Note: Descriptions are shown in the official language in which they were submitted.
PLANTS AND SEEDS OF CANOLA VARIETY 5CV686590
FIELD
The present invention relates to a new and distinctive canola variety,
designated SCV686590.
BACKGROUND
There are numerous steps involving significant technical human intervention in
the development
of any novel, desirable plant germplasm. Plant breeding begins with the
analysis and definition of
problems and weaknesses of the current germplasm, the establishment of program
goals, and the
definition of specific breeding objectives. The next step is selection of
germplasm that possess the traits
to meet the program goals. The goal is to combine in a single variety an
improved combination of
desirable traits from the parental germplasm. These important traits may
include higher seed yield,
resistance to diseases and insects, better stems and roots, tolerance to
drought and heat, better agronomic
quality, resistance to herbicides, and improvements in compositional traits.
Canola, Brassica napus oleifira annua, is an important and valuable field
crop. Thus, a
continuing goal of canola plant breeders is to develop stable, high yielding
canola varieties that are
agronomically sound. The reasons for this goal are generally to maximize the
amount of grain produced
on the land used and to supply food for both animals and humans. The high
quality vegetable oil
extracted from canola grain is a primary reason for canola's commercial value.
Thus, in addition to high
grain yields, increasing the oil content level in the grain can maximize crop
value per acre. To
accomplish these goals, the canola breeder must select and develop canola
plants that have the traits that
result in superior varieties.
Additionally, the components may be used in non food product applications.
Biodiesel
production, lubricants, solvents, cleaners, paints, inks, plastics, adhesives,
and foams are a few examples
of other industrial applications for canola and its components.
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Date Recue/Date Received 2022-09-28
SUMMARY
One aspect of the present invention relates to seed of canola variety
SCV686590. The invention
also relates to plants produced by growing the seed of canola variety
SCV686590, as well as the
derivatives of such plants. Further provided are plant parts, including cells,
plant protoplasts, plant cells
of a tissue culture of cells 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 pollen,
flowers, seeds, pods, leaves, stems,
and the like, from canola variety SCV686590, wherein a sample of seed of said
variety has been
deposited under NCMA Accession No. 202206003. In another aspect, the invention
provides a crushed
non-viable canola seed from canola variety SCV686590, wherein a sample of seed
of said variety has
been deposited under NCMA Accession No. 202206003.
Another aspect of the invention relates to an industrial product produced from
a plant comprising
the plant cell of a canola plant of variety SCV686590, wherein a sample of
seed of said variety has been
deposited under NCMA Accession No. 202206003, and wherein the industrial
product is selected from
the group consisting of crushed canola grain, canola hulls, canola meal, and
canola flour.
Yet another aspect of the current invention relates to the use of seed of a
plant of canola variety
SCV686590, wherein a sample of seed of said variety has been deposited under
NCMA Accession No.
202206003, to produce an industrial product selected from the group consisting
of canola meal, livestock
feed, protein concentrate, unblended canola oil, salad oil, cooking oil,
frying oil, vegetable oil, a blended
oil, and a biofuel.
In a further aspect, the invention provides a composition comprising a seed of
canola variety
SCV686590 comprised in plant seed growth media. In certain embodiments, the
plant seed growth
media is a soil or synthetic cultivation medium. In specific embodiments, the
growth medium may be
comprised in a container or may, for example, be soil in a field. Plant seed
growth media are well known
to those of skill in the art and include, but are in no way limited to, soil
or synthetic cultivation medium.
Advantageously, plant seed growth media can provide adequate physical support
for seeds and can retain
moisture and/or nutritional components. Examples of characteristics for soils
that may be desirable in
certain embodiments can be found, for instance, in U.S. Patent Nos. 3,932,166
and 4,707,176. Synthetic
plant cultivation media are also well known in the art and may, in certain
embodiments, comprise
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Date Recue/Date Received 2022-09-28
polymers or hydrogels. Examples of such compositions are described, for
example, in U.S. Patent No.
4,241,537.
Another aspect of the invention relates to a tissue culture of regenerable
cells of the canola variety
SCV686590, as well as plants regenerated therefrom, wherein the regenerated
canola plant is capable of
expressing all the morphological and physiological characteristics of a plant
grown from the canola seed
designated SCV686590.
Yet another aspect of the current invention is a canola plant comprising a
single locus conversion
of the canola variety 5CV686590, wherein the canola plant is otherwise capable
of expressing all the
morphological and physiological characteristics of the canola variety
5CV686590. In particular
embodiments of the invention, the single locus conversion may comprise a
transgenic gene which has
been introduced by genetic transformation into the canola variety 5CV686590 or
a progenitor thereof
and wherein a sample of seed of canola variety SCV686590 has been deposited
under NCMA Accession
No. 202206003. In still other embodiments of the invention, the single locus
conversion may comprise
a dominant or recessive allele. The single locus conversion may confer
potentially any trait upon the
single locus converted plant, including male fertility or sterility, herbicide
tolerance, insect resistance,
pest resistance, disease resistance, modified fatty acid metabolism, abiotic
stress resistance, modified
seed yield, modified oil percent, modified protein percent, altered seed amino
acid composition,
modified lodging resistance, site specific genetic recombination, modified
carbohydrate metabolism,
resistance to bacterial disease, resistance to fungal disease, resistance to
viral disease, and improved
nutritional quality.
