Register or Login To Download This Patent As A PDF
| United States Patent Application |
20090158453
|
| Kind Code
|
A1
|
|
Pazdernik; David L.
|
June 18, 2009
|
SOYBEAN CULTIVAR SG3870NRR
Abstract
A soybean cultivar designated SG3870NRR is disclosed. The invention
relates to the seeds of soybean cultivar SG3870NRR, to the plants of
soybean SG3870NRR, to plant parts of soybean cultivar SG3870NRR and to
methods for producing a soybean plant produced by crossing soybean
cultivar SG3870NRR with itself or with another soybean variety. The
invention also relates to methods for producing a soybean plant
containing in its genetic material one or more transgenes and to the
transgenic soybean plants and plant parts produced by those methods. This
invention also relates to soybean cultivars or breeding cultivars and
plant parts derived from soybean variety SG3870NRR, to methods for
producing other soybean cultivars, lines or plant parts derived from
soybean cultivar SG3870NRR and to the soybean plants, varieties, and
their parts derived from use of those methods. The invention further
relates to hybrid soybean seeds, plants and plant parts produced by
crossing the cultivar SG3870NRR with another soybean cultivar.
| Inventors: |
Pazdernik; David L.; (O'Fallon, IL)
|
| Correspondence Address:
|
JONDLE & ASSOCIATES P.C.
858 HAPPY CANYON ROAD SUITE 230
CASTLE ROCK
CO
80108
US
|
| Serial No.:
|
954571 |
| Series Code:
|
11
|
| Filed:
|
December 12, 2007 |
| Current U.S. Class: |
800/263; 426/629; 426/630; 435/415; 800/260; 800/264; 800/265; 800/278; 800/279; 800/281; 800/284; 800/300; 800/301; 800/302; 800/312 |
| Class at Publication: |
800/263; 426/629; 426/630; 435/415; 800/260; 800/264; 800/265; 800/278; 800/279; 800/281; 800/284; 800/300; 800/301; 800/302; 800/312 |
| International Class: |
A01H 1/00 20060101 A01H001/00; A01H 5/00 20060101 A01H005/00; A23L 1/20 20060101 A23L001/20; A23L 1/36 20060101 A23L001/36; C12N 15/82 20060101 C12N015/82; C12N 5/02 20060101 C12N005/02 |
Claims
1. A seed of soybean cultivar SG3870NRR, representative sample seed of
said cultivar was deposited under ATCC Accession No. PTA-9656.
2. A soybean plant, or a part thereof, produced by growing the seed of
claim 1.
3. A tissue culture produced from protoplasts or cells from the plant of
claim 2, wherein said cells or protoplasts of the tissue culture are
produced from a plant part selected from the group consisting of leaf,
pollen, embryo, cotyledon, hypocotyl, meristematic cell, root, root tip,
pistil, anther, flower, shoot, stem, pod and petiole.
4. A soybean plant regenerated from the tissue culture of claim 3, wherein
the plant has all of the morphological and physiological characteristics
of cultivar SG3870NRR.
5. A method for producing a soybean seed comprising crossing two soybean
plants and harvesting the resultant soybean seed, wherein at least one
soybean plant is the soybean plant of claim 2.
6. A soybean seed produced by the method of claim 5.
7. A soybean plant, or a part thereof, produced by growing said seed of
claim 6.
8. A method of producing an herbicide resistant soybean plant wherein the
method comprises transforming the soybean plant of claim 2 with a
transgene wherein the transgene confers resistance to an herbicide
selected from the group consisting of imidazolinone, cyclohexanedione,
sulfonylurea, glyphosate, glufosinate, phenoxy proprionic acid,
L-phosphinothricin, triazine and benzonitrile.
9. An herbicide resistant soybean plant produced by the method of claim 8.
10. A method of producing a pest or insect resistant soybean plant wherein
the method comprises transforming the soybean plant of claim 2 with a
transgene that confers pest or insect resistance.
11. A pest or insect resistant soybean plant produced by the method of
claim 10.
12. The soybean plant of claim 11, wherein the transgene encodes a
Bacillus thuringiensis endotoxin.
13. A method of producing a disease resistant soybean plant wherein the
method comprises transforming the soybean plant of claim 2 with a
transgene that confers disease resistance.
14. A disease resistant soybean plant produced by the method of claim 13.
15. A method of producing a soybean plant with modified fatty acid
metabolism or modified carbohydrate metabolism wherein the method
comprises transforming the soybean plant of claim 2 with a transgene
encoding a protein selected from the group consisting of phytase,
fructosyltransferase, levansucrase, .alpha.-amylase, invertase and starch
branching enzyme or encoding an antisense of stearyl-ACP desaturase.
16. A soybean plant having modified fatty acid metabolism or modified
carbohydrate metabolism produced by the method of claim 15.
17. A method of introducing a desired trait into soybean cultivar
SG3870NRR wherein the method comprises:(a) crossing a SG3870NRR plant,
wherein a representative sample of seed was deposited under ATCC
Accession No. PTA-9656, with a plant of another soybean cultivar that
comprises a desired trait to produce progeny plants wherein the desired
trait is selected from the group consisting of male sterility, herbicide
resistance, insect resistance, modified fatty acid metabolism, modified
carbohydrate metabolism, modified seed yield, modified oil percent,
modified protein percent, modified lodging resistance, modified
shattering, modified iron-deficiency chlorosis and resistance to
bacterial disease, fungal disease or viral disease;(b) selecting one or
more progeny plants that have the desired trait to produce selected
progeny plants;(c) crossing the selected progeny plants with the
SG3870NRR plants to produce backcross progeny plants;(d) selecting for
backcross progeny plants that have the desired trait and all of the
physiological and morphological characteristics of soybean cultivar
SG3870NRR listed in Table 1; and(e) repeating steps (c) and (d) three or
more times in succession to produce selected fourth or higher backcross
progeny plants that comprise the desired trait and all of the
physiological and morphological characteristics of soybean cultivar
SG3870NRR listed in Table 1.
18. A soybean plant produced by the method of claim 17, wherein the plant
has the desired trait.
19. The soybean plant of claim 18, wherein the desired trait is herbicide
resistance and the resistance is conferred to an herbicide selected from
the group consisting of imidazolinone, cyclohexanedione, sulfonylurea,
glyphosate, glufosinate, phenoxy proprionic acid, L-phosphinothricin,
triazine and benzonitrile.
20. The soybean plant of claim 18, wherein the desired trait is insect
resistance and the insect resistance is conferred by a transgene encoding
a Bacillus thuringiensis endotoxin.
21. The soybean plant of claim 18, wherein the desired trait is modified
fatty acid metabolism or modified carbohydrate metabolism and said
desired trait is conferred by a nucleic acid encoding a protein selected
from the group consisting of phytase, fructosyltransferase, levansucrase,
a-amylase, invertase and starch branching enzyme or encoding an antisense
of stearyl-ACP desaturase.
22. (canceled)
23. A method of producing a commodity plant product comprising obtaining
the plant of claim 2, or a part thereof, wherein the commodity plant
product is protein concentrate, protein isolate, soybean hulls, meal,
flour or oil.
Description
BACKGROUND OF THE INVENTION
[0001]The present invention relates to a new and distinctive soybean
cultivar, designated SG3870NRR. All publications cited in this
application are herein incorporated by reference.
[0002]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 possesses 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, and
better agronomic quality.
[0003]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 cultivar used commercially (e.g., F.sub.1 hybrid cultivar,
pureline cultivar, 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, and recurrent
selection.
[0004]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 cultivar. This
approach has been used extensively for breeding disease-resistant
cultivars. 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.
[0005]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 cultivars produced per unit of input (e.g., per year, per
dollar expended, etc.).
[0006]Promising advanced breeding lines are thoroughly tested and compared
to appropriate standards in environments representative of the commercial
target area(s) for three or more years. The best lines are candidates for
new commercial cultivars; those still deficient in a few traits may be
used as parents to produce new populations for further selection.
[0007]These processes, which lead to the final step of marketing and
distribution, usually take from eight to twelve years from the time the
first cross is made. Therefore, development of new cultivars is a
time-consuming process that requires precise forward planning, efficient
use of resources, and a minimum of changes in direction.
[0008]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 a widely grown standard
cultivar. If a single observation is inconclusive, replicated
observations provide a better estimate of its genetic worth.
[0009]The goal of soybean plant breeding is to develop new, unique and
superior soybean cultivars and hybrids. The breeder initially selects and
crosses two or more parental lines, followed by repeated selfing and
selection, producing many new genetic combinations. The breeder can
theoretically generate billions of different genetic combinations via
crossing, selfing and mutations. The breeder has no direct control at the
cellular level. Therefore, two breeders will never develop the same line,
or even very similar lines, having the same soybean traits.
[0010]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 cultivars that
are developed are unpredictable 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 cultivar twice by using the 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
cultivars.
[0011]The development of new soybean cultivars requires the development
and selection of soybean varieties, the crossing of these varieties and
selection of superior hybrid crosses. The hybrid seed is produced by
manual crosses between selected male-fertile parents or by using male
sterility systems. These hybrids are selected for certain single gene
traits such as pod color, flower color, pubescence color or herbicide
resistance which indicate that the seed is truly a hybrid. Additional
data on parental lines, as well as the phenotype of the hybrid, influence
the breeder's decision whether to continue with the specific hybrid
cross.
[0012]Pedigree breeding and recurrent selection breeding methods are used
to develop cultivars from breeding populations. Breeding programs combine
desirable traits from two or more cultivars or various broad-based
sources into breeding pools from which cultivars are developed by selfing
and selection of desired phenotypes. The new cultivars are evaluated to
determine which have commercial potential.
[0013]Pedigree breeding is used commonly for the improvement of
self-pollinating crops. Two parents that 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.1s. Selection of the best
individuals may begin in the F.sub.2 population; then, beginning in the
F.sub.3, the best individuals in the best families are selected.
Replicated testing of families can begin in the 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 cultivars.
[0014]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.
[0015]Backcross breeding has been used to transfer genes for a simply
inherited, highly heritable trait into a desirable homozygous cultivar or
inbred line which is the recurrent parent. The source of the trait to be
transferred is called 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 (e.g.,
cultivar) and the desirable trait transferred from the donor parent.
[0016]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.
[0017]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. The procedure has been referred to as modified
single-seed descent or the pod-bulk technique.
[0018]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.
[0019]In addition to phenotypic observations, the genotype of a plant can
also be examined. There are many laboratory-based techniques available
for the analysis, comparison and characterization of plant genotype;
among these are Isozyme Electrophoresis, Restriction Fragment Length
Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs),
Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification
Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),
Amplified Fragment Length polymorphisms (AFLPs), Simple Sequence Repeats
(SSRs--which are also referred to as Microsatellites), and Single
Nucleotide Polymorphisms (SNPs).
[0020]Proper testing should detect any major faults and establish the
level of superiority or improvement over current cultivars. In addition
to showing superior performance, there must be a demand for a new
cultivar that is compatible with industry standards or which creates a
new market. The introduction of a new cultivar 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 cultivar should take into consideration research and development
costs as well as technical superiority of the final cultivar. For
seed-propagated cultivars, it must be feasible to produce seed easily and
economically.
[0021]Soybean, Glycine max (L), is an important and valuable field crop.
Thus, a continuing goal of soybean plant breeders is to develop stable,
high yielding soybean cultivars that are agronomically sound. The reasons
for this goal are obviously 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 traits that result in superior cultivars.
[0022]The foregoing examples of the related art and limitations related
therewith are intended to be illustrative and not exclusive. Other
limitations of the related art will become apparent to those of skill in
the art upon a reading of the specification.
SUMMARY OF THE INVENTION
[0023]The following embodiments and aspects thereof are described in
conjunction with systems, tools and methods which are meant to be
exemplary, not limiting in scope. In various embodiments, one or more of
the above-described problems have been reduced or eliminated, while other
embodiments are directed to other improvements.
[0024]According to the invention, there is provided a new soybean cultivar
designated SG3870NRR. This invention thus relates to the seeds of soybean
cultivar SG3870NRR, to the plants of soybean cultivar SG3870NRR and to
methods for producing a soybean plant produced by crossing the soybean
cultivar SG3870NRR with itself or another soybean cultivar, and the
creation of variants by mutagenesis or transformation of soybean cultivar
SG3870NRR.
[0025]Thus, any such methods using the soybean cultivar SG3870NRR are part
of this invention: selfing, backcrosses, hybrid production, crosses to
populations, and the like. All plants produced using soybean cultivar
SG3870NRR as at least one parent are within the scope of this invention.
Advantageously, the soybean cultivar could be used in crosses with other,
different, soybean plants to produce first generation (F.sub.1) soybean
hybrid seeds and plants with superior characteristics.
[0026]In another aspect, the present invention provides for single or
multiple gene converted plants of soybean cultivar SG3870NRR. The
transferred gene(s) may preferably be a dominant or recessive allele.
Preferably, the transferred gene(s) will confer such traits as herbicide
resistance, insect resistance, resistance for bacterial, fungal, or viral
disease, male fertility, male sterility, enhanced nutritional quality,
modified fatty acid metabolism, modified carbohydrate metabolism,
modified seed yield, modified oil percent, modified protein percent,
modified lodging resistance, modified shattering, modified
iron-deficiency chlorosis and industrial usage. The gene may be a
naturally occurring soybean gene or a transgene introduced through
genetic engineering techniques.
