrated into a
chromosome or stable extra-chromosomal element, so that it is
passed on to successive generations.
[0089] As described above, the present invention teaches
compositions and methods for increasing the imidazolinone
resistance of a wheat plant or seed as compared to a wild-type
variety of the plant or seed. In a preferred embodiment, the
imidazolinone resistance of a wheat plant or seed is increased such
that the plant or seed can withstand an imidazolinone herbicide
application of preferably approximately 10-400 g ai ha.sup.-1, more
preferably 20-160 g ai ha.sup.-1, and most preferably 40-80 g ai
ha.sup.-1. As used herein, to "withstand" an imidazolinone
herbicide application means that the plant is either not killed or
not injured by such application.
[0090] Additionally provided herein is a method of controlling
weeds within the vicinity of a wheat plant, comprising applying an
imidazolinone herbicide to the weeds and to the wheat plant,
wherein the wheat plant has increased resistance to the
imidazolinone herbicide as compared to a wild type variety of the
wheat plant, and wherein the plant comprises one or more IMI
nucleic acids. In one embodiment, the plant comprises multiple IMI
nucleic acids located on or derived from different genomes. In
another embodiment, the plant comprises a non-Mill nucleic acid. By
providing for wheat plants having increased resistance to
imidazolinone, a wide variety of formulations can be employed for
protecting wheat plants from weeds, so as to enhance plant growth
and reduce competition for nutrients. An imidazolinone herbicide
can be used by itself for pre-emergence, post-emergence,
pre-planting and at-planting control of weeds in areas surrounding
the wheat plants described herein or an imidazolinone herbicide
formulation can be used that contains other additives. The
imidazolinone herbicide can also be used as a seed treatment.
Additives found in an imidazolinone herbicide formulation include
other herbicides, detergents, adjuvants, spreading agents, sticking
agents, stabilizing agents, or the like. The imidazolinone
herbicide formulation can be a wet or dry preparation and can
include, but is not limited to, flowable powders, emulsifiable
concentrates and liquid concentrates. The imidazolinone herbicide
and herbicide formulations can be applied in accordance with
conventional methods, for example, by spraying, irrigation,
dusting, or the like.
[0091] Throughout this application, various publications are
referenced. The disclosures of all of these publications and those
references cited within those publications in their entireties are
hereby incorporated by reference into this application in order to
more fully describe the state of the art to which this invention
pertains.
[0092] It should also be understood that the foregoing relates to
preferred embodiments of the present invention and that numerous
changes may be made therein without departing from the scope of the
invention. The invention is further illustrated by the following
examples, which are not to be construed in any way as imposing
limitations upon the scope thereof. On the contrary, it is to be
clearly understood that resort may be had to various other
embodiments, modifications, and equivalents thereof, which, after
reading the description herein, may suggest themselves to those
skilled in the art without departing from the spirit of the present
invention and/or the scope of the appended claims.
EXAMPLES
Example 1
Mutagenesis and Selection of Resistant Wheat Lines
[0093] Approximately 40,000 seeds of Triticum aestivum L. cv CDC
Teal (Hughes and Hucl, 1993 Can. J. Plant Sci. 73:193-197) were
mutagenized using modified procedures described by Washington and
Sears (1970). Seeds were pre-soaked in distilled water for four
hours, followed by treatment with 0.3% EMS for six hours. Seeds
were rinsed continually with tap water for seven hours and allowed
to dry for approximately four hours before being planted in the
field. The M.sub.1 plants were selfed and the seed was harvested in
bulk. Approximately 2.times.10.sup.6 M.sub.2 plants were grown in
the field the following year and were sprayed at the two leaf stage
with imazamox at a rate of 40 g ai ha.sup.-1 in a spray volume of
100 L ha.sup.-1. Merge 0.05% (v/v) adjuvant was added to the spray
solution. Six lines resistant to imazamox were selected and
designated as lines 1A, 9A, 10A, 11A, 15A, and 16A. The M.sub.3 and
M.sub.4 generations were grown in a walk-in growth chamber and
plants resistant to imazamox were selected using rates of 20 g ai
ha.sup.-1. Resistant plants were selected in the M.sub.5 generation
after application of 40 g ai ha.sup.-1 in the field. M.sub.5 seed
was homozygous for the trait, as progeny testing detected no
segregation for resistance to imazamox.
Example 2
Methods Used to Determine Inheritance and Allelism of IMI Genes
[0094] To determine the genetic control of resistance to imazamox
in the six wheat lines, reciprocal crosses between the six
homozygous resistant M.sub.6 lines and CDC Teal (susceptible to
imazamox) were made. Randomly selected F.sub.1 plants from each of
the crosses were backcrossed to CDC Teal to form backcross
(BC)F.sub.1 populations. To investigate allelism, all possible
inter-crosses between the six mutants and SWP965001
(Grandin/3*Fidel-FS-4) were made. SWP965001 is a spring wheat line
that is homozygous for the FS-4 allele. Parental genotypes were
grown in a walk-in growth chamber with a 16 hour photoperiod and a
24.degree. C. day and 16.degree. C. night temperature regime.
Spikes that were 3/4 emerged from the boot were emasculated and
then pollinated 2-3 days after the emasculation date. Randomly
selected F.sub.2 plants from all segregating crosses were selfed to
produce F.sub.2:3 families. Parental, F.sub.1, BCF.sub.1, F.sub.2
plants and F.sub.2:3 families were tested for reaction to imazamox.
All experiments were conducted in a walk-in growth chamber with a
16 hour photoperiod and a 23.degree. C. day and 16.degree. C. night
temperature regime. A completely random design was used for all
experiments. In experiments involving F.sub.2:3 families, effort
was taken to randomize both within and among families. The F.sub.1
and F.sub.2 populations were screened in the same experiment along
with parental genotypes and CDC Teal as controls. Both the
BCF.sub.1 and F.sub.2:3 populations were screened in two separate
experiments along with appropriate parental genotypes as
controls.
[0095] Herbicide treatments were applied to plants growing in
8.times.16 cell flats at the two leaf stage using a traveling cable
sprayer calibrated to spray 100 L ha.sup.-1. Imazamox was applied
to plants at a rate of 20 g ai ha.sup.-1 using an 8001 EVS nozzle
at a pressure of 275 kPa. Merge surfactant (0.05% v/v) was added to
the herbicide solution prior to application. Fifteen days after
herbicide application, plants were rated based on parental
reactions and were considered as resistant, intermediate, or
susceptible. Resistant plants were phenotypically unaffected
following herbicide treatment whereas intermediate plants were
characterized by halted growth of the first two leaves, darkening
(dark-green pigmentation) of the leaves, and the emergence of
coleoptilar tillers. Susceptible plants were characterized by
failure to develop new leaves, extensive leaf chlorosis, and
eventually, plant death. For Mendelian analysis of the segregating
populations, plants were scored into resistant and susceptible
categories and tested for goodness of fit to various 1 gene, 2 gene
and 3 gene models using chi-square analysis. For F.sub.2 and
BCF.sub.1 plant data, intermediate reactions were included in the
resistant reaction category. Yates correction for continuity was
used to adjust the chi-square value when only a single degree of
freedom was used in the chi-square analysis (Steele and Torrie 1980
Principles and procedures of statistics. McGraw-Hill, New York,
N.Y. pp 633).
Example 3
Results Regarding Inheritance of IMI Genes
[0096] All resistant parents produced a similar phenotype when
sprayed with 20 g ai ha.sup.-1 of imazamox. (FIG. 1). Reciprocal
crosses between the resistant lines and the susceptible parent (CDC
Teal) resulted in F.sub.1 plants that survived application of
imazamox (FIG. 1), indicating that resistance to imazamox is a
nuclear and not a cytoplasmic trait. With the exception of cross
15A.times.Teal, the F.sub.1 plants resulting from each of the
resistant lines crossed with CDC Teal displayed an intermediate
reaction (FIG. 1). Since the F.sub.1 plants were phenotypically
intermediate between the two parents, it was concluded that
resistance to imazamox in these lines was a partially dominant
trait (FIG. 1). Genetic analysis of resistance to imidazolinones
and sulfonylureas in Arabidopsis thaliana (Haughn and Somerville,
1986 Mol. Gen. Genet. 204:430-434) Zea mays (Newhouse et al., 1991
Theor. Appl. Genet. 83:65-70), Brassica napus (Swanson et al., 1989
Theor. Appl. Gen. 78:525-530), and Glycine max (Sebastian et al.,
1989 Crop Sci. 29:1403-1408) also indicated the presence of a
single, partially dominant nuclear gene.
[0097] Fourteen F.sub.1 plants resulting from the 15A.times.Teal
cross were rated as resistant (FIG. 1). Evaluation of F.sub.2
populations from this cross indicated that two independently
segregating loci were involved in conferring resistance in this
genotype (FIG. 2). Since the F.sub.1 would carry two heterozygous
resistant loci, one would expect that a resistant reaction would be
observed. If each of these loci alone would confer partial
dominance, additively, two heterozygous loci would produce a
resistant reaction. Swanson et al. (1989) combined two
semi-dominant imidazolinone resistance alleles from Brassica napus,
representing two unlinked genes, to produce a F.sub.1 hybrid that
was superior in imidazolinone resistance than either of the
heterozygous lines alone. The authors concluded that resistance
mechanisms are additive, and a higher level of resistance is
observed in lines carrying more than one resistance allele.
[0098] An analysis of cytoplasmic inheritance was conducted in the
F.sub.2 generation by testing homogeneity of deviations from
segregation ratios between the two reciprocal F.sub.2 populations.
Chi-square analysis revealed no significant deviations between
reciprocal populations, confirming the absence of cytoplasmic
inheritance (FIG. 2). Since cytoplasmic inheritance was absent,
data from the two reciprocal populations was combined and a total
chi-square on pooled F.sub.2 data was calculated (FIG. 2).
[0099] With the exception of Teal.times.15A, all F.sub.2
populations resulting from resistant.times.susceptible crosses gave
a good fit to a 3:1 resistant susceptible ratio indicating
segregation of a single major gene for resistance to imazamox (FIG.
2). When F.sub.1 plants were crossed to the susceptible parent,
resulting BCF.sub.1 populations gave a good fit to a 1:1
resistant:susceptible ratio, confirming the single locus hypothesis
(FIG. 2). The F.sub.2 population data from the cross 15A.times.Teal
fit a 15:1 resistant:susceptible ratio (P=0.08), indicating
segregation of two independent, complementary genes (FIG. 2). The
BCF.sub.1 population gave good fit to a 3:1 resistant:susceptible
ratio with a chi-square P value of 0.35, confirming the results of
the F.sub.2 (FIG. 2).
[0100] Since it is speculated from F.sub.2 data that resistance in
lines 1A, 9A, 10A, 11A, and 16A are controlled by a single major
gene, F.sub.2:3 families should segregate and fit a 1:2:1
homozygous resistance:segregating: homozygous susceptible family
ratio. Evaluation of F.sub.2:3 families indicated that crosses
Teal.times.1A, Teal.times.9A, Teal.times.10A, Teal.times.11A, and
Teal.times.16A all fit a 1:2:1 resistant:segregating:susceptible
F.sub.2:3 family ratio with chi-square P values of 0.64, 0.66,
0.52, 0.40, and 0.94, respectively (FIG. 3). These results confirm
the results of the F.sub.2 and BCF.sub.1 data that resistance in
lines 1A, 10A, 9A, 11A, and resistance in 16A is controlled by a
single major gene. This pattern of inheritance is consistent with
other findings that have reported the genetic control of resistance
to AHAS inhibitor herbicides. To date, nearly all plant mutations
conferring resistance to imidazolinones show that a single,
partially dominant gene controls the resistance trait. In Triticum
aestivum, Zea mays, Glycine max, Arabidopsis thaliana, and
Nicotiana tabacum, resistance to AHAS inhibitors is inherited as a
single partially dominant nuclear gene (Newhouse et al. 1991;
Newhouse et al. 1992; Chaleff and Ray, 1984 Science 223:1148-1151;
Sathasivan et al., 1991 Plant Physiol. 97:1044-1050). Plant
resistance to AHAS inhibitor herbicides occurs mostly because of a
single point mutation to the gene encoding the AHAS enzyme (Harms
et al. 1992, Mol. Gen. Genet. 233:427-435; Winder and Spalding,
1988 Mol. Gen. Genet. 238:394-399).
[0101] The F.sub.2 data resulting from the cross Teal.times.15A
provided a good fit to a 15:1 resistant:susceptible ratio,
suggesting segregation of two, independently segregating loci (FIG.