Still yet another aspect of the invention relates to the use of a plant of
canola variety 5CV686590,
wherein a sample of seed of said variety has been deposited under NCMA
Accession No. 202206003,
and a second canola plant to produce a canola seed or a descendant plant. In a
particular embodiment,
the invention comprises a plant or plant cell or a descendant of a plant or
plant cell of a canola plant
designated variety 5CV686590. In some embodiments, the descendant has the same
desirable traits as
the plant designated variety 5CV686590.
Still yet another aspect of the invention relates to a first generation (F1)
hybrid canola seed
produced by crossing a plant of the canola variety 5CV686590 to a second
canola plant. Also included
in the invention are the F 1 hybrid canola plants grown from the hybrid seed
produced by crossing the
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Date Recue/Date Received 2022-09-28
canola variety SCV686590 to a second canola plant. Still further included in
the invention are the seeds
of an Fi hybrid plant produced with the canola variety SCV686590 as one
parent, the second generation
(F2) canola plant grown from the seed of the Fi hybrid plant, and the seeds of
the F2 plant.
Still yet another aspect of the invention is a method of producing canola
seeds comprising
crossing a plant of the canola variety SCV686590 to any second canola plant,
including itself or another
plant of the variety SCV686590, wherein the variety SCV686590 has been
deposited under NCMA
Accession No. 202206003. In particular embodiments of the invention, the
method of crossing comprises
the steps of a) planting seeds of the canola variety 5CV686590; b) cultivating
canola plants resulting
from said seeds until said plants bear flowers; c) allowing fertilization of
the flowers of said plants; and
d) harvesting seeds produced from said plants.
Still yet another aspect of the invention is a method of producing hybrid
canola seeds comprising
crossing the canola variety 5CV686590 to a second, distinct canola plant which
is nonisogenic to the
canola variety 5CV686590. In particular embodiments of the invention, the
crossing comprises the steps
of a) planting seeds of canola variety 5CV686590 and a second, distinct canola
plant, b) cultivating the
canola plants grown from the seeds until the plants bear flowers; c) cross
pollinating a flower on one of
the two plants with the pollen of the other plant, and d) harvesting the seeds
resulting from the cross
pollinating.
Still yet another aspect of the invention is a method for developing a canola
plant in a canola
breeding program comprising: a) obtaining a canola plant, or its parts, of the
variety 5CV686590; and
b) employing said plant or parts as a source of breeding material using plant
breeding techniques. In the
method, the plant breeding techniques may be selected from the group
consisting of recurrent selection,
bulk selection, backcrossing, pedigree breeding, genetic marker-assisted
selection, genome editing and
genetic transformation. In certain embodiments of the invention, the canola
plant of variety 5CV686590
is used as the male or female parent.
Still yet another aspect of the invention is a method of producing a canola
plant derived from the
canola variety 5CV686590, the method comprising the steps of: (a) preparing a
progeny plant derived
from canola variety 5CV686590 by crossing a plant of the canola variety
5CV686590 with a second
canola plant; and (b) crossing the progeny plant with itself or a second plant
to produce a progeny plant
of a subsequent generation which is derived from a plant of the canola variety
5CV686590. In one
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Date Recue/Date Received 2022-09-28
embodiment of the invention, the method further comprises: (c) crossing the
progeny plant of a
subsequent generation with itself or a second plant; and (d) repeating steps
(b) and (c) for, in some
embodiments, at least 2, 3, 4 or more additional generations to produce an
inbred canola plant derived
from the canola variety SCV686590. Also provided by the invention is a plant
produced by this and the
other methods of the invention.
In another embodiment of the invention, the method of producing a canola plant
derived from
the canola variety SCV686590 further comprises: (a) crossing the canola
variety SCV686590-derived
canola plant with itself or another canola plant to yield additional canola
variety SCV686590-derived
progeny canola seed; (b) growing the progeny canola seed of step (a) under
plant growth conditions to
yield additional canola variety SCV686590-derived canola plants; and (c)
repeating the crossing and
growing steps of (a) and (b) to generate further canola variety SCV686590-
derived canola plants. In
specific embodiments, steps (a) and (b) may be repeated at least 1, 2, 3, 4,
or 5 or more times as desired.
The invention still further provides a canola plant produced by this and the
foregoing methods.
