U.S. patent application number 15/075717 was filed with the patent office on 2017-09-21 for soybean variety xr25al15x.
The applicant listed for this patent is MONSANTO TECHNOLOGY LLC. Invention is credited to Thomas Jury.
Application Number | 20170265422 15/075717 |
Document ID | / |
Family ID | 59828135 |
Filed Date | 2017-09-21 |
United States Patent
Application |
20170265422 |
Kind Code |
A1 |
Jury; Thomas |
September 21, 2017 |
SOYBEAN VARIETY XR25AL15X
Abstract
The invention relates to the soybean variety designated
XR25AL15X. Provided by the invention are the seeds, plants and
derivatives of the soybean variety XR25AL15X. Also provided by the
invention are tissue cultures of the soybean variety XR25AL15X and
the plants regenerated therefrom. Still further provided by the
invention are methods for producing soybean plants by crossing the
soybean variety XR25AL15X with itself or another soybean variety
and plants produced by such methods.
Inventors: |
Jury; Thomas; (Ames,
IA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MONSANTO TECHNOLOGY LLC |
St. Louis |
MO |
US |
|
|
Family ID: |
59828135 |
Appl. No.: |
15/075717 |
Filed: |
March 21, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A01H 5/10 20130101; C12N
15/8271 20130101; A01H 1/02 20130101; C12N 15/8275 20130101; C12N
15/8286 20130101; C12N 15/8289 20130101; C12N 15/8279 20130101;
C12N 15/8247 20130101; C12N 15/8245 20130101; C12N 15/8241
20130101; C12N 15/8251 20130101; C12N 15/8274 20130101; C12N
15/8278 20130101 |
International
Class: |
A01H 5/10 20060101
A01H005/10; C12N 15/82 20060101 C12N015/82; A01H 1/02 20060101
A01H001/02 |
Claims
1. A plant of soybean variety XR25AL15X, wherein a sample of seed
of said variety has been deposited under ATCC Accession No.
PTA-122689.
2. A plant part of the plant of claim 1, wherein the plant part
comprises at least one cell of said plant.
3. The plant part of claim 2, further defined as pollen, a
meristem, a cell, or an ovule.
4. A seed of soybean variety XR25AL15X, wherein a sample of seed of
said variety has been deposited under ATCC Accession No.
PTA-122689.
5. A method of producing soybean seed, wherein the method comprises
crossing the plant of claim 1 with itself or a second soybean
plant.
6. The method of claim 5, wherein the method is further defined as
comprising crossing the plant of soybean variety XR25AL15X with a
second, distinct soybean plant.
7. A soybean seed produced by the method of claim 6.
8. A soybean plant produced by growing the seed of claim 7.
9. A composition comprising the seed of claim 4 comprised in plant
seed growth media, wherein a sample of seed of said variety has
been deposited under ATCC Accession No. PTA-122689.
10. A plant of soybean variety XR25AL15X, further comprising a
single locus conversion, wherein a sample of seed of soybean
variety XR25AL15X has been deposited under ATCC Accession No.
PTA-122689.
11. The plant of claim 10, wherein the single locus conversion
comprises a transgene.
12. A seed that produces the plant of claim 10.
13. The seed of claim 12, wherein the single locus confers a trait
selected from the group consisting of male sterility, herbicide
tolerance, insect resistance, pest resistance, disease resistance,
modified fatty acid metabolism, abiotic stress resistance, altered
seed amino acid composition, site-specific genetic recombination,
and modified carbohydrate metabolism.
14. The seed of claim 12, wherein the single locus confers
tolerance to an herbicide selected from the group consisting of
glyphosate, sulfonylurea, imidazalinone, dicamba, glufosinate,
phenoxy propionic acid, cyclohexanedione, triazine, benzonitrile,
PPO-inhibitor herbicides, and bromoxynil.
15. The seed of claim 12, wherein the single locus conversion
comprises a transgene.
16. The method of claim 6, wherein the method further comprises:
(a) crossing a plant grown from said soybean seed with itself or a
different soybean plant to produce a seed of a progeny plant of a
subsequent generation; (b) growing a progeny plant of a subsequent
generation from said seed of a progeny plant of a subsequent
generation and crossing the progeny plant of a subsequent
generation with itself or a second plant to produce a progeny plant
of a further subsequent generation; and (c) repeating steps (a) and
(b) using said progeny plant of a further subsequent generation
from step (b) in place of the plant grown from said soybean seed in
step (a), wherein steps (a) and (b) are repeated with sufficient
inbreeding to produce an inbred soybean plant derived from the
soybean variety XR25AL15X.
17. The method of claim 16, further comprising crossing said inbred
soybean plant derived from the soybean variety XR25AL15X with a
plant of a different genotype to produce a seed of a hybrid soybean
plant derived from the soybean variety XR25AL15X.
18. A method of producing a commodity plant product comprising
collecting the commodity plant product from the plant of claim
1.
19. The method of claim 18, wherein the commodity plant product is
protein concentrate, protein isolate, grain, soybean hulls, meal,
flour, or oil.
20. A soybean commodity plant product produced by the method of
claim 19, wherein the commodity plant product comprises at least
one cell of soybean variety XR25AL15X.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates generally to the field of
soybean breeding. In particular, the invention relates to the novel
soybean variety XR25AL15X.
[0003] 2. Description of Related Art
[0004] There are numerous steps in the development of any novel,
desirable plant germplasm. Plant breeding begins with the analysis
and definition of problems and weaknesses of the current germplasm,
the establishment of program goals, and the definition of specific
breeding objectives. The next step is selection of germplasm that
possess the traits to meet the program goals. The goal is to
combine in a single variety an improved combination of desirable
traits from the parental germplasm. These important traits may
include higher seed yield, resistance to diseases and insects,
better stems and roots, tolerance to drought and heat, better
agronomic quality, resistance to herbicides, and improvements in
compositional traits.
[0005] Soybean, Glycine max (L.), is a valuable field crop. Thus, a
continuing goal of plant breeders is to develop stable, high
yielding soybean varieties that are agronomically sound. The
reasons for this goal are to maximize the amount of grain produced
on the land used and to supply food for both animals and humans. To
accomplish this goal, the soybean breeder must select and develop
soybean plants that have the traits that result in superior
varieties.
[0006] The oil extracted from soybeans is widely used in food
products, such as margarine, cooking oil, and salad dressings.
Soybean oil is composed of saturated, monounsaturated, and
polyunsaturated fatty acids, with a typical composition of 11%
palmitic, 4% stearic, 25% oleic, 50% linoleic, and 9% linolenic
fatty acid content ("Economic Implications of Modified Soybean
Traits Summary Report," Iowa Soybean Promotion Board & American
Soybean Association Special Report 92S, May 1990).
SUMMARY OF THE INVENTION
[0007] One aspect of the present invention relates to seed of the
soybean variety XR25AL15X. The invention also relates to plants
produced by growing the seed of the soybean variety XR25AL15X, as
well as the derivatives of such plants. Further provided are plant
parts, including cells, plant protoplasts, plant cells of a tissue
culture from which soybean plants can be regenerated, plant calli,
plant clumps, and plant cells that are intact in plants or parts of
plants, such as pollen, flowers, seeds, pods, leaves, stems, and
the like.
[0008] In a further aspect, the invention provides a composition
comprising a seed of soybean variety XR25AL15X comprised in plant
seed growth media. In certain embodiments, the plant seed growth
media is a soil or synthetic cultivation medium. In specific
embodiments, the growth medium may be comprised in a container or
may, for example, be soil in a field. Plant seed growth media are
well known to those of skill in the art and include, but are in no
way limited to, soil or synthetic cultivation medium.
Advantageously, plant seed growth media can provide adequate
physical support for seeds and can retain moisture and/or
nutritional components. Examples of characteristics for soils that
may be desirable in certain embodiments can be found, for instance,
in U.S. Pat. Nos. 3,932,166 and 4,707,176. Synthetic plant
cultivation media are also well known in the art and may, in
certain embodiments, comprise polymers or hydrogels. Examples of
such compositions are described, for example, in U.S. Pat. No.
4,241,537.
[0009] Another aspect of the invention relates to a tissue culture
of regenerable cells of the soybean variety XR25AL15X, as well as
plants regenerated therefrom, wherein the regenerated soybean plant
is capable of expressing all the morphological and physiological
characteristics of a plant grown from the soybean seed designated
XR25AL15X.
[0010] Yet another aspect of the current invention is a soybean
plant further comprising a single locus conversion. In one
embodiment, the soybean plant is defined as comprising the single
locus conversion and otherwise capable of expressing all of the
morphological and physiological characteristics of the soybean
variety XR25AL15X. In particular embodiments of the invention, the
single locus conversion may comprise a transgenic gene which has
been introduced by genetic transformation into the soybean variety
XR25AL15X or a progenitor thereof. In still other embodiments of
the invention, the single locus conversion may comprise a dominant
or recessive allele. The locus conversion may confer potentially
any trait upon the single locus converted plant, including
herbicide resistance, insect resistance, resistance to bacterial,
fungal, or viral disease, male fertility or sterility, and improved
nutritional quality.
[0011] Still yet another aspect of the invention relates to a first
generation (F.sub.1) hybrid soybean seed produced by crossing a
plant of the soybean variety XR25AL15X to a second soybean plant.
Also included in the invention are the F.sub.1 hybrid soybean
plants grown from the hybrid seed produced by crossing the soybean
variety XR25AL15X to a second soybean plant. Still further included
in the invention are the seeds of an F.sub.1 hybrid plant produced
with the soybean variety XR25AL15X as one parent, the second
generation (F.sub.2) hybrid soybean plant grown from the seed of
the F.sub.1 hybrid plant, and the seeds of the F.sub.2 hybrid
plant.
[0012] Still yet another aspect of the invention is a method of
producing soybean seeds comprising crossing a plant of the soybean
variety XR25AL15X to any second soybean plant, including itself or
another plant of the variety XR25AL15X. In particular embodiments
of the invention, the method of crossing comprises the steps of a)
planting seeds of the soybean variety XR25AL15X; b) cultivating
soybean plants resulting from said seeds until said plants bear
flowers, c) allowing fertilization of the flowers of said plants;
and d) harvesting seeds produced from said plants.
[0013] Still yet another aspect of the invention is a method of
producing hybrid soybean seeds comprising crossing the soybean
variety XR25AL15X to a second, distinct soybean plant which is
nonisogenic to the soybean variety XR25AL15X. In particular
embodiments of the invention, the crossing comprises the steps of
a) planting seeds of soybean variety XR25AL15X and a second,
distinct soybean plant, b) cultivating the soybean plants grown
from the seeds until the plants bear flowers; c) cross pollinating
a flower on one of the two plants with the pollen of the other
plant, and d) harvesting the seeds resulting from the cross
pollinating.
[0014] Still yet another aspect of the invention is a method for
developing a soybean plant in a soybean breeding program
comprising: obtaining a soybean plant, or its parts, of the variety
XR25AL15X; and b) employing said plant or parts as a source of
breeding material using plant breeding techniques. In the method,
the plant breeding techniques may be selected from the group
consisting of recurrent selection, mass selection, bulk selection,
backcrossing, pedigree breeding, genetic marker-assisted selection
and genetic transformation. In certain embodiments of the
invention, the soybean plant of variety XR25AL15X is used as the
male or female parent.
[0015] Still yet another aspect of the invention is a method of
producing a soybean plant derived from the soybean variety
XR25AL15X, the method comprising the steps of: (a) preparing a
progeny plant derived from soybean variety XR25AL15X by crossing a
plant of the soybean variety XR25AL15X with a second soybean plant;
and (b) crossing the progeny plant with itself or a second plant to
produce a progeny plant of a subsequent generation which is derived
from a plant of the soybean variety XR25AL15X. In one embodiment of
the invention, the method further comprises: (c) crossing the
progeny plant of a subsequent generation with itself or a second
plant; and (d) repeating steps (b) and (c) for, in some
embodiments, at least 2, 3, 4 or more additional generations to
produce an inbred soybean plant derived from the soybean variety
XR25AL15X. Also provided by the invention is a plant produced by
this and the other methods of the invention.
[0016] In another embodiment of the invention, the method of
producing a soybean plant derived from the soybean variety
XR25AL15X further comprises: (a) crossing the soybean variety
XR25AL15X-derived soybean plant with itself or another soybean
plant to yield additional soybean variety XR25AL15X-derived progeny
soybean seed; (b) growing the progeny soybean seed of step (a)
under plant growth conditions to yield additional soybean variety
XR25AL15X-derived soybean plants; and (c) repeating the crossing
and growing steps of (a) and (b) to generate further soybean
variety XR25AL15X-derived soybean plants. In specific embodiments,
steps (a) and (b) may be repeated at least 1, 2, 3, 4, or 5 or more
times as desired. The invention still further provides a soybean
plant produced by this and the foregoing methods.
DETAILED DESCRIPTION OF THE INVENTION
[0017] The instant invention provides methods and composition
relating to plants, seeds and derivatives of the soybean variety
XR25AL15X. Soybean variety XR25AL15X is adapted to early group II.
Soybean variety XR25AL15X was developed from an initial cross of
AG2831/AW1911F2-B0BAH. The breeding history of the variety can be
summarized as follows:
TABLE-US-00001 Breeding History Generation Year Description Cross
2010 The cross was made near Isabela, PR F.sub.1 2011 Plants were
grown near Isabela, PR and advanced using bulk. F.sub.2 2011 Plants
were grown near Kunia, HI and advanced using sps F.sub.3 2012
Plants were grown near Janesville, WI and advanced using sps
F.sub.4 2012 Plants were grown near Rancagua, Chile in Progeny Rows
and the variety XR25AL15X was selected based on the agronomic
characteristics, general phenotypic appearance, and traits of
interest based on molecular marker information.