[0027]In another aspect, the present invention provides regenerable cells
for use in tissue culture of soybean plant SG3870NRR. The tissue culture
will preferably be capable of regenerating plants having all the
physiological and morphological characteristics of the foregoing soybean
plant, and of regenerating plants having substantially the same genotype
as the foregoing soybean plant. Preferably, the regenerable cells in such
tissue cultures will be embryos, protoplasts, meristematic cells, callus,
pollen, leaves, anthers, cotyledons, hypocotyl, pistils, roots, root
tips, flowers, seeds, petiole, pods or stems. Still further, the present
invention provides soybean plants regenerated from the tissue cultures of
the invention.
[0028]In addition to the exemplary aspects and embodiments described
above, further aspects and embodiments will become apparent by study of
the following descriptions.
DEFINITIONS
[0029]In the description and tables that follow, a number of terms are
used. In order to provide a clear and consistent understanding of the
specification and claims, including the scope to be given such terms, the
following definitions are provided:
[0030]Allele. An allele is any of one or more alternative forms of a gene,
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.
[0031]Backcrossing. Backcrossing is a process in which a breeder
repeatedly crosses hybrid progeny back to one of the parents, for
example, a first generation hybrid F.sub.1 with one of the parental
genotypes of the F.sub.1 hybrid.
[0032]Alter. The utilization of up-regulation, down-regulation, or gene
silencing.
[0033]Brown Stem Rot. This is a visual disease score from 1 to 5 comparing
all genotypes in a given test. The score is based on leaf symptoms of
yellowing and necrosis caused by brown stem rot. Visual scores range from
a score of 1, which indicates no symptoms, to a score of 5 which
indicates severe symptoms of leaf yellowing and necrosis.
[0034]Cell. Cell as used herein includes a plant cell, whether isolated,
in tissue culture or incorporated in a plant or plant part.
[0035]Cotyledon. A cotyledon is a type of seed leaf. The cotyledon
contains the food storage tissues of the seed.
[0036]Embryo. The embryo is the small plant contained within a mature
seed.
[0037]Emergence. This score indicates the ability of the seed to emerge
when planted 3'' deep in sand at a controlled temperature of 25.degree.
C. The number of plants that emerge each day are counted. Based on this
data, each genotype is given a 1 to 5 score based on its rate of
emergence and percent of emergence. A score of 1 indicates an excellent
rate and percent of emergence, an intermediate score of 2.5 indicates
average ratings and a score of 5 indicates a very poor rate and percent
of emergence.
[0038]F.sub.3. This symbol denotes a generation resulting from the selfing
of the F.sub.2 generation along with selection for type and rogueing of
off-types. The "F" number is a term commonly used in genetics, and
designates the number of the filial generation. The "F.sub.3" generation
denotes the offspring resulting from the selfing or self mating of
members of the generation having the next lower "F" number, viz. the
F.sub.2 generation.
[0039]Gene Silencing. The interruption or suppression of the expression of
a gene at the level of transcription or translation.
[0040]Genotype. Refers to the genetic constitution of a cell or organism.
[0041]Hilum. This refers to the scar left on the seed that marks the place
where the seed was attached to the pod prior to the seed being harvested.
[0042]Hypocotyl. A hypocotyl is the portion of an embryo or seedling
between the cotyledons and the root. Therefore, it can be considered a
transition zone between shoot and root.
[0043]Iron Deficiency Chlorosis. Iron deficiency chlorosis (IDC) is a
yellowing of the leaves caused by a lack of iron in the soybean plant.
Iron is essential in the formation of chlorophyll, which gives plants
their green color. In high pH soils iron becomes insoluble and cannot be
absorbed by plant roots. Soybean cultivars differ in their genetic
ability to utilize the available iron. A score of 1 means no stunting of
the plants or yellowing of the leaves and a score of 5 indicates the
plants are dead or dying caused by iron deficiency, a score of 2.5 means
plants have intermediate health with some leaf yellowing.
[0044]Linkage. Refers to a phenomenon wherein alleles on the same
chromosome tend to segregate together more often than expected by chance
if their transmission was independent.
[0045]Linkage Disequilibrium. Refers to a phenomenon wherein alleles tend
to remain together in linkage groups when segregating from parents to
offspring, with a greater frequency than expected from their individual
frequencies.
[0046]Linoleic Acid Percent. Linoleic acid is one of the five most
abundant fatty acids in soybean seeds. It is measured by gas
chromatography and is reported as a percent of the total oil content.
[0047]Locus. A locus confers one or more traits such as, for example, male
sterility, herbicide tolerance, insect resistance, disease resistance,
waxy starch, modified fatty acid metabolism, modified phytic acid
metabolism, modified carbohydrate metabolism and modified protein
metabolism. The trait may be, for example, conferred by a naturally
occurring gene introduced into the genome of the variety by backcrossing,
a natural or induced mutation, or a transgene introduced through genetic
transformation techniques. A locus may comprise one or more alleles
integrated at a single chromosomal location.
[0048]Lodging Resistance. Lodging is rated on a scale of 1 to 5. A score
of 1 indicates erect plants. A score of 2.5 indicates plants are leaning
at a 45.degree. angle in relation to the ground and a score of 5
indicates plants are lying on the ground.
[0049]Maturity Date. Plants are considered mature when 95% of the pods
have reached their mature color. The number of days are calculated either
from August 31 or from the planting date.
[0050]Maturity Group. This refers to an agreed-on industry division of
groups of varieties based on zones in which they are adapted, primarily
according to day length or latitude. They consist of very long day length
varieties (Groups 000, 00, 0), and extend to very short day length
varieties (Groups VII, VIII, IX, X).
[0051]Relative Maturity (RM). The term relative maturity is a numerical
value that is assigned to a soybean variety based on comparisons with the
maturity values of other varieties. The number preceding the decimal
point in the RM refers to the maturity group. The number following the
decimal point refers to the relative earliness or lateness within each
maturity group. For example, a 3.0 is an early group III variety, while a
3.9 is a late group III variety.
[0052]Oil or oil percent. Soybean seeds contain a considerable amount of
oil. Oil is measured by NIR spectrophotometry, and is reported on an as
is percentage basis.
[0053]Oleic Acid Percent. Oleic acid is one of the five most abundant
fatty acids in soybean seeds. It is measured by gas chromatography and is
reported as a percent of the total oil content.
[0054]Palmitic Acid Percent. Palmitic acid is one of the five most
abundant fatty acids in soybean seeds. It is measured by gas
chromatography and is reported as a percent of the total oil content.
[0055]Pedigree Distance. Relationship among generations based on their
ancestral links as evidenced in pedigrees. May be measured by the
distance of the pedigree from a given starting point in the ancestry.
[0056]Percent Identity. Percent identity as used herein refers to the
comparison of the homozygous alleles of two soybean varieties. Percent
identity is determined by comparing a statistically significant number of
the homozygous alleles of two developed varieties. For example, a percent
identity of 90% between soybean variety 1 and soybean variety 2 means
that the two varieties have the same allele at 90% of their loci.
[0057]Percent Similarity. Percent similarity as used herein refers to the
comparison of the homozygous alleles of a soybean variety such as
SG3870NRR with another plant, and if the homozygous allele of SG3870NRR
matches at least one of the alleles from the other plant then they are
scored as similar. Percent similarity is determined by comparing a
statistically significant number of loci and recording the number of loci
with similar alleles as a percentage. A percent similarity of 90% between
SG3870NRR and another plant means that SG3870NRR matches at least one of
the alleles of the other plant at 90% of the loci.
[0058]Phytophthora Tolerance. Tolerance to Phytophthora root rot is rated
on a scale of 1 to 5, with a score of 1 being the best or highest
tolerance ranging to a score of 5 which indicates the plants have no
tolerance to Phytophthora.
[0059]Phenotypic Score. The Phenotypic Score is a visual rating of general
appearance of the variety. All visual traits are considered in the score
including healthiness, standability, appearance and freedom of disease.
Ratings are scored from 1 being excellent to 5 being poor.
[0060]Plant. As used herein, the term "plant" includes reference to an
immature or mature whole plant, including a plant from which seed or
grain or anthers have been removed. Seed or embryo that will produce the
plant is also considered to be the plant.
[0061]Plant Height. Plant height is taken from the top of the
soil to the
top node of the plant and is measured in centimeters.
[0062]Plant Parts. As used herein, the term "plant parts" (or a soybean
plant, or a part thereof) includes protoplasts, leaves, stems, roots,
root tips, anthers, seed, grain, embryo, pollen, ovules, cotyledon,
hypocotyl, pod, flower, shoot, tissue, petiole, cells, meristematic cells
and the like.
[0063]Pod. This refers to the fruit of a soybean plant. It consists of the
hull or shell (pericarp) and the soybean seeds.
[0064]Protein Percent. Soybean seeds contain a considerable amount of
protein. Protein is generally measured by NIR spectrop
hotometry and is
reported on an as is percentage basis.
[0065]Pubescence. This refers to a covering of very fine hairs closely
arranged on the leaves, stems and pods of the soybean plant.
[0066]Quantitative Trait Loci (QTL). Quantitative trait loci (QTL) refer
to genetic loci that control to some degree numerically representable
traits that are usually continuously distributed.
[0067]Regeneration. Regeneration refers to the development of a plant from
tissue culture.
[0068]Sclerotinia White Mold Tolerance. Tolerance to Sclerotinia white
mold is rated on a scale of 1 to 5, with a score of 1 being the best or
highest tolerance ranging to a score of 5 which indicates the plants have
no tolerance to Sclerotinia.
[0069]Seed Protein Peroxidase Activity. Seed protein peroxidase activity
refers to a chemical taxonomic technique to separate cultivars based on
the presence or absence of the peroxidase enzyme in the seed coat. There
are two types of soybean cultivars, those having high peroxidase activity
(dark red color) and those having low peroxidase activity (no color).
[0070]Seed Yield (Bushels/Acre). The yield in bushels/acre is the actual
yield of the grain at harvest.
[0071]Seeds per Pound. Soybean seeds vary in seed 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.
[0072]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 5 comparing all genotypes within a given test. A
score of 1 means pods have not opened and no seeds have fallen out. A
score of 2.5 indicates approximately 50% of the pods have opened, with
seeds falling to the ground and a score of 5 indicates 100% of the pods
are opened.
[0073]Single Gene Converted (Conversion). Single gene converted
(conversion) plants refers to plants which are developed by a plant
breeding technique called backcrossing wherein essentially all of the
desired morphological and physiological characteristics of a variety are
recovered in addition to the single gene transferred into the variety via
the backcrossing technique or via genetic engineering.
DETAILED DESCRIPTION OF THE INVENTION
[0074]Soybean cultivar SG3870NRR is a late maturity group III variety with
resistance to glyphosate herbicides, including ROUNDUP herbicide.
Additionally, soybean cultivar SG3870NRR is resistant to Phytophthora
sojae Races 3 and 7. Soybean cultivar SG3870NRR has higher yield
potential when compared to lines of similar maturity and has excellent
agronomic characteristics including lodging resistance.
[0075]Some of the selection criteria used for various generations include:
seed yield, lodging resistance, emergence, disease tolerance, maturity,
late season plant intactness, plant height and shattering resistance.
[0076]The cultivar has shown uniformity and stability, as described in the
following variety description information. It has been self-pollinated a
sufficient number of generations with careful attention to uniformity of
plant type. The line has been increased with continued observation for
uniformity.
[0077]Soybean cultivar SG3870NRR has the following morphologic and other
characteristics (based primarily on data collected at Breese, Ill.).
TABLE-US-00001
TABLE 1
VARIETY DESCRIPTION INFORMATION
Seed Coat Color (Mature Seed): Yellow
Cotyledon Color (Mature Seed): Yellow
Leaflet Shape: Ovate
Growth Habit: Indeterminate
Flower Color: White
Hilum Color (Mature Seed): Black
Plant Pubescence Color: Tawny
Pod Wall Color: Tan
Maturity Group: III
Relative Maturity: 3.9
Plant Lodging Score: 1.6
Plant Height (cm): 99
Seed Size (# Seeds/lb.): 3000
Seed Content:
% Protein: 34.4
% Oil: 20.4
Physiological Responses: ROUNDUP herbicide resistance
to glyphosate herbicides
Disease resistance: Phytophthora sojae Races 3 and 7
Nematode or insect resistance: Soybean cyst nematode
(Heterodera glycines) Race 3
[0078]This invention is also directed to methods for producing a soybean
plant by crossing a first parent soybean plant with a second parent
soybean plant, wherein the first or second soybean plant is the soybean
plant from cultivar SG3870NRR. Further, both first and second parent
soybean plants may be from cultivar SG3870NRR. Therefore, any methods
using soybean cultivar SG3870NRR are part of this invention: selfing,
backcrosses, hybrid breeding, and crosses to populations. Any plants
produced using soybean cultivar SG3870NRR as at least one parent are
within the scope of this invention.
[0079]Additional methods include, but are not limited to, expression
vectors introduced into plant tissues using a direct gene transfer method
such as microprojectile-mediated delivery, DNA injection, electroporation
and the like. More preferably, expression vectors are introduced into
plant tissues by using either microprojectile-mediated delivery with a
biolistic device or by using Agrobacterium-mediated transformation.
Transformant plants obtained with the protoplasm of the invention are
intended to be within the scope of this invention.
FURTHER EMBODIMENTS OF THE INVENTION
[0080]The advent of new molecular biological techniques has allowed the
isolation and characterization of genetic elements with specific
functions, such as encoding specific protein products. Scientists in the
field of plant biology developed a strong interest in engineering the
genome of plants to contain and express foreign genetic elements, or
additional, or modified versions of native or endogenous genetic elements
in order to alter the traits of a plant in a specific manner. Any DNA
sequences, whether from a different species or from the same species,
which are inserted into the genome using transformation are referred to
herein collectively as "transgenes". In some embodiments of the
invention, a transgenic variant of SG3870NRR may contain at least one
transgene but could contain at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 and/or
no more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2. Over the
last fifteen to twenty years several methods for producing transgenic
plants have been developed, and the present invention also relates to
transgenic variants of the claimed soybean variety SG3870NRR.