In another embodiment of the invention, the method of producing a canola plant
comprises
transforming a canola plant of variety SCV686590 with a transgene that confers
herbicide tolerance,
insect resistance, pest resistance, disease resistance, modified fatty acid
metabolism, abiotic stress
resistance, altered seed amino acid composition, site specific genetic
recombination, or modified
carbohydrate metabolism, wherein a sample of seed of said variety has been
deposited under NCMA
Accession No. 202206003.
An aspect of the invention relates to a plant cell of a canola plant produced
by a method
comprising transforming a canola plant of variety SCV686590 with a transgene
that confers herbicide
tolerance, insect resistance, pest resistance, disease resistance, modified
fatty acid metabolism, abiotic
stress resistance, altered seed amino acid composition, site specific genetic
recombination, or modified
carbohydrate metabolism, wherein the plant cell comprises a full copy of the
genome of canola variety
SCV686590, and wherein a sample of seed of said variety has been deposited
under NCMA Accession
No. 202206003.
Another aspect of the invention relates to a plant cell of a canola plant
comprising a single locus
conversion of the canola variety SCV686590, wherein the locus conversion may
comprise a transgenic
gene which has been introduced by genetic transformation into the canola
variety SCV686590 or a
Date Recue/Date Received 2022-09-28
progenitor thereof, wherein the plant cell comprises a full copy of the genome
of canola variety
SCV686590, and wherein a sample of seed of canola variety SCV686590 has been
deposited under
NCMA Accession No. 202206003.A further aspect of the invention is use of
canola variety SCV686590
or a descendant of canola variety SCV686590, wherein the descendant expressed
the morphological and
physiological characteristics of canola variety SCV686590 listed in Table 1. A
descendant of canola
variety SCV686590 may for instance express the morphological and physiological
characteristics of
canola variety SCV686590 listed in Table 1 as determined at the 5%, 10%, 20%,
25%, 50%, 75%, 80%,
90%, or 95% significance level when grown under substantially similar
environmental conditions. In
certain embodiments, the invention provides the use of canola variety
SCV686590 or a descendant of
canola variety SCV686590, wherein a sample of seed of canola variety SCV686590
has been deposited
under NCMA Accession No. 202206003, for instance to produce a cleaned seed for
subsequent planting,
to breed a canola plant, as a recipient of a single locus conversion, to cross
with another canola plant, as
a recipient of a transgene, for oil or protein production, to grow a crop, or
to produce a genetic marker
profile. In one embodiment, use of canola variety SCV686590 or a descendant of
canola variety
SCV686590 to produce a cleaned seed for subsequent planting comprises treating
the seed with a seed
treatment, and wherein a sample of seed of said variety has been deposited
under NCMA Accession No.
202206003.
DETAILED DESCRIPTION
DEFINITIONS
In the description and tables, a number of terms are used. In order to provide
a clear and
consistent understanding of the specification and claims, the following
definitions are provided:
A: When used in conjunction with the word "comprising" or other open language
in the claims,
the words "a" and "an" denote "one or more."
Allele: Any of one or more alternative forms of a gene locus, all of which
relate to one trait or
characteristic. In a diploid cell or organism, the two alleles of a given gene
occupy corresponding loci
on a pair of homologous chromosomes.
Alter: The utilization of up-regulation, down-regulation, or gene silencing.
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Anther arrangement: The orientation of the anthers in fully opened flowers can
also be useful
as an identifying trait. This can range from introse (facing inward toward
pistil), erect (neither inward
not outward), or extrose (facing outward away from pistil).
Anther dotting: The presence/absence of anther dotting (colored spots on the
tips of anthers)
and if present, the percentage of anther dotting on the tips of anthers in
newly opened flowers is also a
distinguishing trait for varieties.
Anther fertility: This is a measure of the amount of pollen produced on the
anthers of a flower.
It can range from sterile (such as in female parents used for hybrid seed
production) to fertile (all anthers
shedding).
Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny,
for example a
first generation hybrid (F1), back to one of the parents of the hybrid
progeny. Backcrossing can be used
to introduce one or more single locus conversions from one genetic background
into another.
Blackleg (Leptosphaeria maculans): Virulent or severe blackleg of
canola/rapeseed is a fungal
canker or dry rot disease of the actively growing crop that causes stem
girdling and lodging. In heavily
infested crops, up to 100 per cent of the stems may be infected, resulting in
major yield loss. For purposes
of this application, resistance to blackleg is measured using ratings of "R"
(resistant), "MR" (medium
resistant), "MS" (moderately susceptible) or "S" (susceptible).
Cell: Cell as used herein includes a plant cell, whether isolated, in tissue
culture or incorporated
in a plant or plant part.
Chromatography: A technique wherein a mixture of dissolved substances are
bound to a solid
support followed by passing a column of fluid across the solid support and
varying the composition of
the fluid. The components of the mixture are separated by selective elution.