TABLE-US-00002 Yield Testing No. of No. of Generation Year
Locations Rank Entries F.sub.5 2013 7 1 30 F.sub.6 2014 21 13
30
[0018] The soybean variety XR25AL15X has been judged to be uniform
for breeding purposes and testing. The variety XR25AL15X can be
reproduced by planting and growing seeds of the variety under
self-pollinating or sib-pollinating conditions, as is known to
those of skill in the agricultural arts. Variety XR25AL15X shows no
variants other than what would normally be expected due to
environment or that would occur for almost any characteristic
during the course of repeated sexual reproduction.
[0019] The results of an objective evaluation of the variety are
presented below, in Table 1. Those of skill in the art will
recognize that these are typical values that may vary due to
environment and that other values that are substantially equivalent
are within the scope of the invention. An `*` denotes
classifications/scores generated based on greenhouse assays.
TABLE-US-00003 TABLE 1 Phenotypic Description of Variety XR25AL15X
Trait Phenotype Morphology: Relative Maturity 2.3 Flower Color
Purple Pubescence Color Gray Hilum Color Imperfect black Pod Color
Brown Hypocotyl Color Purple Seed Coat Color Yellow Seed Coat
Luster Dull Seed Shape Spherical flattened Cotyledon Color Yellow
Leaf Shape Ovate Leaf Color Green Canopy Intermediate Growth Habit
Indeterminate Disease Reactions Phytophthora Allele* Rps1c
Phytophthora Tolerance* Race 25 Moderately tolerant Soybean Cyst
Nematode Race 1* -- Soybean Cyst Nematode Race 3* Resistant Brown
Stem Rot* -- Sudden Death Syndrome* -- Southern Stem Canker* --
Frog Eye Leaf Spot (Race 3)* -- Southern Root Knot* -- Chloride
Sensitivity* Includer Insect Reactions Soybean Aphid -- Herbicide
Reactions: Glyphosate Resistant, MON89788 Sulfonylurea --
Glufosinate -- Metribuzin -- Dicamba Resistant, MON87708 Fatty
acid: Fatty acid composition Normal
[0020] As disclosed herein above, soybean variety XR25AL15X
contains events MON89788 and MON87708. Event MON89788, also known
as event GM_A19788, confers glyphosate tolerance and is the subject
of U.S. Pat. No. 7,632,985, the disclosure of which is incorporated
herein by reference. Event MON89788 is also covered by one or more
of the following patents: U.S. Pat. Nos. 6,051,753; 6,660,911;
6,949,696; 7,141,722; 7,608,761; 8,053,184; 8,969,006. Event
MON87708 confers dicamba tolerance and is the subject of U.S. Pat.
No. 8,501,407, the disclosure of which is incorporated herein by
reference. Event MON87708 is also covered by one or more of the
following patents: U.S. Pat. Nos. 5,850,019; 7,812,224; 7,838,729;
7,884,262; 7,939,721; 8,119,380; 8,207,092; 8,629,323; 8,754,011;
RE45,048.
[0021] The performance characteristics of soybean variety XR25AL15X
were also analyzed and comparisons were made with selected
varieties. The results of the analysis are presented below, in
Table 2.
TABLE-US-00004 TABLE 2 Exemplary Agronomic Traits of Variety
XR25AL15X and Selected Varieties Entries Compared YLD MAT PHT LDG
EMER PRO SWT XR25AL15X 55.9 29.8 32.3 1.3 2.1 40.3 2,471.5 AG1935
58.4 26.1 32 2.8 1.9 40.2 2,359.1 Deviation -2.5 3.75 0.25 -1.5
0.29 0.14 112.36 Significance + ** + # Obs 21 8 4 2 7 4 4 Years 1 1
1 1 1 1 1 Win Percent 43 0 50 100 25 50 100 Test Mean 55.2 28.2
32.2 1.9 2 40.8 2,505.7 XR25AL15X 56.3 29.3 32.3 2.2 2.1 40.3
2,471.5 AG2031 56.3 27.8 34.3 2.9 1.6 41.6 2,342.9 Deviation 0 1.45
-2.05 -0.74 0.51 -1.26 128.55 Significance * # Obs 26 9 4 3 7 4 4
Years 2 2 1 2 1 1 1 Win Percent 62 22 75 67 0 25 75 Test Mean 53.5
27.7 33.2 2.7 2 40.8 2,533.5 XR25AL15X 55.9 29.8 32.3 1.3 2.1 40.3
2,471.5 AG2035 57.9 27.6 30.5 1.8 1.9 42.2 2,194.5 Deviation -2.06
2.19 1.75 -0.5 0.29 -1.87 276.96 Significance + * * ** # Obs 21 8 4
2 7 4 4 Years 1 1 1 1 1 1 1 Win Percent 38 0 50 100 25 0 100 Test
Mean 55.3 28.4 32.3 1.6 2 40.8 2,508.5 XR25AL15X 56.3 29.8 32.3 2.2
2.1 40.3 2,471.5 AG2134 54.9 26.7 33.1 1.6 1.8 41.3 2,685.8
Deviation 1.42 3.06 -0.87 0.52 0.37 -0.92 -214.33 Significance ** *
** # Obs 25 8 4 3 7 4 4 Years 2 1 1 2 1 1 1 Win Percent 56 0 75 67
17 25 0 Test Mean 53.2 28 33.4 2.7 2 40.7 2,553.9 XR25AL15X 56.6
28.2 33.8 1.9 2.1 40.3 2,471.5 AG2433 55.2 28 34.2 1.8 2 40.4
2,955.5 Deviation 1.45 0.27 -0.43 0.07 0.12 -0.1 -483.98
Significance * # Obs 28 11 5 4 7 4 4 Years 2 2 2 2 1 1 1 Win
Percent 64 36 40 75 25 50 0 Test Mean 53.7 28.4 35.7 2.5 2 40.7
2,614.3 XR25AL15X 57.2 27.9 33.8 1.9 2 41.1 2,333.5 AG2731 56.4
29.6 38.9 2.3 1.8 42 2,381.5 Deviation 0.73 -1.7 -5.13 -0.38 0.23
-0.9 -48 Significance * ** # Obs 26 10 5 4 6 2 2 Years 2 2 2 2 1 1
1 Win Percent 54 90 100 75 25 0 50 Test Mean 54.4 28.4 35.9 2.9 1.9
40.8 2,490.5 XR25AL15X 57 27.9 33.8 1.9 2 40.8 2,412.3 AG2733 55
30.5 33.8 1.2 1.8 40.4 2,688.8 Deviation 2 -2.61 0.02 0.72 0.2 0.36
-276.5 Significance ** # Obs 25 10 5 4 6 3 3 Years 2 2 2 2 1 1 1
Win Percent 64 80 60 33 0 67 0 Test Mean 54.5 29.6 36.9 2.5 1.9
40.9 2,597 XR25AL15X 58.5 26.2 34 1.9 2.2 41 AG2931 56.8 30.4 39
1.8 2.1 40.9 Deviation 1.68 -4.28 -5 0.11 0.1 0.1 Significance **
** # Obs 19 6 4 4 5 1 Years 2 2 2 2 1 1 Win Percent 74 100 100 67
50 100 Test Mean 56.7 29.2 37.1 2.1 2.1 41.5 XR25AL15X 57.8 27.3 34
1.9 2 40.8 2,467.5 AG2933 59.6 32.4 36.6 2.5 1.7 40.8 2,777
Deviation -1.83 -5.12 -2.59 -0.63 0.31 -0.03 -309.5 Significance **
+ # Obs 23 8 4 4 6 3 2 Years 2 2 2 2 1 1 1 Win Percent 48 100 100
75 20 33 0 Test Mean 55.5 30.1 37.7 2.5 1.8 41 2,678.5 XR25AL15X
58.2 26.1 33.7 1.9 2 40.6 2,365 AG3231 56.4 33.1 38.2 1.8 1.8 40.6
2,634.5 Deviation 1.8 -7.01 -4.49 0.05 0.19 -0.05 -269.5
Significance ** # Obs 17 5 3 4 3 2 1 Years 2 2 2 2 1 1 1 Win
Percent 65 100 100 67 33 0 0 Test Mean 55 30.5 37.6 3 1.9 41.2
2,589 XR25AL15X 56.3 29.8 32.3 2.2 2.1 40.4 2,438.7 CR2 2023N 54.8
27.6 32.9 1.8 2.6 40.9 2,290.7 Deviation 1.54 2.2 -0.69 0.37 -0.46
-0.54 147.98 Significance ** # Obs 25 8 4 3 7 3 3 Years 2 1 1 2 1 1
1 Win Percent 60 0 75 67 75 33 67 Test Mean 55.5 29.1 33.5 2.4 2
40.7 2,542.5 XR25AL15X 56.1 29.8 32.3 2.2 2.1 41.1 2,333.5 CR2200
56 27.3 32.8 1.6 1.6 41.4 2,486.5 Deviation 0.12 2.53 -0.5 0.58
0.54 -0.3 -153 Significance ** + # Obs 24 8 4 3 7 2 2 Years 2 1 1 2
1 1 1 Win Percent 50 0 75 0 20 50 0 Test Mean 55.5 28.7 33.1 2.3 2
41.1 2,434.1 XR25AL15X 55.9 29.8 32.3 1.3 2.1 40.4 2,438.7 CR2311N
57.1 27.5 33.3 1.4 2 39.1 2,643.9 Deviation -1.25 2.34 -1 -0.12
0.18 1.28 -205.25 Significance ** * # Obs 21 8 4 2 7 3 3 Years 1 1
1 1 1 1 1 Win Percent 33 0 75 100 40 100 0 Test Mean 54.9 29.3 33.8
1.8 2 40.7 2,548.5 XR25AL15X 55.9 29.8 32.3 1.3 2.1 40.4 2,438.7
CR2322N 54.7 28.9 33.5 1.6 2 40 2,351.8 Deviation 1.16 0.9 -1.2
-0.35 0.17 0.41 86.9 Significance + # Obs 21 8 4 2 7 3 3 Years 1 1
1 1 1 1 1 Win Percent 62 29 75 100 25 67 100 Test Mean 54 30.2 34.7
2.2 2 40.5 2,601.1 **, *, + significant at P < 0.01, 0.05, or
0.10, respectively
I. Breeding Soybean Variety XR25AL15X
[0022] One aspect of the current invention concerns methods for
crossing the soybean variety XR25AL15X with itself or a second
plant and the seeds and plants produced by such methods. These
methods can be used for propagation of the soybean variety
XR25AL15X, or can be used to produce hybrid soybean seeds and the
plants grown therefrom. Hybrid soybean plants can be used by
farmers in the commercial production of soy products or may be
advanced in certain breeding protocols for the production of novel
soybean varieties. A hybrid plant can also be used as a recurrent
parent at any given stage in a backcrossing protocol during the
production of a single locus conversion of the soybean variety
XR25AL15X.
[0023] Soybean variety XR25AL15X is well suited to the development
of new varieties based on the elite nature of the genetic
background of the variety. In selecting a second plant to cross
with XR25AL15X for the purpose of developing novel soybean
varieties, it will typically be desired to choose those plants
which either themselves exhibit one or more selected desirable
characteristics or which exhibit the desired characteristic(s) when
in hybrid combination. Examples of potentially desired
characteristics include seed yield, lodging resistance, emergence,
seedling vigor, disease tolerance, maturity, plant height, high oil
content, high protein content and shattering resistance.
[0024] Choice of breeding or selection methods depends on the mode
of plant reproduction, the heritability of the trait(s) being
improved, and the type of variety used commercially (e.g., F.sub.1
hybrid variety, pureline variety, etc.). For highly heritable
traits, a choice of superior individual plants evaluated at a
single location will be effective, whereas for traits with low
heritability, selection should be based on mean values obtained
from replicated evaluations of families of related plants. Popular
selection methods commonly include pedigree selection, modified
pedigree selection, mass selection, recurrent selection and
backcrossing.
[0025] The complexity of inheritance influences choice of the
breeding method. Backcross breeding is used to transfer one or a
few favorable genes for a highly heritable trait into a desirable
variety. This approach has been used extensively for breeding
disease-resistant varieties (Bowers et al., Crop Sci., 32(1):67-72,
1992; Nickell and Bernard, Crop Sci., 32(3):835, 1992). Various
recurrent selection techniques are used to improve quantitatively
inherited traits controlled by numerous genes. The use of recurrent
selection in self-pollinating crops depends on the ease of
pollination, the frequency of successful hybrids from each
pollination, and the number of hybrid offspring from each
successful cross.
[0026] Each breeding program should include a periodic, objective
evaluation of the efficiency of the breeding procedure. Evaluation
criteria vary depending on the goal and objectives, but should
include gain from selection per year based on comparisons to an
appropriate standard, overall value of the advanced breeding lines,
and number of successful varieties produced per unit of input
(e.g., per year, per dollar expended, etc.).
[0027] Promising advanced breeding lines are thoroughly tested and
compared to appropriate standards in environments representative of
the commercial target area(s) for generally three or more years.
The best lines are candidates for new commercial varieties. Those
still deficient in a few traits may be used as parents to produce
new populations for further selection.
[0028] These processes, which lead to the final step of marketing
and distribution, may take as much as eight to 12 years from the
time the first cross is made. Therefore, development of new
varieties is a time-consuming process that requires precise forward
planning, efficient use of resources, and a minimum of changes in
direction.
[0029] A most difficult task is the identification of individuals
that are genetically superior, because for most traits, the true
genotypic value is masked by other confounding plant traits or
environmental factors. One method of identifying a superior plant
is to observe its performance relative to other experimental plants
and to one or more widely grown standard varieties. Single
observations are generally inconclusive, while replicated
observations provide a better estimate of genetic worth.