2). If this is the case, F.sub.2:3 families should segregate and
fit a 7:8:1 resistant:segregating:susceptible F.sub.2:3 family
ratio. F.sub.2:3 families from the cross 15A.times.Teal did fit the
expected 7:8:1 ratio (FIG. 3), confirming the results of the
F.sub.2 and BCF.sub.1 populations that resistance in 15A is
conferred by two, independent loci. To the inventor's knowledge,
this is the first reported instance were two independently
segregating imidazolinonc resistant alleles were identified in a
single line following seed mutagenesis.
Example 4
Results Regarding Allelism of IMI Genes
[0102] To determine the allelic relationships of resistance genes,
all possible intercrosses between resistant lines were evaluated.
No susceptible plants were observed in the F.sub.2 populations
resulting from the inter-crosses between lines SWP965001, 1A, 9A,
10A, 15A, and 16A (FIG. 4). Since these populations were not
segregating, the resistance genes in these lines are either alleles
at the FS-4 locus, or are very tightly linked. Since these
populations were not segregating in the F.sub.2 generation,
F.sub.2:3 families from these crosses were not evaluated.
[0103] All inter-crosses involving line 11A did segregate in the
F.sub.2 generation, indicating the presence of a unique resistance
gene in 11A (FIG. 4). If two independently segregating resistance
genes are present as the result of crossing two lines, each
carrying a single resistance gene, a 15:1 resistant:susceptible
ratio would be expected in the F.sub.2 generation. In the F.sub.2
generation, crosses SWP965001.times.11A, 1A.times.11A,
10A.times.11A, and 16A.times.11A fit the expected 15:1
resistant:susceptible ratio suggesting independent segregation of
two major resistance genes (FIG. 4). F.sub.2:3 family ratios from
these crosses also gave a good fit to a 7:8:1
resistant:segregating:susceptible ratio, confirming the results
obtained in the F.sub.2 generation (FIG. 5). Cross 11A.times.9A did
produce a segregating F.sub.2 population, but the ratio did not fit
a 15:1 segregation ratio due to an excess of susceptible
segregants. Various other two gene hypotheses were tested, but all
were found to be highly significant (Data not shown). Evaluation of
F.sub.2:3 families from this cross, however did give good fit to a
7:8:1 segregation ratio, indicating segregation of two independent
genes (FIG. 5). These results confirm that the resistance gene in
11A is unique from those in lines SWP96001, 1A, 9A, 10A, and
16A.
[0104] Cross 11A.times.15A did produce a segregating F.sub.2
population. Since 15A is carrying two resistance genes, one allelic
to FS-4, a segregating F.sub.2 population in cross 11A.times.15A
would indicate the presence of three segregating genes. Segregating
generations resulting from cross 15A.times.11A were tested for
segregation of three independent loci. F.sub.2 plants did fit the
expected 63:1 resistant:susceptible ratio, indicating the
segregation of three independent loci (FIG. 4). These results
suggest that the second mutation in 15A is not allelic to the
resistance gene in 11A. F.sub.2:3 families were not screened as
over 330 plants within each family would have to be screened in
order to ensure an adequate power of test (Hanson, 1959 Agron. J.
51:711-716).
[0105] Three independent resistance loci have been identified, each
with an allele conferring resistance to imazamox. Recommended rules
for gene locus and allele symbolization have been published
(McIntosh et al., 1998 Catalogue of Gene Symbols. Volume 5,
Proceedings of the 9.sup.th International Wheat Genetics Symposium.
Saskatoon, Saskatchewan). Non-allelic gene loci of an enzyme that
catalyze the same reaction should be given the same symbol,
corresponding to the trivial name of the enzyme. The trivial name
for AHAS is ALS. Absent data to assign the loci to specific
chromosomes and genomes, they should be designated in sequential
series. The designation of the phenotype observed when changes
occur in the gene resulting in a new allele should reflect that
phenotype. Thus, it is proposed that the FS-4 imidazolinone
resistance allele be designated as Imi1 and the locus it is at
designated as Als1. Imi stands for imidazolinone resistance. This
designation indicates that the gene is a dominant trait and it is
the first allele identified. Segregating F.sub.2 and F.sub.2:3
population data suggests that 15A and 11A carry two new independent
resistance alleles at different loci (FIGS. 2 and 3). The
designations for these alleles are Imi2 for the 11A mutation at the
Als2 locus and Imi3 for the second 15A mutation at the Als3
locus.
[0106] Identified herein are three independently segregating
alleles conferring resistance to imazamox, namely Imi1 (1A, 9A,
10A, 15A and 16A), Imi2 (11A), and Imi3 (15A). It is proposed that
each of the three identified alleles are associated with a
different structural gene coding for herbicide-insensitive forms of
AHAS. Since wheat is a hexaploid, multiple AHAS loci would be
expected. Other polyploid species have been found to have more than
one copy of AHAS. In Nicotiana tabacum, an allotetraploid, two AHAS
genes have been identified and characterized (Mazur et al. 1987).
Chaleff and Ray (1984) identified two independently segregating
sulfonylurea resistance alleles in Nicotiana tabacum, each coding
for an altered form of AHAS. Zea mays possesses two constitutively
expressed identical AHAS genes (Fang et al., 1992 Plant Mol. Biol.
18:1185-1187). In allotetraploid Brassica napus and Gossypium
hirsutum, an AHAS multi-gene family consisting of five and six
members, respectively, is present (Rutledge et al., 1991 Mol. Gen.
Genet. 229:31-40; Grula et al., 1995 Plant Mol. Biol. 28:837-846).
Higher levels of resistance to herbicides have been observed in
polyploid species when multiple resistance alleles are present.
Swanson et al. (1989 Theor. Appl. Gen. 78:525-530) combined two
unique imidazolinone resistance alleles from two homozygous
Brassica napus lines resulting in progeny with a higher level of
resistance than either homozygous line alone. Creason and Chaleff
(1988 Theor. Appl. Genet. 76:177-182) identified Nicotiana tabacum
plants homozygous for two mutations that conferred resistance to
sulfonylureas. Plants homozygous for both mutations were five-fold
more resistant to foliar applications of chlorsulfuron than were
plants homozygous for each single mutation. The present invention
proposes producing increased levels of resistance to an
imidazolinone herbicide in wheat by combining any two or all three
resistance alleles.
Example 5
Tolerance to Imidazolinone Herbicides in Teal11A, Teal15A and
Teal11A/15A Hybrid
[0107] The increased tolerance exhibited by Teal 11A and Teal 15A
to 20 grams per hectare of imazamox has been exemplified in
previous examples by the ability to distinguish tolerant from
susceptible parental and segregant plants in inheritance studies.
Teal 11A has been shown to confer similar levels of tolerance to
imidazolinone herbicides to that conferred by the FS4 mutation in
Fidel in various greenhouse and field comparisons. The similarity
in tolerance is also reflected in comparing the in vitro activity
of AHAS extracted from tolerant plants. This is possible because
the tolerance in Teal 11A, Teal 15A, and FS4 is due to a mutation
in the AHAS enzyme rendering it resistant to inhibition by
imidazolinone herbicides. FIG. 6 indicates that the activity of
AHAS enzyme extracted from Teal 11A and BW755, a line containing
FS4, changes similarly as the rate of imazamox increases, and both
have a higher percentage of active (resistant) enzyme at the
highest concentration of imazamox than does the wild type check,
Teal.
[0108] The presence of two IMI nucleic acids in Teal 15A provides
increased tolerance to imidazolinone herbicides compared to a line
such as BW755 carrying only one IMI nucleic acid. This increased
tolerance is reflected both in less injury at higher herbicide
rates, but in having more uninhibited AHAS enzyme activity. FIG. 7
illustrates that a 10.times.rate of imazamox (200 g/ha), all
treated one gene plants were injured, while no two gene plants were
injured. At all concentrations of imazamox in an in vitro assay of
AHAS activity (FIG. 6), but particularly at the highest
concentrations, Teal ISA had a higher percentage of active
(resistant) enzyme than did either of the single gene lines,
Teal11A and BW755.
[0109] Combining three non-allelic genes each conferring tolerance
to imidazolinone herbicides results in greater tolerance than with
only two non-allelic genes (FIG. 7). At a 30.times. rate, or 600
g/ha of imazamox, over half of plants sustained no injury in a
still-segregating selfed population of Teal15A crossed with Teal
11A, while all plants of the homozygous population of Teal15A
sustained injury.
Sequence CWU 1
1
3511675DNATriticum aestivumCDS(2)..(1672)modified_base(764)unknown
nucleotide 1c gac gtc ttc gcc tac ccc ggc ggc gcc tcc atg gag atc
cac cag gcg 49 Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile
His Gln Ala 1 5 10 15ctg acg cgc tcg ccc gtc atc acc aac cac ctc
ttc cgc cac gag cag 97Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu
Phe Arg His Glu Gln 20 25 30ggg gag gcg ttc gcg gcg tcc ggc tac gcc
cgc gcg tcc ggc cgc gtc 145Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala
Arg Ala Ser Gly Arg Val 35 40 45ggc gtc tgc gtc gcc acc tcc ggc ccg
ggg gcc acc aac ctc gtc tcc 193Gly Val Cys Val Ala Thr Ser Gly Pro
Gly Ala Thr Asn Leu Val Ser 50 55 60gcg ctc gcc gac gcc ctc ctc gac
tcc atc ccc atg gtc gcc atc acg 241Ala Leu Ala Asp Ala Leu Leu Asp
Ser Ile Pro Met Val Ala Ile Thr65 70 75 80ggc cag gtc ccc cgc cgc
atg atc ggc acg gac gcg ttc cag gag acg 289Gly Gln Val Pro Arg Arg
Met Ile Gly Thr Asp Ala Phe Gln Glu Thr 85 90 95ccc ata gtg gag gtc
acg cgc tcc atc acc aag cac aac tac ctg gtc 337Pro Ile Val Glu Val
Thr Arg Ser Ile Thr Lys His Asn Tyr Leu Val 100 105 110ctt gac gtg
gag gat atc ccc cgc gtc atc cag gaa gcc ttc ttc ctt 385Leu Asp Val
Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe Leu 115 120 125gca
tcc tct ggc cgc ccg ggg ccg gtg cta gtt gat atc ccc aag gac 433Ala
Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile Pro Lys Asp 130 135
140atc cag cag cag atg gct gtg ccc gtc tgg gac act cca atg agt ttg
481Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser
Leu145 150 155 160cca ggg tac atc gcc cgc ctg ccc aag cca cca tct
act gaa tcg ctt 529Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser
Thr Glu Ser Leu 165 170 175gag cag gtc ctg cgt ctg gtt ggc gag tca
cgg cgc cca att ctg tat 577Glu Gln Val Leu Arg Leu Val Gly Glu Ser
Arg Arg Pro Ile Leu Tyr 180 185 190gtt ggt ggt ggc tgc gct gcg tct
ggc gag gag ttg cgc cgc ttt gtt 625Val Gly Gly Gly Cys Ala Ala Ser
Gly Glu Glu Leu Arg Arg Phe Val 195 200 205gag ctt act ggg att cca
gtt aca act act ctg atg ggc ctt ggc aac 673Glu Leu Thr Gly Ile Pro
Val Thr Thr Thr Leu Met Gly Leu Gly Asn 210 215 220ttc ccc agc gac
gac cca ctg tct ctg cgc atg ctt ggg atg cat ggc 721Phe Pro Ser Asp
Asp Pro Leu Ser Leu Arg Met Leu Gly Met His Gly225 230 235 240act
gtg tat gca aat tat gca gta gat aag gct gac ctg ttg ntc gca 769Thr
Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu Ile Ala 245 250
255ttt ggt gtg cgg ttt gat gat cgt gtg act ggg aaa atc gag gct ttt
817Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala Phe
260 265 270gca agc agg tcc aag att gng cac att gac att gac cca gct
gag att 865Ala Ser Arg Ser Lys Ile Glu His Ile Asp Ile Asp Pro Ala
Glu Ile 275 280 285ggc aag aac aag cag cca cat gtc tcc att tgt gca
gat gtt aan ctt 913Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala
Asp Val Lys Leu 290 295 300gct tta cag ggg ttg aat gat cta tta aat
ggg agc aaa gca caa cag 961Ala Leu Gln Gly Leu Asn Asp Leu Leu Asn
Gly Ser Lys Ala Gln Gln305 310 315 320ggt ctg gat ttt ggt cca tgg
cac aag gag ttg gat cag can aan agg 1009Gly Leu Asp Phe Gly Pro Trp