Cotyledon width: The cotyledons are leaf structures that form in the
developing seeds of canola
which make up the majority of the mature seed of these species. When the seed
germinates, the
cotyledons are pushed out of the soil by the growing hypocotyls (segment of
the seedling stem below
the cotyledons and above the root) and they unfold as the first photosynthetic
leafs of the plant. The
width of the cotyledons varies by variety and can be classified as narrow,
medium, or wide.
Crossing: The mating of two parent plants.
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Cross-pollination: Fertilization by the union of two gametes from different
plants.
Descendant plant: A plant that is descended from a particular other plant that
is, a descendant
plant is an offspring of a particular other plant
Elite canola line or variety: A canola line or variety, per se, which has been
sold commercially.
Elite canola parent line or variety: A canola line or variety which is a
parent of a canola hybrid
which has been commercially sold.
Emasculate: The removal of plant male sex organs or the inactivation of the
organs with a
cytoplasmic or nuclear genetic factor or a chemical agent conferring male
sterility.
Embryo: The embryo is the small plant contained within a mature seed.
Emergence: The emergence score describes the ability of a seed to emerge from
the soil after
planting. Each genotype is given a 1 to 9 score based on its percent of
emergence. A score of 1 indicates
an excellent rate and percent of emergence, an intermediate score of 5
indicates an average rating and a
9 score indicates a very poor rate and percent of emergence.
Enzymes: Molecules which can act as catalysts in biological reactions.
Essentially all of the morphological and physiological characteristics: The
characteristics of
a plant are recovered that are otherwise present when compared in the same
environment, other than
occasional variant traits that might arise during backcrossing or direct
introduction of a transgene.
Fi Hybrid: The first generation progeny of the cross of two nonisogenic
plants.
FAME analysis: Fatty Acid Methyl Ester analysis is a method that allows for
accurate
quantification of the fatty acids that make up complex lipid classes.
Flower bud location. The location of the unopened flower buds relative to the
adjacent opened
flowers is useful in distinguishing between the canola species. For example,
unopened buds are held
above the most recently opened flowers in B. napus, and they are positioned
below the most recently
opened flower buds in B. rapa.
Flowering date: This is measured by the number of days from planting to the
stage when 50%
of the plants in a population have one or more open flowers. This varies from
variety to variety.
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Fusarium Wilt: Fusarium wilt, largely caused by Fusarium oxysporum, is a
disease of canola
that causes part or all of a plant to wilt, reducing yield by up to 30% or
more on badly affected fields.
For purposes of this application, resistance to Fusarium wilt is measured
using ratings of "R" (resistant),
"MR" (medium resistant), "MS" (moderately susceptible) or "S" (susceptible).
Gene silencing: Gene silencing means the interruption or suppression of the
expression of a
gene at the level of transcription or translation.
Genomic Estimated Breeding Value (GEBV): An estimation of genotyped
populations using
statistical model or models to predict the breeding values of a plant or
plants
Genomic Selection (GS) or Genome-wide selection (GWS): A use of genome-wide
genotypic
data to predict genomic estimated breeding values (GEBV) for selection
purposes in breeding process.
Genotype: The genetic constitution of a cell or organism.
Glucosinolates: These are measured in micromoles (pm) of total alipathic
glucosinolates per
gram of air-dried oil-free meal. The level of glucosinolates is somewhat
influenced by the sulfur fertility
of the soil but is also controlled by the genetic makeup of each variety and
thus can be useful in
characterizing varieties.
Growth habit: At the end of flowering, the angle relative to the ground
surface of the outermost
fully expanded leaf petioles is a variety specific trait. This trait can range
from erect (very upright along
the stem) to prostrate (almost horizontal and parallel with the ground
surface).
Haploid: A cell or organism having one set of the two sets of chromosomes in a
diploid.
Industrial product: Refers to the result of crushing canola seed for oil and
any components
extracted from the leftover oil-free canola meal. Crushing in this connection
refers to the process of
grinding canola grain and extracting oil, leaving protein-rich canola meal as
a by-product. Industrial
products encompass components comprising plant cells or components being
purified plant cell
constituents not comprising plant cells as such. Industrial products comprise
crushed canola grain, canola
hulls, canola meal, canola flour, livestock feed, protein concentrate,
unblended canola oil, salad oil,
cooking oil, frying oil, vegetable oil, a blended oil, and biofuel.
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Leaf attachment to the stem: This trait is especially useful for
distinguishing between the two
canola species. For example, the base of the leaf blade of the upper stem
leaves of B. rapa completely
clasp the stem, whereas those of the B. napus only partially clasp the stem.
Those of the mustard species
do not clasp the stem at all.
Leaf blade color: The color of the leaf blades is variety-specific and can
range from light to
medium dark green to blue green.
Leaf development of lobes: The leaves on the upper portion of the stem can
show varying
degrees of development of lobes, which are disconnected from one another along
the petiole of the leaf.
The degree of lobing is variety specific and can range from absent (no
lobes)/weak through very strong
(abundant lobes).
