U.S. patent application number 12/641804 was filed with the patent office on 2010-06-24 for canola hybrid 45s51.
This patent application is currently assigned to Pioneer Hi-Bred International, Inc.. Invention is credited to Igor Falak, Jayantilal D. Patel.
Application Number | 20100162424 12/641804 |
Document ID | / |
Family ID | 42268130 |
Filed Date | 2010-06-24 |
United States Patent
Application |
20100162424 |
Kind Code |
A1 |
Patel; Jayantilal D. ; et
al. |
June 24, 2010 |
Canola Hybrid 45S51
Abstract
A novel canola variety designated 45S51 and seed, plants and
plant parts thereof, produced by crossing Pioneer Hi-Bred
International, Inc. proprietary inbred canola varieties. Methods
for producing a canola plant that comprises crossing canola variety
45S51 with another canola plant. Methods for producing a canola
plant containing in its genetic material one or more traits
introgressed into 45S51 through backcross conversion and/or
transformation, and to the canola seed, plant and plant part
produced thereby. This invention relates to the canola variety
45S51, the seed, the plant produced from the seed, and variants,
mutants, and minor modifications of canola variety 45S51. This
invention further relates to methods for producing canola varieties
derived from canola variety 45S51.
Inventors: |
Patel; Jayantilal D.;
(Thornhill, CA) ; Falak; Igor; (Guelph,
CA) |
Correspondence
Address: |
PIONEER HI-BRED INTERNATIONAL, INC.
7250 N.W. 62ND AVENUE, P.O. BOX 552
JOHNSTON
IA
50131-0552
US
|
Assignee: |
Pioneer Hi-Bred International,
Inc.
|
Family ID: |
42268130 |
Appl. No.: |
12/641804 |
Filed: |
December 18, 2009 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61138692 |
Dec 18, 2008 |
|
|
|
Current U.S.
Class: |
800/260 ;
800/300; 800/301; 800/302; 800/303; 800/306 |
Current CPC
Class: |
C12N 15/8275 20130101;
C12N 15/8247 20130101; C12N 15/8281 20130101; C12N 15/8278
20130101; C12N 15/8253 20130101; C12N 15/8251 20130101; C12N
15/8273 20130101; C12N 15/8286 20130101; C12N 15/8283 20130101;
C12N 15/8254 20130101; C12N 15/8274 20130101; C12N 15/8245
20130101; A01H 5/10 20130101; C12N 15/8282 20130101; C12N 15/8289
20130101 |
Class at
Publication: |
800/260 ;
800/306; 800/300; 800/301; 800/302; 800/303 |
International
Class: |
A01H 5/00 20060101
A01H005/00; A01H 1/02 20060101 A01H001/02 |
Claims
1. A canola variety 45S51, produced by crossing a first plant of
variety with a second plant of variety, wherein representative seed
of said varieties and have been deposited under ATCC Accession
Number PTA-______ and PTA-______, respectively.
2. A seed of the canola variety of claim 1.
3. The seed of claim 2, further comprising a transgenic event.
4. The seed of claim 3, wherein the transgenic event confers a
trait selected from the group consisting of male sterility,
site-specific recombination, abiotic stress tolerance, altered
phosphorus, altered antioxidants, altered fatty acids, altered
essential amino acids, altered carbohydrates, herbicide resistance,
insect resistance and disease resistance.
5. The seed of claim 2, further comprising a locus conversion.
6. The seed of claim 5, wherein the locus conversion confers a
trait selected from the group consisting of male sterility,
site-specific recombination, abiotic stress tolerance, altered
phosphorus, altered antioxidants, altered fatty acids, altered
essential amino acids, altered carbohydrates, herbicide resistance,
insect resistance and disease resistance.
7. A plant or plant part of the canola variety of claim 1.
8. A method for producing a second canola plant comprising applying
plant breeding techniques to a first canola plant, or parts
thereof, wherein said first canola plant is the canola plant of
claim 7, and wherein application of said techniques results in the
production of said second canola plant.
9. The method of claim 8, further defined as producing an inbred
canola plant derived from canola variety 45S51, the method
comprising the steps of: (a) crossing said first canola plant with
itself or another canola plant to produce seed of a subsequent
generation; (b) harvesting and planting the seed of the subsequent
generation to produce at least one plant of the subsequent
generation; (c) repeating steps (a) and (b) for an additional 2-10
generations to produce an inbred canola plant derived from canola
variety 45S51.
10. The method of claim 8, further defined as producing an inbred
canola plant derived from canola variety 45S51, the method
comprising the steps of: (a) crossing said first canola plant with
an inducer variety to produce haploid seed; and (b) doubling the
haploid seed to produce an inbred canola plant derived from canola
variety 45S51.
11. The plant or plant part of claim 7, further comprising a
transgenic event.
12. The plant of claim 11, wherein the transgenic event confers a
trait selected from the group consisting of selected from the group
consisting of male sterility, site-specific recombination, abiotic
stress tolerance, altered phosphorus, altered antioxidants, altered
fatty acids, altered essential amino acids, altered carbohydrates,
herbicide resistance, insect resistance and disease resistance.
13. A method for producing a second canola plant comprising
applying plant breeding techniques to a first canola plant, or
parts thereof, wherein said first canola plant is the canola plant
of claim 11, and wherein application of said techniques results in
the production of said second canola plant.
14. The method of claim 13, further defined as producing an inbred
canola plant derived from canola variety 45S51, the method
comprising the steps of: (a) crossing said first canola plant with
itself or another canola plant to produce seed of a subsequent
generation; (b) harvesting and planting the seed of the subsequent
generation to produce at least one plant of the subsequent
generation; (c) repeating steps (a) and (b) for an additional 2-10
generations to produce an inbred canola plant derived from canola
variety 45S51.
15. The method of claim 13, further defined as producing an inbred
canola plant derived from canola variety 45S51, the method
comprising the steps of: (a) crossing said first canola plant with
an inducer variety to produce haploid seed; and (b) doubling the
haploid seed to produce an inbred canola plant derived from canola
variety 45S51.
16. The plant or plant part of claim 7, further comprising a locus
conversion.
17. The plant or plant part of claim 16, wherein the locus
conversion confers a trait selected from the group consisting of
male sterility, site-specific recombination, abiotic stress
tolerance, altered phosphorus, altered antioxidants, altered fatty
acids, altered essential amino acids, altered carbohydrates,
herbicide resistance, insect resistance and disease resistance.
18. A method for producing a second canola plant comprising
applying plant breeding techniques to a first canola plant, or
parts thereof, wherein said first canola plant is the canola plant
of claim 16, and wherein application of said techniques results in
the production of said second canola plant.
19. The method of claim 18, further defined as producing an inbred
canola plant derived from canola variety 45S51, the method
comprising the steps of: (a) crossing said first canola plant with
itself or another canola plant to produce seed of a subsequent
generation; (b) harvesting and planting the seed of the subsequent
generation to produce at least one plant of the subsequent
generation; (c) repeating steps (a) and (b) for an additional 2-10
generations to produce an inbred canola plant derived from canola
variety 45S51.
20. The method of claim 18, further defined as producing an inbred
canola plant derived from canola variety 45S51, the method
comprising the steps of: (a) crossing said first canola plant with
an inducer variety to produce haploid seed; and (b) doubling the
haploid seed to produce an inbred canola plant derived from canola
variety 45S51.
Description
CROSS-REFERENCE TO RELATED 5 APPLICATIONS
[0001] This application claims priority under 35 U.S.C. .sctn.119
to provisional application Ser. No. 61/138,692 filed Dec. 18, 2008,
herein incorporated by reference in its entirety.
FIELD OF THE INVENTION
[0002] The invention is in the field of Brassica napus breeding
(i.e., canola breeding), specifically relating to the canola hybrid
designated 45S51.
BACKGROUND OF THE INVENTION
[0003] The present invention relates to a novel rapeseed variety
designated 45S51 which is the result of years of careful breeding
and selection. Since such variety is of high quality and possesses
a relatively low level of erucic acid in the vegetable oil
component and a relatively low level of glucosinolate content in
the meal component, it can be termed "canola" in accordance with
the terminology commonly used by plant scientists.
[0004] The goal of plant breeding is to combine in a single variety
or hybrid various desirable traits. For field crops, these traits
may include resistance to diseases and insects, tolerance to heat
and drought, reducing the time to crop maturity, greater yield, and
better agronomic quality. With mechanical harvesting of many crops,
uniformity of plant characteristics such as germination and stand
establishment, growth rate, maturity, and plant and pod height, is
important.
[0005] Field crops are bred through techniques that take advantage
of the plant's method of pollination. A plant is self-pollinated if
pollen from one flower is transferred to the same or another flower
of the same plant or a genetically identical plant. A plant is
sib-pollinated when individuals within the same family or line are
used for pollination. A plant is cross-pollinated if the pollen
comes from a flower on a genetically different plant from a
different family or line. The term "cross-pollination" used herein
does not include self-pollination or sib-pollination.
[0006] The creation of new superior, agronomically sound, and
stable high-yielding cultivars of many plant types including canola
has posed an ongoing challenge to plant breeders. In the practical
application of a chosen breeding program, the breeder often
initially selects and crosses two or more parental lines, followed
by repeated selfing and selection, thereby producing many unique
genetic combinations. The breeder can theoretically generate
billions of different genetic combinations via crossing, selfing
and mutagenesis. However, the breeder commonly has no direct
control at the cellular level of the plant. Therefore, two breeders
will never independently develop the same variety having the same
canola traits.
[0007] In each cycle of evaluation, the plant breeder selects the
germplasm to advance to the next generation. This germplasm is
grown under chosen geographical, climatic and soil conditions, and
further selections are then made during and at the end of the
growing season. The characteristics of the varieties developed are
incapable of prediction in advance. This unpredictability is
because the 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 cannot predict in advance
the final resulting varieties that are to be developed, except
possibly in a very gross and general fashion. Even the same breeder
is incapable of producing the same variety twice by using the same
original parents and the same selection techniques. This
unpredictability commonly results in the expenditure of large
research monies and effort to develop a new and superior canola
variety.
[0008] Canola breeding programs utilize techniques such as mass and
recurrent selection, backcrossing, pedigree breeding and haploidy.
For a general description of rapeseed and Canola breeding, see,
Downey and Rakow, (1987) "Rapeseed and Mustard" In: Principles of
Cultivar Development, Fehr, (ed.), pp 437-486; New York; Macmillan
and Co.; Thompson, (1983) "Breeding winter oilseed rape Brassica
napus"; Advances in Applied Biology 7:1-104; and Ward, et. al.,
(1985) Oilseed Rape, Farming Press Ltd., Wharfedale Road, Ipswich,
Suffolk, each of which is hereby incorporated by reference.
[0009] Recurrent selection is used to improve populations of either
self- or cross-pollinating Brassica. Through recurrent selection, a
genetically variable population of heterozygous individuals is
created by intercrossing several different parents. The best plants
are selected based on individual superiority, outstanding progeny,
and/or excellent combining ability. The selected plants are
intercrossed to produce a new population in which further cycles of
selection are continued. Various recurrent selection techniques are
used to improve quantitatively inherited traits controlled by
numerous genes.
[0010] Breeding programs use backcross breeding to transfer genes
for a simply inherited, highly heritable trait into another line
that serves as the recurrent parent. The source of the trait to be
transferred is called the donor parent. After the initial cross,
individual plants possessing the desired trait of the donor parent
are selected and are crossed (backcrossed) to the recurrent parent
for several generations. The resulting plant is expected to have
the attributes of the recurrent parent and the desirable trait
transferred from the donor parent. This approach has been used for
breeding disease resistant phenotypes of many plant species, and
has been used to transfer low erucic acid and low glucosinolate
content into lines and breeding populations of Brassica.
[0011] Pedigree breeding and recurrent selection breeding methods
are used to develop varieties from breeding populations. Pedigree
breeding starts with the crossing of two genotypes, each of which
may have one or more desirable characteristics that is lacking in
the other or which complements the other. If the two original
parents do not provide all of the desired characteristics, other
sources can be included in the breeding population. In the pedigree
method, superior plants are selfed and selected in successive
generations. In the succeeding generations the heterozygous
condition gives way to homogeneous lines as a result of
self-pollination and selection. Typically in the pedigree method of
breeding, five or more generations of selfing and selection are
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. For example, two parents that are
believed to possess favorable complementary traits are crossed to
produce an F.sub.1. An F.sub.2 population is produced by selfing
one or several F.sub.1's or by intercrossing two F.sub.1's (i.e.,
sib mating). Selection of the best individuals may begin in the
F.sub.2 population, and 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 commonly are
tested for potential release as new cultivars. Backcrossing may be
used in conjunction with pedigree breeding; for example, a
combination of backcrossing and pedigree breeding with recurrent
selection has been used to incorporate blackleg resistance into
certain cultivars of Brassica napus.