[0081]One embodiment of the invention is a process for producing soybean
variety SG3870NRR further comprising a desired trait, said process
comprising transforming a soybean plant of variety SG3870NRR with a
transgene that confers a desired trait. Another embodiment is the product
produced by this process. In one embodiment the desired trait may be one
or more of herbicide resistance, insect resistance, disease resistance,
decreased phytate, or modified fatty acid or carbohydrate metabolism. The
specific gene may be any known in the art or listed herein, including; a
polynucleotide conferring resistance to imidazolinone, sulfonylurea,
glyphosate, glufosinate, triazine, benzonitrile, cyclohexanedione,
phenoxy proprionic acid and L-phosphinothricin; a polynucleotide encoding
a Bacillus thuringiensis polypeptide, a polynucleotide encoding phytase,
FAD-2, FAD-3, galactinol synthase or a raffinose synthetic enzyme; or a
polynucleotide conferring resistance to soybean cyst nematode, brown stem
rot, Phytophthora root rot, soybean mosaic virus or sudden death
syndrome.
[0082]Numerous methods for plant transformation have been developed,
including biological and physical plant transformation protocols. See,
for example, Miki et al., "Procedures for Introducing Foreign DNA into
Plants" in Methods in Plant Molecular Biology and Biotechnology, Glick,
B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages
67-88 and Armstrong, "The First Decade of Maize Transformation: A Review
and Future Perspective" (Maydica 44:101-109, 1999). In addition,
expression vectors and in vitro culture methods for plant cell or tissue
transformation and regeneration of plants are available. See, for
example, Gruber et al., "Vectors for Plant Transformation" in Methods in
Plant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J.
E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.
[0083]Numerous methods for plant transformation have been developed,
including biological and physical plant transformation protocols. See,
for example, Miki et al., "Procedures for Introducing Foreign DNA into
Plants" in Methods in Plant Molecular Biology and Biotechnology, Glick,
B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages
67-88 and Armstrong, "The First Decade of Maize Transformation: A Review
and Future Perspective" (Maydica 44:101-109, 1999). In addition,
expression vectors and in vitro culture methods for plant cell or tissue
transformation and regeneration of plants are available. See, for
example, Gruber et al., "Vectors for Plant Transformation" in Methods in
Plant Molecular Biology and Biotechnology, Glick, B. R. and Thompson, J.
E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages 89-119.
[0084]A genetic trait which has been engineered into the genome of a
particular soybean plant may then be moved into the genome of another
variety using traditional breeding techniques that are well known in the
plant breeding arts. For example, a backcrossing approach is commonly
used to move a transgene from a transformed soybean variety into an
already developed soybean variety, and the resulting backcross conversion
plant would then comprise the transgene(s).
[0085]Various genetic elements can be introduced into the plant genome
using transformation. These elements include, but are not limited to
genes, coding sequences, inducible, constitutive, and tissue specific
promoters, enhancing sequences, and signal and targeting sequences. For
example, see the traits, genes and transformation methods listed in U.S.
Pat. No. 6,118,055.
[0086]Plant transformation involves the construction of an expression
vector which will function in plant cells. Such a vector comprises DNA
comprising a gene under control of, or operatively linked to, a
regulatory element (for example, a promoter). The expression vector may
contain one or more such operably linked gene/regulatory element
combinations. The vector(s) may be in the form of a plasmid and can be
used alone or in combination with other plasmids to provide transformed
soybean plants using transformation methods as described below to
incorporate transgenes into the genetic material of the soybean plant(s).
Expression Vectors for Soybean Transformation: Marker Genes
[0087]Expression vectors include at least one genetic marker operably
linked to a regulatory element (a promoter, for example) that allows
transformed cells containing the marker to be either recovered by
negative selection, i.e., inhibiting growth of cells that do not contain
the selectable marker gene, or by positive selection, i.e., screening for
the product encoded by the genetic marker. Many commonly used selectable
marker genes for plant transformation are well known in the
transformation arts, and include, for example, genes that code for
enzymes that metabolically detoxify a selective chemical agent which may
be an antibiotic or an herbicide, or genes that encode an altered target
which is insensitive to the inhibitor. A few positive selection methods
are also known in the art.
[0088]One commonly used selectable marker gene for plant transformation is
the neomycin phosphotransferase II (nptII) gene which, when under the
control of plant regulatory signals, confers resistance to kanamycin.
Fraley et al., Proc. Natl. Acad. Sci. USA, 80:4803 (1983). Another
commonly used selectable marker gene is the hygromycin phosp
hotransferase
gene which confers resistance to the antibiotic hygromycin. Vanden Elzen
et al., Plant Mol. Biol., 5:299 (1985).
[0089]Additional selectable marker genes of bacterial origin that confer
resistance to antibiotics include gentamycin acetyl transferase,
streptomycin phosphotransferase and aminoglycoside-3'-adenyl transferase,
the bleomycin resistance determinant (Hayford et al., Plant Physiol.
86:1216 (1988), Jones et al., Mol. Gen. Genet., 210:86 (1987), Svab et
al., Plant Mol. Biol. 14:197 (1990), Hille et al., Plant Mol. Biol. 7:171
(1986)). Other selectable marker genes confer resistance to herbicides
such as glyphosate, glufosinate or bromoxynil (Comai et al., Nature
317:741-744 (1985), Gordon-Kamm et al., Plant Cell 2:603-618 (1990) and
Stalker et al., Science 242:419-423 (1988)).
[0090]Selectable marker genes for plant transformation not of bacterial
origin include, for example, mouse dihydrofolate reductase, plant
5-enolpyruvylshikimate-3-phosphate synthase and plant acetolactate
synthase (Eichholtz et al., Somatic Cell Mol. Genet. 13:67 (1987), Shah
et al., Science 233:478 (1986), Charest et al., Plant Cell Rep. 8:643
(1990)).
[0091]Another class of marker genes for plant transformation requires
screening of presumptively transformed plant cells rather than direct
genetic selection of transformed cells for resistance to a toxic
substance such as an antibiotic. These genes are particularly useful to
quantify or visualize the spatial pattern of expression of a gene in
specific tissues and are frequently referred to as reporter genes because
they can be fused to a gene or gene regulatory sequence for the
investigation of gene expression. Commonly used genes for screening
presumptively transformed cells include .beta.-glucuronidase (GUS),
.beta.-galactosidase, luciferase and chloramphenicol acetyltransferase
(Jefferson, R. A., Plant Mol. Biol. Rep. 5:387 (1987), Teeri et al., EMBO
J. 8:343 (1989), Koncz et al., Proc. Natl. Acad. Sci. USA 84:131 (1987),
DeBlock et al., EMBO J. 3:1681 (1984)).
[0092]In vivo methods for visualizing GUS activity that do not require
destruction of plant tissue are available (Molecular Probes publication
2908, IMAGENE GREEN, p. 1-4 (1993) and Naleway et al., J. Cell Biol.
115:151a (1991)). However, these in vivo methods for visualizing GUS
activity have not proven useful for recovery of transformed cells because
of low sensitivity, high fluorescent backgrounds and limitations
associated with the use of luciferase genes as selectable markers.
[0093]More recently, a gene encoding Green Fluorescent Protein (GFP) has
been utilized as a marker for gene expression in prokaryotic and
eukaryotic cells (Chalfie et al., Science 263:802 (1994)). GFP and
mutants of GFP may be used as screenable markers.
Expression Vectors for Soybean Transformation: Promoters
[0094]Genes included in expression vectors must be driven by a nucleotide
sequence comprising a regulatory element, for example, a promoter.
Several types of promoters are well known in the transformation arts as
are other regulatory elements that can be used alone or in combination
with promoters.
[0095]As used herein, "promoter" includes reference to a region of DNA
upstream from the start of transcription and involved in recognition and
binding of RNA polymerase and other proteins to initiate transcription. A
"plant promoter" is a promoter capable of initiating transcription in
plant cells. Examples of promoters under developmental control include
promoters that preferentially initiate transcription in certain tissues,
such as leaves, roots, seeds, fibers, xylem vessels, tracheids, or
sclerenchyma. Such promoters are referred to as "tissue-preferred".
Promoters that initiate transcription only in a certain tissue are
referred to as "tissue-specific". A "cell-type" specific promoter
primarily drives expression in certain cell types in one or more organs,
for example, vascular cells in roots or leaves. An "inducible" promoter
is a promoter which is under environmental control. Examples of
environmental conditions that may effect transcription by inducible
promoters include anaerobic conditions or the presence of light.
Tissue-specific, tissue-preferred, cell type specific, and inducible
promoters constitute the class of "non-constitutive" promoters. A
"constitutive" promoter is a promoter that is active under most
environmental conditions.
[0096]A. Inducible Promoters--An inducible promoter is operably linked to
a gene for expression in soybean. Optionally, the inducible promoter is
operably linked to a nucleotide sequence encoding a signal sequence which
is operably linked to a gene for expression in soybean. With an inducible
promoter the rate of transcription increases in response to an inducing
agent.
[0097]Any inducible promoter can be used in the instant invention. See
Ward et al., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible
promoters include, but are not limited to, that from the ACEI system
which responds to copper (Mett et al., Proc. Natl. Acad. Sci. USA
90:4567-4571 (1993)); In2 gene from maize which responds to
benzenesulfonamide herbicide safeners (Hershey et al., Mol. Gen. Genetics
227:229-237 (1991) and Gatz et al., Mol. Gen. Genetics 243:32-38 (1994))
or Tet repressor from Tn10 (Gatz et al., Mol. Gen. Genetics 227:229-237
(1991)). A particularly preferred inducible promoter is a promoter that
responds to an inducing agent to which plants do not normally respond. An
exemplary inducible promoter is the inducible promoter from a steroid
hormone gene, the transcriptional activity of which is induced by a
glucocorticosteroid hormone (Schena et al., Proc. Natl. Acad. Sci. USA
88:0421 (1991)).
[0098]B. Constitutive Promoters--A constitutive promoter is operably
linked to a gene for expression in soybean or the constitutive promoter
is operably linked to a nucleotide sequence encoding a signal sequence
which is operably linked to a gene for expression in soybean.
[0099]Many different constitutive promoters can be utilized in the instant
invention. Exemplary constitutive promoters include, but are not limited
to, the promoters from plant viruses such as the 35S promoter from CaMV
(Odell et al., Nature 313:810-812 (1985)) and the promoters from such
genes as rice actin (McElroy et al., Plant Cell 2: 163-171 (1990));
ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) and
Christensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last et
al., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.
3:2723-2730 (1984)) and maize H3 histone (Lepetit et al., Mol. Gen.
Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):
291-300 (1992)). The ALS promoter, Xba1/NcoI fragment 5' to the Brassica
napus ALS3 structural gene (or a nucleotide sequence similarity to said
Xba1/NcoI fragment), represents a particularly useful constitutive
promoter. See PCT application WO 96/30530.
[0100]C. Tissue-specific or Tissue-preferred Promoters--A tissue-specific
promoter is operably linked to a gene for expression in soybean.
Optionally, the tissue-specific promoter is operably linked to a
nucleotide sequence encoding a signal sequence which is operably linked
to a gene for expression in soybean. Plants transformed with a gene of
interest operably linked to a tissue-specific promoter produce the
protein product of the transgene exclusively, or preferentially, in a
specific tissue.
[0101]Any tissue-specific or tissue-preferred promoter can be utilized in
the instant invention. Exemplary tissue-specific or tissue-preferred
promoters include, but are not limited to, a root-preferred promoter such
as that from the phaseolin gene (Murai et al., Science 23:476-482 (1983)
and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci. USA 82:3320-3324
(1985)); a leaf-specific and light-induced promoter such as that from cab
or rubisco (Simpson et al., EMBO J. 4(11):2723-2729 (1985) and Timko et
al., Nature 318:579-582 (1985)); an anther-specific promoter such as that
from LAT52 (Twell et al., Mol. Gen. Genetics 217:240-245 (1989)); a
pollen-specific promoter such as that from Zm13 (Guerrero et al., Mol.
Gen. Genetics 244:161-168 (1993)) or a microspore-preferred promoter such
as that from apg (Twell et al., Sex. Plant Reprod. 6:217-224 (1993)).
Signal Sequences for Targeting Proteins to Subcellular Compartments
[0102]Transport of a protein produced by transgenes to a subcellular
compartment such as the chloroplast, vacuole, peroxisome, glyoxysome,
cell wall or mitochondrion or for secretion into the apoplast, is
accomplished by means of operably linking the nucleotide sequence
encoding a signal sequence to the 5' and/or 3' region of a gene encoding
the protein of interest. Targeting sequences at the 5' and/or 3' end of
the structural gene may determine during protein synthesis and processing
where the encoded protein is ultimately compartmentalized.
[0103]The presence of a signal sequence directs a polypeptide to either an
intracellular organelle or subcellular compartment or for secretion to
the apoplast. Many signal sequences are known in the art. See, for
example, Becker et al., Plant Mol. Biol. 20:49 (1992); Knox, C., et al.,
Plant Mol. Biol. 9:3-17 (1987); Lerner et al., Plant Physiol. 91:124-129
(1989); Frontes et al., Plant Cell 3:483-496 (1991); Matsuoka et al.,
Proc. Natl. Acad. Sci. 88:834 (1991); Gould et al., J. Cell. Biol.