[0030] The goal of plant breeding is to develop new, unique and
superior soybean varieties and hybrids. The breeder initially
selects and crosses two or more parental lines, followed by
repeated selfing and selection, producing many new genetic
combinations. Each year, the plant breeder selects the germplasm to
advance to the next generation. This germplasm is grown under
unique and different geographical, climatic and soil conditions,
and further selections are then made, during and at the end of the
growing season. The varieties which are developed are
unpredictable. This unpredictability is because the breeder's
selection occurs in unique environments, with no control at the DNA
level (using conventional breeding procedures), and with millions
of different possible genetic combinations being generated. A
breeder of ordinary skill in the art cannot predict the final
resulting lines he develops, except possibly in a very gross and
general fashion. The same breeder cannot produce the same variety
twice by using the exact same original parents and the same
selection techniques. This unpredictability results in the
expenditure of large amounts of research monies to develop superior
new soybean varieties.
[0031] Pedigree breeding and recurrent selection breeding methods
are used to develop varieties from breeding populations. Breeding
programs combine desirable traits from two or more varieties or
various broad-based sources into breeding pools from which
varieties are developed by selfing and selection of desired
phenotypes. The new varieties are evaluated to determine which have
commercial potential.
[0032] Pedigree breeding is commonly used for the improvement of
self-pollinating crops. Two parents which possess favorable,
complementary traits are crossed to produce an F.sub.1. An F.sub.2
population is produced by selfing one or several F.sub.1's.
Selection of the best individuals may begin in the F.sub.2
population (or later depending upon the breeder's objectives);
then, beginning in the F.sub.3, the best individuals in the best
families can be selected. Replicated testing of families can begin
in the F.sub.3 or F.sub.4 generation to improve the effectiveness
of selection for traits with low heritability. At an advanced stage
of inbreeding (i.e., F.sub.6 and F.sub.7), the best lines or
mixtures of phenotypically similar lines are tested for potential
release as new varieties.
[0033] Mass and recurrent selections can be used to improve
populations of either self- or cross-pollinating crops. A
genetically variable population of heterozygous individuals is
either identified or created by intercrossing several different
parents. The best plants are selected based on individual
superiority, outstanding progeny, or excellent combining ability.
The selected plants are intercrossed to produce a new population in
which further cycles of selection are continued.
[0034] Backcross breeding has been used to transfer genetic loci
for simply inherited, highly heritable traits into a desirable
homozygous variety which is the recurrent parent. The source of the
trait to be transferred is called the donor or nonrecurrent parent.
The resulting plant is expected to have the attributes of the
recurrent parent (i.e., variety) and the desirable trait
transferred from the donor parent. After the initial cross,
individuals possessing the phenotype of the donor parent are
selected and repeatedly crossed (backcrossed) to the recurrent
parent. The resulting plant is expected to have the attributes of
the recurrent parent (i.e., variety) and the desirable trait
transferred from the donor parent.
[0035] The single-seed descent procedure in the strict sense refers
to planting a segregating population, harvesting a sample of one
seed per plant, and using the one-seed sample to plant the next
generation. When the population has been advanced from the F.sub.2
to the desired level of inbreeding, the plants from which lines are
derived will each trace to different F.sub.2 individuals. The
number of plants in a population declines each generation due to
failure of some seeds to germinate or some plants to produce at
least one seed. As a result, not all of the F.sub.2 plants
originally sampled in the population will be represented by a
progeny when generation advance is completed.
[0036] In a multiple-seed procedure, soybean breeders commonly
harvest one or more pods from each plant in a population and thresh
them together to form a bulk. Part of the bulk is used to plant the
next generation and part is put in reserve. This procedure is also
referred to as modified single-seed descent or the pod-bulk
technique.
[0037] The multiple-seed procedure has been used to save labor at
harvest. It is considerably faster to thresh pods with a machine
than to remove one seed from each by hand for the single-seed
procedure. The multiple-seed procedure also makes it possible to
plant the same number of seeds of a population each generation of
inbreeding. Enough seeds are harvested to make up for those plants
that did not germinate or produce seed.
[0038] Descriptions of other breeding methods that are commonly
used for different traits and crops can be found in one of several
reference books (e.g., Allard, "Principles of plant breeding," John
Wiley & Sons, NY, University of California, Davis, Calif.,
50-98, 1960; Simmonds, "Principles of crop improvement," Longman,
Inc., NY, 369-399, 1979; Sneep and Hendriksen, "Plant breeding
perspectives," Wageningen (ed), Center for Agricultural Publishing
and Documentation, 1979; Fehr, In: Soybeans: Improvement,
Production and Uses," 2d Ed., Manograph 16:249, 1987; Fehr,
"Principles of cultivar development," Theory and Technique (Vol 1)
and Crop Species Soybean (Vol 2), Iowa State Univ., Macmillian Pub.
Co., NY, 360-376, 1987; Poehlman and Sleper, "Breeding Field Crops"
Iowa State University Press, Ames, 1995; Sprague and Dudley, eds.,
Corn and Improvement, 5th ed., 2006).
[0039] Proper testing should detect any major faults and establish
the level of superiority or improvement over current varieties. In
addition to showing superior performance, there must be a demand
for a new variety that is compatible with industry standards or
which creates a new market. The introduction of a new variety will
incur additional costs to the seed producer, the grower, processor
and consumer; for special advertising and marketing, altered seed
and commercial production practices, and new product utilization.
The testing preceding release of a new variety should take into
consideration research and development costs as well as technical
superiority of the final variety. For seed-propagated varieties, it
must be feasible to produce seed easily and economically.
[0040] In addition to phenotypic observations, a plant can also be
identified by its genotype. The genotype of a plant can be
characterized through a molecular marker profile, which can
identify plants of the same variety or a related variety, can
identify plants and plant parts which are genetically superior as a
result of an event comprising a backcross conversion, transgene, or
genetic sterility factor, or can be used to determine or validate a
pedigree. Such molecular marker profiling can be accomplished using
a variety of techniques including, but not limited to, restriction
fragment length polymorphism (RFLP), amplified fragment length
polymorphism (AFLP), sequence-tagged sites (STS), randomly
amplified polymorphic DNA (RAPD), arbitrarily primed polymerase
chain reaction (AP-PCR), DNA amplification fingerprinting (DAF),
sequence characterized amplified regions (SCARs), variable number
tandem repeat (VNTR), short tandem repeat (STR), single feature
polymorphism (SFP), simple sequence length polymorphism (SSLP),
restriction site associated DNA, allozymes, isozyme markers, single
nucleotide polymorphisms (SNPs), or simple sequence repeat (SSR)
markers, also known as microsatellites (Gupta et al., 1999; Korzun
et al., 2001). Various types of these markers, for example, can be
used to identify individual varieties developed from specific
parent varieties, as well as cells or other plant parts thereof.
For example, see Cregan et al. (1999) "An Integrated Genetic
Linkage Map of the Soybean Genome" Crop Science 39:1464-1490, and
Berry et al. (2003) "Assessing Probability of Ancestry Using Simple
Sequence Repeat Profiles: Applications to Maize Inbred Lines and
Soybean Varieties" Genetics 165:331-342, each of which are
incorporated by reference herein in their entirety.
[0041] In some examples, one or more markers may be used to
characterize and/or evaluate a soybean variety. Particular markers
used for these purposes are not limited to any particular set of
markers, but are envisioned to include any type of marker and
marker profile that provides a means for distinguishing varieties.
One method of comparison may to use only homozygous loci for
soybean variety XR25AL15X.
[0042] Primers and PCR protocols for assaying these and other
markers are disclosed in, for example, Soybase (sponsored by the
USDA Agricultural Research Service and Iowa State University)
located on the world wide web at 129.186.26/94/SSR.html. In
addition to being used for identification of soybean variety
XR25AL15X, as well as plant parts and plant cells of soybean
variety XR25AL15X, a genetic profile may be used to identify a
soybean plant produced through the use of soybean variety XR25AL15X
or to verify a pedigree for progeny plants produced through the use
of soybean variety XR25AL15X. A genetic marker profile may also be
useful in breeding and developing backcross conversions.
[0043] In an embodiment, the present invention provides a soybean
plant characterized by molecular and physiological data obtained
from a representative sample of said variety deposited with the
American Type Culture Collection (ATCC). Thus, plants, seeds, or
parts thereof, having all or essentially all of the morphological
and physiological characteristics of soybean variety XR25AL15X are
provided. Further provided is a soybean plant formed by the
combination of the disclosed soybean plant or plant cell with
another soybean plant or cell and comprising the homozygous alleles
of the variety.
[0044] In some examples, a plant, a plant part, or a seed of
soybean variety XR25AL15X may be characterized by producing a
molecular profile. A molecular profile may include, but is not
limited to, one or more genotypic and/or phenotypic profile(s). A
genotypic profile may include, but is not limited to, a marker
profile, such as a genetic map, a linkage map, a trait maker
profile, a SNP profile, an SSR profile, a genome-wide marker
profile, a haplotype, and the like. A molecular profile may also be
a nucleic acid sequence profile, and/or a physical map. A
phenotypic profile may include, but is not limited to, a protein
expression profile, a metabolic profile, an mRNA expression
profile, and the like.
[0045] One means of performing genetic marker profiles is using SSR
polymorphisms that are well known in the art. A marker system based
on SSRs can be highly informative in linkage analysis relative to
other marker systems, in that multiple alleles may be present.
Another advantage of this type of marker is that through use of
flanking primers, detection of SSRs can be achieved, for example,
by using the polymerase chain reaction (PCR), thereby eliminating
the need for labor-intensive Southern hybridization.
[0046] PCR detection may be performed using two oligonucleotide
primers flanking the polymorphic segment of repetitive DNA to
amplify the SSR region.
[0047] Following amplification, markers can be scored by
electrophoresis of the amplification products. Scoring of marker
genotype is based on the size of the amplified fragment, which
correlates to the number of base pairs of the fragment. While
variation in the primer used or in the laboratory procedures can
affect the reported fragment size, relative values should remain
constant regardless of specific primer or laboratory used. When
comparing varieties, it may be beneficial to have all profiles
performed in the same lab. Primers that can be used are publically
available and may be found in, for example, Soybase or Cregan (Crop
Science 39:1464-1490, 1999).
[0048] A genotypic profile of soybean variety XR25AL15X can be used
to identify a plant comprising variety XR25AL15X as a parent, since
such plants will comprise the same homozygous alleles as variety
XR25AL15X. Because the soybean variety is essentially homozygous at
all relevant loci, most loci should have only one type of allele
present. In contrast, a genetic marker profile of an F1 progeny
should be the sum of those parents, e.g., if one parent was
homozygous for allele X at a particular locus, and the other parent
homozygous for allele Y at that locus, then the F1 progeny will be
XY (heterozygous) at that locus. Subsequent generations of progeny
produced by selection and breeding are expected to be of genotype
XX (homozygous), YY (homozygous), or XY (heterozygous) for that
locus position. When the F1 plant is selfed or sibbed for
successive filial generations, the locus should be either X or Y
for that position.
[0049] In addition, plants and plant parts substantially benefiting
from the use of variety XR25AL15X in their development, such as
variety XR25AL15X comprising a backcross conversion, transgene, or
genetic sterility factor, may be identified by having a molecular
marker profile with a high percent identity to soybean variety
XR25AL15X. Such a percent identity might be 90%, 91%, 92%, 93%,
94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to soybean
variety XR25AL15X.
[0050] A genotypic profile of variety XR25AL15X also can be used to
identify essentially derived varieties and other progeny varieties
developed from the use of variety XR25AL15X, as well as cells and
other plant parts thereof. Plants of the invention include any
plant having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,
99%, 99.5%, or 99.9% of the markers in the genotypic profile, and
that retain 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%/0, 99%,
99.5%, or 99.9% of the morphological and physiological
characteristics of variety XR25AL15X when grown under the same
conditions. Such plants may be developed using markers well known
in the art. Progeny plants and plant parts produced using variety
XR25AL15X may be identified, for example, by having a molecular
marker profile of at least 25%, 30%, 35%, 400%, 45%, 50%, 55%, 60%,
65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%,
86%, 87%, 88%, 89%, 900%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,
99%, or 99.5% genetic contribution from soybean variety XR25AL15X,
as measured by either percent identity or percent similarity. Such
progeny may be further characterized as being within a pedigree
distance of variety XR25AL15X, such as within 1, 2, 3, 4, or 5 or
less cross pollinations to a soybean plant other than variety
XR25AL15X, or a plant that has variety XR25AL15X as a progenitor.
Unique molecular profiles may be identified with other molecular
tools, such as SNPs and RFLPs.
[0051] Any time the soybean variety XR25AL15X is crossed with
another, different, variety, first generation (F.sub.1) soybean
progeny are produced. The hybrid progeny are produced regardless of
characteristics of the two varieties produced. As such, an F.sub.1
hybrid soybean plant may be produced by crossing XR25AL5X with any
second soybean plant. The second soybean plant may be genetically
homogeneous (e.g., inbred) or may itself be a hybrid. Therefore,
any F.sub.1 hybrid soybean plant produced by crossing soybean
variety XR25AL15X with a second soybean plant is a part of the
present invention.
[0052] Soybean plants (Glycine max L.) can be crossed by either
natural or mechanical techniques (see, e.g., Fehr, "Soybean," In:
Hybridization of Crop Plants, Fehr and Hadley (eds), Am. Soc.
Agron. and Crop Sci. Soc. Am., Madison, Wis., 590-599, 1980).
Natural pollination occurs in soybeans either by self pollination
or natural cross pollination, which typically is aided by
pollinating organisms. In either natural or artificial crosses,
flowering and flowering time are an important consideration.
Soybean is a short-day plant, but there is considerable genetic
variation for sensitivity to photoperiod (Hamner, "Glycine max(L.)