His Lys Glu Leu Asp Gln Gln Xaa Arg 325 330 335gag ttt cct cta gga
ttc aag act ttt ggc gag gcc atc ccg ccg caa 1057Glu Phe Pro Leu Gly
Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro Gln 340 345 350tat gct atc
cag gta ctg gat gag ctg aca aaa ggg gag gcg atc att 1105Tyr Ala Ile
Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile Ile 355 360 365gcc
act ggt gtt ggg cag cac cag atg tgg gcg gct cag tat tac act 1153Ala
Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln Tyr Tyr Thr 370 375
380tac aag cgg cca cgg cag tgg ctg tct tcg tct ggt ttg ggg gca atg
1201Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala
Met385 390 395 400gga ttt ggg tta cca gct gca gct ggc gct gct gtg
gcc aac cca ggt 1249Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val
Ala Asn Pro Gly 405 410 415gtt aca gtt gtt gac att gat ggt gat ggt
agt ttc ctc atg aac att 1297Val Thr Val Val Asp Ile Asp Gly Asp Gly
Ser Phe Leu Met Asn Ile 420 425 430cag gag ttg gcg ttg atc cgc att
gag aac ctc cca gtg aag gtg atg 1345Gln Glu Leu Ala Leu Ile Arg Ile
Glu Asn Leu Pro Val Lys Val Met 435 440 445ata ttg aac aac cag cat
ctg gga atg gtg gtg cag tgg gag gat agg 1393Ile Leu Asn Asn Gln His
Leu Gly Met Val Val Gln Trp Glu Asp Arg 450 455 460ttt tac aag gcc
aat cgg gcg cac aca tac ctt ggc aac cca gaa aat 1441Phe Tyr Lys Ala
Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu Asn465 470 475 480gag
agt gag ata tat cca gat ttt gtg acg att gct aaa gga ttc aac 1489Glu
Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly Phe Asn 485 490
495gtt cca gca gtt cga gtg acg aag aag agc gaa gtc act gca gca atc
1537Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala Ile
500 505 510aag aag atg ctt gag acc cca ggg cca tac ttg ttg gat atc
ata gtc 1585Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile
Ile Val 515 520 525ccg cat cag gag cac gtg ctg cct atg atc cca aac
ggt ggt gct ttc 1633Pro His Gln Glu His Val Leu Pro Met Ile Pro Asn
Gly Gly Ala Phe 530 535 540aag gac atg atc atg gag ggt gat ggc agg
acc tcg tac tga 1675Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser
Tyr545 550 5552557PRTTriticum aestivum 2Asp Val Phe Ala Tyr Pro Gly
Gly Ala Ser Met Glu Ile His Gln Ala1 5 10 15Leu Thr Arg Ser Pro Val
Ile Thr Asn His Leu Phe Arg His Glu Gln 20 25 30Gly Glu Ala Phe Ala
Ala Ser Gly Tyr Ala Arg Ala Ser Gly Arg Val 35 40 45Gly Val Cys Val
Ala Thr Ser Gly Pro Gly Ala Thr Asn Leu Val Ser 50 55 60Ala Leu Ala
Asp Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile Thr65 70 75 80Gly
Gln Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu Thr 85 90
95Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu Val
100 105 110Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe
Phe Leu 115 120 125Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp
Ile Pro Lys Asp 130 135 140Ile Gln Gln Gln Met Ala Val Pro Val Trp
Asp Thr Pro Met Ser Leu145 150 155 160Pro Gly Tyr Ile Ala Arg Leu
Pro Lys Pro Pro Ser Thr Glu Ser Leu 165 170 175Glu Gln Val Leu Arg
Leu Val Gly Glu Ser Arg Arg Pro Ile Leu Tyr 180 185 190Val Gly Gly
Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe Val 195 200 205Glu
Leu Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly Asn 210 215
220Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His
Gly225 230 235 240Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp
Leu Leu Ile Ala 245 250 255Phe Gly Val Arg Phe Asp Asp Arg Val Thr
Gly Lys Ile Glu Ala Phe 260 265 270Ala Ser Arg Ser Lys Ile Glu His
Ile Asp Ile Asp Pro Ala Glu Ile 275 280 285Gly Lys Asn Lys Gln Pro
His Val Ser Ile Cys Ala Asp Val Lys Leu 290 295 300Ala Leu Gln Gly
Leu Asn Asp Leu Leu Asn Gly Ser Lys Ala Gln Gln305 310 315 320Gly
Leu Asp Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys Arg 325 330
335Glu Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro Gln
340 345 350Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala
Ile Ile 355 360 365Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala
Gln Tyr Tyr Thr 370 375 380Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser
Ser Gly Leu Gly Ala Met385 390 395 400Gly Phe Gly Leu Pro Ala Ala
Ala Gly Ala Ala Val Ala Asn Pro Gly 405 410 415Val Thr Val Val Asp
Ile Asp Gly Asp Gly Ser Phe Leu Met Asn Ile 420 425 430Gln Glu Leu
Ala Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val Met 435 440 445Ile
Leu Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp Arg 450 455
460Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu
Asn465 470 475 480Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala
Lys Gly Phe Asn 485 490 495Val Pro Ala Val Arg Val Thr Lys Lys Ser
Glu Val Thr Ala Ala Ile 500 505 510Lys Lys Met Leu Glu Thr Pro Gly
Pro Tyr Leu Leu Asp Ile Ile Val 515 520 525Pro His Gln Glu His Val
Leu Pro Met Ile Pro Asn Gly Gly Ala Phe 530 535 540Lys Asp Met Ile
Met Glu Gly Asp Gly Arg Thr Ser Tyr545 550 55531672DNATriticum
aestivumCDS(2)..(1669) 3c gac gtc ttc gcc tac cct ggc ggc gcg tcc
atg gag atc cac cag gcg 49 Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser
Met Glu Ile His Gln Ala 1 5 10 15ctg acg cgc tcg cca gtc atc acc
aac cac ctc ttc cgc cac gag cag 97Leu Thr Arg Ser Pro Val Ile Thr
Asn His Leu Phe Arg His Glu Gln 20 25 30ggg gag gcg ttc gcg gcg tcc
ggg tac gcc cgc gcg tcc ggc cgc gtc 145Gly Glu Ala Phe Ala Ala Ser
Gly Tyr Ala Arg Ala Ser Gly Arg Val 35 40 45ggc gtc tgc gtc gcc acc
tcc ggc ccg ggg gcc acc aac ctc gtc tcc 193Gly Val Cys Val Ala Thr
Ser Gly Pro Gly Ala Thr Asn Leu Val Ser 50 55 60gcg ctc gcc gac gct
ctc ctc gac tcc atc ccc atg gtc gcc atc acg 241Ala Leu Ala Asp Ala
Leu Leu Asp Ser Ile Pro Met Val Ala Ile Thr65 70 75 80ggc cag gtc
ccc cgc cgc atg atc ggc acg gat gcg ttc cag gag acg 289Gly Gln Val
Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu Thr 85 90 95ccc atc
gtg gag gtc acg cgc tcc atc acc aag cac aac tac ctg gtc 337Pro Ile
Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu Val 100 105
110ctt gac gtg gag gat atc ccc cgc gtc atc cag gaa gcc ttc ttc ctc
385Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe Leu
115 120 125gca tcc tct ggc cgc ccg ggg ccg gtg ctg gtt gat atc ccc
aag gac 433Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile Pro
Lys Asp 130 135 140atc cag cag cag atg gct gtg cct gtc tgg gac acg
ccg atg agt ttg 481Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr
Pro Met Ser Leu145 150 155 160cca ggg tac atc gcc cgc ctg ccc aag
cca cca tct act gaa tcg ctt 529Pro Gly Tyr Ile Ala Arg Leu Pro Lys
Pro Pro Ser Thr Glu Ser Leu 165 170 175gag cag gtc ctg cgt ctg gtt
ggc gag tca cgg cgc cca att ctg tat 577Glu Gln Val Leu Arg Leu Val
Gly Glu Ser Arg Arg Pro Ile Leu Tyr 180 185 190gtt ggt ggt ggc tgc
gct gca tct ggt gag gag ttg cgc cgc ttt gtt 625Val Gly Gly Gly Cys
Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe Val 195 200 205gag ctc act
ggg att cca gtt aca act act ctt atg ggc ctt ggc aac 673Glu Leu Thr
Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly Asn 210 215 220ttc
ccc agt gac gac cca ctg tct ctg cgc atg ctg ggg atg cat ggc 721Phe
Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His Gly225 230
235 240act gtg tat gca aat tat gca gta gat aag gct gac ctg ttg ctt
gca 769Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu Leu
Ala 245 250 255ttt ggt gtg cgg ttt gat gat cgt gtg acc ggg aaa atc
gag gct ttt 817Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys Ile
Glu Ala Phe 260 265 270gca agc agg tcc aag att gtg cac att gac att
gac cca gct gag att 865Ala Ser Arg Ser Lys Ile Val His Ile Asp Ile
Asp Pro Ala Glu Ile 275 280 285ggc aag aac aag cag cca cat gtc tcc
att tgt gca gat gtt aag ctt 913Gly Lys Asn Lys Gln Pro His Val Ser
Ile Cys Ala Asp Val Lys Leu 290 295 300gct tta cag ggg ttg aat gct
cta tta aat ggg agc aaa gca caa cag 961Ala Leu Gln Gly Leu Asn Ala
Leu Leu Asn Gly Ser Lys Ala Gln Gln305 310 315 320ggt ctg gat ttt
ggt cca tgg cac aag gag ttg gat cag cag aag agg 1009Gly Leu Asp Phe
Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys Arg 325 330 335gag ttt
cct cta gga ttc aag act ttt ggt gag gcc atc ccg ccg caa 1057Glu Phe
Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro Gln 340 345
350tat gct atc cag gta ctg gat gag ctg aca aaa ggg gag gcg atc att
1105Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile Ile
355 360 365gcc acc ggt gtt ggg cag cat cag atg tgg gcg gct cag tat
tac act 1153Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln Tyr
Tyr Thr 370 375 380tac aag cgg cca cgg cag tgg ctg tct tca tcc ggt
ttg ggt gca atg 1201Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser Gly
Leu Gly Ala Met385 390 395 400gga ttt ggg ttg cca gct gca gct ggc
gct gct gtg gcc aac cca ggt 1249Gly Phe Gly Leu Pro Ala Ala Ala Gly
Ala Ala Val Ala Asn Pro Gly 405 410 415gtt aca gtt gtt gac att gat
ggg gat ggt agt ttc ctc atg aac att 1297Val Thr Val Val Asp Ile Asp
Gly Asp Gly Ser Phe Leu Met Asn Ile 420 425 430cag gag ttg gcg ttg
atc cgt att gag aac ctc cca gtg aag gtg atg 1345Gln Glu Leu Ala Leu
Ile Arg Ile Glu Asn Leu Pro Val Lys Val Met 435 440 445ata ttg aac
aac cag cat ctg gga atg gtg gtg cag tgg gag gat agg 1393Ile Leu Asn
Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp Arg 450 455 460ttt
tac aag gcc aac cgg gcg cac aca tac ctt ggc aac cca gaa aat 1441Phe
Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu Asn465 470
475 480gag agt gag ata tat cca gat ttt gtg acg att gct aaa gga ttc
aac 1489Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly Phe
Asn 485 490 495gtt ccg gca gtt cgt gtg acg aag aag agc gaa gtc act
gca gca atc 1537Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val Thr
Ala Ala Ile 500 505 510aag aag atg ctt gag acc cca ggg cca tac ttg
ttg gat atc att gtc 1585Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu
Leu Asp Ile Ile Val 515 520 525ccg cat cag gag cac gtg ctg cct atg
atc cca aac ggt ggt gct ttt 1633Pro His Gln Glu His Val Leu Pro Met
Ile Pro Asn Gly Gly Ala Phe 530 535 540aag gac atg atc atg gag ggt
gat ggc agg acc tcg tac 1672Lys Asp Met Ile Met Glu Gly Asp Gly Arg
Thr Ser545 550 5554556PRTTriticum aestivum 4Asp Val Phe Ala Tyr Pro
Gly Gly Ala Ser Met Glu Ile His Gln Ala1 5 10 15Leu Thr Arg Ser Pro
Val Ile Thr Asn His Leu Phe Arg His Glu Gln 20 25 30Gly Glu Ala Phe
Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly Arg Val 35 40 45Gly Val Cys
Val Ala Thr Ser Gly Pro Gly Ala Thr Asn Leu Val Ser 50 55 60Ala Leu
Ala Asp Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile Thr65 70 75
80Gly Gln Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu Thr
85 90 95Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu
Val
100 105 110Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe
Phe Leu 115 120 125Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp
Ile Pro Lys Asp 130 135 140Ile Gln Gln Gln Met Ala Val Pro Val Trp
Asp Thr Pro Met Ser Leu145 150 155 160Pro Gly Tyr Ile Ala Arg Leu
Pro Lys Pro Pro Ser Thr Glu Ser Leu 165 170 175Glu Gln Val Leu Arg
Leu Val Gly Glu Ser Arg Arg Pro Ile Leu Tyr 180 185 190Val Gly Gly
Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe Val 195 200 205Glu
Leu Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly Asn 210 215
220Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His
Gly225 230 235 240Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp
Leu Leu Leu Ala 245 250 255Phe Gly Val Arg Phe Asp Asp Arg Val Thr
Gly Lys Ile Glu Ala Phe 260 265 270Ala Ser Arg Ser Lys Ile Val His
Ile Asp Ile Asp Pro Ala Glu Ile 275 280 285Gly Lys Asn Lys Gln Pro
His Val Ser Ile Cys Ala Asp Val Lys Leu 290 295 300Ala Leu Gln Gly
Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala Gln Gln305 310 315 320Gly
Leu Asp Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys Arg 325 330
335Glu Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro Gln
340 345 350Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala
Ile Ile 355 360 365Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala
Gln Tyr Tyr Thr 370 375 380Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser
Ser Gly Leu Gly Ala Met385 390 395 400Gly Phe Gly Leu Pro Ala Ala
Ala Gly Ala Ala Val Ala Asn Pro Gly 405 410 415Val Thr Val Val Asp
Ile Asp Gly Asp Gly Ser Phe Leu Met Asn Ile 420 425 430Gln Glu Leu
Ala Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val Met 435 440 445Ile
Leu Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp Arg 450 455
460Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu
Asn465 470 475 480Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala
Lys Gly Phe Asn 485 490 495Val Pro Ala Val Arg Val Thr Lys Lys Ser
Glu Val Thr Ala Ala Ile 500 505 510Lys Lys Met Leu Glu Thr Pro Gly
Pro Tyr Leu Leu Asp Ile Ile Val 515 520 525Pro His Gln Glu His Val
Leu Pro Met Ile Pro Asn Gly Gly Ala Phe 530 535 540Lys Asp Met Ile
Met Glu Gly Asp Gly Arg Thr Ser545 550 55551524DNATriticum aestivum
5gtctgcgtcg ccacctccgg cccgggggcc accaacctcg tctccgcgct cgccgacgcc
60ctcctcgact ccatccccat ggtcgccatc acgggccagg tcccccgccg catgatcggc
120acggacgcgt tccaggagac gcccatagtg gaggtcacgc gctccatcac
caagcacaac 180tacctggtcc ttgacgtgga ggatatcccc cgcgtcatcc
aggaagcctt cttccttgca 240tcctctggcc gcccggggcc ggtgctagtt
gatatcccca aggacatcca gcagcagatg 300gctgtgcccg tctgggacac
tccaatgagt ttgccagggt acatcgcccg cctgcccaag 360ccaccatcta
ctgaatcgct tgagcaggtc ctgcgtctgg ttggcgagtc acggcgccca
420attctgtatg ttggtggtgg ctgcgctgcg tctggcgagg agttgcgccg
ctttgttgag 480cttactggga ttccagttac aactactctg atgggccttg
gcaacttccc cagcgacgac 540ccactgtctc tgcgcatgct tgggatgcat
ggcactgtgt atgcaaatta tgcagtagat 600aaggctgacc tgttgctcgc
atttggtgtg cggtttgatg atcgtgtgac tgggaaaatc 660gaggcttttg
caagcaggtc caagattgtg cacattgaca ttgacccagc tgagattggc
720aagaacaagc agccacatgt ctccatttgt gcagatgtta agcttgcttt
acaggggttg 780aatgatctat taaatgggag caaagcacaa cagggtctgg
attttggtcc atggcacaag 840gagttggatc agcagaagag ggagtttcct
ctaggattca agacttttgg cgaggccatc 900ccgccgcaat atgctatcca
ggtactggat gagctgacaa aaggggaggc gatcattgcc 960actggtgttg
ggcagcacca gatgtgggcg gctcagtatt acacttacaa gcggccacgg
1020cagtggctgt cttcgtctgg tttgggggca atgggatttg ggttaccagc
tgcagctggc 1080gctgctgtgg ccaacccagg tgttacagtt gttgacattg
atggtgatgg tagtttcctc 1140atgaacattc aggagttggc gttgatccgc
attgagaacc tcccagtgaa ggtgatgata 1200ttgaacaacc agcatctggg
aatggtggtg cagtgggagg ataggtttta caaggccaat 1260cgggcgcaca
cataccttgg caacccagaa aatgagagtg agatatatcc agattttgtg
1320acgattgcta aaggattcaa cgttccagca gttcgagtga cgaagaagag
cgaagtcact 1380gcagcaatca agaagatgct tgagacccca gggccatact
tgttggatat catagtcccg 1440catcaggagc acgtgctgcc tatgatccca
agcggtggtg ctttcaagga catgatcatg 1500gagggtgatg gcaggacctc gtac
152461524DNATriticum aestivum 6gtctgcgtcg ccacctccgg cccgggggcc
accaacctcg tctccgcgct cgccgacgct 60ctcctcgact ccatccccat ggtcgccatc
acgggccagg tcccccgccg catgatcggc 120acggatgcgt tccaggagac
gcccatcgtg gaggtcacgc gctccatcac caagcacaac 180tacctggtcc
ttgacgtgga ggatatcccc cgcgtcatcc aggaagcctt cttcctcgca
240tcctctggcc gcccggggcc ggtgctggtt gatatcccca aggacatcca
gcagcagatg 300gctgtgcctg tctgggacac gccgatgagt ttgccagggt
acatcgcccg cctgcccaag 360ccaccatcta ctgaatcgct tgagcaggtc
ctgcgtctgg ttggcgagtc acggcgccca 420attctgtatg ttggtggtgg
ctgcgctgca tctggtgagg agttgcgccg ctttgttgag 480ctcactggga
ttccagttac aactactctt atgggccttg gcaacttccc cagtgacgac
540ccactgtctc tgcgcatgct ggggatgcat ggcactgtgt atgcaaatta
tgcagtagat 600aaggctgacc tgttgcttgc atttggtgtg cggtttgatg
atcgtgtgac cgggaaaatc 660gaggcttttg caagcaggtc caagattgtg
cacattgaca ttgacccagc tgagattggc 720aagaacaagc agccacatgt
ctccatttgt gcagatgtta agcttgcttt acaggggttg 780aatgctctat
taaatgggag caaagcacaa cagggtctgg attttggtcc atggcacaag
840gagttggatc agcagaagag ggagtttcct ctaggattca agacttttgg
tgaggccatc 900ccgccgcaat atgctatcca ggtactggat gagctgacaa
aaggggaggc gatcattgcc 960accggtgttg ggcagcatca gatgtgggcg
gctcagtatt acacttacaa gcggccacgg 1020cagtggctgt cttcatccgg
tttgggtgca atgggatttg ggttgccagc tgcagctggc 1080gctgctgtgg
ccaacccagg tgttacagtt gttgacattg atggggatgg tagtttcctc
1140atgaacattc aggagttggc gttgatccgt attgagaacc tcccagtgaa
ggtgatgata 1200ttgaacaacc agcatctggg aatggtggtg cagtgggagg
ataggtttta caaggccaac 1260cgggcgcaca cataccttgg caacccagaa
aatgagagtg agatatatcc agattttgtg 1320acgattgcta aaggattcaa
cgttccggca gttcgtgtga cgaagaagag cgaagtcact 1380gcagcaatca
agaagatgct tgagacccca gggccatact tgttggatat cattgtcccg
1440catcaggagc acgtgctgcc tatgatccca agcggtggtg cttttaagga
catgatcatg 1500gagggtgatg gcaggacctc gtac 152471524DNATriticum
aestivum 7gtctgcgtcg ccacctccgg cccgggggcc accaacctcg tctccgcgct
cgctgacgcc 60ctcctcgact ccatccccat ggtcgccatc acgggccagg tcccccgccg
catgatcggc 120acggacgcgt tccaggagac gcccatagtg gaggtcacgc
gctccatcac caagcacaac 180tacctggtcc ttgacgtgga ggatatcccc
cgcgtcatcc aggaagcctt cttcctcgcg 240tcctctggcc gcccggggcc
ggtgctggtt gatatcccca aggatatcca gcagcagatg 300gccgtgccta
tctgggacac gccgatgagt ttgccagggt acatcgcccg cctgcccaag
360ccaccatcta ctgaatcgct tgagcaggtc ctgcgtctgg ttggcgagtc
acggcgccca 420attctgtatg ttggtggtgg ctgcgctgca tccggcgagg
agttgcgccg ctttgttgag 480ctcactggga ttccggttac aactactctg
atgggccttg gcaacttccc cagcgacgac 540ccactgtctc tgcgcatgct
tgggatgcat ggcactgtgt atgcaaatta tgcagtcgat 600aaggctgacc
tgttgcttgc atttggtgtg cggtttgatg atcgcgtgac tgggaaaatc
660gaggcctttg caagcaggtc caagattgtg cacattgaca ttgacccagc
tgagattggc 720aagaacaagc agccacatgt ctccatttgt gcagatgtta
agcttgcttt acaggggttg 780aatgctctat taaatgggag caaagcacaa
cagggtctgg attttggtcc atggcacaag 840gagttggatc agcagaagag
ggagtttcct ctaggattca agacttttgg cgaggccatc 900ccgccgcaat
atgctatcca ggtactggat gagctgacaa aaggggaggc gatcattgct
960actggtgttg ggcagcacca gatgtgggcg gctcagtatt acacttacaa
gcggccacgg 1020cagtggctgt cttcgtctgg tttgggggca atgggatttg
ggttaccagc tgcagctggc 1080gctgctgtgg ccaacccagg tgttacagtt
gttgacattg atggagatgg tagtttcctc 1140atgaacattc aggagttggc
attgatccgt attgagaacc tccctgtgaa ggtgatgata 1200ttgaacaacc
agcatctggg aatggtggtg caatgggagg ataggtttta caaggccaat
1260cgggcgcaca cataccttgg caacccagaa aatgagagtg agatatatcc
agattttgtg 1320acgattgcta aaggattcaa cgttccggca gttcgtgtga
cgaagaagag cgaagtcact 1380gcagcaatca agaagatgct tgagacccca
gggccatact tgttggatat catcgtcccg 1440catcaggagc acgtgctgcc
tatgatccca agcggtggtg ctttcaagga catgatcatg 1500gagggtgatg
gcaggacctc gtac 1524813PRTArtificial SequenceDescription of
Artificial Sequence Illustrative conserved peptide sequence 8Ala
Ile Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr1 5
1094PRTArtificial SequenceDescription of Artificial Sequence
Illustrative conserved peptide sequence 9Gln Trp Glu
Asp11019PRTArtificial SequenceDescription of Artificial Sequence
Illustrative conserved peptide sequence 10Val Phe Ala Tyr Pro Gly
Gly Ala Ser Met Glu Ile His Gln Ala Leu1 5 10 15Thr Arg
Ser116PRTArtificial SequenceDescription of Artificial Sequence
Illustrative conserved peptide sequence 11Ala Phe Gln Glu Thr Pro1
5125PRTArtificial SequenceDescription of Artificial Sequence
Illustrative conserved peptide sequence 12Ile Pro Ser Gly Gly1
5131935DNAOryza sp.CDS(1)..(1932) 13atg gct acg acc gcc gcg gcc gcg
gcc gcc gcc ctg tcc gcc gcc gcg 48Met Ala Thr Thr Ala Ala Ala Ala
Ala Ala Ala Leu Ser Ala Ala Ala1 5 10 15acg gcc aag acc ggc cgt aag
aac cac cag cga cac cac gtc ctt ccc 96Thr Ala Lys Thr Gly Arg Lys
Asn His Gln Arg His His Val Leu Pro 20 25 30gct cga ggc cgg gtg ggg
gcg gcg gcg gtc agg tgc tcg gcg gtg tcc 144Ala Arg Gly Arg Val Gly
Ala Ala Ala Val Arg Cys Ser Ala Val Ser 35 40 45ccg gtc acc ccg ccg
tcc ccg gcg ccg ccg gcc acg ccg ctc cgg ccg 192Pro Val Thr Pro Pro
Ser Pro Ala Pro Pro Ala Thr Pro Leu Arg Pro 50 55 60tgg ggg ccg gcc
gag ccc cgc aag ggc gcg gac atc ctc gtg gag gcg 240Trp Gly Pro Ala
Glu Pro Arg Lys Gly Ala Asp Ile Leu Val Glu Ala65 70 75 80ctg gag
cgg tgc ggc gtc agc gac gtg ttc gcc tac ccg ggc ggc gcg 288Leu Glu
Arg Cys Gly Val Ser Asp Val Phe Ala Tyr Pro Gly Gly Ala 85 90 95tcc
atg gag atc cac cag gcg ctg acg cgc tcc ccg gtc atc acc aac 336Ser
Met Glu Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile Thr Asn 100 105
110cac ctc ttc cgc cac gag cag ggc gag gcg ttc gcg gcg tcc ggg tac
384His Leu Phe Arg His Glu Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr
115 120 125gcg cgc gcg tcc ggc cgc gtc ggg gtc tgc gtc gcc acc tcc
ggc ccc 432Ala Arg Ala Ser Gly Arg Val Gly Val Cys Val Ala Thr Ser
Gly Pro 130 135 140ggg gca acc aac ctc gtg tcc gcg ctc gcc gac gcg
ctg ctc gac tcc 480Gly Ala Thr Asn Leu Val Ser Ala Leu Ala Asp Ala
Leu Leu Asp Ser145 150 155 160gtc ccg atg gtc gcc atc acg ggc cag
gtc ccc cgc cgc atg atc ggc 528Val Pro Met Val Ala Ile Thr Gly Gln
Val Pro Arg Arg Met Ile Gly 165 170 175acc gac gcc ttc cag gag acg
ccc ata gtc gag gtc acc cgc tcc atc 576Thr Asp Ala Phe Gln Glu Thr
Pro Ile Val Glu Val Thr Arg Ser Ile 180 185 190acc aag cac aat tac
ctt gtc ctt gat gtg gag gac atc ccc cgc gtc 624Thr Lys His Asn Tyr
Leu Val Leu Asp Val Glu Asp Ile Pro Arg Val 195 200 205ata cag gaa
gcc ttc ttc ctc gcg tcc tcg ggc cgt cct ggc ccg gtg 672Ile Gln Glu
Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val 210 215 220ctg
gtc gac atc ccc aag gac atc cag cag cag atg gcc gtg ccg gtc 720Leu
Val Asp Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val Pro Val225 230
235 240tgg gac acc tcg atg aat cta cca ggg tac atc gca cgc ctg ccc
aag 768Trp Asp Thr Ser Met Asn Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys 245 250 255cca ccc gcg aca gaa ttg ctt gag cag gtc ttg cgt ctg
gtt ggc gag 816Pro Pro Ala Thr Glu Leu Leu Glu Gln Val Leu Arg Leu
Val Gly Glu 260 265 270tca cgg cgc ccg att ctc tat gtc ggt ggt ggc
tgc tct gca tct ggt 864Ser Arg Arg Pro Ile Leu Tyr Val Gly Gly Gly
Cys Ser Ala Ser Gly 275 280 285gac gaa ttg cgc tgg ttt gtt gag ctg
act ggt atc cca gtt aca acc 912Asp Glu Leu Arg Trp Phe Val Glu Leu
Thr Gly Ile Pro Val Thr Thr 290 295 300act ctg atg ggc ctc ggc aat
ttc ccc agt gac gac ccg ttg tcc ctg 960Thr Leu Met Gly Leu Gly Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu305 310 315 320cgc atg ctt ggg
atg cat ggc acg gtg tac gca aat tat gcc gtg gat 1008Arg Met Leu Gly
Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp 325 330 335aag gct
gac ctg ttg ctt gcg ttt ggt gtg cgg ttt gat gat cgt gtg 1056Lys Ala
Asp Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp Arg Val 340 345
350aca ggg aaa att gag gct ttt gca agc agg gcc aag att gtg cac att
1104Thr Gly Lys Ile Glu Ala Phe Ala Ser Arg Ala Lys Ile Val His Ile
355 360 365gac att gat cca gca gag att gga aag aac aag caa cca cat
gtg tca 1152Asp Ile Asp Pro Ala Glu Ile Gly Lys Asn Lys Gln Pro His
Val Ser 370 375 380att tgc gca gat gtt aag ctt gct tta cag ggc ttg
aat gct ctg cta 1200Ile Cys Ala Asp Val Lys Leu Ala Leu Gln Gly Leu
Asn Ala Leu Leu385 390 395 400caa cag agc aca aca aag aca agt tct
gat ttt agt gca tgg cac aat 1248Gln Gln Ser Thr Thr Lys Thr Ser Ser
Asp Phe Ser Ala Trp His Asn 405 410 415gag ttg gac cag cag aag agg
gag ttt cct ctg ggg tac aaa act ttt 1296Glu Leu Asp Gln Gln Lys Arg
Glu Phe Pro Leu Gly Tyr Lys Thr Phe 420 425 430ggt gaa gag atc cca
ccg caa tat gcc att cag gtg ctg gat gag ctg 1344Gly Glu Glu Ile Pro
Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu 435 440 445acg aaa ggt
gag gca atc atc gct act ggt gtt ggg cag cac cag atg 1392Thr Lys Gly
Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met 450 455 460tgg
gcg gca caa tat tac acc tac aag cgg cca cgg cag tgg ctg tct 1440Trp
Ala Ala Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser465 470
475 480tcg gct ggt ctg ggc gca atg gga ttt ggg ctg cct gct gca gct
ggt 1488Ser Ala Gly Leu Gly Ala Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly 485 490 495gct tct gtg gct aac cca ggt gtc aca gtt gtt gat att
gat ggg gat 1536Ala Ser Val Ala Asn Pro Gly Val Thr Val Val Asp Ile
Asp Gly Asp 500 505 510ggt agc ttc ctc atg aac att cag gag ctg gca
ttg atc cgc att gag 1584Gly Ser Phe Leu Met Asn Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu 515 520 525aac ctc cct gtg aag gtg atg gtg ttg
aac aac caa cat ttg ggt atg 1632Asn Leu Pro Val Lys Val Met Val Leu
Asn Asn Gln His Leu Gly Met 530 535 540gtg gtg caa tgg gag gat agg