108:1657 (1989); Creissen et al., Plant J. 2:129 (1991); Kalderon, et
al., Cell 39:499-509 (1984); Steifel, et al., Plant Cell 2:785-793
(1990).
Foreign Protein Genes and Agronomic Genes
[0104]With transgenic plants according to the present invention, a foreign
protein can be produced in commercial quantities. Thus, techniques for
the selection and propagation of transformed plants, which are well
understood in the art, yield a plurality of transgenic plants which are
harvested in a conventional manner, and a foreign protein then can be
extracted from a tissue of interest or from total biomass. Protein
extraction from plant biomass can be accomplished by known methods which
are discussed, for example, by Heney and Orr, Anal. Biochem. 114:92-6
(1981).
[0105]According to a preferred embodiment, the transgenic plant provided
for commercial production of foreign protein is a soybean plant. In
another preferred embodiment, the biomass of interest is seed. For the
relatively small number of transgenic plants that show higher levels of
expression, a genetic map can be generated, primarily via conventional
RFLP, PCR and SSR analysis, which identifies the approximate chromosomal
location of the integrated DNA molecule. For exemplary methodologies in
this regard, see Glick and Thompson, Methods in Plant Molecular Biology
and Biotechnology, CRC Press, Boca Raton 269:284 (1993). Map information
concerning chromosomal location is useful for proprietary protection of a
subject transgenic plant.
[0106]Wang et al. discuss "Large Scale Identification, Mapping and
Genotyping of Single-Nucleotide Polymorphisms in the Human Genome",
Science, 280:1077-1082, 1998, and similar capabilities are becoming
increasingly available for the soybean genome. Map information concerning
chromosomal location is useful for proprietary protection of a subject
transgenic plant. If unauthorized propagation is undertaken and crosses
made with other germplasm, the map of the integration region can be
compared to similar maps for suspect plants to determine if the latter
have a common parentage with the subject plant. Map comparisons would
involve hybridizations, RFLP, PCR, SSR and sequencing, all of which are
conventional techniques. SNPs may also be used alone or in combination
with other techniques.
[0107]Likewise, by means of the present invention, plants can be
genetically engineered to express various phenotypes of agronomic
interest. Through the transformation of soybean the expression of genes
can be altered to enhance disease resistance, insect resistance,
herbicide resistance, agronomic, grain quality and other traits.
Transformation can also be used to insert DNA sequences which control or
help control male-sterility. DNA sequences native to soybean as well as
non-native DNA sequences can be transformed into soybean and used to
alter levels of native or non-native proteins. Various promoters,
targeting sequences, enhancing sequences, and other DNA sequences can be
inserted into the genome for the purpose of altering the expression of
proteins. Reduction of the activity of specific genes (also known as gene
silencing, or gene suppression) is desirable for several aspects of
genetic engineering in plants.
[0108]Many techniques for gene silencing are well known to one of skill in
the art, including but not limited to knock-outs (such as by insertion of
a transposable element such as mu (Vicki Chandler, The Maize Handbook ch.
118 (Springer-Verlag 1994) or other genetic elements such as a FRT, Lox
or other site specific integration site, antisense technology (see, e.g.,
Sheehy et al. (1988) PNAS USA 85:8805-8809; and U.S. Pat. Nos. 5,107,065;
5,453,566; and 5,759,829); co-suppression (e.g., Taylor (1997) Plant Cell
9:1245; Jorgensen (1990) Trends Biotech. 8(12):340-344; Flavell (1994)
PNAS USA 91:3490-3496; Finnegan et al. (1994) Bio/Technology 12: 883-888;
and Neuhuber et al. (1994) Mol. Gen. Genet. 244:230-241); RNA
interference (Napoli et al. (1990) Plant Cell 2:279-289; U.S. Pat. No.
5,034,323; Sharp (1999) Genes Dev. 13:139-141; Zamore et al. (2000) Cell
101:25-33; and Montgomery et al. (1998) PNAS USA 95:15502-15507),
virus-induced gene silencing (Burton, et al. (2000) Plant Cell
12:691-705; and Baulcombe (1999) Curr. Op. Plant Bio. 2:109-113);
target-RNA-specific ribozymes (Haseloff et al. (1988) Nature 334:
585-591); hairpin structures (Smith et al. (2000) Nature 407:319-320; WO
99/53050; and WO 98/53083); MicroRNA (Aukerman & Sakai (2003) Plant Cell
15:2730-2741); ribozymes (Steinecke et al. (1992) EMBO J. 11:1525; and
Perriman et al. (1993) Antisense Res. Dev. 3:253); oligonucleotide
mediated targeted modification (e.g., WO 03/076574 and WO 99/25853);
Zn-finger targeted molecules (e.g., WO 01/52620; WO 03/048345; and WO
00/42219); and other methods or combinations of the above methods known
to those of skill in the art.
[0109]Likewise, by means of the present invention, agronomic genes can be
expressed in transformed plants. More particularly, plants can be
genetically engineered to express various phenotypes of agronomic
interest. Exemplary genes implicated in this regard include, but are not
limited to, those categorized below:
1. Genes that Confer Resistance to Pests or Disease and that Encode:
[0110]A. Plant disease resistance genes. 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 variety can be transformed
with one or more cloned resistance genes to engineer plants that are
resistant to specific pathogen strains. See, for example Jones et al.,
Science 266:789 (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 encodes a protein
kinase); Mindrinos et al. Cell 78:1089 (1994) (Arabidopsis RSP2 gene for
resistance to Pseudomonas syringae), McDowell & Woffenden, (2003) Trends
Biotechnol. 21(4): 178-83 and Toyoda et al., (2002) Transgenic Res. 11
(6):567-82.
[0111]B. A gene conferring resistance to a pest, such as soybean cyst
nematode. See e.g., PCT Application WO 96/30517; PCT Application WO
93/19181.
[0112]C. A Bacillus thuringiensis protein, a derivative thereof or a
synthetic polypeptide modeled thereon. See, for example, Geiser et al.,
Gene 48:109 (1986), who disclose the cloning and nucleotide sequence of a
Bt .delta.-endotoxin gene. Moreover, DNA molecules encoding
.delta.-endotoxin genes can be purchased from American Type Culture
Collection, Manassas, Va., for example, under ATCC Accession Nos. 40098,
67136, 31995 and 31998.
[0113]D. 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.
[0114]E. A vitamin-binding protein such as avidin. See PCT application US
93/06487 which teaches the use of avidin and avidin homologues as
larvicides against insect pests.
[0115]F. 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), Sumitani et
al., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence of
Streptomyces nitrosporeus .alpha.-amylase inhibitor) and U.S. Pat. No.
5,494,813 (Hepher and Atkinson, issued Feb. 27, 1996).
[0116]G. An insect-specific hormone or pheromone such as an ecdysteroid or
juvenile hormone, a variant thereof, a mimetic based thereon, or an
antagonist or agonist thereof. 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.
[0117]H. An insect-specific peptide or neuropeptide which, upon
expression, disrupts the physiology of the affected pest. For example,
see the disclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression
cloning yields DNA coding for insect diuretic hormone receptor), and
Pratt et al., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin
is identified in Diploptera puntata); Chattopadhyay et al. (2004)
Critical Reviews in Microbiology 30 (1): 33-54 2004; Zjawiony (2004) J
Nat Prod 67 (2): 300-310; Carlini & Grossi-de-Sa (2002) Toxicon, 40 (11):
1515-1539; Ussuf et al. (2001) Curr Sci. 80 (7): 847-853; and Vasconcelos
& Oliveira (2004) Toxicon 44 (4): 385-403. See also U.S. Pat. No.
5,266,317 to Tomalski et al., which discloses genes encoding
insect-specific, paralytic neurotoxins.
[0118]I. An insect-specific venom produced in nature by a snake, a wasp,
etc. For example, see Pang et al., Gene 116:165 (1992), for disclosure of
heterologous expression in plants of a gene coding for a scorpion
insectotoxic peptide.
[0119]J. An enzyme responsible for a hyperaccumulation of a monoterpene, a
sesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivative
or another non-protein molecule with insecticidal activity.
[0120]K. An enzyme involved in the modification, including the
post-translational modification, of a biologically active molecule; for
example, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme, a
nuclease, a cyclase, a transaminase, an esterase, a hydrolase, a
phosphatase, a kinase, a phosphorylase, a polymerase, an elastase, a
chitinase and a glucanase, whether natural or synthetic. See PCT
application WO 93/02197 (Scott et al.), which discloses the nucleotide
sequence of a callase gene. DNA molecules which contain
chitinase-encoding sequences can be obtained, for example, from the ATCC
under Accession Nos. 39637 and 67152. See also Kramer et al., Insect
Biochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequence of
a cDNA encoding tobacco hornworm chitinase, and Kawalleck et al., Plant
Molec. Biol. 21:673 (1993), who provide the nucleotide sequence of the
parsley ubi4-2 polyubiquitin gene, U.S. Pat. Nos. 7,145,060, 7,087,810
and 6,563,020.
[0121]L. A molecule that stimulates signal transduction. For example, see
the disclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), of
nucleotide sequences for mung bean calmodulin cDNA clones, and Griess et
al., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequence
of a maize calmodulin cDNA clone.
[0122]M. A hydrophobic moment peptide. See PCT application WO 95/16776 and
U.S. Pat. No. 5,580,852, which disclose peptide derivatives of
tachyplesin which inhibit fungal plant pathogens, and PCT application WO
95/18855 and U.S. Pat. No. 5,607,914 which teaches synthetic
antimicrobial peptides that confer disease resistance.
[0123]N. A membrane permease, a channel former or a channel blocker. For
example, see the disclosure of Jaynes et al., Plant Sci 89:43 (1993), of
heterologous expression of a cecropin-.beta. lytic peptide analog to
render transgenic tobacco plants resistant to Pseudomonas solanacearum.
[0124]O. A viral-invasive protein or a complex toxin derived therefrom.
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
and tobacco mosaic virus.
[0125]P. An insect-specific antibody or an immunotoxin derived therefrom.
Thus, an antibody targeted to a critical metabolic function in the insect
gut would inactivate an affected enzyme, killing the insect. See Taylor
et al., Abstract #497, Seventh Int'l Symposium on Molecular Plant-Microbe
Interactions (Edinburgh, Scotland) (1994) (enzymatic inactivation in
transgenic tobacco via production of single-chain antibody fragments).
[0126]Q. A virus-specific antibody. See, for example, Tavladoraki et al.,
Nature 366:469 (1993), who show that transgenic plants expressing
recombinant antibody genes are protected from virus attack.
[0127]R. A developmental-arrestive protein produced in nature by a
pathogen or a parasite. Thus, fungal
endo-.alpha.-1,4-D-polygalacturonases facilitate fungal colonization and
plant nutrient release by solubilizing plant cell wall
homo-.alpha.-1,4-D-galacturonase. See Lamb et al., Bio/Technology 10:1436
(1992). The cloning and characterization of a gene which encodes a bean
endopolygalacturonase-inhibiting protein is described by Toubart et al.,
Plant J. 2:367 (1992).
[0128]S. A developmental-arrestive protein produced in nature by a plant.
For example, Logemann et al., Bio/Technology 10:305 (1992), have shown
that transgenic plants expressing the barley ribosome-inactivating gene
have an increased resistance to fungal disease.
[0129]T. Genes involved in the Systemic Acquired Resistance (SAR) Response
and/or the pathogenesis-related genes. Briggs, S., Current Biology, 5(2)
(1995); Pieterse & Van Loon (2004) Curr. Opin. Plant Bio. 7(4):456-64 and
Somssich (2003) Cell 113(7):815-6.
[0130]U. Antifungal genes. See Cornelissen and Melchers, Plant Physiol.,
101:709-712 (1993); Parijs et al., Planta 183:258-264 (1991) and Bushnell
et al., Can. J. of Plant Path. 20(2):137-149 (1998). Also see U.S. Pat.
No. 6,875,907.
[0131]V. Detoxification genes, such as for fumonisin, beauvericin,
moniliformin and zearalenone and their structurally related derivatives.
For example, see U.S. Pat. No. 5,792,931.
[0132]W. Cystatin and cysteine proteinase inhibitors. See U.S. Pat. No.
7,205,453.
[0133]X. Defensin genes. See WO 03/000863 and U.S. Pat. No. 6,911,577.
[0134]Y. Genes conferring resistance to nematodes, and in particular
soybean cyst nematodes. See e.g. PCT Application WO 96/30517; PCT
Application WO 93/19181, WO 03/033651 and Urwin et al., Planta
204:472-479 (1998), Williamson (1999) Curr Opin Plant Bio. 2(4):327-31.
[0135]Z. Genes that confer resistance to Phytophthora root rot, such as
the Rps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d, Rps 1-e, Rps 1-k, Rps 2,
Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6, Rps 7 and other Rps
genes. See, for example, Shoemaker et al., Phytophthora Root Rot
Resistance Gene Mapping in Soybean, Plant Genome IV Conference, San
Diego, Calif. (1995).
[0136]AA. Genes that confer resistance to Brown Stem Rot, such as
described in U.S. Pat. No. 5,689,035 and incorporated by reference for
this purpose.
2. Genes that Confer Resistance to an Herbicide, for Example:
[0137]A. An herbicide that inhibits the growing point or meristem, such as
an imidazolinone 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), and Miki et al., Theor. Appl. Genet. 80:449
(1990), respectively.