[0012] Plants that have been self-pollinated and selected for type
for many generations become homozygous at almost all gene loci and
produce a uniform population of true breeding progeny. If desired,
double-haploid methods can also be used to extract homogeneous
lines. A cross between two different homozygous lines produces a
uniform population of hybrid plants that may be heterozygous for
many gene loci. A cross of two plants each heterozygous at a number
of gene loci will produce a population of hybrid plants that differ
genetically and will not be uniform.
[0013] The 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, such as
F.sub.1 hybrid variety or open pollinated variety. A true breeding
homozygous line can also be used as a parental line (inbred line)
in a commercial hybrid. If the line is being developed as an inbred
for use in a hybrid, an appropriate pollination control system
should be incorporated in the line. Suitability of an inbred line
in a hybrid combination will depend upon the combining ability
(general combining ability or specific combining ability) of the
inbred.
[0014] Various breeding procedures are also utilized with these
breeding and selection methods. 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.
[0015] In a multiple-seed procedure, canola 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. 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. If desired, doubled-haploid
methods can be used to extract homogeneous lines.
[0016] Molecular markers, including 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. 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 in the plant's
genome.
[0017] 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 markers 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 or Marker Assisted Selection (MAS).
[0018] The production of doubled haploids can also be used for the
development of inbreds in the breeding program. In Brassica napus,
microspore culture technique is used in producing haploid embryos.
The haploid embryos are then regenerated on appropriate media as
haploid plantlets, doubling chromosomes of which results in doubled
haploid plants. This can be advantageous because the process omits
the generations of selfing needed to obtain a homozygous plant from
a heterozygous source.
[0019] A pollination control system and effective transfer of
pollen from one parent to the other offers improved plant breeding
and an effective method for producing hybrid canola seed and
plants. For example, the ogura cytoplasmic male sterility (cms)
system, developed via protoplast fusion between radish (Raphanus
sativus) and rapeseed (Brassica napus), is one of the most
frequently used methods of hybrid production. It provides stable
expression of the male sterility trait (Ogura, 1968, Pelletier, et
al., 1983) and an effective nuclear restorer gene (Heyn, 1976).
[0020] In developing improved new Brassica hybrid varieties,
breeders may use self-incompatible (SI), cytoplasmic male sterile
(CMS) or nuclear male sterile (NMS) Brassica plants as the female
parent. In using these plants, breeders are attempting to improve
the efficiency of seed production and the quality of the F.sub.1
hybrids and to reduce the breeding costs. When hybridization is
conducted without using SI, CMS or NMS plants, it is more difficult
to obtain and isolate the desired traits in the progeny (F.sub.1
generation) because the parents are capable of undergoing both
cross-pollination and self-pollination. If one of the parents is a
SI, CMS or NMS plant that is incapable of producing pollen, only
cross pollination will occur. By eliminating the pollen of one
parental variety in a cross, a plant breeder is assured of
obtaining hybrid seed of uniform quality, provided that the parents
are of uniform quality and the breeder conducts a single cross.
[0021] In one instance, production of F.sub.1 hybrids includes
crossing a CMS Brassica female parent with a pollen-producing male
Brassica parent. To reproduce effectively, however, the male parent
of the F.sub.1 hybrid must have a fertility restorer gene (Rf
gene). The presence of an Rf gene means that the F.sub.1 generation
will not be completely or partially sterile, so that either
self-pollination or cross pollination may occur. Self pollination
of the F.sub.1 generation to produce several subsequent generations
is important to ensure that a desired trait is heritable and stable
and that a new variety has been isolated.
[0022] An example of a Brassica plant which is cytoplasmic male
sterile and used for breeding is ogura (OGU) cytoplasmic male
sterile (Pellan-Delourme, et al., 1987). A fertility restorer for
ogura cytoplasmic male sterile plants has been transferred from
Raphanus sativus (radish) to Brassica by Instit. National de
Recherche Agricole (INRA) in Rennes, France (Pelletier, et al.,
1987). The restorer gene, Rf1 originating from radish, is described
in WO 92/05251 and in Delourme, et al., (1991). Improved versions
of this restorer have been developed. For example, see WO98/27806,
oilseed brassica containing an improved fertility restorer gene for
ogura cytoplasmic male sterility, which is hereby incorporated by
reference.
[0023] Other sources and refinements of CMS sterility in canola
include the Polima cytoplasmic male sterile plant, as well as those
of U.S. Pat. No. 5,789,566, DNA sequence imparting cytoplasmic male
sterility, mitochondrial genome, nuclear genome, mitochondria and
plant containing said sequence and process for the preparation of
hybrids; U.S. Pat. No. 5,973,233 Cytoplasmic male sterility system
production canola hybrids; and WO97/02737 Cytoplasmic male
sterility system producing canola hybrids; EP Patent Application
Number 0 599042A Methods for introducing a fertility restorer gene
and for producing F1 hybrids of Brassica plants thereby; U.S. Pat.
No. 6,229,072 Cytoplasmic male sterility system production canola
hybrids; U.S. Pat. No. 4,658,085 Hybridization using cytoplasmic
male sterility, cytoplasmic herbicide tolerance, and herbicide
tolerance from nuclear genes; all of which are incorporated herein
for this purpose.
[0024] Promising advanced breeding lines commonly are tested and
compared to appropriate standards in environments representative of
the commercial target area(s). The best lines are candidates for
new commercial lines; and those still deficient in a few traits may
be used as parents to produce new populations for further
selection.
[0025] For most traits the true genotypic value may be masked by
other confounding plant traits or environmental factors. One method
for identifying a superior plant is to observe its performance
relative to other experimental plants and to one or more widely
grown standard varieties. If a single observation is inconclusive,
replicated observations provide a better estimate of the genetic
worth.
[0026] Proper testing should detect any major faults and establish
the level of superiority or improvement over current varieties. In
addition to showing superior performance, there must be a demand
for a new variety that is compatible with industry standards or
which creates a new market. The introduction of a new variety
commonly will incur additional costs to the seed producer, the
grower, the processor and the consumer, for special advertising and
marketing, altered seed and commercial production practices, and
new product utilization. The testing preceding release of a new
variety should take into consideration research and development
costs as well as technical superiority of the final variety. For
seed-propagated varieties, it must be feasible to produce seed
easily and economically.
[0027] These processes, which lead to the final step of marketing
and distribution, usually take from approximately six to twelve
years from the time the first cross is made. Therefore, the
development of new varieties such as that of the present invention
is a time-consuming process that requires precise forward planning,
efficient use of resources, and a minimum of changes in
direction.
[0028] Further, as a result of the advances in sterility systems,
lines are developed that can be used as an open pollinated variety
(i.e., a pureline cultivar sold to the grower for planting) and/or
as a sterile inbred (female) used in the production of F.sub.1
hybrid seed. In the latter case, favorable combining ability with a
restorer (male) would be desirable. The resulting hybrid seed would
then be sold to the grower for planting.
[0029] The development of a canola hybrid in a canola plant
breeding program involves three steps: (1) the selection of plants
from various germplasm pools for initial breeding crosses; (2) the
selfing of the selected plants from the breeding crosses for
several generations to produce a series of inbred lines, which,
although different from each other, breed true and are highly
uniform; and (3) crossing the selected inbred lines with different
inbred lines to produce the hybrids. During the inbreeding process
in canola, the vigor of the lines decreases. Vigor is restored when
two different inbred lines are crossed to produce the hybrid. An
important consequence of the homozygosity and homogeneity of the
inbred lines is that the hybrid between a defined pair of inbreds
will always be the same. Once the inbreds that give a superior
hybrid have been identified, the hybrid seed can be reproduced
indefinitely as long as the homogeneity of the inbred parents is
maintained.
[0030] Combining ability of a line, as well as the performance of
the line per se, is a factor in the selection of improved canola
lines that may be used as inbreds. Combining ability refers to a
line's contribution as a parent when crossed with other lines to
form hybrids. The hybrids formed for the purpose of selecting
superior lines are designated test crosses. One way of measuring
combining ability is by using breeding values. Breeding values are
based on the overall mean of a number of test crosses. This mean is
then adjusted to remove environmental effects and it is adjusted
for known genetic relationships among the lines.
[0031] Hybrid seed production requires inactivation of pollen
produced by the female parent. Incomplete inactivation of the
pollen provides the potential for self-pollination. This
inadvertently self-pollinated seed may be unintentionally harvested
and packaged with hybrid seed. Similarly, because the male parent
is grown next to the female parent in the field, there is also the
potential that the male selfed seed could be unintentionally
harvested and packaged with the hybrid seed. Once the seed from the
hybrid bag is planted, it is possible to identify and select these
self-pollinated plants. These self-pollinated plants will be
genetically equivalent to one of the inbred lines used to produce
the hybrid. Though the possibility of inbreds being included in
hybrid seed bags exists, the occurrence is rare because much care
is taken to avoid such inclusions. These self-pollinated plants can
be identified and selected by one skilled in the art, through
either visual or molecular methods.
[0032] Brassica napus canola plants, absent the use of sterility
systems, are recognized to commonly be self-fertile with
approximately 70 to 90 percent of the seed normally forming as the
result of self-pollination. The percentage of cross pollination may
be further enhanced when populations of recognized insect
pollinators at a given growing site are greater. Thus open
pollination is often used in commercial canola production.
[0033] Currently Brassica napus canola is being recognized as an
increasingly important oilseed crop and a source of meal in many
parts of the world. The oil as removed from the seeds commonly
contains a lesser concentration of endogenously formed saturated
fatty acids than other vegetable oils and is well suited for use in
the production of salad oil or other food products or in cooking or
frying applications. The oil also finds utility in industrial
applications. Additionally, the meal component of the seeds can be
used as a nutritious protein concentrate for livestock.
[0034] Canola oil has the lowest level of saturated fatty acids of
all vegetable oils. "Canola" refers to rapeseed (Brassica) which
(1) has an erucic acid (C.sub.22:1) content of at most 2 percent by
weight based on the total fatty acid content of a seed, preferably
at most 0.5 percent by weight and most preferably essentially 0
percent by weight; and (2) produces, after crushing, an air-dried
meal containing less than 30 micromoles (.mu.mol) glucosinolates
per gram of defatted (oil-free) meal. These types of rapeseed are
distinguished by their edibility in comparison to more traditional
varieties of the species.
[0035] Sclerotinia infects over 100 species of plants, including
numerous economically important crops such as Brassica species,
sunflowers, dry beans, soybeans, field peas, lentils, lettuce, and
potatoes (Boland and Hall, 1994). Sclerotinia sclerotiorum is
responsible for over 99% of Sclerotinia disease, while Sclerotinia
minor produces less than 1% of the disease. Sclerotinia produces
sclerotia, irregularly-shaped, dark overwintering bodies, which can
endure in soil for four to five years. The sclerotia can germinate
carpogenically or myceliogenically, depending on the environmental
conditions and crop canopies. The two types of germination cause
two distinct types of diseases. Sclerotia that germinate
carpogenically produce apothecia and ascospores that infect
above-ground tissues, resulting in stem blight, stalk rot, head
rot, pod rot, white mold and blossom blight of plants. Sclerotia
that germinate myceliogenically produce mycelia that infect root
tissues, causing crown rot, root rot and basal stalk rot.
[0036] Sclerotinia causes Sclerotinia stem rot, also known as white
mold, in Brassica, including canola. Canola is a type of Brassica
having a low level of glucosinolates and erucic acid in the seed.
The sclerotia germinate carpogenically in the summer, producing
apothecia. The apothecia release wind-borne ascospores that travel
up to one kilometer. The disease is favoured by moist soil
conditions (at least 10 days at or near field capacity) and
temperatures of 15-25.degree. C., prior to and during canola
flowering. The spores cannot infect leaves and stems directly; they
must first land on flowers, fallen petals, and pollen on the stems
and leaves. Petal age affects the efficiency of infection, with
older petals more likely to result in infection (Heran, et al.,
1999). The fungal spores use the flower parts as a food source as
they germinate and infect the plant.