ttt tac aag gcg aat agg gcg cat aca 1680Val Val Gln Trp Glu Asp Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr545 550 555 560tac ttg ggc aac
ccg gaa tgt gag agc gag ata tat cca gat ttt gtg 1728Tyr Leu Gly Asn
Pro Glu Cys Glu Ser Glu Ile Tyr Pro Asp Phe Val 565 570 575act att
gct aag ggg ttc aat att cct gca gtc cgt gta aca aag aag 1776Thr Ile
Ala Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr Lys Lys 580 585
590agt gaa gtc cgt gcc gcc atc aag aag atg ctc gag act cca ggg cca
1824Ser Glu Val Arg Ala Ala Ile Lys Lys Met Leu Glu Thr Pro Gly Pro
595 600 605tac ttg ttg gat atc atc gtc ccg cac cag gag cat gtg ctg
cct atg 1872Tyr Leu Leu Asp Ile Ile Val Pro His Gln Glu His Val Leu
Pro Met 610 615 620atc cca agt ggg ggc gca ttc aag gac atg atc ctg
gat ggt gat ggc 1920Ile Pro Ser Gly Gly Ala Phe Lys Asp Met Ile Leu
Asp Gly Asp Gly625 630 635 640agg act gtg tat taa 1935Arg Thr Val
Tyr14644PRTOryza sp. 14Met Ala Thr Thr Ala Ala Ala Ala Ala Ala Ala
Leu Ser Ala Ala Ala1 5 10 15Thr Ala Lys Thr Gly Arg Lys Asn His Gln
Arg His His Val Leu Pro 20 25 30Ala Arg Gly Arg Val Gly Ala Ala Ala
Val Arg Cys Ser Ala Val Ser 35 40 45Pro Val Thr Pro Pro Ser Pro Ala
Pro Pro Ala Thr Pro Leu Arg Pro 50 55 60Trp Gly Pro Ala Glu Pro Arg
Lys Gly Ala Asp Ile Leu Val Glu Ala65 70
75 80Leu Glu Arg Cys Gly Val Ser Asp Val Phe Ala Tyr Pro Gly Gly
Ala 85 90 95Ser Met Glu Ile His Gln Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn 100 105 110His Leu Phe Arg His Glu Gln Gly Glu Ala Phe Ala
Ala Ser Gly Tyr 115 120 125Ala Arg Ala Ser Gly Arg Val Gly Val Cys
Val Ala Thr Ser Gly Pro 130 135 140Gly Ala Thr Asn Leu Val Ser Ala
Leu Ala Asp Ala Leu Leu Asp Ser145 150 155 160Val Pro Met Val Ala
Ile Thr Gly Gln Val Pro Arg Arg Met Ile Gly 165 170 175Thr Asp Ala
Phe Gln Glu Thr Pro Ile Val Glu Val Thr Arg Ser Ile 180 185 190Thr
Lys His Asn Tyr Leu Val Leu Asp Val Glu Asp Ile Pro Arg Val 195 200
205Ile Gln Glu Ala Phe Phe Leu Ala Ser Ser Gly Arg Pro Gly Pro Val
210 215 220Leu Val Asp Ile Pro Lys Asp Ile Gln Gln Gln Met Ala Val
Pro Val225 230 235 240Trp Asp Thr Ser Met Asn Leu Pro Gly Tyr Ile
Ala Arg Leu Pro Lys 245 250 255Pro Pro Ala Thr Glu Leu Leu Glu Gln
Val Leu Arg Leu Val Gly Glu 260 265 270Ser Arg Arg Pro Ile Leu Tyr
Val Gly Gly Gly Cys Ser Ala Ser Gly 275 280 285Asp Glu Leu Arg Trp
Phe Val Glu Leu Thr Gly Ile Pro Val Thr Thr 290 295 300Thr Leu Met
Gly Leu Gly Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu305 310 315
320Arg Met Leu Gly Met His Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp
325 330 335Lys Ala Asp Leu Leu Leu Ala Phe Gly Val Arg Phe Asp Asp
Arg Val 340 345 350Thr Gly Lys Ile Glu Ala Phe Ala Ser Arg Ala Lys
Ile Val His Ile 355 360 365Asp Ile Asp Pro Ala Glu Ile Gly Lys Asn
Lys Gln Pro His Val Ser 370 375 380Ile Cys Ala Asp Val Lys Leu Ala
Leu Gln Gly Leu Asn Ala Leu Leu385 390 395 400Gln Gln Ser Thr Thr
Lys Thr Ser Ser Asp Phe Ser Ala Trp His Asn 405 410 415Glu Leu Asp
Gln Gln Lys Arg Glu Phe Pro Leu Gly Tyr Lys Thr Phe 420 425 430Gly
Glu Glu Ile Pro Pro Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu 435 440
445Thr Lys Gly Glu Ala Ile Ile Ala Thr Gly Val Gly Gln His Gln Met
450 455 460Trp Ala Ala Gln Tyr Tyr Thr Tyr Lys Arg Pro Arg Gln Trp
Leu Ser465 470 475 480Ser Ala Gly Leu Gly Ala Met Gly Phe Gly Leu
Pro Ala Ala Ala Gly 485 490 495Ala Ser Val Ala Asn Pro Gly Val Thr
Val Val Asp Ile Asp Gly Asp 500 505 510Gly Ser Phe Leu Met Asn Ile
Gln Glu Leu Ala Leu Ile Arg Ile Glu 515 520 525Asn Leu Pro Val Lys
Val Met Val Leu Asn Asn Gln His Leu Gly Met 530 535 540Val Val Gln
Trp Glu Asp Arg Phe Tyr Lys Ala Asn Arg Ala His Thr545 550 555
560Tyr Leu Gly Asn Pro Glu Cys Glu Ser Glu Ile Tyr Pro Asp Phe Val
565 570 575Thr Ile Ala Lys Gly Phe Asn Ile Pro Ala Val Arg Val Thr
Lys Lys 580 585 590Ser Glu Val Arg Ala Ala Ile Lys Lys Met Leu Glu
Thr Pro Gly Pro 595 600 605Tyr Leu Leu Asp Ile Ile Val Pro His Gln
Glu His Val Leu Pro Met 610 615 620Ile Pro Ser Gly Gly Ala Phe Lys
Asp Met Ile Leu Asp Gly Asp Gly625 630 635 640Arg Thr Val
Tyr151674DNATriticum
aestivumCDS(1)..(1671)modified_base(681)unknown nucleotide 15gtc
gac gtc ttc gcc tac ccc ggc ggc gcc tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg ccc gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggc tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gcc ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gac gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc ata gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctt gca tcc tct ggc cgc
ccg ggg ccg gtg cta gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg ccc gtc tgg gac act cca atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gcg tct ggc gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctt act ggg att cca gtt aca act act ctg
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccn agc gac gac cca ctg tct ctg
cgc atg ctt ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ctc 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255gca ttt ggt gtg cgg
ttt gat gat cgt gtg act ggg aaa atc gag gct 816Ala Phe Gly Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gca gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gat cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Asp Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggc gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc act ggt gtt ggg
cag cac cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tcg tct ggt ttg ggg gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg tta cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggt gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgc att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aat cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt cca gca gtt
cga gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc ata 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca agc ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535
540ttc aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55516557PRTTriticum aestivum 16Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu
Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230
235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Leu 245 250 255Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn
Asp Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Ser Gly Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu
Gly Asp Gly Arg Thr Ser545 550 555171674DNATriticum
aestivumCDS(1)..(1671)modified_base(718)..(719)unknown nucleotide
17gtc gac gtc ttc gcc tac ccc ggc ggc gcc tcc atg gag atc cac cag
48Val Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1
5 10 15gcg ctg acg cgc tcg ccc gtc atc acc aac cac ctc ttc cgc cac
gag 96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His
Glu 20 25 30cag ggg gag gcg ttc gcg gcg tcc ggc tac gcc cgc gcg tcc
ggc cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser
Gly Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc
aac ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr
Asn Leu Val 50 55 60tcc gcg ctc gcc gac gcc ctc ctc gac tcc atc ccc
atg gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro
Met Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc
acg gac gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly
Thr Asp Ala Phe Gln Glu 85 90 95acg ccc ata gtg gag gtc acg cgc tcc
atc acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser
Ile Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc
ccc cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile
Pro Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctt gca tcc tct ggc
cgc ccg ggg ccg gtg cta gtt gat atc ccc aag 432Leu Ala Ser Ser Gly
Arg Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag
cag cag atg gct gtg ccc gtc tgg gac act cca atg agt 480Asp Ile Gln
Gln Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155
160ttg cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg
528Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser
165 170 175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca
att ctg 576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro
Ile Leu 180 185 190tat gtt ggt ggt ggc tgc gct gcg tct ggc gag gag
ttg cgc cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu
Leu Arg Arg Phe 195 200 205gtt gag ctt act ggg att cca gtt aca act
act ctg atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr
Thr Leu Met Gly Leu Gly 210 215 220aac ttc ccc agc gac gac cca ctg
tct ctg cgc atg ctt ggg atg nnt 720Asn Phe Pro Ser Asp Asp Pro Leu
Ser Leu Arg Met Leu Gly Met Asn225 230 235 240ggc act gtg tat gca
aat tat gca gta gat aag gct gac ctg ttg ctc 768Gly Thr Val Tyr Ala
Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255gca ttt ggt
gtg cgg ttt gat gat cgt gtg act ggg aaa atc gag gct 816Ala Phe Gly
Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gca gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gat cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Asp Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggc gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc act ggt gtt ggg
cag cac cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tcg tct ggt ttg ggg gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg tta cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggt gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgc att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aat cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt cca gca gtt
cga gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc ata 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca aac ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Asn Gly Gly Ala 530 535
540ttc aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55518557PRTTriticum aestivum 18Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu
Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met Asn225 230
235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Leu 245 250 255Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn
Asp Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Asn Gly Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu
Gly Asp Gly Arg Thr Ser545 550 555191674DNATriticum
aestivumCDS(1)..(1671)modified_base(643)unknown nucleotide 19gtc
gac gtc ttc gcc tac ccc ggc ggc gcc tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg ccc gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggc tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gcc ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gac gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc ata gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctt gca tcc tct ggc cgc
ccg ggg ccg gtg cta gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg ccc gtc tgg gac act cca atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gcg tct ggc gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctt act ggg att nca gtt aca act act ctg
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Thr Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccc agc gac gac cca ctg tct ctg
cgc atg ctt ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ctc 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255gca ttt gnt gtg cgg
ttt gat gat cgt gtg act ggg aaa atc gag gct 816Ala Phe Asp Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aan aac aan cag cca cat gtc tcc att tgt gca gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gat cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Asp Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggc gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc act ggt gtt ggg
cag cac cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tcg tct ggt ttg ggg gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg tta cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggt gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgc att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aat cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt cca gca gtt