[0138]B. Glyphosate (resistance conferred by mutant
5-enolpyruvlshikimate-3-phosphate synthase (EPSPS) and aroA genes,
respectively) and other phosphono compounds such as glufosinate
(phosphinothricin acetyl transferase (PAT) and Streptomyces hygroscopicus
PAT bar genes), and pyridinoxy or phenoxy proprionic acids and
cyclohexanediones (ACCase inhibitor-encoding genes). 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. U.S.
Pat. No. 5,627,061 to Barry et al. also describes genes encoding EPSPS
enzymes. See also U.S. Pat. Nos. 6,566,587; 6,338,961; 6,248,876 B1;
6,040,497; 5,804,425; 5,633,435; 5,145,783; 4,971,908; 5,312,910;
5,188,642; 4,940,835; 5,866,775; 6,225,114 B1; 6,130,366; 5,310,667;
4,535,060; 4,769,061; 5,633,448; 5,510,471; Re. 36,449; RE 37,287 E; and
5,491,288; and international publications EP1173580; WO 01/66704;
EP1173581 and EP1173582, which are incorporated herein by reference for
this purpose. Glyphosate resistance is also imparted to plants that
express a gene that encodes a glyphosate oxido-reductase enzyme as
described more fully in U.S. Pat. Nos. 5,776,760 and 5,463,175, which are
incorporated herein by reference for this purpose. In addition glyphosate
resistance can be imparted to plants by the over expression of genes
encoding glyphosate N-acetyltransferase. See, for example, U.S.
application Ser. No. 10/427,692. 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. 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 PAT gene is provided in European application No. 0 242 246
to Leemans et al. DeGreef et al., Bio/Technology 7:61 (1989) describe the
production of transgenic plants that express chimeric bar genes coding
for phosphinothricin acetyl transferase activity. Exemplary of genes
conferring resistance to phenoxy proprionic acids and cyclohexones, such
as sethoxydim and haloxyfop are the Acc1-S1, Acc1-S2, and Acc2-S3 genes
described by Marshall et al., Theor. Appl. Genet. 83:435 (1992).
[0139]C. An herbicide that inhibits p
hotosynthesis, 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. 285:173 (1992).
[0140]D. Acetohydroxy acid synthase, which has been found to make plants
that express this enzyme resistant to multiple types of herbicides, has
been introduced into a variety of plants. See Hattori et al., Mol. Gen.
Genet. 246:419, 1995. Other genes that confer tolerance to herbicides
include a gene encoding a chimeric protein of rat cytochrome P4507A1 and
yeast NADPH-cytochrome P450 oxidoreductase (Shiota et al., Plant
Physiol., 106:17, 1994), genes for glutathione reductase and superoxide
dismutase (Aono et al., Plant Cell Physiol. 36:1687, 1995), and genes for
various phosphotransferases (Datta et al., Plant Mol. Biol. 20:619,
1992).
[0141]E. Protoporphyrinogen oxidase (protox) is necessary for the
production of chlorophyll, which is necessary for all plant survival. The
protox enzyme serves as the target for a variety of herbicidal compounds.
These herbicides also inhibit growth of all the different species of
plants present, causing their total destruction. The development of
plants containing altered protox activity which are resistant to these
herbicides are described in U.S. Pat. Nos. 6,288,306; 6,282,837;
5,767,373; and international publication WO 01/12825.
3. Genes that Confer or Contribute to a Value-Added Trait, Such as:
[0142]A. Modified fatty acid metabolism, for example, by transforming a
plant with an antisense gene of stearyl-ACP desaturase to increase
stearic acid content of the plant. See Knultzon et al., Proc. Natl. Acad.
Sci. USA 89:2625 (1992).
[0143]B. Decreased phytate content--1) Introduction of a phytase-encoding
gene enhances 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. 2) Up-regulation of a gene that reduces phytate
content. In maize, this, for example, could be accomplished, by cloning
and then re-introducing DNA associated with one or more of the alleles,
such as the LPA alleles, identified in maize mutants characterized by low
levels of phytic acid, such as in Raboy et al., Maydica 35: 383 (1990)
and/or by altering inositol kinase activity as in WO 02/059324,
US2003/000901 1, WO 03/027243, US2003/0079247, WO 99/05298, U.S. Pat. No.
6,197,561, U.S. Pat. No. 6,291,224, U.S. Pat. No. 6,391,348, WO
2002/059324, U.S. Pat. No. 2003/0079247, WO98/45448, WO99/55882,
WO01/04147.
[0144]C. Modified carbohydrate composition effected, for example, by
transforming plants with a gene coding for an enzyme that alters the
branching pattern of starch, or, a gene altering thioredoxin such as NTR
and/or TRX (See U.S. Pat. No. 6,531,648 which is incorporated by
reference for this purpose) and/or a gamma zein knock out or mutant such
as cs27 or TUSC27 or en27 (See U.S. Pat. No. 6,858,778 and
US2005/0160488, US2005/0204418; which are incorporated by reference for
this purpose). See Shiroza et al., J. Bacteriol. 170: 810 (1988)
(nucleotide sequence of Streptococcus mutans fructosyltransferase gene),
Steinmetz et al., Mol. Gen. Genet. 200: 220 (1985) (nucleotide sequence
of Bacillus subtilis levansucrase gene), Pen et al., Bio/Technology 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), Sogaard 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 11), WO 99/10498 (improved
digestibility and/or starch extraction through modification of
UDP-D-xylose 4-epimerase, Fragile 1 and 2, Ref 1, HCHL, C4H), U.S. Pat.
No. 6,232,529 (method of producing high oil seed by modification of
starch levels (AGP)). The fatty acid modification genes mentioned above
may also be used to affect starch content and/or composition through the
interrelationship of the starch and oil pathways.
[0145]D. Elevated oleic acid via FAD-2 gene modification and/or decreased
linolenic acid via FAD-3 gene modification. See U.S. Pat. Nos. 6,063,947;
6,323,392; and international publication WO 93/11245.
[0146]E. Altering conjugated linolenic or linoleic acid content, such as
in WO 01/12800. Altering LEC1, AGP, Dek1, Superal1, mi1ps, various lpa
genes such as Ipa1, Ipa3, hpt or hggt. For example, see WO 02/42424, WO
98/22604, WO 03/011015, U.S. Pat. No. 6,423,886, U.S. Pat. Nos.
6,197,561, 6,825,397, 7,157,621 and US 2003/0079247, WO 02/057439, WO
03/011015 and Rivera-Madrid, R. et al. Proc. Natl. Acad. Sci.
92:5620-5624 (1995).
[0147]F. Altered antioxidant content or composition, such as alteration of
tocopherol or tocotrienols. For example, see U.S. Pat. Nos. 6,787,683 and
7,154,029 and WO 00/68393 involving the manipulation of antioxidant
levels through alteration of a phytl prenyl transferase (ppt), WO
03/082899 through alteration of a homogentisate geranyl geranyl
transferase (hggt).
[0148]G. Altered essential seed amino acids. For example, see U.S. Pat.
No. 6,127,600 (method of increasing accumulation of essential amino acids
in seeds), U.S. Pat. No. 6,080,913 (binary methods of increasing
accumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389
(high lysine), WO 99/40209 (alteration of amino acid compositions in
seeds), WO 99/29882 (methods for altering amino acid content of
proteins), U.S. Pat. No. 5,850,016 (alteration of amino acid compositions
in seeds), WO 98/20133 (proteins with enhanced levels of essential amino
acids), U.S. Pat. No. 5,885,802 (high methionine), U.S. Pat. No.
5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plant amino acid
biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increased lysine and
threonine), U.S. Pat. No. 6,441,274 (plant tryptophan synthase beta
subunit), U.S. Pat. No. 6,346,403 (methionine metabolic enzymes), U.S.
Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414 (increased
methionine), WO 98/56935 (plant amino acid biosynthetic enzymes), WO
98/45458 (engineered seed protein having higher percentage of essential
amino acids), WO 98/42831 (increased lysine), U.S. Pat. No. 5,633,436
(increasing sulfur amino acid content), U.S. Pat. No. 5,559,223
(synthetic storage proteins with defined structure containing
programmable levels of essential amino acids for improvement of the
nutritional value of plants), WO 96/01905 (increased threonine), WO
95/15392 (increased lysine), U.S. Pat. Nos. 6,930,225, 7,179,955, U.S.
Publication No. 2004/0068767, U.S. Pat. No. 6,803,498, WO 01/79516, and
WO 00/09706 (Ces A: cellulose synthase), U.S. Pat. No. 6,194,638
(hemicellulose), U.S. Pat. No. 6,399,859 and U.S. Pat. No. 7,098,381
(UDPGdH), U.S. Pat. No. 6,194,638 (RGP).
4. Genes that Control Male Sterility
[0149]There are several methods of conferring genetic male sterility
available, such as multiple mutant genes at separate locations within the
genome that confer male sterility, as disclosed in U.S. Pat. Nos.
4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations as
described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. In
addition to these methods, Albertsen et al., U.S. Pat. No. 5,432,068,
describe a system of nuclear male sterility which includes: identifying a
gene which is critical to male fertility; silencing this native gene
which is critical to male fertility; removing the native promoter from
the essential male fertility gene and replacing it with an inducible
promoter; inserting this genetically engineered gene back into the plant;
and thus creating a plant that is male sterile because the inducible
promoter is not "on" resulting in the male fertility gene not being
transcribed. Fertility is restored by inducing, or turning "on", the
promoter, which in turn allows the gene that confers male fertility to be
transcribed.
[0150]A. Introduction of a deacetylase gene under the control of a
tapetum-specific promoter and with the application of the chemical
N-Ac-PPT. See international publication WO 01/29237.
[0151]B. Introduction of various stamen-specific promoters. See
international publications WO 92/13956 and WO 92/13957.
[0152]C. Introduction of the barnase and the barstar genes. See Paul et
al., Plant Mol. Biol. 19:611-622, 1992).
[0153]For additional examples of nuclear male and female sterility systems
and genes, see also, U.S. Pat. No. 5,859,341; U.S. Pat. No. 6,297,426;
U.S. Pat. No. 5,478,369; U.S. Pat. No. 5,824,524; U.S. Pat. No.
5,850,014; and U.S. Pat. No. 6,265,640; all of which are hereby
incorporated by reference.
5. Genes that Create a Site for Site Specific DNA Integration.
[0154]This includes the introduction of FRT sites that may be used in the
FLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.
For example, see Lyznik, et al., Site-Specific Recombination for Genetic
Engineering in Plants, Plant Cell Rep (2003) 21:925-932 and WO 99/25821,
which are hereby incorporated by reference. Other systems that may be
used include the Gin recombinase of phage Mu (Maeser et al., 1991; Vicki
Chandler, The Maize Handbook ch. 118 (Springer-Verlag 1994), the Pin
recombinase of E. coli (Enomoto et al., 1983), and the R/RS system of the
pSR1 plasmid (Araki et al., 1992).
6. Genes that Affect Abiotic Stress Resistance.
[0155]Genes that affect abiotic stress resistance (including but not
limited to flowering, pod and seed development, enhancement of nitrogen
utilization efficiency, altered nitrogen responsiveness, drought
resistance or tolerance, cold resistance or tolerance, and salt
resistance or tolerance) and increased yield under stress. For example,
see: WO 00/73475 where water use efficiency is altered through alteration
of malate; U.S. Pat. No. 5,892,009, U.S. Pat. No. 5,965,705, U.S. Pat.
No. 5,929,305, U.S. Pat. No. 5,891,859, U.S. Pat. No. 6,417,428, U.S.
Pat. No. 6,664,446, U.S. Pat. No. 6,706,866, U.S. Pat. No. 6,717,034,
U.S. Pat. No. 6,801,104, WO 2000/060089, WO 2001/026459, WO 2001/035725,
WO 2001/034726, WO 2001/035727, WO 2001/036444, WO 2001/036597, WO
2001/036598, WO 2002/015675, WO 2002/017430, WO 2002/077185, WO
2002/079403, WO 2003/013227, WO 2003/013228, WO 2003/014327, WO
2004/031349, WO 2004/076638, WO 98/09521, and WO 99/38977 describing
genes, including CBF genes and transcription factors effective in
mitigating the negative effects of freezing, high salinity, and drought
on plants, as well as conferring other positive effects on plant
phenotype; US 2004/0148654 and WO 01/36596 where abscisic acid is altered
in plants resulting in improved plant phenotype such as increased yield
and/or increased tolerance to abiotic stress; WO 2000/006341, WO
04/090143, U.S. application Ser. No. 10/817,483 and U.S. Pat. No.
6,992,237 where cytokinin expression is modified resulting in plants with
increased stress tolerance, such as drought tolerance, and/or increased
yield. Also see WO 02/02776, WO 2003/052063, JP2002281975, U.S. Pat. No.
6,084,153, WO 01/64898, U.S. Pat. Nos. 6,177,275 and 6,107,547
(enhancement of nitrogen utilization and altered nitrogen
responsiveness). For ethylene alteration, see US 20040128719, US
20030166197 and WO 2000/32761. For plant transcription factors or
transcriptional regulators of abiotic stress, see e.g. US 20040098764 or
US 20040078852.
[0156]Other genes and transcription factors that affect plant growth and
agronomic traits such as yield, flowering, plant growth and/or plant
structure, can be introduced or introgressed into plants, see e.g. WO
97/49811 (LHY), WO 98/56918 (ESD4), WO 97/10339 and U.S. Pat. No.