Leaf glaucosity: This refers to the waxiness of the leaves and is
characteristic of specific
varieties, although environment can have some effect on the degree of
waxiness. This trait can range
from absent (no waxiness)/weak through very strong. The degree of waxiness can
be best determined
by rubbing the leaf surface and noting the degree of wax present.
Leaf indentation of margin: The leaves on the upper portion of the stem can
also show varying
degrees of serration along the leaf margins. The degree of serration or
indentation of the leaf margins
can vary from absent (smooth margin)/weak to strong (heavy saw-tooth like
margin).
Leaf pubescence: The leaf pubescence is the degree of hairiness of the leaf
surface and is
especially useful for distinguishing between the canola species. There are two
main classes of
pubescence, which are glabrous (smooth/not hairy), and pubescent (hairy),
which mainly differentiate
between the B. napus and B. rapa species, respectively.
Leaf surface: The leaf surface can also be used to distinguish between
varieties. The surface
can be smooth or rugose (lumpy), with varying degrees between the two
extremes.
Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate
together
more often than expected by chance if their transmission was independent.
Linkage disequilibrium: Refers to a phenomenon wherein alleles tend to remain
together in
linkage groups when segregating from parents to offspring, with a greater
frequency than expected from
their individual frequencies.
Date Recue/Date Received 2022-09-28
Locus: A locus confers one or more traits such as, for example, male
sterility, herbicide
tolerance, insect resistance, disease resistance, modified fatty acid
metabolism, modified phytic acid
metabolism, modified carbohydrate metabolism and modified protein metabolism.
The trait may be, for
example, conferred by a naturally occurring gene introduced into the genome of
the variety by
backcrossing, a natural or induced mutation, or a transgene introduced through
genetic transformation
techniques. A locus may comprise one or more alleles integrated at a single
chromosomal location.
Lodging resistance: Lodging is rated on a scale of 1 to 5. A score of 1
indicates erect plants.
A score of 5 indicates plants are lying on the ground.
Marker: A readily detectable phenotype, preferably inherited in codominant
fashion (both
alleles at a locus in a diploid heterozygote are readily detectable), with no
environmental variance
component, i.e., heritability of 1.
Marker assisted breeding or marker assisted selection (MAS): A process of
selecting a
desired trait or desired traits in a plant or plants by detecting one or more
markers from the plant, where
the marker is associated with the desired trait.
Maturity: The maturity of a variety is measured as the number of days between
planting and
physiological maturity. This is useful trait in distinguishing varieties
relative to one another.
Moisture: The average percentage moisture in the seeds of the variety.
Oil content: This is measured as percent of the whole dried seed and is
characteristic of different
varieties. It can be determined using various analytical techniques such as
NMR, NIR, and Soxhlet
extraction.
Oil or Oil Percent: Seed oil content is measured and reported on a percentage
basis.
Percent linolenic acid: Percent oil of the seed that is linolenic acid.
Percent oleic acid (OLE): Percent oil of the seed that is oleic acid.
Percentage of total fatty acids: This is determined by extracting a sample of
oil from seed,
producing the methyl esters of fatty acids present in that oil sample and
analyzing the proportions of the
various fatty acids in the sample using gas chromatography. The fatty acid
composition can also be a
distinguishing characteristic of a variety.
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Petal color: The petal color on the first day a flower opens can be a
distinguishing characteristic
for a variety. It can be white, varying shades of yellow, or orange.
Phenotype: The detectable characteristics of a cell or organism, the
characteristics of which are
the manifestation of gene expression.
Plant: As used herein, the term "plant" includes reference to an immature or
mature whole plant,
including a plant from which seed or grain or anthers have been removed. Seed
or embryo that will
produce the plant is also considered to be the plant.
Plant height: This is the height of the plant at the end of flowering if the
floral branches are
extended upright (i.e., not lodged). This varies from variety to variety and
although it can be influenced
by environment, relative comparisons between varieties grown side by side are
useful for variety
identification.
Plant parts: As used herein, the term "plant parts" (or a canola plant, or a
part thereof) includes
protoplasts, leaves, stems, roots, root tips, anthers, pistils, seed, grain,
embryo, pollen, ovules, cotyledon,
hypocotyl, pod, flower, shoot, tissue, petiole, cells, meristematic cells, and
the like.
Protein content: This is measured as percent of whole dried seed and is
characteristic of
different varieties. This can be determined using various analytical
techniques such as NIR and Kjeldahl.
Quantitative trait loci (QTL): Quantitative trait loci (QTL) refer to genetic
loci that control to
some degree numerically representable traits that are usually continuously
distributed.
Regeneration: The development of a plant from tissue culture.
Resistance to lodging: This measures the ability of a variety to stand up in
the field under high
yield conditions and severe environmental factors. A variety can have good
(remains upright), fair, or
poor (falls over) resistance to lodging. The degree of resistance to lodging
is not expressed under all
conditions but is most meaningful when there is some degree of lodging in a
field trial.