cga gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc ata 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca aac ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Asn Gly Gly Ala 530 535
540ttc aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55520557PRTTriticum aestivum 20Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu
Thr Gly Ile Thr Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230
235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Leu 245 250 255Ala Phe Asp Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn
Asp Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Asn Gly
Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr
Ser545 550 555211674DNATriticum
aestivumCDS(1)..(1671)modified_base(766)unknown nucleotide 21gtc
gac gtc ttc gcc tac ccc ggc ggc gcc tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg ccc gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggc tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gcc ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gac gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc ata gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctt gca tcc tct ggc cgc
ccg ggg ccg gtg cta gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg ccc gtc tgg gac act cca atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gcg tct ggc gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctt act ggg att cca gtt aca act act ctg
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccc agc gac gac cca ctg tct ctg
cgc atg ctt ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ntc 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Ile 245 250 255nca ttt ggt gtg cgg
ttt gat gat cgt gtg act ggg aaa atc gag gct 816Thr Phe Gly Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gcc gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gat cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Asp Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggc gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc act ggt gtt ggg
cag cac cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tcg tct ggt ttg ggg gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg tta cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggt gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgc att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aat cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt cca gca gtt
cga gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc ata 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca agc ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535
540ttc aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55522557PRTTriticum aestivum 22Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu
Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230
235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Ile 245 250 255Thr Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn
Asp Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Ser Gly Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu
Gly Asp Gly Arg Thr Ser545 550 555231677DNATriticum
aestivumCDS(1)..(1674)modified_base(766)unknown nucleotide 23gtc
gac gtc ttc gcc tac ccc ggc ggc gcc tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg ccc gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggc tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gcc ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gac gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc ata gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctt gca tcc tct ggc cgc
ccg ggg ccg gtg cta gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg ccc gtc tgg gac act cca atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gcg tct ggc gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctt act ggg att cca gtt aca act act ctg
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccc agc gac gac cca ctg tct ctg
cgc atg ctt ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ntc 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Ile 245 250 255gca ttt ggt gtg cgg
ttt gat gat cgt gtg act ggg aaa atc gag gct 816Ala Phe Gly Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gng cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Glu His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gca gat gtt aan 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gat cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Asp Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag can aan 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Thr Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggc gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc act ggt gtt ggg
cag cac cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tcg tct ggt ttg ggg gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg tta cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggt gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgc att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aat cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt cca gca gtt
cga gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc ata 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca aac ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Asn Gly Gly Ala 530 535
540ttc aag gac atg atc atg gag ggt gat ggc agg acc tcg tac tga
1677Phe Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser Tyr545 550
55524558PRTTriticum aestivum 24Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro
Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro Ile Val
Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105 110Val Leu
Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe 115 120
125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile Pro Lys
130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp Thr Pro
Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro
Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg Leu Val Gly
Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly Cys Ala
Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu Thr Gly
Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn Phe Pro
Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230 235
240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu Ile
245 250 255Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys Ile
Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Glu His Ile Asp Ile
Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val Ser
Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn Asp
Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp Phe
Gly Pro Trp His Lys Glu Leu Asp Gln Thr Lys 325 330 335Arg Glu Phe
Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345 350Gln
Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile 355 360
365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln Tyr Tyr
370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu
Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala
Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile Asp Gly
Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala Leu Ile
Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu Asn Asn
Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg Phe Tyr
Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470 475
480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly Phe
485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val Thr
Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr Leu
Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro Met
Ile Pro Asn Gly Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu Gly
Asp Gly Arg Thr Ser Tyr545 550 555251674DNATriticum
aestivumCDS(1)..(1671)modified_base(681)unknown nucleotide 25gtc
gac gtc ttc gcc tac cct ggc ggc gcg tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg cca gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggg tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gct ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gat gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc atc gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctc gca tcc tct ggc cgc
ccg ggg ccg gtg ctg gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg cct gtc tgg gac acg ccg atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gca tct ggt gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctc act ggg att cca gtt aca act act ctt
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccn agt gac gac cca ctg tct ctg
cgc atg ctg ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ntt 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Ile 245 250 255gca ttt ggt gtg cgg
ttt gat gat cgt gtg acc ggg aaa atc gag gct 816Ala Phe Gly Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gca gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gct cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggt gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc acc ggt gtt ggg
cag cat cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tca tcc ggt ttg ggt gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg ttg cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggg gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgt att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aac cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt ccg gca gtt
cgt gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc att 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca agc ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535
540ttt aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55526557PRTTriticum aestivum 26Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu
Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230
235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Ile 245 250 255Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn
Ala Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Ser Gly Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu
Gly Asp Gly Arg Thr Ser545 550 555271674DNATriticum
aestivumCDS(1)..(1671)modified_base(644)unknown nucleotide 27gtc
gac gtc ttc gcc tac cct ggc ggc gcg tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg cca gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggg tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gct ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gat gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc atc gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctc gca tcc tct ggc cgc
ccg ggg ccg gtg ctg gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg cct gtc tgg gac acg ccg atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gca tct ggt gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctc act ggg att cna gtt aca act act ctt
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Xaa Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccc agt gac gac cca ctg tct ctg
cgc atg ctg ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ctt 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255gca ttt ggt gtg cgg
ttt gat gat cgt gtg acc ggg aaa atc gag gct 816Ala Phe Gly Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gca gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gct cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335agg gag
ttt cct cta gga ttc aag act ttt ggt gag gcc atc ccg ccg 1056Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350caa tat gct atc cag gta ctg gat gag ctg aca aaa ggg gag gcg atc
1104Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365att gcc acc ggt gtt ggg cag cat cag atg tgg gcg gct cag
tat tac 1152Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380act tac aag cgg cca cgg cag tgg ctg tct tca tcc
ggt ttg ggt gca 1200Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400atg gga ttt ggg ttg cca gct gca gct
ggc gct gct gtg gcc aac cca 1248Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415ggt gtt aca gtt gtt gac att