6,573,430 (TFL), U.S. Pat. No. 6,713,663 (FT), WO 96/14414 (CON), WO
96/38560, WO 01/21822 (VRN1), WO 00/44918 (VRN2), WO 99/49064 (GI), WO
00/46358 (FR1), WO 97/29123, U.S. Pat. No. 6,794,560, U.S. Pat. No.
6,307,126 (GAI), WO 99/09174 (D8 and Rht), and WO 2004/076638 and WO
2004/031349 (transcription factors).
Methods for Soybean Transformation
[0157]Numerous methods for plant transformation have been developed
including biological and physical plant transformation protocols. See,
for example, Miki et al., "Procedures for Introducing Foreign DNA into
Plants" in Methods in Plant Molecular Biology and Biotechnology, Glick,
B. R. and Thompson, J. E. Eds. (CRC Press, Inc. Boca Raton, 1993) pages
67-88. In addition, expression vectors and in-vitro culture methods for
plant cell or tissue transformation and regeneration of plants are
available. See, for example, Gruber et al., "Vectors for Plant
Transformation" in Methods in Plant Molecular Biology and Biotechnology,
Glick, B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993)
pages 89-119.
[0158]A. Agrobacterium-mediated Transformation--One method for introducing
an expression vector into plants is based on the natural transformation
system of Agrobacterium. See, for example, Horsch et al., Science
227:1229 (1985). A. tumefaciens and A. rhizogenes are plant pathogenic
soil bacteria which genetically transform plant cells. The Ti and Ri
plasmids of A. tumefaciens and A. rhizogenes, respectively, carry genes
responsible for genetic transformation of the plant. See, for example,
Kado, C. I., Crit. Rev. Plant Sci. 10:1 (1991). Descriptions of
Agrobacterium vector systems and methods for Agrobacterium-mediated gene
transfer are provided by Gruber et al., supra, Miki et al., supra and
Moloney et al., Plant Cell Reports 8:238 (1989). See also, U.S. Pat. No.
5,563,055 (Townsend and Thomas), issued Oct. 8, 1996.
[0159]B. Direct Gene Transfer--Several methods of plant transformation,
collectively referred to as direct gene transfer, have been developed as
an alternative to Agrobacterium-mediated transformation. A generally
applicable method of plant transformation is microprojectile-mediated
transformation where DNA is carried on the surface of microprojectiles
measuring 1 to 4 .mu.m. The expression vector is introduced into plant
tissues with a biolistic device that accelerates the microprojectiles to
speeds of 300 to 600 m/s which is sufficient to penetrate plant cell
walls and membranes. Sanford et al., Part. Sci. Technol. 5:27 (1987);
Sanford, J. C., Trends Biotech. 6:299 (1988); Klein et al., Bio/Tech.
6:559-563 (1988); Sanford, J. C. Physiol Plant 7:206 (1990); Klein et
al., Biotechnology 10:268 (1992). See also U.S. Pat. No. 5,015,580
(Christou, et al.), issued May 14, 1991 and U.S. Pat. No. 5,322,783
(Tomes, et al.), issued Jun. 21, 1994.
[0160]Another method for physical delivery of DNA to plants is sonication
of target cells. Zhang et al., Bio/Technology 9:996 (1991).
Alternatively, liposome and spheroplast fusion have been used to
introduce expression vectors into plants. Deshayes et al., EMBO J.,
4:2731 (1985); Christou et al., Proc Natl. Acad. Sci. USA 84:3962 (1987).
Direct uptake of DNA into protoplasts using CaCl.sub.2 precipitation,
polyvinyl alcohol or poly-L-ornithine have also been reported. Hain et
al., Mol. Gen. Genet. 199:161 (1985) and Draper et al., Plant Cell
Physiol. 23:451 (1982). Electroporation of protoplasts and whole cells
and tissues have also been described (Donn et al., In Abstracts of VIIth
International Congress on Plant Cell and Tissue Culture IAPTC, A2-38, p
53 (1990); D'Halluin et al., Plant Cell 4:1495-1505 (1992) and Spencer et
al., Plant Mol. Biol. 24:51-61 (1994)).
[0161]Following transformation of soybean target tissues, expression of
the above-described selectable marker genes allows for preferential
selection of transformed cells, tissues and/or plants, using regeneration
and selection methods well known in the art.
[0162]The foregoing methods for transformation would typically be used for
producing a transgenic variety. The transgenic variety could then be
crossed with another (non-transformed or transformed) variety in order to
produce a new transgenic variety. Alternatively, a genetic trait that has
been engineered into a particular soybean line using the foregoing
transformation techniques could be moved into another line using
traditional backcrossing techniques that are well known in the plant
breeding arts. For example, a backcrossing approach could be used to move
an engineered trait from a public, non-elite variety into an elite
variety, or from a variety containing a foreign gene in its genome into a
variety or varieties that do not contain that gene. As used herein,
"crossing" can refer to a simple X by Y cross or the process of
backcrossing depending on the context.
Genetic Marker Profile Through SSR and First Generation Progeny
[0163]In addition to phenotypic observations, a plant can also be
identified by its genotype. The genotype of a plant can be characterized
through a genetic marker profile which can identify plants of the same
variety or a related variety or be used to determine or validate a
pedigree. Genetic marker profiles can be obtained by techniques such as
Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified
Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction
(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized
Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms
(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to as
Microsatellites, and Single Nucleotide Polymorphisms (SNPs). For example,
see Cregan et. al, "An Integrated Genetic Linkage Map of the Soybean
Genome" Crop Science 39:1464-1490 (1999), and Berry et al., Assessing
Probability of Ancestry Using Simple Sequence Repeat Profiles:
Applications to Maize Inbred Lines and Soybean Varieties" Genetics
165:331-342 (2003), each of which are incorporated by reference herein in
their entirety.
[0164]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 which provides a means of distinguishing
varieties. One method of comparison is to use only homozygous loci for
SG3870NRR.
[0165]Primers and PCR protocols for assaying these and other markers are
disclosed in the Soybase (sponsored by the USDA Agricultural Research
Service and Iowa State University). In addition to being used for
identification of soybean variety SG3870NRR and plant parts and plant
cells of variety SG3870NRR, the genetic profile may be used to identify a
soybean plant produced through the use of SG3870NRR or to verify a
pedigree for progeny plants produced through the use of SG3870NRR. The
genetic marker profile is also useful in breeding and developing
backcross conversions.
[0166]The present invention comprises a soybean plant characterized by
molecular and physiological data obtained from the representative sample
of said variety deposited with the American Type Culture Collection
(ATCC). Further provided by the invention 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.
[0167]Means of performing genetic marker profiles using SSR polymorphisms
are well known in the art. SSRs are genetic markers based on
polymorphisms in repeated nucleotide sequences, such as microsatellites.
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 the
polymerase chain reaction (PCR), thereby eliminating the need for
labor-intensive Southern hybridization. The PCR detection is done by use
of two oligonucleotide primers flanking the polymorphic segment of
repetitive DNA. Repeated cycles of heat denaturation of the DNA followed
by annealing of the primers to their complementary sequences at low
temperatures, and extension of the annealed primers with DNA polymerase,
comprise the major part of the methodology.
[0168]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 may be measured by the number of
base pairs of the fragment. While variation in the primer used or in
laboratory procedures can affect the reported fragment size, relative
values should remain constant regardless of the specific primer or
laboratory used. When comparing varieties it is preferable if all SSR
profiles are performed in the same lab.
[0169]Primers used are publicly available and may be found in the Soybase
or Cregan supra. See also, PCT Publication No. WO 99/31964 Nucleotide
Polymorphisms in Soybean, U.S. Pat. No. 6,162,967 Positional Cloning of
Soybean Cyst Nematode Resistance Genes, and U.S. application Ser. No.
09/954,773 Soybean Sudden Death Syndrome Resistant Soybeans and Methods
of Breeding and Identifying Resistant Plants, the disclosure of which are
incorporated herein by reference.
[0170]The SSR profile of soybean plant SG3870NRR can be used to identify
plants comprising SG3870NRR as a parent, since such plants will comprise
the same homozygous alleles as SG3870NRR. 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
F.sub.1 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 F.sub.1 progeny will be
xy (heterozygous) at that locus. Subsequent generations of progeny
produced by selection and breeding are expected to be of genotype x
(homozygous), y (homozygous), or xy (heterozygous) for that locus
position. When the F.sub.1 plant is selfed or sibbed for successive
filial generations, the locus should be either x or y for that position.
[0171]In addition, plants and plant parts substantially benefiting from
the use of SG3870NRR in their development, such as SG3870NRR comprising a
backcross conversion, transgene, or genetic sterility factor, may be
identified by having a molecular marker profile with a high percent
identity to SG3870NRR. Such a percent identity might be 95%, 96%, 97%,
98%, 99%, 99.5% or 99.9% identical to SG3870NRR.
[0172]The SSR profile of SG3870NRR also can be used to identify
essentially derived varieties and other progeny varieties developed from
the use of SG3870NRR, as well as cells and other plant parts thereof.
Such plants may be developed using the markers identified in WO 00/31964,
U.S. Pat. No. 6,162,967 and U.S. application Ser. No. 09/954,773. Progeny
plants and plant parts produced using SG3870NRR may be identified by
having a molecular marker profile of at least 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,
84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99% or 99.5% genetic contribution from soybean variety, as measured
by either percent identity or percent similarity. Such progeny may be
further characterized as being within a pedigree distance of SG3870NRR,
such as within 1, 2, 3, 4 or 5 or less cross-pollinations to a soybean
plant other than SG3870NRR or a plant that has SG3870NRR as a progenitor.
Unique molecular profiles may be identified with other molecular
tools
such as SNPs and RFLPs.
[0173]While determining the SSR genetic marker profile of the plants
described supra, several unique SSR profiles may also be identified which
did not appear in either parent of such plant. Such unique SSR profiles
may arise during the breeding process from recombination or mutation. A
combination of several unique alleles provides a means of identifying a
plant variety, an F.sub.1 progeny produced from such variety, and progeny
produced from such variety.
Single-Gene Conversions
[0174]When the term "soybean plant" is used in the context of the present
invention, this also includes any single gene conversions of that
variety. The term single gene converted plant as used herein refers to
those soybean plants which are developed by a plant breeding technique
called backcrossing wherein essentially all of the desired morphological
and physiological characteristics of a variety are recovered in addition
to the single gene transferred into the variety via the backcrossing
technique. Backcrossing methods can be used with the present invention to
improve or introduce a characteristic into the variety. The term
"backcrossing" as used herein refers to the repeated crossing of a hybrid
progeny back to the recurrent parent, i.e., backcrossing 1, 2, 3, 4, 5,
6, 7, 8 or more times to the recurrent parent. The parental soybean plant
that contributes the gene for the desired characteristic is termed the
nonrecurrent or donor parent. This terminology refers to the fact that
the nonrecurrent parent is used one time in the backcross protocol and
therefore does not recur. The parental soybean plant to which the gene or
genes from the nonrecurrent parent are transferred is known as the
recurrent parent as it is used for several rounds in the backcrossing
protocol (Poehlman & Sleper, 1994; Fehr, Principles of Cultivar
Development pp. 261-286 (1987)). In a typical backcross protocol, the
original variety of interest (recurrent parent) is crossed to a second
variety (nonrecurrent parent) that carries the single gene 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 desired morphological
and physiological characteristics of the recurrent parent are recovered
in the converted plant, in addition to the single transferred gene from
the nonrecurrent parent.
[0175]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 single trait or characteristic in the
original variety. To accomplish this, a single gene of the recurrent
variety is modified or substituted with the desired gene from the
nonrecurrent parent, while retaining essentially all of the rest of the
desired genetic, and therefore the desired physiological and
morphological, 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 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.
[0176]Many single gene traits have been identified that are not regularly
selected for in the development of a new variety but that can be improved
by backcrossing techniques. Single gene traits may or may not be
transgenic; examples of these traits include but are not limited to, male
sterility, waxy starch, herbicide resistance, resistance for bacterial,
fungal, or viral disease, insect resistance, male fertility, enhanced
nutritional quality, industrial usage, yield stability and yield
enhancement. These genes are generally inherited through the nucleus.
Several of these single gene traits are described in U.S. Pat. Nos.
5,959,185; 5,973,234 and 5,977,445; the disclosures of which are
specifically hereby incorporated by reference.
Introduction of a New Trait or Locus into SG3870NRR
[0177]Variety SG3870NRR represents a new base genetic variety into which a
new locus or trait may be introgressed. Direct transformation and
backcrossing represent two important methods that can be used to
accomplish such an introgression. The term backcross conversion and
single locus conversion are used interchangeably to designate the product
of a backcrossing program.
Backcross Conversions of SG3870NRR
[0178]A backcross conversion of SG3870NRR occurs when DNA sequences are
introduced through backcrossing (Hallauer et al, 1988, "Corn Breeding"
Corn and Corn Improvements, No. 18, pp. 463-481), with SG3870NRR utilized
as the recurrent parent. Both naturally occurring and transgenic DNA
sequences may be introduced through backcrossing techniques. A backcross
conversion may produce a plant with a trait or locus conversion in at
least two or more backcrosses, including at least 2 crosses, at least 3
crosses, at least 4 crosses, at least 5 crosses and the like. Molecular
marker assisted breeding or selection may be utilized to reduce the
number of backcrosses necessary to achieve the backcross conversion. For
example, see Openshaw, S. J. et al., Marker-assisted Selection in
Backcross Breeding. In: Proceedings Symposium of the Analysis of
Molecular Data, August 1994, Crop Science Society of America, Corvallis,
Oreg., where it is demonstrated that a backcross conversion can be made
in as few as two backcrosses.