Seed coat color: The color of the seed coat can be variety specific and can
range from black
through brown through yellow. Color can also be mixed for some varieties.
Seed coat mucilage: This is useful for differentiating between the two species
of canola, with
B. rapa varieties having mucilage present in their seed coats, while B. napus
varieties do not have this
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Date Recue/Date Received 2022-09-28
present. It is detected by imbibing seeds with water and monitoring the
mucilage that is exuded by the
seed.
Seed Weight (SWT): Canola seeds vary in size; therefore, the number of seeds
required to make
up one pound also varies. This affects the pounds of seed required to plant a
given area and can also
impact end uses. Seed weight may be expressed as grams per 1000 seeds.
Seedling growth habit: The rosette consists of the first 2-8 true leaves and a
variety can be
characterized as having a strong rosette (closely packed leaves) or a weak
rosette (loosely arranged
leaves).
Self-pollination: The transfer of pollen from the anther to the stigma of the
same plant.
Silique (pod) habit: This trait is variety-specific and is a measure of the
orientation of the pods
along the racemes (flowering stems). This trait can range from erect (pods
angled close to racemes)
through horizontal (pods perpendicular to racemes) through arching (pods show
distinct arching habit).
Silique (pod) length of beak: The beak is the segment at the end of the pod
that does not contain
seed (it is a remnant of the stigma and style for the flower). The length of
the beak can be variety specific
and can range from short through medium through long.
Silique (pod) length of pedicel: The pedicel is the stem that attaches the pod
to the raceme of
flowering shoot. The length of the pedicel can be variety specific and can
vary from short through
medium through long.
Silique (pod) length: This is the length of the fully developed pods and can
range from short to
medium to long. It is best used by making comparisons relative to reference
varieties.
Silique (pod) type: This is typically a bilateral single pod for both species
of canola and is not
really useful for variety identification within these species.
Silique (pod) width: This is the width of the fully developed pods and can
range from narrow
to medium to wide. It is best used by making comparisons relative to reference
varieties.
Single gene converted (conversion): Single gene converted (conversion) plant
refers to plants
which are developed by a plant breeding technique called backcrossing, or via
genetic engineering,
wherein essentially all of the desired morphological and physiological
characteristics of a variety are
13
Date Recue/Date Received 2022-09-28
recovered, in addition to the single gene transferred into the variety via the
backcrossing technique or
via genetic transformation.
Single Locus Converted (Conversion) Plant: Plants which are developed by a
plant breeding
technique called backcrossing, wherein essentially all of the morphological
and physiological
characteristics of a canola variety are recovered in addition to the
characteristics of the single locus
transferred into the variety via the backcrossing technique and/or by genetic
transformation.
Stem intensity of anthocyanin coloration: The stems and other organs of canola
plants can
have varying degrees of purple coloration which is due to the presence of
anthocyanin (purple) pigments.
The degree of coloration is somewhat subject to growing conditions, but
varieties typically show varying
degrees of coloration ranging from: absent (no purple)/very weak to very
strong (deep purple coloration).
Substantially Equivalent: A characteristic that, when compared, does not show
a statistically
significant difference (e.g., p = 0.05) from the mean.
Tissue Culture: A composition comprising isolated cells of the same or a
different type or a
collection of such cells organized into parts of a plant.
Total saturated (TOTSAT): Total percent oil of the seed of the saturated fats
in the oil
including C12:0, C14:0, C16:0, C18:0, C20:0, C22:0 and C24Ø
Transgene: A genetic locus comprising a sequence which has been introduced
into the genome
of a canola plant by transformation.
CANOLA VARIETY 5CV686590
Spring canola variety 5CV686590 was developed from the initial cross between
two proprietary
non-public lines.
Generation Year Description
Cross 2008 Cross was made in Lethbridge, Alberta.
Fl 2009 Fl seed was planted in Lethbridge, Alberta May and selfed to
produce
F2 seed.
F2 2009 F2 seed was planted in Temuco, Chile and selfed to produce F3
seed.
F3 2010 F3 seed was planted in Lethbridge, Alberta and selfed to
produce F4
seed.
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Date Recue/Date Received 2022-09-28
F4 2011 F4 seed was planted in Lethbridge, Alberta and selfed to
produce F5
seed.
F5 2011 F5 seed was planted in Lethbridge, Alberta and selfed to
produce F6
seed.
F6 2012 F6 seed was planted in Lethbridge, Alberta and selfed to
produce F7
seed.
F7 2013 F7 seed was planted in Lethbridge, Alberta and selfed to
produce F8
seed.
F8 2013 F8 seed was planted in Cranbrook, British Columbia and selfed
to
produce F9 seed.
F9 2014 F9 seed was planted in Winnipeg, Manitoba and selfed to
produce F10
seed.
The results of an objective evaluation of Canola variety SCV686590 are
presented below, in
Table 1. Those of skill in the art will recognize that these are typical
values that may vary due to
environment and that other values that are substantially equivalent are within
the scope of the invention.