Merrill," In: The Induction of Flowering: Some Case Histories,
Evans (ed), Cornell Univ. Press, Ithaca, N.Y., 62-89, 1969;
Criswell and Hume, Crop Sci., 12:657-660, 1972). The critical day
length for flowering ranges from about 13 h for genotypes adapted
to tropical latitudes to 24 h for photoperiod-insensitive genotypes
grown at higher latitudes (Shibles et al., "Soybean," In: Crop
Physiology, Some Case Histories, Evans (ed), Cambridge Univ. Press,
Cambridge, England, 51-189, 1975). Soybeans seem to be insensitive
to day length for 9 days after emergence. Photoperiods shorter than
the critical day length are required for 7 to 26 days to complete
flower induction (Borthwick and Parker, Bot. Gaz., 100:374-387,
1938; Shanmugasundaram and Tsou, Crop Sci., 18:598-601, 1978).
[0053] Sensitivity to day length is an important consideration when
genotypes are grown outside of their area of adaptation. When
genotypes adapted to tropical latitudes are grown in the field at
higher latitudes, they may not mature before frost occurs. Plants
can be induced to flower and mature earlier by creating
artificially short days or by grafting (Fehr, "Soybean," In:
Hybridization of Crop Plants, Fehr and Hadley (eds), Am. Soc.
Agron. and Crop Sci. Soc. Am., Madison, Wis., 590-599, 1980).
Soybeans frequently are grown in winter nurseries located at sea
level in tropical latitudes where day lengths are much shorter than
their critical photoperiod. The short day lengths and warm
temperatures encourage early flowering and seed maturation, and
genotypes can produce a seed crop in 90 days or fewer after
planting. Early flowering is useful for generation advance when
only a few self-pollinated seeds per plant are needed, but not for
artificial hybridization because the flowers self-pollinate before
they are large enough to manipulate for hybridization. Artificial
lighting can be used to extend the natural day length to about 14.5
h to obtain flowers suitable for hybridization and to increase
yields of self-pollinated seed.
[0054] The effect of a short photoperiod on flowering and seed
yield can be partly offset by altitude, probably due to the effects
of cool temperature (Major et al., Crop Sci., 15:174-179, 1975). At
tropical latitudes, varieties adapted to the northern U.S. perform
more like those adapted to the southern U.S. at high altitudes than
they do at sea level.
[0055] The light level required to delay flowering is dependent on
the quality of light emitted from the source and the genotype being
grown. Blue light with a wavelength of about 480 nm requires more
than 30 times the energy to inhibit flowering as red light with a
wavelength of about 640 nm (Parker et al., Bot. Gaz., 108:1-26,
1946).
[0056] Temperature can also play a significant role in the
flowering and development of soybean (Major et al., Crop Sci.,
15:174-179, 1975). It can influence the time of flowering and
suitability of flowers for hybridization. Temperatures below
21.degree. C. or above 32.degree. C. can reduce floral initiation
or seed set (Hamner, "Glycine max(L.) Merrill," In: The Induction
of Flowering: Some Case Histories, Evans (ed), Cornell Univ. Press,
Ithaca, N.Y., 62-89, 1969, van Schaik and Probst, Agron. J.,
50:192-197, 1958). Artificial hybridization is most successful
between 26.degree. C. and 32.degree. C. because cooler temperatures
reduce pollen shed and result in flowers that self-pollinate before
they are large enough to manipulate. Warmer temperatures frequently
are associated with increased flower abortion caused by moisture
stress; however, successful crosses are possible at about
35.degree. C. if soil moisture is adequate.
[0057] Soybeans have been classified as indeterminate,
semi-determinate, and determinate based on the abruptness of stem
termination after flowering begins (Bernard and Weiss, "Qualitative
genetics," In: Soybeans: Improvement, Production, and Uses,
Caldwell (ed), Am. Soc. of Agron., Madison, Wis., 117-154, 1973).
When grown at their latitude of adaptation, indeterminate genotypes
flower when about one-half of the nodes on the main stem have
developed. They have short racemes with few flowers, and their
terminal node has only a few flowers. Semi-determinate genotypes
also flower when about one-half of the nodes on the main stem have
developed, but node development and flowering on the main stem
stops more abruptly than on indeterminate genotypes. Their racemes
are short and have few flowers, except for the terminal one, which
may have several times more flowers than those lower on the plant.
Determinate varieties begin flowering when all or most of the nodes
on the main stem have developed. They usually have elongated
racemes that may be several centimeters in length and may have a
large number of flowers. Stem termination and flowering habit are
reported to be controlled by two major genes (Bernard and Weiss,
"Qualitative genetics," In: Soybeans: Improvement, Production, and
Uses, Caldwell (ed), Am. Soc. of Agron., Madison, Wis., 117-154,
1973).
[0058] Soybean flowers typically are self-pollinated on the day the
corolla opens. The amount of natural crossing, which is typically
associated with insect vectors such as honeybees, is approximately
1% for adjacent plants within a row and 0.5% between plants in
adjacent rows (Boerma and Moradshahi, Crop Sci., 15:858-861, 1975).
The structure of soybean flowers is similar to that of other legume
species and consists of a calyx with five sepals, a corolla with
five petals, 10 stamens, and a pistil (Carlson, "Morphology", In:
Soybeans: Improvement. Production, and Uses, Caldwell (ed), Am.
Soc. of Agron., Madison, Wis., 17-95, 1973). The calyx encloses the
corolla until the day before anthesis. The corolla emerges and
unfolds to expose a standard, two wing petals, and two keel petals.
An open flower is about 7 mm long from the base of the calyx to the
tip of the standard and 6 mm wide across the standard. The pistil
consists of a single ovary that contains one to five ovules, a
style that curves toward the standard, and a club-shaped stigma.
The stigma is receptive to pollen about 1 day before anthesis and
remains receptive for 2 days after anthesis, if the flower petals
are not removed. Filaments of nine stamens are fused, and the one
nearest the standard is free. The stamens form a ring below the
stigma until about 1 day before anthesis, then their filaments
begin to elongate rapidly and elevate the anthers around the
stigma. The anthers dehisce on the day of anthesis, pollen grains
fall on the stigma, and within 10 h the pollen tubes reach the
ovary and fertilization is completed (Johnson and Bernard, "Soybean
genetics and breeding," In: The Soybean, Norman (ed), Academic
Press, NY, 1-73, 1963).
[0059] Self-pollination occurs naturally in soybean with no
manipulation of the flowers. For the crossing of two soybean
plants, it is often beneficial, although not required, to utilize
artificial hybridization. In artificial hybridization, the flower
used as a female in a cross is manually cross pollinated prior to
maturation of pollen from the flower, thereby preventing self
fertilization, or alternatively, the male parts of the flower are
emasculated using a technique known in the art. Techniques for
emasculating the male parts of a soybean flower include, for
example, physical removal of the male parts, use of a genetic
factor conferring male sterility, and application of a chemical
gametocide to the male parts.
[0060] For artificial hybridization employing emasculation, flowers
that are expected to open the following day are selected on the
female parent. The buds are swollen and the corolla is just visible
through the calyx or has begun to emerge. The selected buds on a
parent plant are prepared, and all self-pollinated flowers or
immature buds are removed. Special care is required to remove
immature buds that are hidden under the stipules at the leaf axil,
and which could develop into flowers at a later date. To remove
flowers, the flower is grasped and the location of the stigma
determined by examining the sepals. A long, curvy sepal covers the
keel, and the stigma is on the opposite side of the flower. The
calyx is removed by pulling each sepal down and around the flower,
and the exposed corolla is removed just above the calyx scar,
taking care to remove the keel petals without injuring the stigma.
The ring of anthers is visible after the corolla is removed, unless
the anthers were removed with the petals. Cross-pollination can
then be carried out using, for example, petri dishes or envelopes
in which male flowers have been collected. Desiccators containing
calcium chloride crystals are used in some environments to dry male
flowers to obtain adequate pollen shed.
[0061] It has been demonstrated that emasculation is unnecessary to
prevent self-pollination (Walker et al., Crop Sci., 19:285-286,
1979). When emasculation is not used, the anthers near the stigma
frequently are removed to make it clearly visible for pollination.
The female flower usually is hand-pollinated immediately after it
is prepared; although a delay of several hours does not seem to
reduce seed set. Pollen shed typically begins in the morning and
may end when temperatures are above 30.degree. C., or may begin
later and continue throughout much of the day with more moderate
temperatures.
[0062] Pollen is available from a flower with a recently opened
corolla, but the degree of corolla opening associated with pollen
shed may vary during the day. In many environments, it is possible
to collect male flowers and use them immediately without storage.
In the southern U.S. and other humid climates, pollen shed occurs
in the morning when female flowers are more immature and difficult
to manipulate than in the afternoon, and the flowers may be damp
from heavy dew. In those circumstances, male flowers may be
collected into envelopes or petri dishes in the morning and the
open container placed in a desiccator for about 4 h at a
temperature of about 25.degree. C. The desiccator may be taken to
the field in the afternoon and kept in the shade to prevent
excessive temperatures from developing within it. Pollen viability
can be maintained in flowers for up to 2 days when stored at about
5.degree. C. In a desiccator at 3.degree. C., flowers can be stored
successfully for several weeks; however, varieties may differ in
the percentage of pollen that germinates after long-term storage
(Kuehl, "Pollen viability and stigma receptivity of Glycine max
(L.) Merrill," Thesis, North Carolina State College, Raleigh, N.C.,
1961).
[0063] Either with or without emasculation of the female flower,
hand pollination can be carried out by removing the stamens and
pistil with a forceps from a flower of the male parent and gently
brushing the anthers against the stigma of the female flower.
Access to the stamens can be achieved by removing the front sepal
and keel petals, or piercing the keel with closed forceps and
allowing them to open to push the petals away. Brushing the anthers
on the stigma causes them to rupture, and the highest percentage of
successful crosses is obtained when pollen is clearly visible on
the stigma. Pollen shed can be checked by tapping the anthers
before brushing the stigma. Several male flowers may have to be
used to obtain suitable pollen shed when conditions are
unfavorable, or the same male may be used to pollinate several
flowers with good pollen shed.
[0064] When male flowers do not have to be collected and dried in a
desiccator, it may be desired to plant the parents of a cross
adjacent to each other. Plants usually are grown in rows 65 to 100
cm apart to facilitate movement of personnel within the field
nursery. Yield of self-pollinated seed from an individual plant may
range from a few seeds to more than 1,000 as a function of plant
density. A density of 30 plants/m of row can be used when 30 or
fewer seeds per plant is adequate, 10 plants/m can be used to
obtain about 100 seeds/plant, and 3 plants/m usually results in
maximum seed production per plant. Densities of 12 plants/m or less
commonly are used for artificial hybridization.
[0065] Multiple planting dates about 7 to 14 days apart usually are
used to match parents of different flowering dates. When
differences in flowering dates are extreme between parents,
flowering of the later parent can be hastened by creating an
artificially short day or flowering of the earlier parent can be
delayed by use of artificially long days or delayed planting. For
example, crosses with genotypes adapted to the southern U.S. are
made in northern U.S. locations by covering the late genotype with
a box, large can, or similar container to create an artificially
short photoperiod of about 12 h for about 15 days beginning when
there are three nodes with trifoliate leaves on the main stem.
Plants induced to flower early tend to have flowers that
self-pollinate when they are small and can be difficult to prepare
for hybridization.
[0066] Grafting can be used to hasten the flowering of late
flowering genotypes. A scion from a late genotype grafted on a
stock that has begun to flower will begin to bloom up to 42 days
earlier than normal (Kiihl et al., Crop Sci., 17:181-182, 1977).
First flowers on the scion appear from 21 to 50 days after the
graft.
[0067] Observing pod development 7 days after pollination generally
is adequate to identify a successful cross. Abortion of pods and
seeds can occur several weeks after pollination, but the percentage
of abortion usually is low if plant stress is minimized (Shibles et
al., "Soybean," In: Crop Physiology, Some Case Histories, Evans
(ed), Cambridge Univ. Press, Cambridge, England, 51-189, 1975).
Pods that develop from artificial hybridization can be
distinguished from self-pollinated pods by the presence of the
calyx scar, caused by removal of the sepals. The sepals begin to
fall off as the pods mature; therefore, harvest should be completed
at or immediately before the time the pods reach their mature
color. Harvesting pods early also avoids any loss by
shattering.
[0068] Once harvested, pods are typically air-dried at not more
than 38.degree. C. until the seeds contain 13% moisture or less,
then the seeds are removed by hand. Seed can be stored
satisfactorily at about 25.degree. C. for up to a year if relative
humidity is 50% or less. In humid climates, germination percentage
declines rapidly unless the seed is dried to 7% moisture and stored
in an air-tight container at room temperature. Long-term storage in
any climate is best accomplished by drying seed to 7% moisture and
storing it at 10.degree. C. or less in a room maintained at 50%
relative humidity or in an air-tight container.
II. Further Embodiments of the Invention
[0069] In certain aspects of the invention, plants of soybean
variety XR25AL15X are modified to include at least a first desired
heritable trait. Such plants may, in one embodiment, be developed
by a plant breeding technique called backcrossing, wherein
essentially all of the morphological and physiological
characteristics of a variety are recovered in addition to a genetic
locus transferred into the plant via the backcrossing technique. By
essentially all of the morphological and physiological
characteristics, it is meant that the characteristics of a plant
are recovered that are otherwise present when compared in the same
environment, other than occasional variant traits that might arise
during backcrossing or direct introduction of a transgene. It is
understood that a locus introduced by backcrossing may or may not
be transgenic in origin, and thus the term backcrossing
specifically includes backcrossing to introduce loci that were
created by genetic transformation.