[0179]The complexity of the backcross conversion method depends on the
type of trait being transferred (single genes or closely linked genes as
vs. unlinked genes), the level of expression of the trait, the type of
inheritance (cytoplasmic or nuclear) and the types of parents included in
the cross. It is understood by those of ordinary skill in the art that
for single gene traits that are relatively easy to classify, the
backcross method is effective and relatively easy to manage. (See
Hallauer et al. in Corn and Corn Improvement, Sprague and Dudley, Third
Ed. 1998). Desired traits that may be transferred through backcross
conversion include, but are not limited to, sterility (nuclear and
cytoplasmic), fertility restoration, nutritional enhancements, drought
tolerance, nitrogen utilization, altered fatty acid profile, low phytate,
industrial enhancements, disease resistance (bacterial, fungal or viral),
insect resistance and herbicide resistance. In addition, an introgression
site itself, such as an FRT site, Lox site or other site specific
integration site, may be inserted by backcrossing and utilized for direct
insertion of one or more genes of interest into a specific plant variety.
In some embodiments of the invention, the number of loci that may be
backcrossed into SG3870NRR is at least 1, 2, 3, 4, or 5 and/or no more
than 6, 5, 4, 3, or 2. A single locus may contain several transgenes,
such as a transgene for disease resistance that, in the same expression
vector, also contains a transgene for herbicide resistance. The gene for
herbicide resistance may be used as a selectable marker and/or as a
phenotypic trait. A single locus conversion of site specific integration
system allows for the integration of multiple genes at the converted
loci.
[0180]The backcross conversion may result from either the transfer of a
dominant allele or a recessive allele. Selection of progeny containing
the trait of interest is accomplished by direct selection for a trait
associated with a dominant allele. Transgenes transferred via
backcrossing typically function as a dominant single gene trait and are
relatively easy to classify. Selection of progeny for a trait that is
transferred via a recessive allele requires growing and selfing the first
backcross generation to determine which plants carry the recessive
alleles. Recessive traits may require additional progeny testing in
successive backcross generations to determine the presence of the locus
of interest. The last backcross generation is usually selfed to give pure
breeding progeny for the gene(s) being transferred, although a backcross
conversion with a stably introgressed trait may also be maintained by
further backcrossing to the recurrent parent with selection for the
converted trait.
[0181]Along with selection for the trait of interest, progeny are selected
for the phenotype of the recurrent parent. The backcross is a form of
inbreeding, and the features of the recurrent parent are automatically
recovered after successive backcrosses. Poehiman, Breeding Field Crops,
P. 204 (1987). Poehlman suggests from one to four or more backcrosses,
but as noted above, the number of backcrosses necessary can be reduced
with the use of molecular markers. Other factors, such as a genetically
similar donor parent, may also reduce the number of backcrosses
necessary. As noted by Poehlman, backcrossing is easiest for simply
inherited, dominant and easily recognized traits.
[0182]One process for adding or modifying a trait or locus in soybean
variety SG3870NRR comprises crossing SG3870NRR plants grown from
SG3870NRR seed with plants of another soybean variety that comprise the
desired trait or locus, selecting F.sub.1 progeny plants that comprise
the desired trait or locus to produce selected F.sub.1 progeny plants,
crossing the selected progeny plants with the SG3870NRR plants to produce
backcross progeny plants, selecting for backcross progeny plants that
have the desired trait or locus and the morphological characteristics of
soybean variety SG3870NRR to produce selected backcross progeny plants;
and backcrossing to SG3870NRR three or more times in succession to
produce selected fourth or higher backcross progeny plants that comprise
said trait or locus. The modified SG3870NRR may be further characterized
as having the physiological and morphological characteristics of soybean
variety SG3870NRR listed in Table 1 as determined at the 5% significance
level when grown in the same environmental conditions and/or may be
characterized by percent similarity or identity to SG3870NRR as
determined by SSR markers. The above method may be utilized with fewer
backcrosses in appropriate situations, such as when the donor parent is
highly related or markers are used in the selection step. Desired traits
that may be used include those nucleic acids known in the art, some of
which are listed herein, that will affect traits through nucleic acid
expression or inhibition. Desired loci include the introgression of FRT,
Lox and other sites for site specific integration, which may also affect
a desired trait if a functional nucleic acid is inserted at the
integration site.
[0183]In addition, the above process and other similar processes described
herein may be used to produce first generation progeny soybean seed by
adding a step at the end of the process that comprises crossing SG3870NRR
with the introgressed trait or locus with a different soybean plant and
harvesting the resultant first generation progeny soybean seed.
Tissue Culture
[0184]Further reproduction of the variety can occur by tissue culture and
regeneration. Tissue culture of various tissues of soybeans and
regeneration of plants therefrom is well known and widely published. For
example, reference may be had to Komatsuda, T. et al., Crop Sci.
31:333-337 (1991); Stephens, P. A., et al., Theor. Appl. Genet. (1991)
82:633-635; Komatsuda, T. et al., Plant Cell, Tissue and Organ Culture,
28:103-113 (1992); Dhir, S. et al., Plant Cell Reports (1992) 11:285-289;
Pandey, P. et al., Japan J. Breed. 42:1-5 (1992); and Shetty, K., et al.,
Plant Science 81:245-251 (1992); as well as U.S. Pat. No. 5,024,944
issued Jun. 18, 1991 to Collins et al., and U.S. Pat. No. 5,008,200
issued Apr. 16, 1991 to Ranch et al. Thus, another aspect of this
invention is to provide cells which upon growth and differentiation
produce soybean plants having the physiological and morphological
characteristics of soybean cultivar SG3870NRR.
[0185]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, plant clumps, and plant cells that can
generate tissue culture that are intact in plants or parts of plants,
such as embryos, pollen, flowers, seeds, pods, petioles, leaves, stems,
roots, root tips, anthers, pistils and the like. Means for preparing and
maintaining plant tissue culture are well known in the art. By way of
example, a tissue culture comprising organs has been used to produce
regenerated plants. U.S. Pat. Nos. 5,959,185; 5,973,234 and 5,977,445
describe certain techniques, the disclosures of which are incorporated
herein by reference.
Using SG3870NRR to Develop Other Soybean Varieties
[0186]Soybean varieties such as SG3870NRR are typically developed for use
in seed and grain production. However, soybean varieties such as
SG3870NRR also provide a source of breeding material that may be used to
develop new soybean varieties. Plant breeding techniques known in the art
and used in a soybean plant breeding program include, but are not limited
to, recurrent selection, mass selection, bulk selection, mass selection,
backcrossing, pedigree breeding, open pollination breeding, restriction
fragment length polymorphism enhanced selection, genetic marker enhanced
selection, making double haploids, and transformation. Often combinations
of these techniques are used. The development of soybean varieties in a
plant breeding program requires, in general, the development and
evaluation of homozygous varieties. There are many analytical methods
available to evaluate a new variety. The oldest and most traditional
method of analysis is the observation of phenotypic traits but genotypic
analysis may also be used.
Additional Breeding Methods
[0187]This invention is directed to methods for producing a soybean plant
by crossing a first parent soybean plant with a second parent soybean
plant wherein either the first or second parent soybean plant is variety
SG3870NRR. The other parent may be any other soybean plant, such as a
soybean plant that is part of a synthetic or natural population. Any such
methods using soybean variety SG3870NRR are part of this invention:
selfing, sibbing, backcrosses, mass selection, pedigree breeding, bulk
selection, hybrid production, crosses to populations, and the like. These
methods are well known in the art and some of the more commonly used
breeding methods are described below. Descriptions of breeding methods
can be found in one of several reference books (e.g., Allard, Principles
of Plant Breeding, 1960; Simmonds, Principles of Crop Improvement, 1979;
Sneep et al., 1979; Fehr, "Breeding Methods for Cultivar Development",
Chapter 7, Soybean Improvement, Production and Uses, 2.sup.nd ed., Wilcox
editor, 1987).
[0188]The following describes breeding methods that may be used with
soybean cultivar SG3870NRR in the development of further soybean plants.
One such embodiment is a method for developing a cultivar SG3870NRR
progeny soybean plant in a soybean plant breeding program comprising:
obtaining the soybean plant, or a part thereof, of cultivar SG3870NRR
utilizing said plant or plant part as a source of breeding material and
selecting a soybean cultivar SG3870NRR progeny plant with molecular
markers in common with cultivar SG3870NRR and/or with morphological
and/or physiological characteristics selected from the characteristics
listed in Tables 1 or 2. Breeding steps that may be used in the soybean
plant breeding program include pedigree breeding, backcrossing, mutation
breeding, and recurrent selection. In conjunction with these steps,
techniques such as RFLP-enhanced selection, genetic marker enhanced
selection (for example SSR markers) and the making of double haploids may
be utilized.
[0189]Another method involves producing a population of soybean cultivar
SG3870NRR progeny soybean plants, comprising crossing cultivar SG3870NRR
with another soybean plant, thereby producing a population of soybean
plants, which, on average, derive 50% of their alleles from soybean
cultivar SG3870NRR. A plant of this population may be selected and
repeatedly selfed or sibbed with a soybean cultivar resulting from these
successive filial generations. One embodiment of this invention is the
soybean cultivar produced by this method and that has obtained at least
50% of its alleles from soybean cultivar SG3870NRR.
[0190]One of ordinary skill in the art of plant breeding would know how to
evaluate the traits of two plant varieties to determine if there is no
significant difference between the two traits expressed by those
varieties. For example, see Fehr and Walt, Principles of Cultivar
Development, p 261-286 (1987). Thus the invention includes soybean
cultivar SG3870NRR progeny soybean plants comprising a combination of at
least two cultivar SG3870NRR traits selected from the group consisting of
those listed in Tables 1 and 2 or the cultivar SG3870NRR combination of
traits listed in the Summary of the Invention, so that said progeny
soybean plant is not significantly different for said traits than soybean
cultivar SG3870NRR as determined at the 5% significance level when grown
in the same environmental conditions. Using techniques described herein,
molecular markers may be used to identify said progeny plant as a soybean
cultivar SG3870NRR progeny plant. Mean trait values may be used to
determine whether trait differences are significant, and preferably the
traits are measured on plants grown under the same environmental
conditions. Once such a variety is developed its value is substantial
since it is important to advance the germplasm base as a whole in order
to maintain or improve traits such as yield, disease resistance, pest
resistance, and plant performance in extreme environmental conditions.
[0191]Progeny of soybean cultivar SG3870NRR may also be characterized
through their filial relationship with soybean cultivar SG3870NRR, as for
example, being within a certain number of breeding crosses of soybean
cultivar SG3870NRR. A breeding cross is a cross made to introduce new
genetics into the progeny, and is distinguished from a cross, such as a
self or a sib cross, made to select among existing genetic alleles. The
lower the number of breeding crosses in the pedigree, the closer the
relationship between soybean cultivar SG3870NRR and its progeny. For
example, progeny produced by the methods described herein may be within
1, 2, 3, 4 or 5 breeding crosses of soybean cultivar SG3870NRR.
[0192]As used herein, the term "plant" includes plant cells, plant
protoplasts, plant cell tissue cultures 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 embryos, pollen, ovules, flowers,
pods, leaves, roots, root tips, anthers, cotyledons, hypocotyls,
meristematic cells, stems, pistils, petiole, and the like.
Pedigree Breeding
[0193]Pedigree breeding starts with the crossing of two genotypes, such as
SG3870NRR and another soybean variety having one or more desirable
characteristics that is lacking or which complements SG3870NRR. If the
two original parents do not provide all the desired characteristics,
other sources can be included in the breeding population. In the pedigree
method, superior plants are selfed and selected in successive filial
generations. In the succeeding filial generations the heterozygous
condition gives way to homogeneous varieties as a result of
self-pollination and selection. Typically in the pedigree method of
breeding, five or more successive filial generations of selfing and
selection is practiced: F.sub.1 to F.sub.2; F.sub.2 to F.sub.3; F.sub.3
to F.sub.4; F.sub.4 to F.sub.5, etc. After a sufficient amount of
inbreeding, successive filial generations will serve to increase seed of
the developed variety. Preferably, the developed variety comprises
homozygous alleles at about 95% or more of its loci.
[0194]In addition to being used to create a backcross conversion,
backcrossing can also be used in combination with pedigree breeding. As
discussed previously, backcrossing can be used to transfer one or more
specifically desirable traits from one variety, the donor parent, to a
developed variety called the recurrent parent, which has overall good
agronomic characteristics yet lacks that desirable trait or traits.
However, the same procedure can be used to move the progeny toward the
genotype of the recurrent parent but at the same time retain many
components of the non-recurrent parent by stopping the backcrossing at an
early stage and proceeding with selfing and selection. For example, a
soybean variety may be crossed with another variety to produce a first
generation progeny plant. The first generation progeny plant may then be
backcrossed to one of its parent varieties to create a BC1 or BC2.
Progeny are selfed and selected so that the newly developed variety has
many of the attributes of the recurrent parent and yet several of the
desired attributes of the non-recurrent parent. This approach leverages
the value and strengths of the recurrent parent for use in new soybean
varieties.
[0195]Therefore, an embodiment of this invention is a method of making a
backcross conversion of soybean variety SG3870NRR, comprising the steps
of crossing a plant of soybean variety SG3870NRR with a donor plant
comprising a desired trait, selecting an F.sub.1 progeny plant comprising
the desired trait, and backcrossing the selected F.sub.1 progeny plant to
a plant of soybean variety SG3870NRR. This method may further comprise
the step of obtaining a molecular marker profile of soybean variety
SG3870NRR and using the molecular marker profile to select for a progeny
plant with the desired trait and the molecular marker profile of
SG3870NRR. In one embodiment the desired trait is a mutant gene or
transgene present in the donor parent.