[0037] The severity of Sclerotinia in Brassica is variable, and is
dependent on the time of infection and climatic conditions (Heran,
et al., 1999). The disease is favored by cool temperatures and
prolonged periods of precipitation. Temperatures between 20 and
25.degree. C. and relative humidities of greater than 80% are
required for optimal plant infection (Heran, et al., 1999). Losses
ranging from 5 to 100% have been reported for individual fields
(Manitoba Agriculture, Food and Rural Initiatives, 2004). On
average, yield losses are estimated to be 0.4 to 0.5 times the
Sclerotinia Sclerotiorum Field Severity score, a rating based on
both percentage infection and disease severity. More information is
provided herein at Example 2. For example, if a field has 20%
infection (20/100 plants infected), then the yield loss would be
about 10% provided plants are dying prematurely due to the
infection of the main stem (rating 5--SSFS=20')/0). If the plants
are affected much less (rating 1--SSFS=4%), yield loss is reduced
accordingly. Further, Sclerotinia can cause heavy losses in wet
swaths. Sclerotinia sclerotiorum caused economic losses to canola
growers in Minnesota and North Dakota of 17.3, 20.8, and 16.8
million dollars in 1999, 2000 and 2001, respectively (Bradley, et
al. 2006). In Canada, this disease is extremely important in
Southern Manitoba, parts of South Central Alberta and also in
Eastern areas of Saskatchewan. Since weather plays an important
role in development of this disease, its occurrence is irregular
and unpredictable. Certain reports estimate about 0.8 to 1.3
million acres of canola being sprayed with fungicide in Southern
Manitoba annually. The fungicide application costs about $25 per
acre, which represents a significant cost for canola producers.
Moreover, producers may decide to apply fungicide based on the
weather forecast, while later changes in the weather pattern
discourage disease development, resulting in wasted product, time,
and fuel. Creation of sclerotinia tolerant canola cultivars has
been an important goal for many of the Canadian canola breeding
organizations.
[0038] No canola cultivar carrying an improved level of genetic
resistance to Sclerotinia has previously been released in Canada.
45S51 is the first canola hybrid cultivar having this improved
level of sclerotinia tolerance. The sclerotinia tolerance in 45S51
comes from parents developed by conventional plant breeding
techniques of crossing and selection. The parental lines were
developed and screened in a field screening nursery which was
inoculated to ensure high and consistent Sclerotinia disease
pressure. See, for example, the methods described in PCT
publication WO2006/135717.
[0039] Since canola variety 45S51 is a hybrid produced from
substantially homogeneous parents, it can be reproduced by planting
seeds of such parents, growing the resulting canola plants under
controlled pollination conditions with adequate isolation so that
cross-pollination occurs between the parents, and harvesting the
resulting hybrid seed using conventional agronomic practices.
[0040] The symptoms of Sclerotinia infection usually develop
several weeks after flowering begins. The plants develop pale-grey
to white lesions, at or above the soil line and on upper branches
and pods. The infections often develop where the leaf and the stem
join because the infected petals lodge there. Once plants are
infected, the mold continues to grow into the stem and invade
healthy tissue. Infected stems appear bleached and tend to shred.
Hard black fungal sclerotia develop within the infected stems,
branches, or pods. Plants infected at flowering produce little or
no seed. Plants with girdled stems wilt and ripen prematurely.
Severely infected crops frequently lodge, shatter at swathing, and
make swathing more time consuming. Infections can occur in all
above-ground plant parts, especially in dense or lodged stands,
where plant-to-plant contact facilitates the spread of infection.
New sclerotia carry the disease over to the next season.
[0041] Conventional methods for control of Sclerotinia diseases
include (a) chemical control, (b) disease resistance and (c)
cultural control, each of which is described below.
[0042] (a) Fungicides such as benomyl, vinclozolin and iprodione
remain the main method of control of Sclerotinia disease (Morall,
et al., 1985; Tu, 1983). Recently, additional fungicidal
formulations have been developed for use against Sclerotinia,
including azoxystrobin, prothioconazole, and boscalid. (Johnson,
2005) However, use of fungicide is expensive and can be harmful to
the user and environment. Further, resistance to some fungicides
has occurred due to repeated use.
[0043] (b) In certain cultivars of bean, safflower, sunflower and
soybean, some progress has been made in developing partial
(incomplete) resistance. Partial resistance is often referred to as
tolerance. However, success in developing partial resistance has
been very limited, probably because partial physiological
resistance is a multigene trait as demonstrated in bean (Fuller, et
al., 1984). In addition to partial physiological resistance, some
progress has been made to breed for morphological traits to avoid
Sclerotinia infection, such as upright growth habit, lodging
resistance and narrow canopy. For example, bean plants with partial
physiological resistance and with an upright stature, narrow canopy
and indeterminate growth habit were best able to avoid Sclerotinia
(Saindon, et al., 1993). Early maturing cultivars of safflower
showed good field resistance to Sclerotinia. Finally, in soybean,
cultivar characteristics such as height, early maturity and great
lodging resistance result in less disease, primarily because of a
reduction of favorable microclimate conditions for the disease.
(Boland and Hall, 1987; Buzzell, et al. 1993)
[0044] (c) Cultural practices, such as using pathogen-free or
fungicide-treated seed, increasing row spacing, decreasing seeding
rate to reduce secondary spread of the disease, and burying
sclerotia to prevent carpogenic germination, may reduce Sclerotinia
disease but not effectively control the disease.
[0045] All Canadian canola genotypes are susceptible to Sclerotinia
stem rot (Manitoba Agriculture, Food and Rural Initiatives, 2004).
This includes all known spring petalled genotypes of canola
quality. There is also no resistance to Sclerotinia in Australian
canola varieties. (Hind-Lanoiselet, et al. 2004). Some varieties
with certain morphological traits are better able to withstand
Sclerotinia infection. For example, Polish varieties (Brassica
rapa) have lighter canopies and seem to have much lower infection
levels. In addition, petal-less varieties (apetalous varieties)
avoid Sclerotinia infection to a greater extent (Okuyama, et al.,
1995; Fu, 1990). Other examples of morphological traits which
confer a degree of reduced field susceptibility in Brassica
genotypes include increased standability, reduced petal retention,
branching (less compact and/or higher), and early leaf abscission.
Jurke and Fernando, (2003) screened eleven canola genotypes for
Sclerotinia disease incidence. Significant variation in disease
incidence was explained by plant morphology, and the difference in
petal retention was identified as the most important factor.
However, these morphological traits alone do not confer resistance
to Sclerotinia, and all canola products in Canada are considered
susceptible to Sclerotinia.
[0046] Winter canola genotypes are also susceptible to Sclerotinia.
In Germany, for example, no Sclerotinia-resistant varieties are
available. (Specht, 2005) The widely-grown German variety Express
is considered susceptible to moderately susceptible and belongs to
the group of less susceptible varieties/hybrids.
[0047] Spraying with fungicide is the only means of controlling
Sclerotinia in canola crops grown under disease-favorable
conditions at flowering. Typical fungicides used for controlling
Sclerotinia on Brassica include Rovral.TM./Proline.TM. from Bayer
and Ronilan.TM./Lance.TM. from BASF. The active ingredient in
Lance.TM. is Boscalid, and it is marketed as Endura.TM. in the
United States. The fungicide should be applied before symptoms of
stem rot are visible and usually at the 20-30% bloom stage of the
crop. If infection is already evident, there is no use in applying
fungicide as it is too late to have an effect. Accordingly, growers
must assess their fields for disease risk to decide whether to
apply a fungicide. This can be done by using a government provided
checklist or by using a petal testing kit. Either method is
cumbersome and prone to errors. (Hind-Lanoiselet, 2004; Johnson,
2005)
[0048] Numerous efforts have been made to develop Sclerotinia
resistant Brassica plants. Built-in resistance would be more
convenient, economical, and environmentally-friendly than
controlling Sclerotinia by application of fungicides. Since the
trait is polygenic it would be stable and not prone to loss of
efficacy, as fungicides may be.
SUMMARY OF THE INVENTION
[0049] According to the present invention, there is provided a
novel Brassica napus hybrid designated 45S51. This invention thus
relates to the seeds of the 45S51 hybrid, to plants of the 45S51
hybrid, and to methods for producing a canola plant by crossing the
45S51 hybrid with itself or another canola plant (whether by use of
male sterility or open pollination), and to methods for producing a
canola plant containing in its genetic material one or more
transgenes, and to transgenic plants produced by that method. This
invention also relates to canola seeds and plants produced by
crossing the hybrid 45S51 with another line.
DEFINITIONS
[0050] In the description and tables which follow, a number of
terms are used. In order to aid in a clear and consistent
understanding of the specification, the following definitions and
evaluation criteria are provided.
[0051] Anther Fertility. The ability of a plant to produce pollen
1=sterile, 2=all anthers shedding pollen (vs. Pollen Formation
which is amount of pollen produced).
[0052] Anther Arrangement. The general disposition of the anthers
in typical fully opened flowers is observed.
[0053] Chlorophyll Content. The typical chlorophyll content of the
mature seeds is determined by using methods recommended by the
Western Canada Canola/Rapeseed Recommending Committee (WCC/RRC) and
is considered to be low if <8 ppm, medium if 8 to 15 ppm, and
high if >15 ppm. Also, chlorophyll could be analyzed using NIR
(Near Infra Red spectroscopy) as long as the instrument is
calibrated according to the manufacturer's specifications.
[0054] Cotyledon. A cotyledon is a part of the embryo within the
seed of a plant; it is also referred to as a seed leaf. Upon
germination, the cotyledon may become the embryonic first leaf of a
seedling.
[0055] Cotyledon Length. The distance between the indentation at
the top of the cotyledon and the point where the width of the
petiole is approximately 4 mm.
[0056] Cotyledon Width. The width at the widest point of the
cotyledon when the plant is at the two- to three-leaf stage of
development.
[0057] Disease Resistance: Resistance to various diseases is
evaluated and is expressed on a scale of 0=highly resistant,
5=highly susceptible. The WCC/RRC blackleg classification is based
on % severity index described as follows: [0058] 0-30%=Resistant
[0059] 30%-50%=Moderately Resistant [0060] 50%-70%=Moderately
Susceptible [0061] 70%-90%=Susceptible [0062] >90%=Highly
susceptible. The % severity index=blackleg rating on 0-5 for a
variety/blackleg rating for HS variety Westar. Sclerotinia scoring
is described in Example 2 herein.
[0063] Erucic Acid Content: The percentage of the fatty acids in
the form of C22:1. as determined by one of the methods recommended
by the WCC/RRC, being AOCS Official Method Ce 2-66 Preparation of
Methyl esters of Long-Chain Fatty Acids or AOCS Official Method Ce
1-66 Fatty Acid Composition by Gas Chromatography.
[0064] Fatty Acid Content: The typical percentages by weight of
fatty acids present in the endogenously formed oil of the mature
whole dried seeds are determined. During such determination the
seeds are crushed and are extracted as fatty acid methyl esters
following reaction with methanol and sodium methoxide. Next the
resulting ester is analyzed for fatty acid content by gas liquid
chromatography using a capillary column which allows separation on
the basis of the degree of unsaturation and fatty acid chain
length. This procedure is described in the work of Daun, et al.,
(1983) J. Amer. Oil Chem. Soc. 60:1751 to 1754 which is herein
incorporated by reference.
[0065] Flower Bud Location. A determination is made whether typical
buds are disposed above or below the most recently opened
flowers.
[0066] Flower Date 50%. (Same as Time to flowering) The number of
days from planting until 50% of the plants in a planted area have
at least one open flower.
[0067] Flower Petal Coloration. The coloration of open exposed
petals on the first day of flowering is observed.
[0068] Frost Tolerance (Spring Type Only). The ability of young
plants to withstand late spring frosts at a typical growing area is
evaluated and is expressed on a scale of 1 (poor) to 5
(excellent).
[0069] Gene Silencing. The interruption or suppression of the
expression of a gene at the level of transcription or
translation.
[0070] Genotype. Refers to the genetic constitution of a cell or
organism.
[0071] Glucosinolate Content. The total glucosinolates of seed at
8.5% moisture, as measured by AOCS Official Method AK-1-92
(determination of glucosinolates content in rapeseed-colza by
HPLC), is expressed as micromoles per gram of defatted, oil-free
meal. Capillary gas chromatography of the trimethylsityl
derivatives of extracted and purified desulfoglucosinolates with
optimization to obtain optimum indole glucosinolate detection is
described in "Procedures of the Western Canada Canola/Rapeseed
Recommending Committee Incorporated for the Evaluation and
Recommendation for Registration of Canola/Rapeseed Candidate
Cultivars in Western Canada". Also, glucosinolates could be
analyzed using NIR (Near Infra Red spectroscopy) as long as the
instrument is calibrated according to the manufacturer's
specifications.
[0072] Grain. Seed produced by the plant or a self or sib of the
plant that is intended for food or feed use.
[0073] Green Seed. The number of seeds that are distinctly green
throughout as defined by the Canadian Grain Commission. Expressed
as a percentage of seeds tested.