gat ggg gat ggt agt ttc ctc atg aac 1296Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430att cag gag ttg gcg
ttg atc cgt att gag aac ctc cca gtg aag gtg 1344Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445atg ata ttg
aac aac cag cat ctg gga atg gtg gtg cag tgg gag gat 1392Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460agg
ttt tac aag gcc aac cgg gcg cac aca tac ctt ggc aac cca gaa 1440Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480aat gag agt gag ata tat cca gat ttt gtg acg att gct aaa gga
ttc 1488Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495aac gtt ccg gca gtt cgt gtg acg aag aag agc gaa gtc
act gca gca 1536Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510atc aag aag atg ctt gag acc cca ggg cca tac
ttg ttg gat atc att 1584Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525gtc ccg cat cag gag cac gtg ctg cct
atg atc cca agc ggt ggt gct 1632Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Ser Gly Gly Ala 530 535 540ttt aag gac atg atc atg gag
ggt gat ggc agg acc tcg tac 1674Phe Lys Asp Met Ile Met Glu Gly Asp
Gly Arg Thr Ser545 550 55528557PRTTriticum
aestivumMOD_RES(215)unknown amino acid 28Val Asp Val Phe Ala Tyr
Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser
Pro Val Ile Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala
Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val
Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala
Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75
80Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu
85 90 95Thr Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr
Leu 100 105 110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu
Ala Phe Phe 115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu
Val Asp Ile Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro
Val Trp Asp Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala
Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val
Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val
Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200
205Val Glu Leu Thr Gly Ile Xaa Val Thr Thr Thr Leu Met Gly Leu Gly
210 215 220Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly
Met His225 230 235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys
Ala Asp Leu Leu Leu 245 250 255Ala Phe Gly Val Arg Phe Asp Asp Arg
Val Thr Gly Lys Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile
Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys
Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu
Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala Gln305 310 315
320Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys
325 330 335Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile
Pro Pro 340 345 350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys
Gly Glu Ala Ile 355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met
Trp Ala Ala Gln Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp
Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu
Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr
Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile
Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440
445Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp
450 455 460Arg Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn
Pro Glu465 470 475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr
Ile Ala Lys Gly Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys
Lys Ser Glu Val Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr
Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu
His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535 540Phe Lys Asp
Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550 555291674DNATriticum
aestivumCDS(1)..(1671)modified_base(782)unknown nucleotide 29gtc
gac gtc ttc gcc tac cct ggc ggc gcg tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg cca gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggg tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gct ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gat gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc atc gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctc gca tcc tct ggc cgc
ccg ggg ccg gtg ctg gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg cct gtc tgg gac acg ccg atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gca tct ggt gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctc act ggg att cca gtt aca act act ctt
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccc agt gac gac cca ctg tct ctg
cgc atg ctg ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggc act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ctt 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255gca ttt ggt gtg cng
gtt gat gat cgt gtg anc ggg aaa atc gan gct 816Ala Phe Gly Val Gln
Val Asp Asp Arg Val Asn Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gng cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Glu His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cnt gtc tcc att tgt gca gat gtt aan 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Xaa 290 295
300ctt gct tta cag ggg ttg aat gcn cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggn ctg gat ttt ggt cca tgg cnc aag gag ttg
gat cag caa aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335ang gag ttt cct cta gga ttc aan act ttt
ggn gan gcc atc ccg ccg 1056Lys Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350cca tat gct atc cag gta ctg gat
gag ctg aca aaa ggg gag gcg atc 1104Pro Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc acc ggn gtt ggg
cag cat can atg tgg gcg gct cag tat tac 1152Ile Ala Thr Xaa Val Gly
Gln His Thr Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
ccn cgg cag tgg ctg tct tca tcc ggt ttg ggt gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg ttg cca gct gca gct ggc ggc nct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Gly Xaa Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggg gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgt att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aac cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt ccg gca gtt
cgt gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc att 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca agc ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535
540ttt aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55530557PRTTriticum aestivumMOD_RES(412)unknown amino acid 30Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255Ala Phe Gly Val Gln
Val Asp Asp Arg Val Asn Gly Lys Ile Glu Ala 260 265 270Phe Ala Ser
Arg Ser Lys Ile Glu His Ile Asp Ile Asp Pro Ala Glu 275 280 285Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335Lys Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350Pro Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365Ile Ala Thr Xaa Val Gly
Gln His Thr Met Trp Ala Ala Gln Tyr Tyr 370 375 380Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Gly Xaa Val Ala Asn Pro 405 410
415Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535
540Phe Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
555311674DNATriticum aestivumCDS(1)..(1671) 31gtc gac gtc ttc gcc
tac cct ggc ggc gcg tcc atg gag atc cac cag 48Val Asp Val Phe Ala
Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10 15gcg ctg acg cgc
tcg cca gtc atc acc aac cac ctc ttc cgc cac gag 96Ala Leu Thr Arg
Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu 20 25 30cag ggg gag
gcg ttc gcg gcg tcc ggg tac gcc cgc gcg tcc ggc cgc 144Gln Gly Glu
Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45gtc ggc gtc tgc gtc gcc
acc tcc ggc ccg ggg gcc acc aac ctc gtc 192Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60tcc gcg ctc gcc gac
gct ctc ctc gac tcc atc ccc atg gtc gcc atc 240Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80acg ggc cag
gtc ccc cgc cgc atg atc ggc acg gat gcg ttc cag gag 288Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95acg ccc
atc gtg gag gtc acg cgc tcc atc acc aag cac aac tac ctg 336Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110gtc ctt gac gtg gag gat atc ccc cgc gtc atc cag gaa gcc ttc ttc
384Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125ctc gca tcc tct ggc cgc ccg ggg ccg gtg ctg gtt gat atc
ccc aag 432Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140gac atc cag cag cag atg gct gtg cct gtc tgg gac
acg ccg atg agt 480Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160ttg cca ggg tac atc gcc cgc ctg ccc
aag cca cca tct act gaa tcg 528Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175ctt gag cag gtc ctg cgt ctg
gtt ggc gag tca cgg cgc cca att ctg 576Leu Glu Gln Val Leu Arg Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190tat gtt ggt ggt ggc
tgc gct gca tct ggt gag gag ttg cgc cgc ttt 624Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205gtt gag ctc
act ggg att cca gtt aca act act ctt atg ggc ctt ggc 672Val Glu Leu
Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220aac
ttc ccc agt gac gac cca ctg tct ctg cgc atg ctg ggg atg cat 720Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230
235 240ggc act gtg tat gca aat tat gca gta gat aag gct gac ctg ttg
ctt 768Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Leu 245 250 255gca ttt ggt gtg cgg ttt gat gat cgt gtg acc ggg aaa
atc gag gct 816Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270ttt gca agc agg tcc aag att gtg cac att gac
att gac cca gct gag 864Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285att ggc aag aac aag cag cca cat gtc
tcc att tgt gca gat gtt aag 912Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300ctt gct tta cag ggg ttg aat
gct cta tta aat ggg agc aaa gca caa 960Leu Ala Leu Gln Gly Leu Asn
Ala Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320cag ggt ctg gat
ttt ggt cca tgg cac aag gag ttg gat cag cag aag 1008Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335agg gag
ttt cct cta gga ttc aag act ttt ggt gag gcc atc ccg ccg 1056Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350caa tat gct atc cag gta ctg gat gag ctg aca aaa ggg gag gcg atc
1104Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365att gcc acc ggt gtt ggg cag cat cag atg tgg gcg gct cag
tat tac 1152Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380act tac aag cgg cca cgg cag tgg ctg tct tca tcc
ggt ttg ggt gca 1200Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400atg gga ttt ggg ttg cca gct gca gct
ggc gct gct gtg gcc aac cca 1248Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415ggt gtt aca gtt gtt gac att
gat ggg gat ggt agt ttc ctc atg aac 1296Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430att cag gag ttg gcg
ttg atc cgt att gag aac ctc cca gtg aag gtg 1344Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445atg ata ttg
aac aac cag cat ctg gga atg gtg gtg cag tgg gag gat 1392Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460agg
ttt tac aag gcc aac cgg gcg cac aca tac ctt ggc aac cca gaa 1440Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480aat gag agt gag ata tat cca gat ttt gtg acg att gct aaa gga
ttc 1488Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495aac gtt ccg gca gtt cgt gtg acg aag aag agc gaa gtc
act gca gca 1536Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510atc aag aag atg ctt gag acc cca ggg cca tac
ttg ttg gat atc att 1584Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525gtc ccg cat cag gag cac gtg ctg cct
atg atc cca aac ggt ggt gct 1632Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Asn Gly Gly Ala 530 535 540ttt aag gac atg atc atg gag
ggt gat ggc agg acc tcg tac 1674Phe Lys Asp Met Ile Met Glu Gly Asp
Gly Arg Thr Ser545 550 55532557PRTTriticum aestivum 32Val Asp Val
Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu
Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu 20 25 30Gln
Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40
45Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn Leu Val