[0070] In a typical backcross protocol, the original variety of
interest (recurrent parent) is crossed to a second variety
(nonrecurrent parent) that carries the single locus of interest to
be transferred. The resulting progeny from this cross are then
crossed again to the recurrent parent and the process is repeated
until a soybean plant is obtained wherein essentially all of the
morphological and physiological characteristics of the recurrent
parent are recovered in the converted plant, in addition to the
transferred locus from the nonrecurrent parent.
[0071] The selection of a suitable recurrent parent is an important
step for a successful backcrossing procedure. The goal of a
backcross protocol is to alter or substitute a trait or
characteristic in the original variety. To accomplish this, a locus
of the recurrent variety is modified or substituted with the
desired locus from the nonrecurrent parent, while retaining
essentially all of the rest of the genetic, and therefore the
morphological and physiological constitution of the original
variety. The choice of the particular nonrecurrent parent will
depend on the purpose of the backcross; one of the major purposes
is to add some commercially desirable, agronomically important
trait to the plant. The exact backcrossing protocol will depend on
the characteristic or trait being altered to determine an
appropriate testing protocol. Although backcrossing methods are
simplified when the characteristic being transferred is a dominant
allele, a recessive allele may also be transferred. In this
instance it may be necessary to introduce a test of the progeny to
determine if the desired characteristic has been successfully
transferred.
[0072] Soybean varieties can also be developed from more than two
parents (Fehr, In: "Soybeans: Improvement, Production and Uses,"
2nd Ed., Manograph 16:249, 1987). The technique, known as modified
backcrossing, uses different recurrent parents during the
backcrossing. Modified backcrossing may be used to replace the
original recurrent parent with a variety having certain more
desirable characteristics or multiple parents may be used to obtain
different desirable characteristics from each.
[0073] Many traits have been identified that are not regularly
selected for in the development of a new inbred but that can be
improved by backcrossing techniques. Traits may or may not be
transgenic; examples of these traits include, but are not limited
to, male sterility, herbicide resistance, resistance to bacterial,
fungal, or viral disease, insect and pest resistance, restoration
of male fertility, enhanced nutritional quality, yield stability,
and yield enhancement. These comprise genes generally inherited
through the nucleus.
[0074] Direct selection may be applied where the locus acts as a
dominant trait. An example of a dominant trait is the herbicide
resistance trait. For this selection process, the progeny of the
initial cross are sprayed with the herbicide prior to the
backcrossing. The spraying eliminates any plants which do not have
the desired herbicide resistance characteristic, and only those
plants which have the herbicide resistance gene are used in the
subsequent backcross. This process is then repeated for all
additional backcross generations.
[0075] Selection of soybean plants for breeding is not necessarily
dependent on the phenotype of a plant and instead can be based on
genetic investigations. For example, one may utilize a suitable
genetic marker which is closely associated with a trait of
interest. One of these markers may therefore be used to identify
the presence or absence of a trait in the offspring of a particular
cross, and hence may be used in selection of progeny for continued
breeding. This technique may commonly be referred to as marker
assisted selection. Any other type of genetic marker or other assay
which is able to identify the relative presence or absence of a
trait of interest in a plant may also be useful for breeding
purposes. Procedures for marker assisted selection applicable to
the breeding of soybeans are well known in the art. Such methods
will be of particular utility in the case of recessive traits and
variable phenotypes, or where conventional assays may be more
expensive, time consuming or otherwise disadvantageous. Types of
genetic markers which could be used in accordance with the
invention include, but are not necessarily limited to, Simple
Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic
Acids Res., 18:6531-6535, 1990), Randomly Amplified Polymorphic
DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence
Characterized Amplified Regions (SCARs), Arbitrary Primed
Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length
Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein
by reference in its entirety), and Single Nucleotide Polymorphisms
(SNPs) (Wang et al., Science, 280:1077-1082, 1998).
[0076] Many qualitative characters also have potential use as
phenotype-based genetic markers in soybeans; however, some or many
may not differ among varieties commonly used as parents (Bernard
and Weiss, "Qualitative genetics," In: Soybeans: Improvement,
Production, and Uses, Caldwell (ed), Am. Soc. of Agron., Madison,
Wis., 117-154, 1973). The most widely used genetic markers are
flower color (purple dominant to white), pubescence color (brown
dominant to gray), and pod color (brown dominant to tan). The
association of purple hypocotyl color with purple flowers and green
hypocotyl color with white flowers is commonly used to identify
hybrids in the seedling stage. Differences in maturity, height,
hilum color, and pest resistance between parents can also be used
to verify hybrid plants.
[0077] Many useful traits that can be introduced by backcrossing,
as well as directly into a plant, are those which are introduced by
genetic transformation techniques. Genetic transformation may
therefore be used to insert a selected transgene into the soybean
variety of the invention or may, alternatively, be used for the
preparation of transgenes which can be introduced by backcrossing.
Methods for the transformation of many economically important
plants, including soybeans, are well known to those of skill in the
art. Techniques which may be employed for the genetic
transformation of soybeans include, but are not limited to,
electroporation, microprojectile bombardment,
Agrobacterium-mediated transformation and direct DNA uptake by
protoplasts.
[0078] To effect transformation by electroporation, one may employ
either friable tissues, such as a suspension culture of cells or
embryogenic callus or alternatively one may transform immature
embryos or other organized tissue directly. In this technique, one
would partially degrade the cell walls of the chosen cells by
exposing them to pectin-degrading enzymes (pectolyases) or
mechanically wound tissues in a controlled manner.
[0079] Protoplasts may also be employed for electroporation
transformation of plants (Bates, Mol. Biotechnol., 2(2):135-145,
1994; Lazzeri, Methods Mol. Biol., 49:95-106, 1995). For example,
the generation of transgenic soybean plants by electroporation of
cotyledon-derived protoplasts was described by Dhir and Widholm in
Intl. Pat. App. Publ. No. WO 92/17598, the disclosure of which is
specifically incorporated herein by reference.
[0080] A particularly efficient method for delivering transforming
DNA segments to plant cells is microprojectile bombardment. In this
method, particles are coated with nucleic acids and delivered into
cells by a propelling force. Exemplary particles include those
comprised of tungsten, platinum, and often, gold. For the
bombardment, cells in suspension are concentrated on filters or
solid culture medium. Alternatively, immature embryos or other
target cells may be arranged on solid culture medium. The cells to
be bombarded are positioned at an appropriate distance below the
macroprojectile stopping plate.
[0081] An illustrative embodiment of a method for delivering DNA
into plant cells by acceleration is the Biolistics Particle
Delivery System, which can be used to propel particles coated with
DNA or cells through a screen, such as a stainless steel or Nytex
screen, onto a surface covered with target soybean cells. The
screen disperses the particles so that they are not delivered to
the recipient cells in large aggregates. It is believed that a
screen intervening between the projectile apparatus and the cells
to be bombarded reduces the size of the projectile aggregate and
may contribute to a higher frequency of transformation by reducing
the damage inflicted on the recipient cells by projectiles that are
too large.
[0082] Microprojectile bombardment techniques are widely
applicable, and may be used to transform virtually any plant
species. The application of microprojectile bombardment for the
transformation of soybeans is described, for example, in U.S. Pat.
No. 5,322,783, the disclosure of which is specifically incorporated
herein by reference in its entirety.
[0083] Agrobacterium-mediated transfer is another widely applicable
system for introducing gene loci into plant cells. An advantage of
the technique is that DNA can be introduced into whole plant
tissues, thereby bypassing the need for regeneration of an intact
plant from a protoplast. Modern Agrobacterium transformation
vectors are capable of replication in E. coli as well as
Agrobacterium, allowing for convenient manipulations (Klee et al.,
Bio. Tech., 3(7):637-642, 1985). Moreover, recent technological
advances in vectors for Agrobacterium-mediated gene transfer have
improved the arrangement of genes and cloning sites in the vectors
to facilitate the construction of vectors capable of expressing
various polypeptide coding genes. Vectors can have convenient
multiple-cloning sites (MCS) flanked by a promoter and a
polyadenylation site for direct expression of inserted polypeptide
coding genes. Other vectors can comprise site-specific
recombination sequences, enabling insertion of a desired DNA
sequence without the use of restriction enzymes (Curtis et al.,
Plant Physiology 133:462-469, 2003). Additionally, Agrobacterium
containing both armed and disarmed Ti genes can be used for
transformation.
[0084] In those plant strains where Agrobacterium-mediated
transformation is efficient, it is the method of choice because of
the facile and defined nature of the gene locus transfer. The use
of Agrobacterium-mediated plant integrating vectors to introduce
DNA into plant cells is well known in the art (Fraley et al., Bio.
Tech., 3(7):629-635, 1985; U.S. Pat. No. 5,563,055). Use of
Agrobacterium in the context of soybean transformation has been
described, for example, by Chee and Slightom (Methods Mol. Biol.,
44:101-119, 1995) and in U.S. Pat. No. 5,569,834, the disclosures
of which are specifically incorporated herein by reference in their
entirety.
[0085] Transformation of plant protoplasts also can be achieved
using methods based on calcium phosphate precipitation,
polyethylene glycol treatment, electroporation, and combinations of
these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet.,
199(2):169-177, 1985; Omirulleh et al., Plant Mol. Biol.,
21(3):415-428, 1993; Fromm et al., Nature, 319(6056):791-793, 1986;
Uchimiya et al., Mol. Gen. Genet., 204(2):204-207, 1986; Marcotte
et al., Nature, 335(6189):454-457, 1988). The demonstrated ability
to regenerate soybean plants from protoplasts makes each of these
techniques applicable to soybean (Dhir et al., Plant Cell Rep.,
10(2):97-101, 1991).
[0086] Many hundreds if not thousands of different genes are known
and could potentially be introduced into a soybean plant according
to the invention. Non-limiting examples of particular genes and
corresponding phenotypes one may choose to introduce into a soybean
plant are presented below.
[0087] A. Herbicide Resistance
[0088] Numerous herbicide resistance genes are known and may be
employed with the invention. An example is a gene conferring
resistance to a herbicide that inhibits the growing point or
meristem, such as an imidazalinone or a sulfonylurea. Exemplary
genes in this category code for mutant ALS and AHAS enzyme as
described, for example, by Lee et al., EMBO J., 7:1241, 1988; Gleen
et al., Plant Molec. Biology, 18:1185-1187, 1992; and Miki et al.,
Theor. Appl. Genet., 80:449, 1990.
[0089] Resistance genes for glyphosate (resistance conferred by
mutant 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and aroA
genes, respectively) and other phosphono compounds such as
glufosinate (phosphinothricin acetyltransferase (PAT) and
Streptomyces hygroscopicus phosphinothricin-acetyl transferase
(bar) genes) may also be used. See, for example, U.S. Pat. No.
4,940,835 to Shah et al., which discloses the nucleotide sequence
of a form of EPSPS which can confer glyphosate resistance. Examples
of specific EPSPS transformation events conferring glyphosate
resistance are provided by U.S. Pat. Nos. 6,040,497 and 7,632,985.
The MON89788 event disclosed in U.S. Pat. No. 7,632,985 in
particular is beneficial in conferring glyphosate tolerance in
combination with an increase in average yield relative to prior
events.
[0090] A DNA molecule encoding a mutant aroA gene can be obtained
under ATCC Accession Number 39256, and the nucleotide sequence of
the mutant gene is disclosed in U.S. Pat. No. 4,769,061 to Comai. A
hygromycin B phosphotransferase gene from E. coli which confers
resistance to glyphosate in tobacco callus and plants is described
in Penaloza-Vazquez et al., Plant Cell Reports, 14:482-487, 1995.
European Patent Application No. 0 333 033 to Kumada et al., and
U.S. Pat. No. 4,975,374 to Goodman et al., disclose nucleotide
sequences of glutamine synthetase genes which confer resistance to
herbicides such as L-phosphinothricin. The nucleotide sequence of a
phosphinothricin acetyltransferase gene is provided in European
Patent Application No. 0 242 246 to Leemans et al. DeGreef et al.
(Biotechnology, 7:61, 1989), describe the production of transgenic
plants that express chimeric bar genes coding for phosphinothricin
acetyl transferase activity. Exemplary genes conferring resistance
to phenoxy propionic acids and cyclohexanediones, such as
sethoxydim and haloxyfop are the Acct-S1, Acct-S2 and Acct-S3 genes
described by Marshall et al., (Theor. Appl. Genet., 83:4:35,
1992).
[0091] Genes are also known conferring resistance to a herbicide
that inhibits photosynthesis, such as a triazine (psbA and gs+
genes) and a benzonitrile (nitrilase gene). Przibila et al. (Plant
Cell, 3:169, 1991) describe the transformation of Chlamydomonas
with plasmids encoding mutant psbA genes. Nucleotide sequences for
nitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to
Stalker, and DNA molecules containing these genes are available
under ATCC Accession Nos. 53435, 67441, and 67442. Cloning and
expression of DNA coding for a glutathione S-transferase is
described by Hayes et al. (Biochem. J., 285(Pt 1): 173-180, 1992).
Protoporphyrinogen oxidase (PPO) is the target of the PPO-inhibitor
class of herbicides; a PPO-inhibitor resistant PPO gene was
recently identified in Amaranthus tuberculatus (Patzoldt et al.,
PNAS, 103(33):12329-2334, 2006). The herbicide methyl viologen
inhibits CO.sub.2 assimilation. Foyer et al. (Plant Physiol.,
109:1047-1057, 1995) describe a plant overexpressing glutathione
reductase (GR) which is resistant to methyl viologen treatment.