Recurrent Selection and Mass Selection
[0196]Recurrent selection is a method used in a plant breeding program to
improve a population of plants. SG3870NRR is suitable for use in a
recurrent selection program. The method entails individual plants cross
pollinating with each other to form progeny. The progeny are grown and
the superior progeny selected by any number of selection methods, which
include individual plant, half-sib progeny, full-sib progeny and selfed
progeny. The selected progeny are cross pollinated with each other to
form progeny for another population. This population is planted and again
superior plants are selected to cross pollinate with each other.
Recurrent selection is a cyclical process and therefore can be repeated
as many times as desired. The objective of recurrent selection is to
improve the traits of a population. The improved population can then be
used as a source of breeding material to obtain new varieties for
commercial or breeding use, including the production of a synthetic
cultivar. A synthetic cultivar is the resultant progeny formed by the
intercrossing of several selected varieties.
[0197]Mass selection is a useful technique when used in conjunction with
molecular marker enhanced selection. In mass selection seeds from
individuals are selected based on phenotype or genotype. These selected
seeds are then bulked and used to grow the next generation. Bulk
selection requires growing a population of plants in a bulk plot,
allowing the plants to self-pollinate, harvesting the seed in bulk and
then using a sample of the seed harvested in bulk to plant the next
generation. Also, instead of self pollination, directed pollination could
be used as part of the breeding program.
Mutation Breeding
[0198]Mutation breeding is another method of introducing new traits into
soybean variety SG3870NRR. Mutations that occur spontaneously or are
artificially induced can be useful sources of variability for a plant
breeder. The goal of artificial mutagenesis is to increase the rate of
mutation for a desired characteristic. Mutation rates can be increased by
many different means including temperature, long-term seed storage,
tissue culture conditions, radiation; such as X-rays, Gamma rays (e.g.
cobalt 60 or cesium 137), neutrons, (product of nuclear fission by
uranium 235 in an atomic reactor), Beta radiation (emitted from
radioisotopes such as phosphorus 32 or carbon 14), or ultraviolet
radiation (preferably from 2500 to 2900 nm), or chemical mutagens (such
as base analogues (5-bromo-uracil), related compounds (8-ethoxy
caffeine), antibiotics (streptonigrin), alkylating agents (sulfur
mustards, nitrogen mustards, epoxides, ethylenamines, sulfates,
sulfonates, sulfones, lactones), azide, hydroxylamine, nitrous acid, or
acridines. Once a desired trait is observed through mutagenesis the trait
may then be incorporated into existing germplasm by traditional breeding
techniques. Details of mutation breeding can be found in "Principles of
Cultivar Development" Fehr, 1993 Macmillan Publishing Company. In
addition, mutations created in other soybean plants may be used to
produce a backcross conversion of soybean cultivar SG3870NRR that
comprises such mutation.
Breeding with Molecular Markers
[0199]Molecular markers, which includes markers identified through the use
of techniques such as Isozyme Electrophoresis, Restriction Fragment
Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs
(RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA
Amplification Fingerprinting (DAF), Sequence Characterized Amplified
Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), Simple
Sequence Repeats (SSRs) and Single Nucleotide Polymorphisms (SNPs), may
be used in plant breeding methods utilizing soybean cultivar SG3870NRR.
[0200]Isozyme Electrophoresis and RFLPs have been widely used to determine
genetic composition. Shoemaker and Olsen, ((1993) Molecular Linkage Map
of Soybean (Glycine max L. Merr.). p. 6.131-6.138. In S. J. O'Brien (ed.)
Genetic Maps: Locus Maps of Complex Genomes. Cold Spring Harbor
Laboratory Press. Cold Spring Harbor, N.Y.), developed a molecular
genetic linkage map that consisted of 25 linkage groups with about 365
RFLP, 11 RAPD (random amplified polymorphic DNA), three classical
markers, and four isozyme loci. See also, Shoemaker R. C. 1994 RFLP Map
of Soybean. P. 299-309 In R. L. Phillips and I. K. Vasil (ed.) DNA-based
markers in plants. Kluwer Academic Press Dordrecht, the Netherlands.
[0201]SSR technology is currently the most efficient and practical marker
technology; more marker loci can be routinely used and more alleles per
marker locus can be found using SSRs in comparison to RFLPs. For example
Diwan and Cregan, described a highly polymorphic microsatellite loci in
soybean with as many as 26 alleles. (Diwan, N., and P. B. Cregan 1997
Automated sizing of fluorescent-labeled simple sequence repeat (SSR)
markers to assay genetic variation in Soybean Theor. Appl. Genet.
95:220-225.) Single Nucleotide Polymorphisms may also be used to identify
the unique genetic composition of the invention and progeny varieties
retaining that unique genetic composition. Various molecular marker
techniques may be used in combination to enhance overall resolution.
[0202]Soybean DNA molecular marker linkage maps have been rapidly
constructed and widely implemented in genetic studies. One such study is
described in Cregan et. al, "An Integrated Genetic Linkage Map of the
Soybean Genome" Crop Science 39:1464-1490 (1999). Sequences and PCR
conditions of SSR Loci in Soybean as well as the most current genetic map
may be found in Soybase on the world wide web.
[0203]One use of molecular markers is Quantitative Trait Loci (QTL)
mapping. QTL mapping is the use of markers, which are known to be closely
linked to alleles that have measurable effects on a quantitative trait.
Selection in the breeding process is based upon the accumulation of
markers linked to the positive effecting alleles and/or the elimination
of the markers linked to the negative effecting alleles from the plant's
genome.
[0204]Molecular markers can also be used during the breeding process for
the selection of qualitative traits. For example, markers closely linked
to alleles or markers containing sequences within the actual alleles of
interest can be used to select plants that contain the alleles of
interest during a backcrossing breeding program. The markers can also be
used to select for the genome of the recurrent parent and against the
genome of the donor parent. Using this procedure can minimize the amount
of genome from the donor parent that remains in the selected plants. It
can also be used to reduce the number of crosses back to the recurrent
parent needed in a backcrossing program. The use of molecular markers in
the selection process is often called genetic marker enhanced selection.
Molecular markers may also be used to identify and exclude certain
sources of germplasm as parental varieties or ancestors of a plant by
providing a means of tracking genetic profiles through crosses.
Production of Double Haploids
[0205]The production of double haploids can also be used for the
development of plants with a homozygous phenotype in the breeding
program. For example, a soybean plant for which soybean cultivar
SG3870NRR is a parent can be used to produce double haploid plants.
Double haploids are produced by the doubling of a set of chromosomes (1
N) from a heterozygous plant to produce a completely homozygous
individual. For example, see Wan et al., "Efficient Production of Doubled
Haploid Plants Through Colchicine Treatment of Anther-Derived Maize
Callus", Theoretical and Applied Genetics, 77:889-892, 1989 and U.S. Pat.
No. 7,135,615. This can be advantageous because the process omits the
generations of selfing needed to obtain a homozygous plant from a
heterozygous source.
[0206]Haploid induction systems have been developed for various plants to
produce haploid tissues, plants and seeds. The haploid induction system
can produce haploid plants from any genotype by crossing a selected line
(as female) with an inducer line. Such inducer lines for maize include
Stock 6 (Coe, 1959, Am. Nat. 93:381-382; Sharkar and Coe, 1966, Genetics
54:453-464), KEMS (Deimling, Roeber, and Geiger, 1997, Vortr.
Pflanzenzuchtg 38:203-224), or KMS and ZMS (Chalyk, Bylich & Chebotar,
1994, MNL 68:47; Chalyk & Chebotar, 2000, Plant Breeding 119:363-364),
and indeterminate gametophyte (ig) mutation (Kermicle 1969 Science
166:1422-1424). The disclosures of which are incorporated herein by
reference.
[0207]Methods for obtaining haploid plants are also disclosed in
Kobayashi, M. et al., Journ. of Heredity 71(1):9-14, 1980, Pollacsek, M.,
Agronomie (Paris) 12(3):247-251, 1992; Cho-Un-Haing et al., Journ. of
Plant Biol., 1996, 39(3):185-188; Verdoodt, L., et al., February 1998,
96(2):294-300; Genetic Manipulation in Plant Breeding, Proceedings
International Symposium Organized by EUCARPIA, Sep. 8-13, 1985, Berlin,
Germany; Chalyk et al., 1994, Maize Genet Coop. Newsletter 68:47; Chalyk,
S.
[0208]Thus, an embodiment of this invention is a process for making a
substantially homozygous SG3870NRR progeny plant by producing or
obtaining a seed from the cross of SG3870NRR and another soybean plant
and applying double haploid methods to the F.sub.1 seed or F.sub.1 plant
or to any successive filial generation. Based on studies in maize and
currently being conducted in soybean, such methods would decrease the
number of generations required to produce a variety with similar genetics
or characteristics to SG3870NRR. See Bernardo, R. and Kahler, A. L.,
Theor. Appl. Genet. 102:986-992, 2001.
[0209]In particular, a process of making seed retaining the molecular
marker profile of soybean variety SG3870NRR is contemplated, such process
comprising obtaining or producing F.sub.1 seed for which soybean variety
SG3870NRR is a parent, inducing doubled haploids to create progeny
without the occurrence of meiotic segregation, obtaining the molecular
marker profile of soybean variety SG3870NRR, and selecting progeny that
retain the molecular marker profile of SG3870NRR.
[0210]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, 1960; Simmonds, 1979; Sneep et al., 1979; Fehr, 1987).
INDUSTRIAL USES
[0211]The seed of soybean cultivar SG3870NRR, the plant produced from the
seed, the hybrid soybean plant produced from the crossing of the variety
with any other soybean plant, hybrid seed, and various parts of the
hybrid soybean plant can be utilized for human food, livestock feed, and
as a raw material in industry. The soybean seed can be crushed or a
component of the soybean seed can be extracted in order to comprise a
component for a food or feed product.
[0212]The soybean is the world's leading source of vegetable oil and
protein meal. The oil extracted from soybeans is used for cooking oil,
margarine, and salad dressings. Soybean oil is composed of saturated,
monounsaturated and polyunsaturated fatty acids. It has 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 and American Soybean
Association Special Report 92S, May 1990). Changes in fatty acid
composition for improved oxidative stability and nutrition are constantly
sought after. Industrial uses of soybean oil which is subjected to
further processing include ingredients for paints, plastics, fibers,
detergents, cosmetics, lubricants and biodiesel fuel. Soybean oil may be
split, inter-esterified, sulfurized, epoxidized, polymerized,
ethoxylated, or cleaved. Designing and producing soybean oil derivatives
with improved functionality and improved oliochemistry is a rapidly
growing field. The typical mixture of triglycerides is usually split and
separated into pure fatty acids, which are then combined with
petroleum-derived alcohols or acids, nitrogen, sulfonates, chlorine, or
with fatty alcohols derived from fats and oils.
[0213]Soybean is also used as a food source for both animals and humans.
Soybean is widely used as a source of protein for animal feeds for
poultry, swine and cattle. During processing of whole soybeans, the
fibrous hull is removed and the oil is extracted. The remaining soybean
meal is a combination of carbohydrates and approximately 50% protein.
[0214]For human consumption soybean meal is made into soybean flour which
is processed to protein concentrates used for meat extenders or specialty
pet foods. Production of edible protein ingredients from soybean offers a
healthier, less expensive replacement for animal protein in meats as well
as in dairy-type products.
Tables
[0215]In Table 2 that follows, traits and characteristics of soybean
cultivar SG3870NRR are compared to two commercial cultivars, taken over
an average of 25 locations. Column 1 shows the variety, column 2 shows
the relative maturity, column 3 shows the lodging resistance score (on a
scale from 1 to 5 with 1 indicating erect plants), column 4 shows the
height in centimeters, column 5 shows the seed yield in Bushels/Acre,
column 6 shows the Soybean Cyst Nematode disease score (where R indicates
Resistant and MR indicates Moderately Resistant) and column 6 shows the
Phytophthora sojae Root Rot Resistance score (where the Resistance genes
are listed).
TABLE-US-00002
TABLE 2
PAIRED COMPARISONS
Variety Maturity Lodging Height Yield SCN PRR
SG3870NRR 3.9 1.6 99 52.5 R 1C
AG3905 4.1 1.4 100 51.6 MR 1C
P93M90 3.8 1.4 101 50.3 R S
AG3705 3.7 1.5 94 49.6 R 1C
Deposit Information
[0216]A deposit of the soybean seed of this invention is maintained by
Soygenetics, LLC, 4846 E. 450 N., Lafayette, Ind. 47905 Attn: Steve
Baluch. Access to this deposit will be available during the pendency of
this application to persons determined by the Commissioner of Patents and
Trademarks to be entitled thereto under 37 CFR .sctn.1.14 and 35 USC
.sctn.122. Upon allowance of any claims in this application, all
restrictions on the availability to the public of the variety will be
irrevocably removed by affording access to a deposit of at least 2,500
seeds of the same variety with the American Type Culture Collection,
Manassas, Va. or National Collections of Industrial, Food and Marine
Bacteria (NCIMB), 23 St Machar Drive, Aberdeen, Scotland, AB24 3RY,
United Kingdom.
[0217]While a number of exemplary aspects and embodiments have been
discussed above, those of skill in the art will recognize certain
modifications, permutations, additions and sub-combinations thereof. It
is therefore intended that the following appended claims and claims
hereafter introduced are interpreted to include all such modifications,
permutations, additions and sub-combinations as are within their true
spirit and scope.
* * * * *