[0074] Herbicide Resistance: Resistance to various herbicides when
applied at standard recommended application rates is expressed on a
scale of 1 (resistant), 2 (tolerant), or 3 (susceptible).
[0075] Leaf Anthocyanin Coloration. The presence or absence of leaf
anthocyanin coloration, and the degree thereof if present, are
observed when the plant has reached the 9- to 11-leaf stage.
[0076] Leaf Attachment to Stem. The presence or absence of clasping
where the leaf attaches to the stem, and when present the degree
thereof, are observed.
[0077] Leaf Attitude. The disposition of typical leaves with
respect to the petiole is observed when at least 6 leaves of the
plant are formed.
[0078] Leaf Color. The leaf blade coloration is observed when at
least 6 leaves of the plant are completely developed.
[0079] Leaf Glaucosity. The presence or absence of a fine whitish
powdery coating on the surface of the leaves, and the degree
thereof when present, are observed.
[0080] Leaf Length. The length of the leaf blades and petioles are
observed when at least 6 leaves of the plant are completely
developed.
[0081] Leaf Lobes. The fully developed upper stem leaves are
observed for the presence or absence of leaf lobes when at least 6
leaves of the plant are completely developed.
[0082] Leaf Margin Depth. A rating of the depth of the dentations
along the upper third of the margin of the largest leaf. 1=very
shallow, 9=very deep.
[0083] Leaf Margin Hairiness. The leaf margins of the first leaf
are observed for the presence or absence of pubescence, and the
degree thereof, when the plant is at the two leaf-stage.
[0084] Leaf Margin Type. A visual rating of the dentations along
the upper third of the margin of the largest leaf. 1=undulating,
2=rounded, 3=sharp.
[0085] Leaf Surface. The leaf surface is observed for the presence
or absence of wrinkles when at least 6 leaves of the plant are
completely developed.
[0086] Leaf Tip Reflexion. The presence or absence of bending of
typical leaf tips and the degree thereof, if present, are observed
at the 6 to 11 leaf-stage.
[0087] Leaf Upper Side Hairiness. The upper surfaces of the leaves
are observed for the presence or absence of hairiness, and the
degree thereof if present, when at least 6 leaves of the plant are
formed.
[0088] Leaf Width. The width of the leaf blades is observed when at
least 6 leaves of the plant are completely developed.
[0089] Length of Beak. The typical length of the silique beak when
mature is observed and is expressed on a scale of 1 (very short) to
9 (very long).
[0090] Locus. A defined segment of DNA.
[0091] Locus Conversion. A locus conversion refers to plants within
a variety that have been modified in a manner that retains the
overall genetics of the variety and further comprises one or more
loci with a specific desired trait, such as male sterility, insect,
disease or herbicide resistance. Examples of single locus
conversions include mutant genes, transgenes and native traits
finely mapped to a single locus. One or more locus conversion
traits may be introduced into a single canola variety.
[0092] Maturity. The number of days from planting to maturity is
observed, with maturity being defined as the plant stage when pods
with seed change color, occurring from green to brown or black, on
the bottom third of the pod-bearing area of the main stem.
[0093] Number of Leaf Lobes. The frequency of leaf lobes, when
present, is observed when at least 6 leaves of the plant are
completely developed.
[0094] Oil Content: The typical percentage by weight oil present in
the mature whole dried seeds is determined by ISO 10565:1993
Oilseeds Simultaneous determination of oil and water--Pulsed NMR
method. Also, oil could be analyzed using NIR (Near Infra Red
spectroscopy) as long as the instrument is calibrated according to
the manufacturer's specifications, reference AOCS Procedure Am 1-92
Determination of Oil, Moisture and Volatile Matter, and Protein by
Near-Infrared Reflectance.
[0095] Pedicel Length. The typical length of the silique stem when
mature is observed and is expressed on a scale of 1 (very short) to
9 (very long).
[0096] Petal Length. The lengths of typical petals of fully opened
flowers are observed.
[0097] Petal Width. The widths of typical petals of fully opened
flowers are observed.
[0098] Petiole Length. The length of the petioles is observed, in a
line forming lobed leaves, when at least 6 leaves of the plant are
completely developed.
[0099] Plant Height. The overall plant height at the end of
flowering is observed.
[0100] Ploidy. This refers to the number of chromosomes exhibited
by the line, for example diploid or tetraploid.
[0101] Pod Anthocyanin Coloration. The presence or absence at
maturity of silique anthocyanin coloration, and the degree thereof
if present, are observed.
[0102] Pod Habit. The typical manner in which the siliques are
borne on the plant at maturity is observed.
[0103] Pod Length. The typical silique length is observed and is
expressed on a scale of 1 (very short) to 9 (very long).
[0104] Pod Attitude. A visual rating of the angle joining the
pedicel to the pod at maturity. 1=erect, 3=semi-erect,
5=horizontal, 7=semi-drooping and 9=drooping.
[0105] Pod Type. The overall configuration of the silique is
observed.
[0106] Pod Width. The typical pod width when mature is observed and
is expressed on a scale of 1 (very narrow) to 9 (very wide).
[0107] Pollen Formation. The relative level of pollen formation is
observed at the time of dehiscence.
[0108] Protein Content: The typical percentage by weight of protein
in the oil free meal of the mature whole dried seeds is determined
by AOCS Official Method Ba 4e-93 Combustion Method for the
Determination of Crude Protein. Also, protein could be analyzed
using NIR (Near Infra Red spectroscopy) as long as the instrument
is calibrated according to the manufacturer's specifications,
reference AOCS Procedure Am 1-92 Determination of Oil, Moisture and
Volatile Matter, and Protein by Near-Infrared Reflectance.
[0109] Resistance. The ability of a plant to withstand exposure to
an insect, disease, herbicide or other condition. A resistant plant
variety or hybrid will have a level of resistance higher than a
comparable wild-type variety or hybrid. "Tolerance" is a term
commonly used in crops affected by Sclerotinia, such as canola,
soybean, and sunflower, and is used to describe an improved level
of field resistance.
[0110] Resistant to Lodging. Resistance to lodging at maturity is
expressed on a scale of 1 (weak) to 9 (strong).
[0111] Resistance to Shattering. Resistance to silique shattering
is observed at seed maturity and is expressed on a scale of 1
(poor) to 9 (excellent).
[0112] Root Anthocyanin Coloration. The presence or absence of
anthocyanin coloration in the skin at the top of the root is
observed when the plant has reached at least the six-leaf
stage.
[0113] Root Anthocyanin Expression. When anthocyanin coloration is
present in skin at the top of the root, it further is observed for
the exhibition of a reddish or bluish cast within such coloration
when the plant has reached at least the six-leaf stage.
[0114] Root Anthocyanin Streaking. When anthocyanin coloration is
present in the skin at the top of the root, it further is observed
for the presence or absence of streaking within such coloration
when the plant has reached at least the six-leaf stage.
[0115] Root Chlorophyll Coloration. The presence or absence of
chlorophyll coloration in the skin at the top of the root is
observed when the plant has reached at least the six-leaf
stage.
[0116] Root Coloration Below Ground. The coloration of the root
skin below ground is observed when the plant has reached at least
the six-leaf stage.
[0117] Root Depth in Soil. The typical root depth is observed when
the plant has reached at least the six-leaf stage.
[0118] Root Flesh Coloration. The internal coloration of the root
flesh is observed when the plant has reached at least the six-leaf
stage.
[0119] Seedling Growth Habit. The growth habit of young seedlings
is observed for the presence of a weak (1) or strong (9) rosette
character and is expressed on a scale of 1 to 9.
[0120] Seeds Per Pod. The average number of seeds per pod is
observed.
[0121] Seed Coat Color. The seed coat color of typical mature seeds
is observed.
[0122] Seed Coat Mucilage. The presence or absence of mucilage on
the seed coat is determined and is expressed on a scale of 1
(absent) to 9 (heavy). During such determination a petri dish is
filled to a depth of 0.3 cm. with tap water provided at room
temperature. Seeds are added to the petri dish and are immersed in
water where they are allowed to stand for five minutes. The
contents of the petri dish containing the immersed seeds next is
examined under a stereo microscope equipped with transmitted light.
The presence of mucilage and the level thereof is observed as the
intensity of a halo surrounding each seed.
[0123] Seed Size. The weight in grams of 1,000 typical seeds is
determined at maturity while such seeds exhibit a moisture content
of approximately 5 to 6 percent by weight.
[0124] Speed of Root Formation. The typical speed of root formation
is observed when the plant has reached the 4- to 11-leaf stage.
[0125] Stem Anthocyanin Coloration. The presence or absence of leaf
anthocyanin coloration and the intensity thereof, if present, are
observed when the plant has reached the 9- to 11-leaf stage.
[0126] Stem Lodging at Maturity. A visual rating of a plant's
ability to resist stem lodging at maturity. 1=very weak (lodged),
9=very strong (erect).
[0127] Time to Flowering. A determination is made of the number of
days when at least 50 percent of the plants have one or more open
buds on a terminal raceme in the year of sowing.
[0128] Seasonal Type. This refers to whether the new line is
considered to be primarily a Spring or Winter type of canola.
[0129] Winter Survival (Winter Type Only). The ability to withstand
winter temperatures at a typical growing area is evaluated and is
expressed on a scale of 1 (poor) to 5 (excellent).
DETAILED DESCRIPTION OF THE INVENTION
[0130] 45S51 is a fully restored spring Brassica napus hybrid. The
hybrid 45S51 is produced by crossing NS5902FR and NS5870MC.
NS5902FR is glyphosate tolerant and is an OGU CMS female. NS5870MC
is an OGU Restorer male. The female line NS5902FR is a sterile
version of NS5902BR. When the sterile NS5902FR is pollinated by the
male line NS5870MC, that carries a gene for the restoration of
fertility, it results in the fertile hybrid, 45S51.
[0131] NS5902BR is the maintainer line (B-line) for NS5902FR
(A-line). The female parent seed for hybrid 45S51 is produced by
crossing the male sterile inbred-A line by the maintainer inbred-B
line, where A and B lines are genetically alike except A line
carries the OGU INRA cytoplasm, while B line carries the normal B.
napus cytoplasm. The maintainer--B line was developed from a cross
between non-registered proprietary breeding lines
(NS3965BR.times.03SN40441) followed by pedigree selection. The last
crossing was completed in 2003 fall and the F1 generation was grown
in greenhouse to produce F2 during winter months of 2004. The F2
populations were evaluated in breeding nursery in Ontario and were
screened for general vigor, maturity, oil, protein and total
glucosinolates. The remnant F2 seeds were then grown in the
greenhouse and subjected to glyphosate screening. Individual F2
plants were screened for sclerotinia tolerance at physiological
maturity, susceptible plants were discarded and the partially
resistant plants were harvested individually. The F3s were then
planted again in the next greenhouse project and were subjected to
glyphosate screening. These plants when harvested individually to
produce F4 progenies. The F4 progenies were evaluated in Ontario
agronomic evaluation nursery for vigor, general uniformity,
glyphosate tolerance, maturity, oil, protein, total glucosinolates
etc. At the same time the F4 lines were also screened in the
sclerotinia misting nursery (field) where artificial conditions
along with spore inoculation created perfect conditions for
sclerotinia disease to appear in full force. Based on agronomic
data, quality data and disease data, F4 line 05SN-5165 was
selected. The F5 seed harvested from 05SN-5165 in greenhouse with
the concurrently planted F4 generation was assigned NS5902BR as a
breeder code.
[0132] NS5870MC is an OGU Restorer male developed at Georgetown
Research Centre of Pioneer Hi-Bred Production Limited through
backcrossing followed by selfing and selection. The original cross
was produced between two non-registered proprietary lines
NS4304MC.times.02SN41269 and then backcrossed to NS4304MC. The
breeding line 02SN41269 carried partial resistance against
Sclerotinia while NS4304MC carried restorer gene for INRA OGU male
sterility and also carried OGU cytoplasm. BC1 was completed in 2003
and the BC1 plants were screened in the greenhouse for Sclerotinia
tolerance and selfed to produce BC1S1. The fertile BC1S1 plants,
also grown in the greenhouse and subjected to sclerotinia tolerance
and were harvested individually to produce BC1S2 after susceptible
plants were discarded. The BC1S2s were evaluated in breeding and
agronomy nursery where selection was applied for uniformity,
homozygosity for restorer gene, general agronomy, lodging
resistance, oil, protein, glucosinolates and total saturates. The
BC1S2s were also screened in the sclerotinia-misting nursery where
disease scores were recorded on each BC1S2 lines. After combining
information from breeding and disease nurseries, the final
selection was carried out for the BC1S2 lines. The selfed seed from
the agronomy/breeding nursery BC1S3 was then sent to Chile for
hybrid seed production as well as for increase in small cages. The
BC1S4 seed was then used for increasing Breeder seed for
NS5870MC.