[0092] Siminszky (Phytochemistry Reviews, 5:445-458, 2006)
describes plant cytochrome P450-mediated detoxification of
multiple, chemically unrelated classes of herbicides. Modified
bacterial genes have been successfully demonstrated to confer
resistance to atrazine, a herbicide that binds to the
plastoquinone-binding membrane protein Q.sub.B in photosystem II to
inhibit electron transport. See, for example, studies by Cheung et
al. (PNAS, 85(2):391-395, 1988), describing tobacco plants
expressing the chloroplast psbA gene from an atrazine-resistant
biotype of Amaranthus hybridus fused to the regulatory sequences of
a nuclear gene, and Wang et al. (Plant Biotech. J., 3:475-486,
2005), describing transgenic alfalfa, Arabidopsis, and tobacco
plants expressing the atzA gene from Pseudomonas sp. that were able
to detoxify atrazine.
[0093] Bayley et al. (Theor. Appl. Genet., 83:645-649, 1992)
describe the creation of 2,4-D-resistant transgenic tobacco and
cotton plants using the 2,4-D monooxygenase gene tfdA from
Alcaligenes eutrophus plasmid pJP5. U.S. Pat. App. Pub. No.
20030135879 describes the isolation of a gene for dicamba
monooxygenase (DMO) from Psueodomonas maltophilia that is involved
in the conversion of dicamba to a non-toxic 3,6-dichlorosalicylic
acid and thus may be used for producing plants tolerant to this
herbicide.
[0094] Other examples of herbicide resistance have been described,
for instance, in U.S. Pat. Nos. 6,803,501; 6,448,476; 6,248,876;
6,225,114; 6,107,549; 5,866,775; 5,804,425; 5,633,435;
5,463,175.
[0095] B. Disease and Pest Resistance
[0096] Plant defenses are often activated by specific interaction
between the product of a disease resistance gene (R) in the plant
and the product of a corresponding avirulence (Avr) gene in the
pathogen. A plant line can be transformed with cloned resistance
gene to engineer plants that are resistant to specific pathogen
strains. See, for example Jones et al. (Science, 266:7891, 1994)
(cloning of the tomato Cf-9 gene for resistance to Cladosporium
fulvum); Martin et al. (Science, 262: 1432, 1993) (tomato Pto gene
for resistance to Pseudomonas syringae pv. tomato); and Mindrinos
et al. (Cell, 78(6): 1089-1099, 1994) (Arabidopsis RPS2 gene for
resistance to Pseudomonas syringae).
[0097] A viral-invasive protein or a complex toxin derived
therefrom may also be used for viral disease resistance. For
example, the accumulation of viral coat proteins in transformed
plant cells imparts resistance to viral infection and/or disease
development effected by the virus from which the coat protein gene
is derived, as well as by related viruses. See Beachy et al. (Ann.
Rev. Phytopathol., 28:451, 1990). Coat protein-mediated resistance
has been conferred upon transformed plants against alfalfa mosaic
virus, cucumber mosaic virus, tobacco streak virus, potato virus X,
potato virus Y, tobacco etch virus, tobacco rattle virus and
tobacco mosaic virus. Id.
[0098] A virus-specific antibody may also be used. See, for
example, Tavladoraki et al. (Nature, 366:469, 1993), who show that
transgenic plants expressing recombinant antibody genes are
protected from virus attack. Virus resistance has also been
described in, for example, U.S. Pat. Nos. 6,617,496; 6,608,241;
6,015,940; 6,013,864; 5,850,023 and 5,304,730. Additional means of
inducing whole-plant resistance to a pathogen include modulation of
the systemic acquired resistance (SAR) or pathogenesis related (PR)
genes, for example genes homologous to the Arabidopsis thaliana
NIMI1/NPR1/SAI1, and/or by increasing salicylic acid production
(Ryals et al., Plant Cell, 8:1809-1819, 1996).
[0099] Logemann et al. (Biotechnology, 10:305, 1992), for example,
disclose transgenic plants expressing a barley
ribosome-inactivating gene that have an increased resistance to
fungal disease. Plant defensins may be used to provide resistance
to fungal pathogens (Thomma et al., Planta, 216:193-202, 2002).
Other examples of fungal disease resistance are provided in U.S.
Pat. Nos. 6,653,280; 6,573,361; 6,506,962; 6,316,407; 6,215,048;
5,516,671; 5,773,696; 6,121,436; and 6,316,407.
[0100] Nematode resistance has been described, for example, in U.S.
Pat. No. 6,228,992, and bacterial disease resistance has been
described in U.S. Pat. No. 5,516,671.
[0101] The use of the herbicide glyphosate for disease control in
soybean plants containing event MON89788, which confers glyphosate
tolerance, has also been described in U.S. Pat. No. 7,608,761.
[0102] C. Insect Resistance
[0103] One example of an insect resistance gene includes a Bacillus
thuringiensis protein, a derivative thereof or a synthetic
polypeptide modeled thereon. See, for example, Geiser et al. (Gene,
48(1):109-118, 1986), who disclose the cloning and nucleotide
sequence of a Bacillus thuringiensis .delta.-endotoxin gene.
Moreover, DNA molecules encoding .delta.-endotoxin genes can be
purchased from the American Type Culture Collection, Manassas, Va.,
for example, under ATCC Accession Nos. 40098, 67136, 31995 and
31998. Another example is a lectin. See, for example, Van Damme et
al., (Plant Molec. Biol., 24:25, 1994), who disclose the nucleotide
sequences of several Clivia miniata mannose-binding lectin genes. A
vitamin-binding protein may also be used, such as avidin. See PCT
Application No. US93/06487, the contents of which are hereby
incorporated by reference. This application teaches the use of
avidin and avidin homologues as larvicides against insect
pests.
[0104] Yet another insect resistance gene is an enzyme inhibitor,
for example, a protease or proteinase inhibitor or an amylase
inhibitor. See, for example, Abe et al. (J. Biol. Chem., 262:16793,
1987) (nucleotide sequence of rice cysteine proteinase inhibitor),
Huub et al. (Plant Molec. Biol., 21:985, 1993) (nucleotide sequence
of cDNA encoding tobacco proteinase inhibitor I), and Sumitani et
al. (Biosci. Biotech. Biochem., 57:1243, 1993) (nucleotide sequence
of Streptomyces nitrosporeus .alpha.-amylase inhibitor).
[0105] An insect-specific hormone or pheromone may also be used.
See, for example, the disclosure by Hammock et al. (Nature,
344:458, 1990), of baculovirus expression of cloned juvenile
hormone esterase, an inactivator of juvenile hormone; Gade and
Goldsworthy (Eds. Physiological System in Insects, Elsevier
Academic Press, Burlington, Mass., 2007), describing allostatins
and their potential use in pest control; and Palli et al. (Vitam.
Horm., 73:59-100, 2005), disclosing use of ecdysteroid and
ecdysteroid receptor in agriculture. The diuretic hormone receptor
(DHR) was identified in Price et al. (Insect Mol. Biol.,
13:469-480, 2004) as a candidate target of insecticides.
[0106] Still other examples include an insect-specific antibody or
an immunotoxin derived therefrom and a developmental-arrestive
protein. See Taylor et al. (Seventh Int'l Symposium on Molecular
Plant-Microbe Interactions, Edinburgh, Scotland, Abstract W97,
1994), who described enzymatic inactivation in transgenic tobacco
via production of single-chain antibody fragments. Numerous other
examples of insect resistance have been described. See, for
example, U.S. Pat. Nos. 6,809,078; 6,713,063; 6,686,452; 6,657,046;
6,645,497; 6,642,030; 6,639,054; 6,620,988; 6,593,293; 6,555,655;
6,538,109; 6,537,756; 6,521,442; 6,501,009; 6,468,523; 6,326,351;
6,313,378; 6,284,949; 6,281,016; 6,248,536; 6,242,241; 6,221,649;
6,177,615; 6,156,573; 6,153,814; 6,110,464; 6,093,695; 6,063,756;
6,063,597; 6,023,013; 5,959,091; 5,942,664; 5,942,658, 5,880,275;
5,763,245 and 5,763,241.
[0107] D. Male Sterility
[0108] Genetic male sterility is available in soybeans and,
although not required for crossing soybean plants, can increase the
efficiency with which hybrids are made, in that it can eliminate
the need to physically emasculate the soybean plant used as a
female in a given cross. (Brim and Stuber, Crop Sci., 13:528-530,
1973). Herbicide-inducible male sterility systems have also been
described. (U.S. Pat. No. 6,762,344).
[0109] Where one desires to employ male-sterility systems, it may
be beneficial to also utilize one or more male-fertility restorer
genes. For example, where cytoplasmic male sterility (CMS) is used,
hybrid seed production requires three inbred lines: (1) a
cytoplasmically male-sterile line having a CMS cytoplasm; (2) a
fertile inbred with normal cytoplasm, which is isogenic with the
CMS line for nuclear genes ("maintainer line"); and (3) a distinct,
fertile inbred with normal cytoplasm, carrying a fertility
restoring gene ("restorer" line). The CMS line is propagated by
pollination with the maintainer line, with all of the progeny being
male sterile, as the CMS cytoplasm is derived from the female
parent. These male sterile plants can then be efficiently employed
as the female parent in hybrid crosses with the restorer line,
without the need for physical emasculation of the male reproductive
parts of the female parent.
[0110] The presence of a male-fertility restorer gene results in
the production of fully fertile F.sub.1 hybrid progeny. If no
restorer gene is present in the male parent, male-sterile hybrids
are obtained. Such hybrids are useful where the vegetative tissue
of the soybean plant is utilized, but in many cases the seeds will
be deemed the most valuable portion of the crop, so fertility of
the hybrids in these crops must be restored. Therefore, one aspect
of the current invention concerns plants of the soybean variety
XR25AL15X comprising a genetic locus capable of restoring male
fertility in an otherwise male-sterile plant. Examples of
male-sterility genes and corresponding restorers which could be
employed with the plants of the invention are well known to those
of skill in the art of plant breeding (see, e.g., U.S. Pat. Nos.
5,530,191 and 5,684,242, the disclosures of which are each
specifically incorporated herein by reference in their
entirety).
[0111] E. Modified Fatty Acid, Phytate and Carbohydrate
Metabolism
[0112] Genes may be used conferring modified fatty acid metabolism.
For example, stearyl-ACP desaturase genes may be used. See Knutzon
et al. (Proc. Natl. Acad. Sci. USA, 89:2624, 1992). Various fatty
acid desaturases have also been described. McDonough et al.
describe a Saccharomyces cerevisiae OLE1 gene encoding
.DELTA.9-fatty acid desaturase, an enzyme which forms the
monounsaturated palmitoleic (16:1) and oleic (18:1) fatty acids
from palmitoyl (16:0) or stearoyl (18:0) CoA (J. Biol. Chem.,
267(9):5931-5936, 1992). Fox et al. describe a gene encoding a
stearoyl-acyl carrier protein delta-9 desaturase from castor (Proc.
Natl. Acad. Sci. USA, 90(6):2486-2490, 1993). Reddy et al. describe
.DELTA.6- and .DELTA.12-desaturases from the cyanobacteria
Synechocystis responsible for the conversion of linoleic acid
(18:2) to gamma-linolenic acid (18:3 gamma) (Plant Mol. Biol.,
22(2):293-300, 1993). A gene from Arabidopsis thaliana that encodes
an omega-3 desaturase has been identified (Arondel et al. Science,
258(5086):1353-1355, 1992). Plant .DELTA.9-desaturases (PCT
Application Publ. No. WO 91/13972) and soybean and Brassica
.DELTA.15-desaturases (European Patent Application Publ. No. EP
0616644) have also been described. U.S. Pat. No. 7,622,632
describes fungal .DELTA.15-desaturases and their use in plants. EP
Patent No. 1656449 describes .DELTA.6-desaturases from Primula as
well as soybean plants having an increased stearidonic acid (SDA,
18:4) content. U.S. Pat. App. Pub. No. 2008-0260929 describes
expression of transgenic desaturase enzymes in corn plants, and
improved fatty acid profiles resulting therefrom.
[0113] Modified oil production is disclosed, for example, in U.S.
Pat. Nos. 6,444,876; 6,426,447 and 6,380,462. High oil production
is disclosed, for example, in U.S. Pat. Nos. 6,495,739; 5,608,149;
6,483,008 and 6,476,295. Modified fatty acid content is disclosed,
for example, in U.S. Pat. Nos. 6,828,475; 6,822,141; 6,770,465;
6,706,950; 6,660,849; 6,596,538; 6,589,767; 6,537,750; 6,489,461
and 6,459,018.
[0114] Phytate metabolism may also be modified by introduction of a
phytase-encoding gene to enhance breakdown of phytate, adding more
free phosphate to the transformed plant. For example, see Van
Hartingsveldt et al. (Gene, 127:87, 1993), for a disclosure of the
nucleotide sequence of an Aspergillus niger phytase gene. In
soybean, this, for example, could be accomplished by cloning and
then reintroducing DNA associated with the single allele which is
responsible for soybean mutants characterized by low levels of
phytic acid. See Raboy et al. (Plant Physiol., 124(1):355-368,
2000).
[0115] A number of genes are known that may be used to alter
carbohydrate metabolism. For example, plants may be transformed
with a gene coding for an enzyme that alters the branching pattern
of starch. See Shiroza et al. (J. Bacteriol., 170:810, 1988)
(nucleotide sequence of Streptococcus mutans fructosyltransferase
gene), Steinmetz et al. (Mol. Gen. Genet., 20:220, 1985)
(nucleotide sequence of Bacillus subtilis levansucrase gene), Pen
el al. (Biotechnology, 10:292, 1992) (production of transgenic
plants that express Bacillus licheniformis .alpha.-amylase), Elliot
et al. (Plant Molec. Biol., 21:515, 1993) (nucleotide sequences of
tomato invertase genes), Sergaard et al. (J. Biol. Chem.,
268:22480, 1993) (site-directed mutagenesis of barley
.alpha.-amylase gene), and Fisher et al. (Plant Physiol., 102:1045,
1993) (maize endosperm starch branching enzyme II). The Z10 gene
encoding a 10 kD zein storage protein from maize may also be used
to alter the quantities of 10 kD zein in the cells relative to
other components (Kirihara et al., Gene, 71(2):359-370, 1988).