TABLE 1: Phenotypic Description of Canola Variety SCV686590
Trait Phenotype
Classification:
Species Brassica napus
L.
Season Type Spring habit
Type of pollination control Cytoplasmic male sterility: INRA Ogura
type
Characteristics of Plants Before Flowering:
Cotyledon width Narrow/medium
Seedling growth habit (leaf rosette) Medium/wide
Stem anthocyanin intensity Medium/low
Leaf type Petiolate/intermediate
Leaf shape Wide elliptic
Date Recue/Date Received 2022-09-28
Trait Phenotype
Leaf color Medium
Leaf waxiness Medium
Leaf lobing Weak/medium
Leaf margin indentation Weak/medium
Leaf attachment to the stem Partial clasping
Characteristics of Plants After Flowering:
Time to flowering 54 days
Plant height at maturity Tall/medium
Plant growth habit Medium
Flower bud location Buds
above recently opened flowers
Petal color Medium yellow
Pod length Medium/long
Pod angle Semi-erect
Time to maturity 102 days
Seed Characteristics
Seed coat color Black
Disease & Pest Reactions:
Blackleg Moderately resistant
Club Root Susceptible
Fusarium wilt Resistant
Herbicide Reactions:
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Date Recue/Date Received 2022-09-28
Trait Phenotype
Glyphosate Resistant
Glufosinate ammonium Susceptible
This invention is also directed to methods for producing a canola plant by
crossing a first parent
canola plant with a second parent canola plant, wherein the first or second
canola plant is the canola
plant from the variety SCV686590. Further, both first and second parent canola
plants may be from the
variety SCV686590. Therefore, any methods using the variety SCV686590 are part
of this invention:
selfing, backcrosses, hybrid breeding, and crosses to populations. Any plants
produced using variety
SCV686590 as a parent are within the scope of this invention.
Additional methods include, but are not limited to, expression vectors
introduced into plant
tissues using a direct gene transfer method such as microprojectile-mediated
delivery, DNA injection,
electroporation, and the like. More preferably, expression vectors may be
introduced into plant tissues
by using either microprojectile-mediated delivery with a ballistic device or
by using Agrobacterium-
mediated transformation. Transformant plants obtained with the protoplasm of
the invention are
intended to be within the scope of this invention.
The advent of new molecular biological techniques has allowed the isolation
and characterization
of genetic elements with specific functions, such as encoding specific protein
products. Scientists in the
field of plant biology developed a strong interest in engineering the genome
of plants to contain and
express foreign genetic elements, or additional, or modified versions of
native or endogenous genetic
elements in order to alter the traits of a plant in a specific manner. Any DNA
sequences, whether from
a different species or from the same species which are inserted into the
genome using transformation,
are referred to herein collectively as "transgenes." In some embodiments of
the invention, a transgenic
variant of 5CV686590 may contain at least one transgene but could contain at
least 1, 2, 3, 4, 5, 6, 7, 8,
9, 10 and/or no more than 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, or 2.
Over the last fifteen to twenty
17
Date Recue/Date Received 2022-09-28
years, several methods for producing transgenic plants have been developed,
and the present invention
also relates to transgenic variants of the claimed canola variety SCV686590.
One embodiment of the invention is a process for producing canola variety
SCV686590 further
comprising a desired trait, said process comprising transforming a canola
plant of variety SCV686590
with a transgene that confers a desired trait. Another embodiment is the
product produced by this
process. In one embodiment the desired trait may be one or more of herbicide
resistance, insect
resistance, disease resistance, modified seed yield, modified oil percent,
modified protein percent,
modified lodging resistance or modified fatty acid or carbohydrate metabolism.
The specific gene may
be any known in the art or listed herein, including; a polynucleotide
conferring resistance to
imidazolinone, sulfonylurea, glyphosate, glufosinate, triazine,
hydroxyphenylpyruvate dioxygenase
inhibitor, protoporphyrinogen oxidase inhibitor and benzonitrile; a
polynucleotide encoding a Bacillus
thuringiensis polypeptide, a polynucleotide encoding phytase, FAD-2, FAD-3,
galactinol synthase or a
raffinose synthetic enzyme; or a polynucleotide conferring resistance to
blackleg, white rust or other
common canola diseases.
Numerous methods for plant transformation have been developed, including
biological and
physical plant transformation protocols, all of which may be used with this
invention. In addition,
expression vectors and in vitro culture methods for plant cell or tissue
transformation and regeneration
of plants are available and may be used in conjunction with the invention.
In an embodiment, a genetic trait which has been engineered into the genome of
a particular
canola plant may be moved into the genome of another variety using traditional
breeding techniques that
are well known in the plant breeding arts. For example, a backcrossing
approach may be used to move
a transgene from a transformed canola variety into an already developed canola
variety, and the resulting
backcross conversion plant would then comprise the transgene(s).