[0133] The hybrid 45551 was evaluated in the Private Co-op Trials
of Pioneer Hi-Bred in 2006. Testing was conducted in 2007 under the
code 06N718R. Checks were 46A65 and Q2.
Varietal Characteristics (See also Tables 1, 4, 5, 6 and 7)
[0134] Seed Yield: Fourteen percent higher than WCC/RRC checks.
[0135] Disease Reaction: Classified as Resistant (R=same class as
46A65) to blackleg (Leptosphaeria maculans) according to WCC/RRC
guidelines. Based on Pioneer Hi-Bred trials, 45S51 is also
resistant (R) to Fusarium wilt. [0136] Also provisionally
classified as MR--Moderately Resistant against white mold
(Sclerotinia sclerotiorum Lib). This classification is provisional
because WCC/RRC does not have any guidelines for Sclerotinia
tolerance classification; therefore, this classification needs
further verification in future following the approved WCC/RRC
protocol. [0137] Plant Height: Approximately 1.5 cm taller than
WCC/RRC checks [0138] Maturity: Similar maturing as WCC/RRC checks.
[0139] Lodging: Slightly better than the checks
Seed Characteristics:
[0139] [0140] Seed color: Dark brown [0141] Grain size: 1000 seed
weight is slightly larger than mean of the checks [0142] Seed oil
content: 1.0% higher than mean of the checks. [0143] Seed protein
content: 0.5% lower than mean of the checks. [0144] Erucic acid:
Less than 0.5% (maximum allowable limit). [0145] Total saturates:
0.27% higher than mean of the checks (required set aside of
"minimum requirements") [0146] Total glucosinolates: 4.8 .mu.mol/g
lower than mean of the checks [0147] Chlorophyll: Lower than mean
of the checks. [0148] Summary: 45S51 (06N718R) is a medium
maturing, high yielding Roundup.RTM. resistant Brassica napus
canola hybrid having resistant "R" rating for blackleg, resistant
"R" rating for Fusarium wilt and moderately resistant (MR) rating
for white mold (Sclerotinia). Its oil content is higher than mean
of checks and protein is slightly lower than the checks. Its seed
size is bit larger and has lower chlorophyll than the checks.
[0149] Inbred maintenance: Pioneer Hi-Bred Production Limited
[0150] Canadian distributor: Pioneer Hi-Bred Limited. [0151]
Status: It has completed two years in Western Canadian Co-op trials
(2006 Private Co-op and 2007 Public Co-op). It meets all minimum
requirements set by WCC/RRC except it failed to meet the total
saturated fatty acid requirements by 0.17%. It meets all acceptable
criteria. The WCC/RRC supported this hybrid by setting aside the
minimum requirements for total saturated fatty acid.
[0152] A canola hybrid needs to be homogenous and reproducible to
be useful for the production of a commercial crop on a reliable
basis. There are a number of analytical methods available to
determine the phenotypic stability of a canola hybrid.
[0153] The oldest and most traditional method of analysis is the
observation of phenotypic traits. The data are usually collected in
field experiments over the life of the canola plants to be
examined. Phenotypic characteristics most often are observed for
traits associated with seed yield, seed oil content, seed protein
content, fatty acid composition of oil, glucosinolate content of
meal, growth habit, lodging resistance, plant height, shattering
resistance, etc.
[0154] In addition to phenotypic observations, the genotype of a
plant can also be examined. A plant's genotype can be used to
identify plants of the same variety or a related variety. For
example, the genotype can be used to determine the pedigree of a
plant. 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).
[0155] The variety of the present invention has shown uniformity
and stability for all traits, as described in the following variety
description information. The variety has been increased with
continued observation for uniformity.
[0156] 45S51 is a medium-maturity, high-yielding, Roundup.RTM.
resistant Brassica napus canola hybrid having resistant "R" rating
for blackleg, resistant "R" rating for Fusarium wilt and moderately
resistant (MR) rating for white mold (Sclerotinia). Its oil content
is higher than the mean of checks and protein is slightly lower
than the WCC/RRC checks 46A65 and Q2. Its seed size is a bit larger
and has lower chlorophyll than the checks.
[0157] Table 1 provides data on morphological, agronomic, and
quality traits for 45S51 and two publicly-available canola
varieties, 45H21 and 45H73. When preparing the detailed phenotypic
information that follows, plants of the new 45S51 variety were
observed while being grown using conventional agronomic practices.
For comparative purposes, canola plants of two publicly-available
canola varieties, 45H21 and 45H73, were similarly grown in a
replicated experiment.
[0158] Observations were recorded on various morphological traits
for the hybrid 45S51 and comparative check cultivars.
TABLE-US-00001 TABLE 1 VARIETY DESCRIPTIONS BASED ON MORPHOLOGICAL,
AGRONOMIC AND QUALITY TRAITS.sup.1 Trait 45S51 45H21 Code Trait
Mean Description Mean Description 2.0 Seasonal Type 1 Spring 1
Spring 3.1 Cot width (1-9) 5.00 Medium 6.60 Medium to wide 3.6
Blade colour 2.0 Medium 2.0 Medium (1 = lgt. grn 4 = blue. grn)
Leaf: 100% 100% percentage of lobed leaves (%) 3.4 Lobe 2.00
Present 2.00 Present development (1 = absent (entire)-2 = present
(lobed)) Number of 4.00 Few to medium 3.00 Few lobes (1 = v. few-9
= v. many) Number of 3.77 Few to medium 3.53 Few lobes (count)
Margin type 2.0 Rounded 2.0 Rounded (1 = undulating- 3 = sharp) 3.5
Indentation of 4.2 Shallow to 4.0 Shallow to medium margin medium
Leaf length 17.16 Medium 16.36 Medium (cm) Leaf width (cm) 8.29
Medium to wide 8.29 Medium to wide Leaf 2.07 1.95 length:width
ratio Petiole length 6.89 Medium to long 6.50 Medium (cm) 3.3 Stem
1.0 Absent 1.7 Weak to absent anthocyanin 3.8 Leaf glaucosity 3.2
Weak to medium 3.5 Weak to medium 4.1 Flower date 47.0 Medium 47.4
Medium 50% 4.5 Petal colour 3.00 Medium yellow 3.00 Medium yellow
(1 = white- 4 = orange, 5 = other Petal length 13.50 Medium to long
13.10 Medium (mm) Petal width 6.30 Medium 6.30 Medium (mm) Petal
2.10 2.10 length:width ratio 4.8 Anther fertility 2.00 Shedding
pollen 2.00 Shedding pollen (1 = sterile, 2 = shedding pollen) 4.12
Silique attitude 3.0 Semi-erect 3.0 Semi-erect (1 = erect- 9 =
drooping) 4.10 Silique length 6.50 Medium to long 5.50 Medium (1 =
v. short- 9 = v. long) Silique length 64.20 Medium long 61.30
Medium (mm) 4.11 Silique width 6.00 Medium to wide 5.50 Medium (1 =
v. narrow- 9 = v. wide) Silique width 5.20 Medium to wide 5.00
Medium (mm) 4.13 Beak length 5.50 Medium 6.00 Medium (1 = v. short-
9 = v. long) Beak length 8.50 Medium 8.70 Medium (mm) 4.14 Pedicel
length 6.00 Medium to long 6.00 Medium to long (1 = v. short- 9 =
v. long) Pedicel length 22.10 Medium to long 22.10 Medium to long
(mm) 4.15 Maturity (days 93.1 Medium 92.9 Medium from planting) 4.2
Plant height 3.00 Short 6.00 Medium (1 = v. short- 9 = v. tall)
Plant height 109.88 Short 114.25 Medium (cm) 5.1 Seed coat 1.50
Black to brown 1.50 Black to brown colour (1 = blk, 2 = bm, 3 =
yel, 4 = mix, 5 = oth) 5.3 Seed weight 3.46 Heavy 3.36 Medium
(grams per 1000 seeds) 6.1 Resistance to 8.00 Very good 8.00 Very
good shattering (3 = poor, 7 = good) 6.2 Resistance to 8.55 Very
good 8.44 Very good lodging (3 = poor, 7 = good) 10.2 Percentage of
0.00 Absent 0.01 Very low total fatty acids - erucic (C22:1) 10.3
Maximum allowable erucic in foundation seed is 0.5% 10.7
Glucosinolates 10.50 Low 10.44 Low (.mu.mole total aliphatic
glucs/g air - dried meal) - very low (<10), low (10-15), med
(15-20), high (>20) 10.9 Chlorophyll 13.84 Medium 12.72 Medium
content (ppm) Low (<8), med (8-15), high (>15) (ppm) 10.1 Oil
percentage 48.59 49.00 (whole dry seed) 10.5 Protein 46.52 46.71
percentage (whole dry seed) 7.4 Blackleg 1.8 Resistant 2.4
Resistant to moderate resistance (0 = not tested, 1 = resistant, 9
= highly susceptible) 7.10 White rust (2 V 0.00 Not tested 0.00 Not
tested and 7 V) (0 = not tested, 1 = resistant, 9 = highly
susceptible) 8.3 Glyphosate 1.00 Resistant 1.00 Resistant (0 = not
tested, 1 = resistant, 5 = tolerant, 9 = susceptible) Imidazolinone
9.00 Susceptible 9.00 Susceptible (0 = not tested, 1 = resistant, 5
= tolerant, 9 = susceptible) Trait 45H73 Code Trait Mean
Description 2.0 Seasonal Type 1 Spring 3.1 Cot width (1-9) 7.00
Medium to wide 3.6 Blade colour (1 = lgt. grn 4 = blue. grn) 2.0
Medium Leaf: percentage of lobed leaves (%) 93% 3.4 Lobe
development (1 = absent (entire)- 2.00 Present 2 = present (lobed))
Number of lobes (1 = v. few-9 = 4.00 Few to medium v. many) Number
of lobes (count) 4.03 Few to medium Margin type (1 = undulating-3 =
sharp) 2.0 Rounded 3.5 Indentation of margin 4.3 Shallow to medium
Leaf length (cm) 18.70 Medium to long Leaf width (cm) 9.77 Wide
Leaf length:width ratio 1.91 Petiole length (cm) 6.56 Medium 3.3
Stem anthocyanin 1.3 Weak to absent 3.8 Leaf glaucosity 2.5 Weak
4.1 Flower date 50% 47.4 Medium 4.5 Petal colour (1 = white-4 =
orange, 5 = 3.00 Medium yellow other Petal length (mm) 13.20 Medium
Petal width (mm) 6.30 Medium Petal length:width ratio 2.10 4.8
Anther fertility (1 = sterile, 2 = shedding 2.00 Shedding pollen
pollen) 4.12 Silique attitude (1 = erect-9 = drooping) 3.3
Semi-erect to horizontal 4.10 Silique length (1 = v. short-9 = v.
long) 8.50 Very long Silique length (mm) 67.70 Very long 4.11
Silique width (1 = v. narrow-9 = v. wide) 3.00 Narrow Silique width
(mm) 4.50 Narrow 4.13 Beak length (1 = v. short-9 = v. long) 3.50
Short to medium Beak length (mm) 7.80 Short to medium 4.14 Pedicel
length (1 = v. short-9 = v. long) 6.00 Medium to long Pedicel
length (mm) 21.50 Medium to long 4.15 Maturity (days from planting)
93.1 Medium 4.2 Plant height (1 = v. short-9 = v. tall) 8.50 Very
tall Plant height (cm) 120.72 Very tall 5.1 Seed coat colour (1 =
blk, 2 = bm, 3 = yel, 1.50 Black to brown 4 = mix, 5 = oth) 5.3
Seed weight (grams per 1000 seeds) 3.10 Light 6.1 Resistance to
shattering (3 = poor, 8.00 Very good 7 = good) 6.2 Resistance to
lodging (3 = poor, 8.64 Very good 7 = good) 10.2 Percentage of
total fatty acids - erucic 0.00 Absent (C22:1) 10.3 Maximum
allowable erucic in foundation seed is 0.5% 10.7 Glucosinolates
(.mu.mole total aliphatic 9.15 Very low glucs/g air - dried meal) -
very low (<10), low (10-15), med (15-20), high (>20) 10.9
Chlorophyll content (ppm) Low (<8), 17.52 High med (8-15), high
(>15) (ppm) 10.1 Oil percentage (whole dry seed) 49.41 10.5
Protein percentage (whole dry seed) 47.68 7.4 Blackleg resistance
(0 = not tested, 1.7 Resistant 1 = resistant, 9 = highly
susceptible) 7.10 White rust (2 V and 7 V) (0 = not tested, 0.00
Not tested 1 = resistant, 9 = highly susceptible) 8.3 Glyphosate (0
= not tested, 1 = resistant, 9.00 Susceptible 5 = tolerant, 9 =
susceptible) Imidazolinone (0 = not tested, 1.00 Resistant 1 =
resistant, 5 = tolerant, 9 = susceptible) .sup.1Morphological data
from 2007 Ontario; Agronomic and quality data from 2007 W.