[0116] F. Resistance to Abiotic Stress
[0117] Abiotic stress includes dehydration or other osmotic stress,
salinity, high or low light intensity, high or low temperatures,
submergence, exposure to heavy metals, and oxidative stress.
Delta-pyrroline-5-carboxylate synthetase (P5CS) from mothbean has
been used to provide protection against general osmotic stress.
Mannitol-1-phosphate dehydrogenase (mt1D) from E. coli has been
used to provide protection against drought and salinity. Choline
oxidase (codA from Arthrobactor globiformis) can protect against
cold and salt. E. coli choline dehydrogenase (betA) provides
protection against salt. Additional protection from cold can be
provided by omega-3-fatty acid desaturase (fad7) from Arabidopsis
thaliana. Trehalose-6-phosphate synthase and levan sucrase (SacB)
from yeast and Bacillus subtilis, respectively, can provide
protection against drought (summarized from Annex II Genetic
Engineering for Abiotic Stress Tolerance in Plants, Consultative
Group On International Agricultural Research Technical Advisory
Committee). Overexpression of superoxide dismutase can be used to
protect against superoxides, as described in U.S. Pat. No.
5,538,878 to Thomas et al.
[0118] G. Additional Traits
[0119] Additional traits can be introduced into the soybean variety
of the present invention. A non-limiting example of such a trait is
a coding sequence which decreases RNA and/or protein levels. The
decreased RNA and/or protein levels may be achieved through RNAi
methods, such as those described in U.S. Pat. No. 6,506,559 to Fire
and Mellow.
[0120] Another trait that may find use with the soybean variety of
the invention is a sequence which allows for site-specific
recombination. Examples of such sequences include the FRT sequence,
used with the FLP recombinase (Zhu and Sadowski, J. Biol. Chem.,
270:23044-23054, 1995); and the LOX sequence, used with CRE
recombinase (Sauer, Mol. Cell. Biol., 7:2087-2096, 1987). The
recombinase genes can be encoded at any location within the genome
of the soybean plant, and are active in the hemizygous state.
[0121] It may also be desirable to make soybean plants more
tolerant to or more easily transformed with Agrobacterium
tumefaciens. Expression of p53 and lap, two baculovirus cell-death
suppressor genes, inhibited tissue necrosis and DNA cleavage.
Additional targets can include plant-encoded proteins that interact
with the Agrobacterium Vir genes; enzymes involved in plant cell
wall formation; and histones, histone acetyltransferases and
histone deacetylases (reviewed in Gelvin, Microbiology & Mol.
Biol. Reviews, 67:16-37, 2003).
[0122] In addition to the modification of oil, fatty acid or
phytate content described above, it may additionally be beneficial
to modify the amounts or levels of other compounds. For example,
the amount or composition of antioxidants can be altered. See, for
example, U.S. Pat. No. 6,787,618; U.S. Pat. App. Pub. No.
20040034886 and International Patent App. Pub. No. WO 00/68393,
which disclose the manipulation of antioxidant levels, and
International Patent App. Pub. No. WO 03/082899, which discloses
the manipulation of a antioxidant biosynthetic pathway.
[0123] Additionally, seed amino acid content may be manipulated.
U.S. Pat. No. 5,850,016 and International Patent App. Pub. No. WO
99/40209 disclose the alteration of the amino acid compositions of
seeds. U.S. Pat. Nos. 6,080,913 and 6,127,600 disclose methods of
increasing accumulation of essential amino acids in seeds.
[0124] U.S. Pat. No. 5,559,223 describes synthetic storage proteins
in which the levels of essential amino acids can be manipulated.
International Patent App. Pub. No. WO 99/29882 discloses methods
for altering amino acid content of proteins. International Patent
App. Pub. No. WO 98/20133 describes proteins with enhanced levels
of essential amino acids. International Patent App. Pub. No. WO
98/56935 and U.S. Pat. Nos. 6,346,403, 6,441,274 and 6,664,445
disclose plant amino acid biosynthetic enzymes. International
Patent App. Pub. No. WO 98/45458 describes synthetic seed proteins
having a higher percentage of essential amino acids than
wild-type.
[0125] U.S. Pat. No. 5,633,436 discloses plants comprising a higher
content of sulfur-containing amino acids; U.S. Pat. No. 5,885,801
discloses plants comprising a high threonine content; U.S. Pat.
Nos. 5,885,802 and 5,912,414 disclose plants comprising a high
methionine content; U.S. Pat. No. 5,990,389 discloses plants
comprising a high lysine content; U.S. Pat. No. 6,459,019 discloses
plants comprising an increased lysine and threonine content;
International Patent App. Pub. No. WO 98/42831 discloses plants
comprising a high lysine content; International Patent App. Pub.
No. WO 96/01905 discloses plants comprising a high threonine
content and International Patent App. Pub. No. WO 95/15392
discloses plants comprising a high lysine content.
III. Origin and Breeding History of an Exemplary Single Locus
Converted Plant
[0126] It is known to those of skill in the art that, by way of the
technique of backcrossing, one or more traits may be introduced
into a given variety while otherwise retaining essentially all of
the traits of that variety. An example of such backcrossing to
introduce a trait into a starting variety is described in U.S. Pat.
No. 6,140,556, the entire disclosure of which is specifically
incorporated herein by reference. The procedure described in U.S.
Pat. No. 6,140,556 can be summarized as follows: The soybean
variety known as Williams '82 [Glycine max L. Merr.] (Reg. No. 222,
PI 518671) was developed using backcrossing techniques to transfer
a locus comprising the Rps.sub.1 gene to the variety Williams
(Bernard and Cremeens, Crop Sci., 28:1027-1028, 1988). Williams '82
is a composite of four resistant lines from the BC.sub.6F.sub.3
generation, which were selected from 12 field-tested resistant
lines from Williams x Kingwa. The variety Williams was used as the
recurrent parent in the backcross and the variety Kingwa was used
as the source of the Rps.sub.1 locus. This gene locus confers
resistance to 19 of the 24 races of the fungal agent phytopthora
root rot.
[0127] The F.sub.1 or F.sub.2 seedlings from each backcross round
were tested for resistance to the fungus by hypocotyl inoculation
using the inoculum of race 5. The final generation was tested using
inoculum of races 1 to 9. In a backcross such as this, where the
desired characteristic being transferred to the recurrent parent is
controlled by a major gene which can be readily evaluated during
the backcrossing, it is common to conduct enough backcrosses to
avoid testing individual progeny for specific traits such as yield
in extensive replicated tests. In general, four or more backcrosses
are used when there is no evaluation of the progeny for specific
traits, such as yield. As in this example, lines with the phenotype
of the recurrent parent may be composited without the usual
replicated tests for traits such as yield, protein or oil
percentage in the individual lines.
[0128] The variety Williams '82 is comparable to the recurrent
parent variety Williams in its traits except resistance to
phytopthora rot. For example, both varieties have a relative
maturity of 38, indeterminate stems, white flowers, brown
pubescence, tan pods at maturity and shiny yellow seeds with black
to light black hila.
IV. Tissue Cultures and In Vitro Regeneration of Soybean Plants
[0129] A further aspect of the invention relates to tissue cultures
of the soybean variety designated XR25AL15X. As used herein, the
term "tissue culture" indicates a composition comprising isolated
cells of the same or a different type or a collection of such cells
organized into parts of a plant. Exemplary types of tissue cultures
are protoplasts, calli and plant cells that are intact in plants or
parts of plants, such as embryos, pollen, flowers, leaves, roots,
root tips, anthers, and the like. In one embodiment, the tissue
culture comprises embryos, protoplasts, meristematic cells, pollen,
leaves or anthers.
[0130] Exemplary procedures for preparing tissue cultures of
regenerable soybean cells and regenerating soybean plants
therefrom, are disclosed in U.S. Pat. Nos. 4,992,375; 5,015,580;
5.024,944 and 5,416,011, each of the disclosures of which is
specifically incorporated herein by reference in its entirety.
[0131] An important ability of a tissue culture is the capability
to regenerate fertile plants. This allows, for example,
transformation of the tissue culture cells followed by regeneration
of transgenic plants. For transformation to be efficient and
successful, DNA must be introduced into cells that give rise to
plants or germ-line tissue.
[0132] Soybeans typically are regenerated via two distinct
processes: shoot morphogenesis and somatic embryogenesis (Finer,
Cheng, Verma, "Soybean transformation: Technologies and progress,"
In: Soybean: Genetics. Molecular Biology and Biotechnology, CAB
Intl, Verma and Shoemaker (ed), Wallingford, Oxon, UK, 250-251,
1996). Shoot morphogenesis is the process of shoot meristem
organization and development. Shoots grow out from a source tissue
and are excised and rooted to obtain an intact plant. During
somatic embryogenesis, an embryo (similar to the zygotic embryo),
containing both shoot and root axes, is formed from somatic plant
tissue. An intact plant rather than a rooted shoot results from the
germination of the somatic embryo.
[0133] Shoot morphogenesis and somatic embryogenesis are different
processes and the specific route of regeneration is primarily
dependent on the explant source and media used for tissue culture
manipulations. While the systems are different, both systems show
variety-specific responses where some lines are more responsive to
tissue culture manipulations than others. A line that is highly
responsive in shoot morphogenesis may not generate many somatic
embryos. Lines that produce large numbers of embryos during an
`induction` step may not give rise to rapidly-growing proliferative
cultures. Therefore, it may be desired to optimize tissue culture
conditions for each soybean line. These optimizations may readily
be carried out by one of skill in the art of tissue culture through
small-scale culture studies. In addition to line-specific
responses, proliferative cultures can be observed with both shoot
morphogenesis and somatic embryogenesis. Proliferation is
beneficial for both systems, as it allows a single, transformed
cell to multiply to the point that it will contribute to germ-line
tissue.
[0134] Shoot morphogenesis was first reported by Wright et al.
(Plant Cell Reports, 5:150-154, 1986) as a system whereby shoots
were obtained de novo from cotyledonary nodes of soybean seedlings.
The shoot meristems were formed subepidermally and morphogenic
tissue could proliferate on a medium containing benzyl adenine
(BA). This system can be used for transformation if the
subepidermal, multicellular origin of the shoots is recognized and
proliferative cultures are utilized. The idea is to target tissue
that will give rise to new shoots and proliferate those cells
within the meristematic tissue to lessen problems associated with
chimerism. Formation of chimeras, resulting from transformation of
only a single cell in a meristem, are problematic if the
transformed cell is not adequately proliferated and does not does
not give rise to germ-line tissue. Once the system is well
understood and reproduced satisfactorily, it can be used as one
target tissue for soybean transformation.
[0135] Somatic embryogenesis in soybean was first reported by
Christianson et al. (Science, 222:632-634, 1983) as a system in
which embryogenic tissue was initially obtained from the zygotic
embryo axis. These embryogenic cultures were proliferative but the
repeatability of the system was low and the origin of the embryos
was not reported. Later histological studies of a different
proliferative embryogenic soybean culture showed that proliferative
embryos were of apical or surface origin with a small number of
cells contributing to embryo formation. The origin of primary
embryos (the first embryos derived from the initial explant) is
dependent on the explant tissue and the auxin levels in the
induction medium (Hartweck et al., In Vitro Cell. Develop. Bio.,
24:821-828, 1988). With proliferative embryonic cultures, single
cells or small groups of surface cells of the `older` somatic
embryos form the `newer` embryos.
[0136] Embryogenic cultures can also be used successfully for
regeneration, including regeneration of transgenic plants, if the
origin of the embryos is recognized and the biological limitations
of proliferative embryogenic cultures are understood. Biological
limitations include the difficulty in developing proliferative
embryogenic cultures and reduced fertility problems
(culture-induced variation) associated with plants regenerated from
long-term proliferative embryogenic cultures. Some of these
problems are accentuated in prolonged cultures. The use of more
recently cultured cells may decrease or eliminate such
problems.
V. Definitions
[0137] In the description and tables, a number of terms are used.
In order to provide a clear and consistent understanding of the
specification and claims, the following definitions are
provided:
[0138] A: When used in conjunction with the word "comprising" or
other open language in the claims, the words "a" and "an" denote
"one or more."
[0139] About: Refers to embodiments or values that include the
standard deviation of the mean for a given item being measured.
[0140] Allele: Any of one or more alternative forms of a gene
locus, all of which relate to one trait or characteristic. In a
diploid cell or organism, the two alleles of a given gene occupy
corresponding loci on a pair of homologous chromosomes.
[0141] Aphids: Aphid resistance in greenhouse screening is scored
on a scale from 1 to 9 based on foliar symptoms and number of
aphids; Resistant (R) corresponds to a rating of 1-3.9, moderately
resistant (MR) 4.0-5.9, moderately susceptible to moderately
resistant (MS-MR) 6.0-6.9, and susceptible (S) 7.0-9.0.
[0142] Asian Soybean Rust (ASR): ASR may be visually scored from 1
to 5, where 1=immune; 2=leaf exhibits red/brown lesions over less
than 50% of surface; 3=leaf exhibits red/brown lesions over greater
than 50% of surface; 4=leaf exhibits tan lesions over less than
50%0 of surface; and 5=leaf exhibits tan lesions over greater than
50% of surface. Resistance to ASR may be characterized
phenotypically as well as genetically. Soybean plants
phenotypically characterized as resistant to ASR typically exhibit
red brown lesions covering less than 25% of the leaf. Genetic
characterization of ASR resistance may be carried out, for example,
by identifying the presence in a soybean plant of one or more
genetic markers linked to the ASR resistance.