50 55 60Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met Val Ala
Ile65 70 75 80Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr Asp Ala
Phe Gln Glu 85 90 95Thr Pro Ile Val Glu Val Thr Arg Ser Ile Thr Lys
His Asn Tyr Leu 100 105 110Val Leu Asp Val Glu Asp Ile Pro Arg Val
Ile Gln Glu Ala Phe Phe 115 120 125Leu Ala Ser Ser Gly Arg Pro Gly
Pro Val Leu Val Asp Ile Pro Lys 130 135 140Asp Ile Gln Gln Gln Met
Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160Leu Pro Gly
Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu
Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185
190Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe
195 200 205Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly
Leu Gly 210 215 220Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met
Leu Gly Met His225 230 235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val
Asp Lys Ala Asp Leu Leu Leu 245 250 255Ala Phe Gly Val Arg Phe Asp
Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser
Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys
Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295 300Leu
Ala Leu Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala Gln305 310
315 320Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln
Lys 325 330 335Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala
Ile Pro Pro 340 345 350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr
Lys Gly Glu Ala Ile 355 360 365Ile Ala Thr Gly Val Gly Gln His Gln
Met Trp Ala Ala Gln Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln
Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly
Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val
Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425
430Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val
435 440 445Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val Gln Trp
Glu Asp 450 455 460Arg Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu
Gly Asn Pro Glu465 470 475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe
Val Thr Ile Ala Lys Gly Phe 485 490 495Asn Val Pro Ala Val Arg Val
Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510Ile Lys Lys Met Leu
Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525Val Pro His
Gln Glu His Val Leu Pro Met Ile Pro Asn Gly Gly Ala 530 535 540Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
555331674DNATriticum
aestivumCDS(1)..(1671)modified_base(723)unknown nucleotide 33gtc
gac gtc ttc gcc tac cct ggc ggc gcg tcc atg gag atc cac cag 48Val
Asp Val Phe Ala Tyr Pro Gly Gly Ala Ser Met Glu Ile His Gln1 5 10
15gcg ctg acg cgc tcg cca gtc atc acc aac cac ctc ttc cgc cac gag
96Ala Leu Thr Arg Ser Pro Val Ile Thr Asn His Leu Phe Arg His Glu
20 25 30cag ggg gag gcg ttc gcg gcg tcc ggg tac gcc cgc gcg tcc ggc
cgc 144Gln Gly Glu Ala Phe Ala Ala Ser Gly Tyr Ala Arg Ala Ser Gly
Arg 35 40 45gtc ggc gtc tgc gtc gcc acc tcc ggc ccg ggg gcc acc aac
ctc gtc 192Val Gly Val Cys Val Ala Thr Ser Gly Pro Gly Ala Thr Asn
Leu Val 50 55 60tcc gcg ctc gcc gac gct ctc ctc gac tcc atc ccc atg
gtc gcc atc 240Ser Ala Leu Ala Asp Ala Leu Leu Asp Ser Ile Pro Met
Val Ala Ile65 70 75 80acg ggc cag gtc ccc cgc cgc atg atc ggc acg
gat gcg ttc cag gag 288Thr Gly Gln Val Pro Arg Arg Met Ile Gly Thr
Asp Ala Phe Gln Glu 85 90 95acg ccc atc gtg gag gtc acg cgc tcc atc
acc aag cac aac tac ctg 336Thr Pro Ile Val Glu Val Thr Arg Ser Ile
Thr Lys His Asn Tyr Leu 100 105 110gtc ctt gac gtg gag gat atc ccc
cgc gtc atc cag gaa gcc ttc ttc 384Val Leu Asp Val Glu Asp Ile Pro
Arg Val Ile Gln Glu Ala Phe Phe 115 120 125ctc gca tcc tct ggc cgc
ccg ggg ccg gtg ctg gtt gat atc ccc aag 432Leu Ala Ser Ser Gly Arg
Pro Gly Pro Val Leu Val Asp Ile Pro Lys 130 135 140gac atc cag cag
cag atg gct gtg cct gtc tgg gac acg ccg atg agt 480Asp Ile Gln Gln
Gln Met Ala Val Pro Val Trp Asp Thr Pro Met Ser145 150 155 160ttg
cca ggg tac atc gcc cgc ctg ccc aag cca cca tct act gaa tcg 528Leu
Pro Gly Tyr Ile Ala Arg Leu Pro Lys Pro Pro Ser Thr Glu Ser 165 170
175ctt gag cag gtc ctg cgt ctg gtt ggc gag tca cgg cgc cca att ctg
576Leu Glu Gln Val Leu Arg Leu Val Gly Glu Ser Arg Arg Pro Ile Leu
180 185 190tat gtt ggt ggt ggc tgc gct gca tct ggt gag gag ttg cgc
cgc ttt 624Tyr Val Gly Gly Gly Cys Ala Ala Ser Gly Glu Glu Leu Arg
Arg Phe 195 200 205gtt gag ctc act ggg att cca gtt aca act act ctt
atg ggc ctt ggc 672Val Glu Leu Thr Gly Ile Pro Val Thr Thr Thr Leu
Met Gly Leu Gly 210 215 220aac ttc ccc agt gac gac cca ctg tct ctg
cgc atg ctg ggg atg cat 720Asn Phe Pro Ser Asp Asp Pro Leu Ser Leu
Arg Met Leu Gly Met His225 230 235 240ggn act gtg tat gca aat tat
gca gta gat aag gct gac ctg ttg ctt 768Gly Thr Val Tyr Ala Asn Tyr
Ala Val Asp Lys Ala Asp Leu Leu Leu 245 250 255gca ttt ggt gtg cgg
ttt gat gat cgt gtg acc ggg aaa atc gag gct 816Ala Phe Gly Val Arg
Phe Asp Asp Arg Val Thr Gly Lys Ile Glu Ala 260 265 270ttt gca agc
agg tcc aag att gtg cac att gac att gac cca gct gag 864Phe Ala Ser
Arg Ser Lys Ile Val His Ile Asp Ile Asp Pro Ala Glu 275 280 285att
ggc aag aac aag cag cca cat gtc tcc att tgt gca gat gtt aag 912Ile
Gly Lys Asn Lys Gln Pro His Val Ser Ile Cys Ala Asp Val Lys 290 295
300ctt gct tta cag ggg ttg aat gct cta tta aat ggg agc aaa gca caa
960Leu Ala Leu Gln Gly Leu Asn Ala Leu Leu Asn Gly Ser Lys Ala
Gln305 310 315 320cag ggt ctg gat ttt ggt cca tgg cac aag gag ttg
gat cag cag aag 1008Gln Gly Leu Asp Phe Gly Pro Trp His Lys Glu Leu
Asp Gln Gln Lys 325 330 335agg gag ttt cct cta gga ttc aag act ttt
ggt gag gcc atc ccg ccg 1056Arg Glu Phe Pro Leu Gly Phe Lys Thr Phe
Gly Glu Ala Ile Pro Pro 340 345 350caa tat gct atc cag gta ctg gat
gag ctg acn aaa ggg gag gcg atc 1104Gln Tyr Ala Ile Gln Val Leu Asp
Glu Leu Thr Lys Gly Glu Ala Ile 355 360 365att gcc acc ggt gtt ggg
cag cat cag atg tgg gcg gct cag tat tac 1152Ile Ala Thr Gly Val Gly
Gln His Gln Met Trp Ala Ala Gln Tyr Tyr 370 375 380act tac aag cgg
cca cgg cag tgg ctg tct tca tcc ggt ttg ggt gca 1200Thr Tyr Lys Arg
Pro Arg Gln Trp Leu Ser Ser Ser Gly Leu Gly Ala385 390 395 400atg
gga ttt ggg ttg cca gct gca gct ggc gct gct gtg gcc aac cca 1248Met
Gly Phe Gly Leu Pro Ala Ala Ala Gly Ala Ala Val Ala Asn Pro 405 410
415ggt gtt aca gtt gtt gac att gat ggg gat ggt agt ttc ctc atg aac
1296Gly Val Thr Val Val Asp Ile Asp Gly Asp Gly Ser Phe Leu Met Asn
420 425 430att cag gag ttg gcg ttg atc cgt att gag aac ctc cca gtg
aag gtg 1344Ile Gln Glu Leu Ala Leu Ile Arg Ile Glu Asn Leu Pro Val
Lys Val 435 440 445atg ata ttg aac aac cag cat ctg gga atg gtg gtg
cag tgg gag gat 1392Met Ile Leu Asn Asn Gln His Leu Gly Met Val Val
Gln Trp Glu Asp 450 455 460agg ttt tac aag gcc aac cgg gcg cac aca
tac ctt ggc aac cca gaa 1440Arg Phe Tyr Lys Ala Asn Arg Ala His Thr
Tyr Leu Gly Asn Pro Glu465 470 475 480aat gag agt gag ata tat cca
gat ttt gtg acg att gct aaa gga ttc 1488Asn Glu Ser Glu Ile Tyr Pro
Asp Phe Val Thr Ile Ala Lys Gly Phe 485 490 495aac gtt ccg gca gtt
cgt gtg acg aag aag agc gaa gtc act gca gca 1536Asn Val Pro Ala Val
Arg Val Thr Lys Lys Ser Glu Val Thr Ala Ala 500 505 510atc aag aag
atg ctt gag acc cca ggg cca tac ttg ttg gat atc att 1584Ile Lys Lys
Met Leu Glu Thr Pro Gly Pro Tyr Leu Leu Asp Ile Ile 515 520 525gtc
ccg cat cag gag cac gtg ctg cct atg atc cca agc ggt ggt gct 1632Val
Pro His Gln Glu His Val Leu Pro Met Ile Pro Ser Gly Gly Ala 530 535
540ttt aag gac atg atc atg gag ggt gat ggc agg acc tcg tac 1674Phe
Lys Asp Met Ile Met Glu Gly Asp Gly Arg Thr Ser545 550
55534557PRTTriticum aestivum 34Val Asp Val Phe Ala Tyr Pro Gly Gly
Ala Ser Met Glu Ile His Gln1 5 10 15Ala Leu Thr Arg Ser Pro Val Ile
Thr Asn His Leu Phe Arg His Glu 20 25 30Gln Gly Glu Ala Phe Ala Ala
Ser Gly Tyr Ala Arg Ala Ser Gly Arg 35 40 45Val Gly Val Cys Val Ala
Thr Ser Gly Pro Gly Ala Thr Asn Leu Val 50 55 60Ser Ala Leu Ala Asp
Ala Leu Leu Asp Ser Ile Pro Met Val Ala Ile65 70 75 80Thr Gly Gln
Val Pro Arg Arg Met Ile Gly Thr Asp Ala Phe Gln Glu 85 90 95Thr Pro
Ile Val Glu Val Thr Arg Ser Ile Thr Lys His Asn Tyr Leu 100 105
110Val Leu Asp Val Glu Asp Ile Pro Arg Val Ile Gln Glu Ala Phe Phe
115 120 125Leu Ala Ser Ser Gly Arg Pro Gly Pro Val Leu Val Asp Ile
Pro Lys 130 135 140Asp Ile Gln Gln Gln Met Ala Val Pro Val Trp Asp
Thr Pro Met Ser145 150 155 160Leu Pro Gly Tyr Ile Ala Arg Leu Pro
Lys Pro Pro Ser Thr Glu Ser 165 170 175Leu Glu Gln Val Leu Arg
Leu
Val Gly Glu Ser Arg Arg Pro Ile Leu 180 185 190Tyr Val Gly Gly Gly
Cys Ala Ala Ser Gly Glu Glu Leu Arg Arg Phe 195 200 205Val Glu Leu
Thr Gly Ile Pro Val Thr Thr Thr Leu Met Gly Leu Gly 210 215 220Asn
Phe Pro Ser Asp Asp Pro Leu Ser Leu Arg Met Leu Gly Met His225 230
235 240Gly Thr Val Tyr Ala Asn Tyr Ala Val Asp Lys Ala Asp Leu Leu
Leu 245 250 255Ala Phe Gly Val Arg Phe Asp Asp Arg Val Thr Gly Lys
Ile Glu Ala 260 265 270Phe Ala Ser Arg Ser Lys Ile Val His Ile Asp
Ile Asp Pro Ala Glu 275 280 285Ile Gly Lys Asn Lys Gln Pro His Val
Ser Ile Cys Ala Asp Val Lys 290 295 300Leu Ala Leu Gln Gly Leu Asn
Ala Leu Leu Asn Gly Ser Lys Ala Gln305 310 315 320Gln Gly Leu Asp
Phe Gly Pro Trp His Lys Glu Leu Asp Gln Gln Lys 325 330 335Arg Glu
Phe Pro Leu Gly Phe Lys Thr Phe Gly Glu Ala Ile Pro Pro 340 345
350Gln Tyr Ala Ile Gln Val Leu Asp Glu Leu Thr Lys Gly Glu Ala Ile
355 360 365Ile Ala Thr Gly Val Gly Gln His Gln Met Trp Ala Ala Gln
Tyr Tyr 370 375 380Thr Tyr Lys Arg Pro Arg Gln Trp Leu Ser Ser Ser
Gly Leu Gly Ala385 390 395 400Met Gly Phe Gly Leu Pro Ala Ala Ala
Gly Ala Ala Val Ala Asn Pro 405 410 415Gly Val Thr Val Val Asp Ile
Asp Gly Asp Gly Ser Phe Leu Met Asn 420 425 430Ile Gln Glu Leu Ala
Leu Ile Arg Ile Glu Asn Leu Pro Val Lys Val 435 440 445Met Ile Leu
Asn Asn Gln His Leu Gly Met Val Val Gln Trp Glu Asp 450 455 460Arg
Phe Tyr Lys Ala Asn Arg Ala His Thr Tyr Leu Gly Asn Pro Glu465 470
475 480Asn Glu Ser Glu Ile Tyr Pro Asp Phe Val Thr Ile Ala Lys Gly
Phe 485 490 495Asn Val Pro Ala Val Arg Val Thr Lys Lys Ser Glu Val
Thr Ala Ala 500 505 510Ile Lys Lys Met Leu Glu Thr Pro Gly Pro Tyr
Leu Leu Asp Ile Ile 515 520 525Val Pro His Gln Glu His Val Leu Pro
Met Ile Pro Ser Gly Gly Ala 530 535 540Phe Lys Asp Met Ile Met Glu
Gly Asp Gly Arg Thr Ser545 550 555351524DNAArtificial
SequenceDescription of Artificial Sequence Consensus sequence
35gtctgcgtcg ccacctccgg cccgggggcc accaacctcg tctccgcgct cgccgacgcc
60ctcctcgact ccatccccat ggtcgccatc acgggccagg tcccccgccg catgatcggc
120acggacgcgt tccaggagac gcccatagtg gaggtcacgc gctccatcac
caagcacaac 180tacctggtcc ttgacgtgga ggatatcccc cgcgtcatcc
aggaagcctt cttcctcgca 240tcctctggcc gcccggggcc ggtgctggtt
gatatcccca aggacatcca gcagcagatg 300gctgtgcctg tctgggacac
gccgatgagt ttgccagggt acatcgcccg cctgcccaag 360ccaccatcta
ctgaatcgct tgagcaggtc ctgcgtctgg ttggcgagtc acggcgccca
420attctgtatg ttggtggtgg ctgcgctgca tctggcgagg agttgcgccg
ctttgttgag 480ctcactggga ttccagttac aactactctg atgggccttg
gcaacttccc cagcgacgac 540ccactgtctc tgcgcatgct tgggatgcat
ggcactgtgt atgcaaatta tgcagtagat 600aaggctgacc tgttgcttgc
atttggtgtg cggtttgatg atcgtgtgac tgggaaaatc 660gaggcttttg
caagcaggtc caagattgtg cacattgaca ttgacccagc tgagattggc
720aagaacaagc agccacatgt ctccatttgt gcagatgtta agcttgcttt
acaggggttg 780aatgctctat taaatgggag caaagcacaa cagggtctgg
attttggtcc atggcacaag 840gagttggatc agcagaagag ggagtttcct
ctaggattca agacttttgg cgaggccatc 900ccgccgcaat atgctatcca
ggtactggat gagctgacaa aaggggaggc gatcattgcc 960actggtgttg
ggcagcacca gatgtgggcg gctcagtatt acacttacaa gcggccacgg
1020cagtggctgt cttcgtctgg tttgggggca atgggatttg ggttaccagc
tgcagctggc 1080gctgctgtgg ccaacccagg tgttacagtt gttgacattg
atggngatgg tagtttcctc 1140atgaacattc aggagttggc gttgatccgt
attgagaacc tcccagtgaa ggtgatgata 1200ttgaacaacc agcatctggg
aatggtggtg cagtgggagg ataggtttta caaggccaat 1260cgggcgcaca
cataccttgg caacccagaa aatgagagtg agatatatcc agattttgtg
1320acgattgcta aaggattcaa cgttccggca gttcgtgtga cgaagaagag
cgaagtcact 1380gcagcaatca agaagatgct tgagacccca gggccatact
tgttggatat catngtcccg 1440catcaggagc acgtgctgcc tatgatccca
agcggtggtg ctttcaagga catgatcatg 1500gagggtgatg gcaggacctc gtac
1524
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