Canada.
[0159] Hybrid 45S51 can be advantageously used in accordance with
the breeding methods described herein and those known in the art to
produce hybrids and other progeny plants retaining desired trait
combinations of 45S51. This invention is thus also directed to
methods for producing a canola plant by crossing a first parent
canola plant with a second parent canola plant wherein either the
first or second parent canola plant is canola variety 45S51.
Further, both first and second parent canola plants can come from
the canola variety 45S51. Either the first or the second parent
plant may be male sterile.
[0160] Still further, this invention also is directed to methods
for producing a 45S51-derived canola plant by crossing canola
variety 45S51 with a second canola plant and growing the progeny
seed, and repeating the crossing and growing steps with the canola
45S51-derived plant from 1 to 2 times, 1 to 3 times, 1 to 4 times,
or 1 to 5 times. Thus, any such methods using the canola variety
45S51 are part of this invention: open pollination, selfing,
backcrosses, hybrid production, crosses to populations, and the
like. All plants produced using canola variety 45S51 as a parent
are within the scope of this invention, including plants derived
from canola variety 45S51. This includes canola lines derived from
45S51 which include components for either male sterility or for
restoration of fertility. Advantageously, the canola variety is
used in crosses with other, different, canola plants to produce
first generation (F.sub.1) canola hybrid seeds and plants with
superior characteristics.
[0161] The invention also includes a single-gene conversion of
45S51. A single-gene conversion occurs when DNA sequences are
introduced through traditional (non-transformation) breeding
techniques, such as backcrossing. DNA sequences, whether naturally
occurring or transgenes, may be introduced using these traditional
breeding techniques. Desired traits transferred through this
process include, but are not limited to, fertility restoration,
fatty acid profile modification, other nutritional enhancements,
industrial enhancements, disease resistance, insect resistance,
herbicide resistance and yield enhancements. The trait of interest
is transferred from the donor parent to the recurrent parent, in
this case, the canola plant disclosed herein. Single-gene traits
may result from the transfer of either a dominant allele or a
recessive allele. Selection of progeny containing the trait of
interest is done by direct selection for a trait associated with a
dominant allele. Selection of progeny for a trait that is
transferred via a recessive allele will require growing and selfing
the first backcross 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
gene of interest.
[0162] It should be understood that the canola variety of the
invention can, through routine manipulation by cytoplasmic genes,
nuclear genes, or other factors, be produced in a male-sterile or
restorer form as described in the references discussed earlier.
Such embodiments are also within the scope of the present claims.
Canola variety 45S51 can be manipulated to be male sterile by any
of a number of methods known in the art, including by the use of
mechanical methods, chemical methods, SI, CMS (either ogura or
another system) or NMS. The term "manipulated to be male sterile"
refers to the use of any available techniques to produce a male
sterile version of canola variety 45S51. The male sterility may be
either partial or complete male sterility. This invention is also
directed to F1 hybrid seed and plants produced by the use of Canola
variety 45S51. Canola variety 45S51 can also further comprise a
component for fertility restoration of a male sterile plant, such
as an Rf restorer gene. In this case, canola variety 45S51 could
then be used as the male plant in hybrid seed production.
[0163] This invention is also directed to the use of 45S51 in
tissue culture. As used herein, the term plant includes plant
protoplasts, plant cell tissue cultures from which canola 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, seeds, flowers, kernels, ears, cobs, leaves, husks, stalks,
roots, root tips, anthers, silk and the like. Pauls, et al., (2006)
(Canadian J of Botany 84(4):668-678) confirmed that tissue culture
as well as microspore culture for regeneration of canola plants can
be accomplished successfully. Chuong, et al., (1985) "A Simple
Culture Method for Brassica Hypocotyl Protoplasts", Plant Cell
Reports 4:4-6; Barsby, et al., (Spring 1996) "A Rapid and Efficient
Alternative Procedure for the Regeneration of Plants from Hypocotyl
Protoplasts of Brassica napus", Plant Cell Reports; Kartha, et al.,
(1974) "In vitro Plant Formation from Stem Explants of Rape",
Physiol. Plant 31:217-220; Narasimhulu, et al., (Spring 1988)
"Species Specific Shoot Regeneration Response of Cotyledonary
Explants of Brassicas", Plant Cell Reports; Swanson, (1990)
"Microspore Culture in Brassica", Methods in Molecular Biology
6(17):159; "Cell Culture techniques and Canola improvement" J. Am.
Oil Chem. Soc. 66(4):455-56 (1989). Thus, it is clear from the
literature that the state of the art is such that these methods of
obtaining plants are, and were, "conventional" in the sense that
they are routinely used and have a very high rate of success.
[0164] The utility of canola variety 45S51 also extends to crosses
with other species. Commonly, suitable species will be of the
family Brassicae.
[0165] 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, that are inserted into
the genome using transformation are referred to herein collectively
as "transgenes". Over the last fifteen to twenty years several
methods for producing transgenic plants have been developed, and
the present invention, in particular embodiments, also relates to
transformed versions of the claimed canola variety 45S51.
[0166] 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, and Genetic Transformation for
the improvement of Canola World Conf, Biotechnol. Fats and Oils
Ind. 43-46 (1988). 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 and Thompson, Eds. (CRC
Press, Inc., Boca Raton, 1993) pages 89-119.
[0167] The most prevalent types of plant transformation involve the
construction of an expression vector. Such a vector comprises a DNA
sequence that contains a gene under the control of or operatively
linked to a regulatory element, for example a promoter. The vector
may contain one or more genes and one or more regulatory
elements.
[0168] A genetic trait which has been engineered into a particular
canola plant using transformation techniques, could be moved into
another line using traditional breeding techniques that are well
known in the plant breeding arts. For example, a backcrossing
approach could be used to move a transgene from a transformed
canola plant to an elite inbred line and the resulting progeny
would comprise a transgene. Also, if an inbred line was used for
the transformation then the transgenic plants could be crossed to a
different line in order to produce a transgenic hybrid canola
plant. As used herein, "crossing" can refer to a simple X by Y
cross, or the process of backcrossing, depending on the context.
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. See, U.S. Pat. No. 6,222,101 which is herein
incorporated by reference.
[0169] 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.
[0170] Protein extraction from plant biomass can be accomplished by
known methods which are discussed, for example, by Heney and Orr,
(1981) Anal. Biochem. 114:92-96.
[0171] A genetic map can be generated, primarily via conventional
Restriction Fragment Length Polymorphisms (RFLP), Polymerase Chain
Reaction (PCR) analysis, and Simple Sequence Repeats (SSR), which
identifies the approximate chromosomal location of the integrated
DNA molecule coding for the foreign protein. For exemplary
methodologies in this regard, see, Glick and Thompson, METHODS IN
PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 269-284 (CRC Press, Boca
Raton, 1993). 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.
[0172] Likewise, by means of the present invention, plants can be
genetically engineered to express various phenotypes of agronomic
interest. Exemplary transgenes 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:
[0173] (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 cloned resistance gene to engineer plants that are
resistant to specific pathogen strains. See, for example Jones, et
al., (1994) Science 266:789 (cloning of the tomato Cf-9 gene for
resistance to Cladosporium fulvum); Martin, et al., (1993) Science
262:1432 (tomato Pto gene for resistance to Pseudomonas syringae
pv. tomato encodes a protein kinase); Mindrinos, et al., (1994)
Cell 78:1089 (Arabidopsis RSP2 gene for resistance to Pseudomonas
syringae); McDowell and Woffenden, (2003) Trends Biotechnol.
21(4):178-83 and Toyoda, et al., (2002) Transgenic Res.
11(6):567-82. A plant resistant to a disease is one that is more
resistant to a pathogen as compared to the wild type plant.
[0174] (B) A gene conferring resistance to fungal pathogens, such
as oxalate oxidase or oxalate decarboxylase (Zhou, et al., (1998)
Pl. Physiol. 117(1):33-41).
[0175] (C) A Bacillus thuringiensis protein, a derivative thereof
or a synthetic polypeptide modeled thereon. See, for example,
Geiser, et al., (1986) Gene 48:109, 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 Numbers. 40098, 67136, 31995 and 31998. Other
examples of Bacillus thuringiensis transgenes being genetically
engineered are given in the following patents and patent
applications and hereby are incorporated by reference for this
purpose: 5,188,960; 5,689,052; 5,880,275; WO 91/114778; WO
99/31248; WO 01/12731; WO 99/24581; WO 97/40162 and U.S.
application Ser. Nos. 10/032,717; 10/414,637; and 10/606,320.
[0176] (D) An insect-specific hormone or pheromone such as an
ecdysteroid and juvenile hormone, a variant thereof, a mimetic
based thereon, or an antagonist or agonist thereof. See, for
example, the disclosure by Hammock, et al., (1990) Nature 344:458,
of baculovirus expression of cloned juvenile hormone esterase, an
inactivator of juvenile hormone.
[0177] (E) An insect-specific peptide which, upon expression,
disrupts the physiology of the affected pest. For example, see the
disclosures of Regan, (1994) J. Biol. Chem. 269:9 (expression
cloning yields DNA coding for insect diuretic hormone receptor) and
Pratt, et al., (1989) Biochem. Biophys. Res. Comm. 163:1243 (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 and
Grossi-de-Sa, (2002) Toxicon 40(11):1515-1539; Ussuf, et al.,
(2001) Curr Sci. 80(7):847-853 and Vasconcelos and Oliveira, (2004)
Toxicon 44(4):385-403. See also, U.S. Pat. No. 5,266,317 to
Tomalski, et al., who disclose genes encoding insect-specific,
paralytic neurotoxins.
[0178] (F) An enzyme responsible for a hyperaccumulation of a
monterpene, a sesquiterpene, a steroid, hydroxamic acid, a
phenylpropanoid derivative or another non-protein molecule with
insecticidal activity.
[0179] (G) 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 Number WO 93/02197 in the name of
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 Numbers
39637 and 67152. See also, Kramer, et al., (1993) Insect Biochem.
Molec. Biol. 23:691, who teach the nucleotide sequence of a cDNA
encoding tobacco hookworm chitinase, and Kawalleck et al., (1993)
Plant Molec. Biol. 21:673, who provide the nucleotide sequence of
the parsley ubi4-2 polyubiquitin gene, U.S. patent application Ser.
Nos. 10/389,432, 10/692,367 and U.S. Pat. No. 6,563,020.
[0180] (H) A molecule that stimulates signal transduction. For
example, see the disclosure by Botella, et al., (1994) Plant Molec.
Biol. 24:757, of nucleotide sequences for mung bean calmodulin cDNA
clones, and Griess, et al., (1994) Plant Physiol. 104:1467, who
provide the nucleotide sequence of a maize calmodulin cDNA
clone.
[0181] (I) A hydrophobic moment peptide. See, PCT Application
Number WO95/16776 and U.S. Pat. No. 5,580,852 (disclosure of
peptide derivatives of Tachyplesin which inhibit fungal plant
pathogens) and PCT Application Number WO95/18855 and U.S. Pat. No.
5,607,914 (teaches synthetic antimicrobial peptides that confer
disease resistance), the respective contents of which are hereby
incorporated by reference for this purpose.
[0182] (J) A membrane permease, a channel former or a channel
blocker. For example, see the disclosure by Jaynes, et al., (1993)
Plant Sci. 89:43, of heterologous expression of a cecropin-beta
lytic peptide analog to render transgenic tobacco plants resistant
to Pseudomonas solanacearum.
[0183] (K) 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., (1990) Ann. Rev. Phytopathol. 28:451. Coat
protein-mediated resistance has been conferred upon transformed
plants against alfalfa mosaic virus, cucumber mosaic virus, tobacco
streak virus, potato virus X, potato virus Y, tobacco etch virus,
tobacco rattle virus and tobacco mosaic virus. Id.
[0184] (L) 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. Cf. 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).
[0185] (M) A virus-specific antibody. See, for example,
Tavladoraki, et al., (1993) Nature 366:469, who show that
transgenic plants expressing recombinant antibody genes are
protected from virus attack.
[0186] (N) 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., (1992)
Bio/Technology 10:1436. The cloning and characterization of a gene
which encodes a bean endopolygalacturonase-inhibiting protein is
described by Toubart, et al., (1992) Plant J. 2:367.