[0143] Backcrossing: A process in which a breeder repeatedly
crosses hybrid progeny, for example a first generation hybrid
(F.sub.1), back to one of the parents of the hybrid progeny.
Backcrossing can be used to introduce one or more single locus
conversions from one genetic background into another.
[0144] Brown Stem Rot (BSR): The greenhouse score is based on the
incidence and severity of pith discoloration. Scores are converted
to a 1-9 scale where Resistant (R) corresponds to a rating<3.1,
moderately resistant (MR) 3.1-5.0, moderately susceptible (MS)
5.1-6.9, susceptible (S) 7.0-7.9, and highly susceptible
(HS)>7.9.
[0145] Chloride Sensitivity: Plants may be categorized as
"includers" or "excluders" with respect to chloride sensitivity.
Excluders tend to partition chloride in the root systems and reduce
the amount of chloride transported to more sensitive, aboveground
tissues. Therefore excluders may display increased tolerance to
elevated soil chloride levels compared to includers. Greenhouse
screening Chloride tolerance is reported on a 1-9 scale where a
rating less than 3 is considered and excluder and 4-9 is considered
an includer.
[0146] Chromatography: A technique wherein a mixture of dissolved
substances are bound to a solid support followed by passing a
column of fluid across the solid support and varying the
composition of the fluid. The components of the mixture are
separated by selective elution.
[0147] Crossing: The mating of two parent plants.
[0148] Cross-pollination: Fertilization by the union of two gametes
from different plants.
[0149] Emasculate: The removal of plant male sex organs or the
inactivation of the organs with a cytoplasmic or nuclear genetic
factor or a chemical agent conferring male sterility.
[0150] Emergence (EMR): The emergence score describes the ability
of a seed to emerge from the soil after planting. Each genotype is
given a 1 to 9 score based on its percent of emergence. A score of
I indicates an excellent rate and percent of emergence, an
intermediate score of 5 indicates an average rating and a 9 score
indicates a very poor rate and percent of emergence.
[0151] Enzymes: Molecules which can act as catalysts in biological
reactions.
[0152] F.sub.1 Hybrid: The first generation progeny of the cross of
two nonisogenic plants.
[0153] Fatty Acids: Are measured and reported as a percent of the
total oil content. In addition to the typical composition of fatty
acids in commodity soybeans, some soybean varieties have modified
profiles. Low linolenic acid soybean oil as defined herein contains
3% or less linolenic acid. Mid oleic acid soybean oil as defined
herein contains typically 50-60% oleic acid. High oleic soybean oil
as defined herein contains typically 75% or greater oleic acid.
Stearidonic acid levels are typically 0 percent in commodity
soybeans.
[0154] Frog Eye Leaf Spot (FELS): Greenhouse assay reaction scores
are based on foliar symptom severity, measured using a 1-9 scale.
Resistant (R) corresponds to a rating<3, moderately resistant
(MR) 3.0-4.9, moderately susceptible (MS) 5.0-6.9, and susceptible
(S)>6.9.
[0155] Genotype: The genetic constitution of a cell or
organism.
[0156] Haploid: A cell or organism having one set of the two sets
of chromosomes in a diploid.
[0157] Iron-Deficiency Chlorosis (IDE=early; IDL=late):
Iron-deficiency chlorosis is scored in a system ranging from 1 to 9
based on visual observations. A score of 1 means no stunting of the
plants or yellowing of the leaves and a score of 9 indicates the
plants are dead or dying caused by iron-deficiency chlorosis; a
score of 5 means plants have intermediate health with some leaf
yellowing.
[0158] Linkage: A phenomenon wherein alleles on the same chromosome
tend to segregate together more often than expected by chance if
their transmission was independent.
[0159] Linolenic Acid Content (LLN): Low-linolenic acid soybean oil
contains three percent or less linolenic acid, compared to eight
percent linolenic acid for traditional soybeans.
[0160] Lodging Resistance (LDG): Lodging is rated on a scale of 1
to 9. A score of I indicates erect plants. A score of 5 indicates
plants are leaning at a 45 degree(s) angle in relation to the
ground and a score of 9 indicates plants are lying on the
ground.
[0161] Marker: A readily detectable phenotype, preferably inherited
in codominant fashion (both alleles at a locus in a diploid
heterozygote are readily detectable), with no environmental
variance component, i.e., heritability of 1.
[0162] Maturity Date (MAT): Plants are considered mature when 95%
of the pods have reached their mature color. The maturity date is
typically described in measured days after August 31 in the
northern hemisphere.
[0163] Moisture (MST): The average percentage moisture in the seeds
of the variety.
[0164] Oil or Oil Percent: Seed oil content is measured and
reported on a percentage basis.
[0165] Or: As used herein is meant to mean "and/or" and be
interchangeable therewith unless explicitly indicated to refer to
the alternative only.
[0166] Phenotype: The detectable characteristics of a cell or
organism, which characteristics are the manifestation of gene
expression.
[0167] Phenotypic Score (PSC): The phenotypic score is a visual
rating of the general appearance of the variety. All visual traits
are considered in the score, including healthiness, standability,
appearance and freedom from disease. Ratings are scored as 1 being
poor to 9 being excellent.
[0168] Phytophthora Root Rot (PRR): Disorder in which the most
recognizable symptom is stem rot. Brown discoloration ranges below
the soil line and up to several inches above the soil line. Leaves
often turn yellow, dull green and/or gray and may become brown and
wilted, but remain attached to the plant.
[0169] Phytophthora Allele: Susceptibility or resistance to
Phytophthora root rot races is affected by alleles such as Rps1a
(denotes resistance to Races 1, 2, 10, 11, 13-18, 24, 26, 27, 31,
32, and 36); Rps1c (denotes resistance to Races 1-3, 6-11, 13, 15,
17, 21, 23, 24, 26, 28-30, 32, 34 and 36); Rps1k (denotes
resistance to Races 1-11, 13-15, 17, 18, 21-24, 26, 36 and 37);
Rps2 (denotes resistance to Races 1-5, 9-29, 33, 34 and 36-39);
Rps3a (denotes resistance to Races 1-5, 8, 9, 11, 13, 14, 16, 18,
23, 25, 28, 29, 31-35); Rps6 (denotes resistance to Races 1-4, 10,
12, 14-16, 18-21 and 25); and Rps7 (denotes resistance to Races 2,
12, 16, 18, 19, 33, 35 and 36).
[0170] Phytophthora Tolerance: Tolerance to Phytophthora root rot
is rated on a scale of 1 to 9 in the greenhouse assay, where a
rating less than 3.5 is considered tolerant, between 3.5-6 is
considered moderately tolerant, and greater than 6 indicates
sensitivity to Phytophthora. (Note that a score in the 1-2 range
may indicate resistance and therefore not be a true reflection of
high tolerance to Phytophthora).
[0171] Plant Height (PHT): Plant height is taken from the top of
soil to the top node of the plant and is measured in inches.
[0172] Predicted Relative Maturity (PRM): The maturity grouping
designated by the soybean industry over a given growing area. This
figure is generally divided into tenths of a relative maturity
group. Within narrow comparisons, the difference of a tenth of a
relative maturity group equates very roughly to a day difference in
maturity at harvest.
[0173] Protein (PRO), or Protein Percent: Seed protein content is
measured and reported on a percentage basis.
[0174] Regeneration: The development of a plant from tissue
culture.
[0175] Relative Maturity: The maturity grouping designated by the
soybean industry over a given growing area. This figure is
generally divided into tenths of a relative maturity group. Within
narrow comparisons, the difference of a tenth of a relative
maturity group equates very roughly to a day difference in maturity
at harvest.
[0176] Seed Protein Peroxidase Activity: Seed protein peroxidase
activity is defined as a chemical taxonomic technique to separate
varieties based on the presence or absence of the peroxidase enzyme
in the seed coat. There are two types of soybean varieties, those
having high peroxidase activity (dark red color) and those having
low peroxidase activity (no color).
[0177] Seed Weight (SWT): Soybean seeds vary in size; therefore,
the number of seeds required to make up one pound also varies. This
affects the pounds of seed required to plant a given area, and can
also impact end uses. (SW100=weight in grams of 100 seeds.)
[0178] Seed Yield (Bushels/Acre): The yield in bushels/acre is the
actual yield of the grain at harvest.
[0179] Seedling Vigor Rating (SDV): General health of the seedling,
measured on a scale of 1 to 9, where 1 is best and 9 is worst.
[0180] Seeds per Pound: Soybean seeds vary in size; therefore, the
number of seeds required to make up one pound also varies. This
affects the pounds of seed required to plant a given area, and can
also impact end uses.
[0181] Selection index (SELIN): The percentage of the test
mean.
[0182] Self-pollination: The transfer of pollen from the anther to
the stigma of the same plant.
[0183] Shattering: The amount of pod dehiscence prior to harvest.
Pod dehiscence involves seeds falling from the pods to the soil.
This is a visual score from 1 to 9 comparing all genotypes within a
given test. A score of I means pods have not opened and no seeds
have fallen out. A score of 5 indicates approximately 50%.degree.
of the pods have opened, with seeds falling to the ground and a
score of 9 indicates 100% of the pods are opened.
[0184] Single Locus Converted (Conversion) Plant: Plants which are
developed by a plant breeding technique called backcrossing and/or
by genetic transformation to introduce a given locus that is
transgenic in origin, wherein essentially all of the morphological
and physiological characteristics of a soybean variety are
recovered in addition to the characteristics of the locus
transferred into the variety via the backcrossing technique or by
genetic transformation. It is understood that once introduced into
any soybean plant genome, a locus that is transgenic in origin
(transgene), can be introduced by backcrossing as with any other
locus.
[0185] Southern Root Knot Nematode (SRKN): Greenhouse assay
reaction scores are based on severity, measured using a 1-9 scale.
Resistant (R) corresponds to a rating<6.1, moderately resistant
(MR) to 6.1<6.6, moderately resistant to moderately susceptible
(MR-MS) 6.6<7.4, and susceptible (S)>7.4.
[0186] Southern Stem Canker (STC): Greenhouse assay scoring is
based on percentage of dead plants (DP). This percentage is
converted to a 1-9 scale: l=no DP, 2=<10% DP, 3=10-30% DP,
4=31-40% DP, 5=41-50%/o DP, 6=51-60% DP, 7=61-70% DP, 8=71-90% DP,
9=91-100% DP. Resistant (R) corresponds to a rating<3.9,
moderately resistant (MR) 4-5.9%, moderately susceptible (MS)
6-7.9, susceptible (S) 8-8.9, and highly susceptible
(HS)>8.9.
[0187] Soybean Cyst Nematode (SCN): Greenhouse screening scores are
based on a female index % of Lee 74. Resistant (R) corresponds to a
rating<10%, moderately resistant (MR) 10-21.9%, moderately
resistant to moderately susceptible (MR-MS) 22-39.9%, and
susceptible (S)>39.9%.
[0188] Stearate: A fatty acid in soybean seeds measured and
reported as a percent of the total oil content.
[0189] Substantially Equivalent: A characteristic that, when
compared, does not show a statistically significant difference
(e.g., p=0.05) from the mean.
[0190] Sudden Death Syndrome: Leaf symptoms appear first as bright
yellow chlorotic spots with progressive development of brown
necrotic areas and eventual leaflet drop. Greenhouse screening
plants are scored on a 1-9 scale based on foliar symptom severity,
measured using a 1-9 scale. Resistant (R) corresponds to a
rating<3, moderately resistant (MR) 3.0-4.9, moderately
susceptible (MS) 5.0-6.9, susceptible (S) 7.0-8.0 and highly
susceptible (HS)>8.
[0191] Tissue Culture: A composition comprising isolated cells of
the same or a different type or a collection of such cells
organized into parts of a plant.
[0192] Transgene: A genetic locus comprising a sequence which has
been introduced into the genome of a soybean plant by
transformation.
[0193] Yield Best Estimate (YLD_BE): The adjusted yield of a plot
in bushels/acre. Plot yields are adjusted using the nearest
neighbor spatial covariate method first described by Papadakis
(Methode statistique pour des experiences sur champ, Thessaloniki
Plant Breeding Institute Bulletin No. 23, Thessaloniki, London,
1937).
[0194] Yield Count (YLD COUNT): The number of evaluated plots.
VI. Deposit Information
[0195] A deposit of the soybean variety XR25AL15X, which is
disclosed herein above and referenced in the claims, will be made
with the American Type Culture Collection (ATCC), 10801 University
Blvd., Manassas, Va. 20110-2209. The date of deposit is Dec. 2,
2015, and the accession number for those deposited seeds of soybean
variety XR25AL15X is ATCC Accession No. PTA-122689. All
restrictions upon the deposit have been removed, and the deposit is
intended to meet all of the requirements of 37 C.F.R.
.sctn.1.801-1.809. The deposit will be maintained in the depository
for a period of 30 years, or 5 years after the last request, or for
the effective life of the patent, whichever is longer, and will be
replaced if necessary during that period.
[0196] All of the compositions and methods disclosed and claimed
herein can be made and executed without undue experimentation in
light of the present disclosure. While the compositions and methods
of this invention have been described in terms of the foregoing
illustrative embodiments, it will be apparent to those of skill in
the art that variations, changes, modifications, and alterations
may be applied to the composition, methods, and in the steps or in
the sequence of steps of the methods described herein, without
departing from the true concept, spirit, and scope of the
invention. More specifically, it will be apparent that certain
agents that are both chemically and physiologically related may be
substituted for the agents described herein while the same or
similar results would be achieved. All such similar substitutes and
modifications apparent to those skilled in the art are deemed to be
within the spirit, scope, and concept of the invention as defined
by the appended claims.
[0197] The references cited herein, to the extent that they provide
exemplary procedural or other details supplementary to those set
forth herein, are specifically incorporated herein by
reference.
* * * * *