[0187] (O) A developmental-arrestive protein produced in nature by
a plant. For example, Logemann, et al., (1992) Bio/Technology
10:305, have shown that transgenic plants expressing the barley
ribosome-inactivating gene have an increased resistance to fungal
disease.
[0188] (P) Genes involved in the Systemic Acquired Resistance (SAR)
Response and/or the pathogenesis related genes. Briggs, (1995)
Current Biology 5(2):128-131, Pieterse and Van Loon, (2004) Curr.
Opin. Plant Bio 7(4):456-64 and Somssich, (2003) Cell
113(7):815-6.
[0189] (Q) Antifungal genes (Cornelissen and Melchers, (1993) Pl.
Physiol. 101:709-712 and Parijs, et al., (1991) Planta 183:258-264
and Bushnell, et al., (1998) Can. J. of Plant Path. 20(2):137-149.
Also see, U.S. patent application Ser. No. 09/950,933.
[0190] (R) 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.
[0191] (S) Cystatin and cysteine proteinase inhibitors. See, U.S.
patent application Ser. No. 10/947,979.
[0192] (T) Defensin genes. See, WO03/000863 and U.S. patent
application Ser. No. 10/178,213.
[0193] (U) Genes that confer resistance to Phytophthora Root Rot,
such as the Brassica equivalents of 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, (1995) Phytophthora Root Rot Resistance
Gene Mapping in Soybean, Plant Genome IV Conference, San Diego,
Calif.
2. Genes that Confer Resistance to a Herbicide, for Example:
[0194] (A) A herbicide that inhibits the growing point or meristem,
such as an imidazalinone or a sulfonylurea. Exemplary genes in this
category code for mutant ALS and AHAS enzyme as described, for
example, by Lee, et al., (1988) EMBO J. 7:1241, and Miki, et al.,
(1990) Theor. Appl. Genet. 80:449, respectively. See also, U.S.
Pat. Nos. 5,605,011; 5,013,659; 5,141,870; 5,767,361; 5,731,180;
5,304,732; 4,761,373; 5,331,107; 5,928,937 and 5,378,824; and
international publication WO 96/33270, which are incorporated
herein by reference for this purpose.
[0195] (B) Glyphosate (resistance imparted by mutant
5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,
respectively) and other phosphono compounds such as glufosinate
(phosphinothricin acetyl transferase, PAT) and Streptomyces
hygroscopicus phosphinothricin-acetyl transferase, bar, genes), and
pyridinoxy or phenoxy propionic acids and cycloshexones (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 EPSP which can confer glyphosate resistance. See also,
U.S. Pat. No. 7,405,074, and related applications, which disclose
compositions and means for providing 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. 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 Number 0 333 033 to Kumada, et al., and
U.S. Pat. No. 4,975,374 to Goodman, et al., disclose nucleotide
sequences of glutamine synthetase genes which confer resistance to
herbicides such as L-phosphinothricin. The nucleotide sequence of a
phosphinothricin-acetyl-transferase gene is provided in European
Application Number 0 242 246 to Leemans, et al., De Greef, et al.,
(1989) Bio/Technology 7:61, describe the production of transgenic
plants that express chimeric bar genes coding for phosphinothricin
acetyl transferase activity. See also, U.S. Pat. Nos. 5,969,213;
5,489,520; 5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477;
5,646,024; 6,177,616 B1 and 5,879,903, which are incorporated
herein by reference for this purpose. Exemplary of genes conferring
resistance to phenoxy propionic acids and cycloshexones, such as
sethoxydim and haloxyfop, are the Acc1-S1, Acc1-S2 and Acc1-S3
genes described by Marshall, et al., (1992) Theor. Appl. Genet.
83:435. See also, U.S. Pat. Nos. 5,188,642; 5,352,605; 5,530,196;
5,633,435; 5,717,084; 5,728,925; 5,804,425 and Canadian Patent
Number 1,313,830, which are incorporated herein by reference for
this purpose.
[0196] (C) A herbicide that inhibits photosynthesis, such as a
triazine (psbA and gs+genes) and a benzonitrile (nitrilase gene).
Przibilla, et al., (1991) Plant Cell 3:169, 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 Numbers 53435, 67441
and 67442. Cloning and expression of DNA coding for a glutathione
S-transferase is described by Hayes, et al., (1992) Biochem. J.
285:173.
[0197] (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,
e.g., Hattori, et al., (1995) Mol Gen Genet. 246:419). 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., (1994) Plant
Physiol 106:17), genes for glutathione reductase and superoxide
dismutase (Aono, et al., (1995) Plant Cell Physiol 36:1687, and
genes for various phosphotransferases (Datta, et al., (1992) Plant
Mol Biol 20:619).
[0198] (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 B1; 6,282,837 B1; and 5,767,373; and
international publication WO 01/12825, which are incorporated
herein by reference for this purpose.
3. Transgenes that Confer or Contribute to an Altered Grain
Characteristic, Such as:
[0199] (A) Altered fatty acids, for example, by [0200] (1)
Down-regulation of stearoyl-ACP desaturase to increase stearic acid
content of the plant. See, Knultzon, et al., (1992) Proc. Natl.
Acad. Sci. USA 89:2624 and WO99/64579 (Genes for Desaturases to
Alter Lipid Profiles in Corn), [0201] (2) Elevating oleic acid via
FAD-2 gene modification and/or decreasing linolenic acid via FAD-3
gene modification (see, U.S. Pat. Nos. 6,063,947; 6,323,392;
6,372,965 and WO 93/11245), [0202] (3) Altering conjugated
linolenic or linoleic acid content, such as in WO 01/12800, [0203]
(4) Altering LEC1, AGP, Dek1, Superal1, mi1ps, various Ipa genes
such as Ipa1, Ipa3, hpt or hggt. For example, see WO 02/42424, WO
98/22604, WO 03/011015, U.S. Pat. Nos. 6,423,886, 6,197,561,
6,825,397, US Patent Application Publication Numbers 2003/0079247,
2003/0204870, WO02/057439, WO03/011015 and Rivera-Madrid, et al.,
(1995) Proc. Natl. Acad. Sci. 92:5620-5624.
[0204] (B) Altered phosphorus content, for example, by the [0205]
(1) Introduction of a phytase-encoding gene would enhance breakdown
of phytate, adding more free phosphate to the transformed plant.
For example, see, Van Hartingsveldt, et al., (1993) Gene 127:87,
for a disclosure of the nucleotide sequence of an Aspergillus niger
phytase gene. [0206] (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., (1990) Maydica 35:383 and/or by
altering inositol kinase activity as in WO 02/059324, US Patent
Application Publication Number 2003/0009011, WO 03/027243, US
Patent Application Publication Number 2003/0079247, WO 99/05298,
U.S. Pat. Nos. 6,197,561, 6,291,224, 6,391,348, WO2002/059324, US
Patent Application Publication Number 2003/0079247, WO98/45448,
WO99/55882, WO01/04147.
[0207] (C) Altered carbohydrates effected, for example, by altering
a gene for an enzyme that affects the branching pattern of starch,
a gene altering thioredoxin. (See, U.S. Pat. No. 6,531,648). See,
Shiroza, et al., (1988) J. Bacteriol 170:810 (nucleotide sequence
of Streptococcus mutans fructosyltransferase gene), Steinmetz, et
al., (1985) Mol. Gen. Genet. 200:220 (nucleotide sequence of
Bacillus subtilis levansucrase gene), Pen, et al., (1992)
Bio/Technology 10:292 (production of transgenic plants that express
Bacillus licheniformis alpha-amylase), Elliot, et al., (1993) Plant
Molec Biol 21:515 (nucleotide sequences of tomato invertase genes),
Sogaard, et al., (1993) J. Biol. Chem. 268:22480 (site-directed
mutagenesis of barley alpha-amylase gene) and Fisher, et al.,
(1993) Plant Physiol 102:1045 (maize endosperm starch branching
enzyme II), WO 99/10498 (improved digestibility and/or starch
extraction through modification of UDP-D-xylose 4-epimerase,
Fragile 1 and 2, Ref1, 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.
[0208] (D) Altered antioxidant content or composition, such as
alteration of tocopherol or tocotrienols. For example, see, U.S.
Pat. No. 6,787,683, US Patent Application Publication Number
2004/0034886 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).
[0209] (E) 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), WO99/40209
(alteration of amino acid compositions in seeds), WO99/29882
(methods for altering amino acid content of proteins), U.S. Pat.
No. 5,850,016 (alteration of amino acid compositions in seeds),
WO98/20133 (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), WO98/56935 (plant amino
acid biosynthetic enzymes), WO98/45458 (engineered seed protein
having higher percentage of essential amino acids), WO98/42831
(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), WO96/01905 (increased threonine), WO95/15392 (increased
lysine), US Patent Application Publication Number 2003/0163838, US
Patent Application Publication Number 2003/0150014, US Patent
Application Publication Number 2004/0068767, U.S. Pat. No.
6,803,498, WO01/79516, and WO00/09706 (Ces A: cellulose synthase),
U.S. Pat. No. 6,194,638 (hemicellulose), U.S. Pat. No. 6,399,859
and US Patent Application Publication Number 2004/0025203 (UDPGdH),
U.S. Pat. No. 6,194,638 (RGP).
4. Genes that Control Pollination, Hybrid Seed Production or
Male-Sterility:
[0210] 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.
[0211] (A) Introduction of a deacetylase gene under the control of
a tapetum-specific promoter and with the application of the
chemical N-Ac-PPT (WO 01/29237).
[0212] (B) Introduction of various stamen-specific promoters (WO
92/13956, WO 92/13957).
[0213] (C) Introduction of the barnase and the barstar gene (Paul,
et al., (1992) Plant Mol. Biol. 19:611-622).
[0214] For additional examples of nuclear male and female sterility
systems and genes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426;
5,478,369; 5,824,524; 5,850,014 and 6,265,640; all of which are
hereby incorporated by reference.
[0215] Also see, U.S. Pat. No. 5,426,041 (invention relating to a
method for the preparation of a seed of a plant comprising crossing
a male sterile plant and a second plant which is male fertile),
U.S. Pat. No. 6,013,859 (molecular methods of hybrid seed
production) and U.S. Pat. No. 6,037,523 (use of male
tissue-preferred regulatory region in mediating fertility), all of
which are hereby incorporated by reference for this purpose.
5. Genes that Create a Site for Site Specific DNA Integration.
[0216] 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., (2003)
"Site-Specific Recombination for Genetic Engineering in Plants",
Plant Cell Rep 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), 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 (Including but not
Limited to Flowering, ear 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.
[0217] For example, see, WO 00/73475 where water use efficiency is
altered through alteration of malate; U.S. Pat. Nos. 5,892,009,
5,965,705, 5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866,
6,717,034, 6,801,104, WO2000060089, WO2001026459, WO2001035725,
WO2001034726, WO2001035727, WO2001036444, WO2001036597,
WO2001036598, WO2002015675, WO2002017430, WO2002077185,
WO2002079403, WO2003013227, WO2003013228, WO2003014327,
WO2004031349, WO2004076638, WO9809521 and WO9938977 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 Patent Application Publication Number
2004/0148654 and WO01/36596 where abscisic acid is altered in
plants resulting in improved plant phenotype such as increased
yield and/or increased tolerance to abiotic stress; WO2000/006341,
WO04/090143, U.S. patent application Ser. Nos. 10/817,483 and
09/545,334 where cytokinin expression is modified resulting in
plants with increased stress tolerance, such as drought tolerance,
and/or increased yield. Also see WO0202776, WO03052063,
JP2002281975, U.S. Pat. No. 6,084,153, WO0164898, U.S. Pat. No.
6,177,275 and U.S. Pat. No. 6,107,547 (enhancement of nitrogen
utilization and altered nitrogen responsiveness). For ethylene
alteration, see, US Patent Application Publication Numbers
2004/0128719, 2003/0166197 and WO200032761. For plant transcription
factors or transcriptional regulators of abiotic stress, see e.g.,
US Patent Application Publication Number 2004/0098764 or US Patent
Application Publication Number 2004/0078852.
[0218] 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., WO97/49811 (LHY), WO98/56918 (ESD4), WO97/10339
and U.S. Pat. No. 6,573,430 (TFL), U.S. Pat. No. 6,713,663 (FT),
WO96/14414 (CON), WO96/38560, WO01/21822 (VRN1), WO00/44918 (VRN2),
WO99/49064 (GI), WO00/46358 (FRI), WO97/29123, U.S. Pat. Nos.
6,794,560, 6,307,126 (GAI), WO99/09174 (D8 and Rht), and
WO2004076638 and WO2004031349 (transcription factors).
INDUSTRIAL APPLICABILITY
[0219] The seed of the 45S51 variety, the plant produced from such
seed, various parts of the 45S51 hybrid canola plant or its
progeny, a canola plant produced from the crossing of the 45S51
variety, and the resulting seed, can be utilized in the production
of an edible vegetable oil or other food products in accordance
with known techniques. The remaining solid meal component derived
from seeds can be used as a nutritious livestock feed.
Deposits
Deposits
[0220] Applicant has made a deposit of at least 2500 seeds of
canola hybrid 45S51 with the American Type Culture Collection
(ATCC), Manassas, Va. 20110 USA, ATCC Deposit No. ______. The seeds
deposited with the ATCC on ______ were taken from the deposit
maintained by Pioneer Hi-Bred International, Inc., 7250 NW
62.sup.nd Avenue, Johnston, Iowa 50131-1000 since prior to the
filing date of this application. Applicant has also made a deposit
of at least 2500 seeds of parental canola varieties NS5870MC and
NS5902BR with the American Type Culture Collection (ATCC),
Manassas, Va. 20110 USA, ATCC Deposit Nos. PTA-______ and
PTA-______, respectively. The seeds deposited with the ATCC on
______ for PTA-______ and on ______ for PTA-______, respectively
were taken from the seed stock maintained by Pioneer Hi-Bred
International, Inc., 7250 NW 62.sup.nd Avenue, Johnston, Iowa
50131-1000 since prior to the filing date of this application.
Access to these deposits will be available during the pendency of
the application to the Commissioner of Patents and Trademarks and
persons determined by the Commissioner to be entitled thereto upon
request. Upon allowance of any claims in the application, the
Applicant will make available to the public, pursuant to 37 C.F.R.
.sctn.1.808, sample(s) of the deposit of at least 2500 seeds of
canola hybrid 45S51 and 2500 seeds of parental canola varieties
NS5870MC and NS5902BR all which are with the American Type Culture
Collection (ATCC), 10801 University Boulevard, Manassas, Va.
20110-2209. This deposits of seed of canola hybrid 45S51 and of
parental canola varieties NS5870MC and NS5902BR will be maintained
in the ATCC depository, which is a public depository, for a period
of 30 years, or 5 years after the most recent request, or for the
enforceable life of the patent, whichever is longer, and will be
replaced if it becomes nonviable during that period. Additionally,
Applicant has satisfied all the requirements of 37 C.F.R.
.sctn..sctn.1.801-1.809, including providing an indication of the
viability of the sample upon deposit. Applicant has no authority to
waive any restrictions imposed by law on the transfer of biological
material or its transportation in commerce. Applicant does not
waive any infringement of their rights granted under this patent or
rights applicable to canola hybrid 45S51 or of parental canola
varieties NS5870MC and NS5902BR under the Plant Variety Protection
Act (7 USC 2321 et seq.).
[0221] The foregoing invention has been described in detail by way
of illustration and example for purposes of exemplification.
However, it will be apparent that changes and modifications such as
single gene modifications and mutations, somaclonal variants,
variant individuals selected from populations of the plants of the
instant variety, and the like, are considered to be within the
scope of the present invention. All references disclosed herein
whether to journal, patents, published applications and the like
are hereby incorporated in their entirety by reference.
Example 1
Herbicide Resistance
[0222] Appropriate field tests in two growing seasons have shown
that 45S51 tolerates the recommended rate (1.5 L/ha) of
Roundup.RTM. (glyphosate) herbicide without showing plant injury or
any significant effect on yield, agronomic, or quality traits. This
hybrid exhibits less than 1500/10,000 (<15%)
glyphosate-susceptible plants.
Example 2
Sclerotinia Field Tolerance
[0223] In order to demonstrate the level of field tolerance to
Sclerotinia of 45S51, two sets of replicated field experiments were
carried out in consecutive years. Prior to physiological maturity,
observations on disease severity (Table 2) and frequency
(incidence) of infected plants were recorded. These two
observations were combined into one Sclerotinia sclerotiorum Field
Severity (SSFS) number using the formula of Bradley, et al., 2004
(2003 Evaluations of Fungicides for Control of Sclerotinia Stem Rot
of Canola in North Dakota and Minnesota. NDSU Extension Service
Report. PP-1263), where SSFS=((% disease incidence.times.disease
severity)/5).
TABLE-US-00002 TABLE 2 Disease severity rating utilized in
collecting field data on individual plants Symptoms Disease Primary
Secondary severity Main Branches Branches rating Stem (Off main
stem) (Off primary) 5 Prematurely ripened or dying plant 4 Girdled
stem, plant More than two dead not ripened* or dying branches 3
Incomplete girdling Two dead or dying branches 2 Large non-girdling
One dead or dying More than two lesion branch affected branches 1
Small non-girdling Girdling or lesion One to two lesion affected
branches 0 No symptoms *Individual and/or combined symptoms
quantified to produce disease severity rating
[0224] According to the SSFS calculation, a canola field with all
plants infected (100% incidence) and a severity score of 1 will
have SSFS=20. That is the same outcome as in another field where
20% of the plants are infected (20% incidence), and each with a
rating of 5 (SSFS=20). Yield losses at field level can be estimated
to be approximately half of the SSFS score. Accordingly, reduction
in either incidence or severity, or both, can result in improved
tolerance and prevent yield losses.
[0225] All commercial canola varieties previously sold in Canada
are susceptible to Sclerotinia. The SSFS value can be a predictor
of yield loss at field scale. Also, the SSFS allows direct
comparison from one field to another as it is ultimately translated
as yield loss. With different degrees of SSFS, it is possible to
classify the cultivar into various classes such as highly
susceptible--HS; susceptible--S; Moderately susceptible--MS;
Moderately Resistant--MR; Resistant--R; and Highly Resistant--HR.
See, Table 3. This is similar to classifications made for other
diseases, such as blackleg.
TABLE-US-00003 TABLE 3 Possible classification of canola varieties
for tolerance under severe Sclerotinia disease pressure Field
Disease severity Yield loss Class Incidence Severity* SSFS Estimate
Highly susceptible (HS) 80-100 5 80-100 40-50 Susceptible (S) 70-79
5 70-79 35-39.5 Moderately susceptible 50-69 5 50-69 25-34.5 (MS)
Moderately resistant (MR) 30-49 5 30-49 15-24.5 Resistant (R) 10-29
5 1-29 0.5-14.5 *See Table 2.
Experiment A. Hybrid 45S51 and two commercial hybrids with similar
days to flowering and maturity were tested with and without
fungicide application (Lance.RTM. or Proline.TM.). A split-plot
experimental design was used with treatments (fungicide application
versus no fungicide application) randomized to the main plots while
varieties were randomized to sub-plots. Six replications were
planted at each location. In Year 1, the experiment was planted at
four locations: Westlock and Leduc in Alberta and Carman and
Winkler in Manitoba. In Year 2, the experiment was planted at six
locations: Fergus and Rockwood in Ontario and Carman, Winkler,
Rosebank and Crystal City in Manitoba. Prior to physiological
maturity, disease incidence and severity were recorded on each plot
in each replicate; the scores were then averaged over all the
replicates and were transformed into SSFS for each site in each
year (Table 4 and Table 5). The overall average over two years is
presented in Table 6.
TABLE-US-00004 TABLE 4 Sclerotinia sclerotiorum Field Severity
(SSFS) calculated for 45S51, 45H26 and 45H73 with and without
fungicide applications at four locations in Year 1. % Westlock,
Leduc, Carman, Winkler, 2007 45H26 Alberta Alberta Manitoba
Manitoba** Average NF Class*** 45H26-NF* 6.3 16.9 30.1 32 21.3 100
HS 45H73-NF 9.8 13.8 28 26.4 19.5 92 HS 45S51-NF 1.9 7.3 7.5 16 8.2
38 MR 45H26-F 1.1 1.1 6.7 17 6.5 30 MR 45H73-F 2.7 1.3 3 11.6 4.7
22 R 45S51-F 0 0.9 1 7.3 2.3 11 R Loc. Mean 3.6 6.9 12.7 18.4 10.4
LSD 2.9 3.1 3.9 5.8 Disease Low Low-Mid Mid-High Mid-High Pressure
*NF = No fungicide application and F = fungicide application as
recommended (Proline .TM. or Lance .RTM.). **high wind caused
lodging resulting in infection due to contact. ***Classification is
based on SSFS value expressed as % of most susceptible check
(45H26-NF).
TABLE-US-00005 TABLE 5 Sclerotinia sclerotiorum Field Severity
(SSFS) calculated for 45S51, 45H26 and 45H73 with and without
fungicide applications at six locations in Year 2. Crystal % Fergus
Rockwoood Carman Winkler Rosebank City 2008 45H26 Ontario Ontario
Manitoba Manitoba Manitoba Manitoba Average NF Class** 45H26-NF*
18.7 18.3 19.8 10.3 20.7 26.8 19.1 100 HS 45H73-NF 11.5 16.3 25.4
11.8 16.3 38.6 20.0 105 HS 45S51-NF 8.5 7.8 13.9 4.9 5.3 25.3 11.0
57 MS 45H26-F 1.6 4.5 2.2 0.4 1 7.5 2.9 15 R 45H73-F 0.8 5.2 3 0.6
0.3 5.5 2.6 13 R 45S51-F 1.9 1.2 1.7 0.8 0 5.2 1.8 9 R Loc. 7.2 8.9
11.0 4.8 7.3 18.2 9.5 Mean LSD 3.6 3.2 4.3 2.0 2.4 0.9 Disease
Low-Mid Low-Mid Mid-High Low Low-Mid Mid-High Pressure *NF = No
fungicide application and F = fungicide application as recommended
(Proline .TM. or Lance .RTM.). **Classification is based on SSFS
value expressed as % of most susceptible check (45H26-NF).
TABLE-US-00006 TABLE 6 Sclerotinia sclerotiorum Field Severity
(SSFS) calculated for 45S51, 45H26 and 45H73 with and without
fungicide at ten locations over two years. Two % 2007 2008 year
45H26 Average Average Avg NF Class 45H26-NF* 21.3 19.1 20.0 100 HS
45H73-NF 19.5 20.0 19.8 99 HS 45S51-NF 8.2 11.0 9.9 49 MR 45H26-F
6.5 2.9 4.3 22 R 45H73-F 4.7 2.6 3.4 17 R 45S51-F 2.3 1.8 2.0 10 R
Loc. Mean 10.4 9.5 9.9 No of 4 6 10 locations *NF = No fungicide
application and F = fungicide application as recommended (Proline
.TM. or Lance .RTM.).
Experiment B. This experiment, also conducted over two years,
involved planting of 45S51 and 45H26 in large, farm-scale strips.
The strip length and width depended on planting equipment used by
farmers. At each site, two strips of 45H26 and two strips of 45S51
were planted; one strip of each cultivar was sprayed with fungicide
(Lance.RTM. or Proline.TM. or Rovral.TM. or Quadris.TM. depending
on availability) at the recommended rate. In Year 1, this
experiment had adequate level of disease (>10% SSFS) at two
sites in Manitoba. In Year 2, this experiment had adequate level of
disease at 6 sites (Kamsack, Elm Creek, Russell, Canora, Ashville,
Somerset) in Southern Manitoba and Saskatchewan.
[0226] Prior to physiological maturity, disease incidence and
severity were recorded on each plot at each location and the
numbers were transformed into SSFS for each site in each year and
then averaged over two years (Table 7).
TABLE-US-00007 TABLE 7 Two year Sclerotinia sclerotiorum Field
Severity (SSFS) calculated for 45S51, 45H26 with and without
fungicide in strip trials planted at 8 locations over two years.
Two year Weighted % of 2007 Avg 2008 Avg Avg 45H26-NF Class
45H26-NF* 17 21.6 20.5 100 HS 45S51-NF 7.3 7.2 7.2 35 MR 45H26-F
2.6 7.9 6.6 32 MR 45S51-F 0.6 4.6 3.6 18 R Loc 2 6 8 *NF = No
fungicide application and F = fungicide application as recommended
(Proline .TM. or Lance .RTM. or Rovral .TM. or Quadris .TM.
depending on availability).
[0227] These experiments show that 45S51 performs better than the
commercial checks 45H26 and 45H73 under both treatment conditions
(i.e., with or without fungicide application). Untreated 45S51
would have a disease rating of moderately resistant (MR) while
fungicide-treated 45S51 can produce a resistance rating equal to
that of a resistant (R) variety. Since the development of disease
is environmentally dependent, more confidence can be placed in data
collected at ten test sites over two years. Experiment B
demonstrated that untreated 45S51 had performance similar to
fungicide-treated 45H26.
* * * * *