In embodiments, various genetic elements can be introduced into the plant
genome using
transformation. These elements include any known in the art, specifically
including, but not limited to
genes, coding sequences, inducible, constitutive, and tissue specific
promoters, enhancing sequences,
and signal and targeting sequences.
Plant transformation involves the construction of an expression vector which
will function in
plant cells. Such a vector comprises DNA comprising a gene under control of or
operatively linked to a
18
Date Recue/Date Received 2022-09-28
regulatory element (for example, a promoter). The expression vector may
contain one or more of such
operably linked gene/regulatory element combinations. The vector(s) may be in
the form of a plasmid
and can be used alone or in combination with other plasmids, to provide
transformed canola plants, using
transformation methods as described below to incorporate transgenes into the
genetic material of the
canola plant(s).
Included among various plant transformation techniques are methods permitting
the site-specific
modification of a plant genome. These modifications can include, but are not
limited to, site-specific
mutations, deletions, insertions, and replacements of nucleotides. These
modifications can be made
anywhere within the genome of a plant, for example, in genomic elements,
including, among others,
coding sequences, regulatory elements, and non-coding DNA sequences. Such
methods may be used to
modify a particular trait conferred by a locus. The techniques for making such
modifications by genome
editing are well known in the art and include, for example, use of CRISPR-Cas
systems, zinc-finger
nucleases (ZFNs), and transcription activator-like effector nucleases
(TALENs), among others. A site-
specific nuclease provided herein may be selected from the group consisting of
a zinc-finger nuclease
(ZFN), a meganuclease, an RNA-guided endonuclease, a TALE-endonuclease
(TALEN), a recombinase,
a transposase, or any combination thereof. See, e.g., Khandagale, K. et al.,
"Genome editing for targeted
improvement in plants," Plant Biotechnol Rep 10: 327-343 (2016); and Gaj, T.
et al., "ZFN, TALEN
and CRISPR/Cas-based methods for genome engineering," Trends Biotechnol.
31(7): 397-405 (2013).
A recombinase may be a serine recombinase attached to a DNA recognition motif,
a tyrosine
recombinase attached to a DNA recognition motif or other recombinase enzyme
known in the art. A
recombinase or transposase may be a DNA transposase or recombinase attached to
a DNA binding
domain. A tyrosine recombinase attached to a DNA recognition motif may be
selected from the group
consisting of a Cre recombinase, a Flp recombinase, and a Tnpl recombinase.
According to some
embodiments, a Cre recombinase or a Gin recombinase provided herein is
tethered to a zinc-finger DNA
binding domain. In another embodiment, a serine recombinase attached to a DNA
recognition motif
provided herein is selected from the group consisting of a PhiC31 integrase,
an R4 integrase, and a TP-
901 integrase. In another embodiment, a DNA transposase attached to a DNA
binding domain provided
herein is selected from the group consisting of a TALE-piggyBac and TALE-
Mutator. An RNA-guided
endonuclease may be selected from the group consisting of Casl, Cas1B, Cas2,
Cas3, Cas4, Cas5, Cas6,
Cas7, Cas8, Cas9 (also known as Csnl and Csx12), Cas10, Csyl, Csy2, Csy3,
Csel, Cse2, Cscl, Csc2,
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Date Recue/Date Received 2022-09-28
Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr 1, Cmr3, Cmr4, Cmr5, Cmr6, Csb
1, Csb2, Csb3,
Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csxl, Csx15, Csfl, Csf2, Csf3, Csf4,
Cpfl, CasX, CasY,
and homologs or modified versions thereof, Argonaute (non-limiting examples of
Argonaute proteins
include Thermus thermophilus Argonaute (TtAgo), Pyrococcus furiosus Argonaute
(PfAgo),
Natronobacterium gregoryi Argonaute (NgAgo) and homologs or modified versions
thereof. According
to some embodiments, an RNA-guided endonuclease may be a Cas9 or Cpfl enzyme.
For example, the
CRISPR/Cas9 system allows targeted cleavage of genomic sequences guided by a
small noncoding RNA
in plants (WO 2015026883A1). As another example, Cpfl(Cas12a) acts as an
endoribonuclease to
process crRNA and an endodeoxyribonuclease to cleave targeted genomic
sequences. The CRISPR/Cpfl
system enables gene deletion, insertion, base editing, and locus tagging in
monocot and dicot plants
(Alok et al., Frontiers in Plant Science, 31 March 2020).
EXPRESSION VECTORS FOR CANOLA TRANSFORMATION: MARKER GENES
Expression vectors include at least one genetic marker operably linked to a
regulatory element (a
promoter, for example) that allows transformed cells containing the marker to
be either recovered by
negative selection, i.e., inhibiting growth of cells that do not contain the
selectable marker gene, or by
positive selection, i.e., screening for the product encoded by the genetic
marker. Many commonly used
selectable marker genes for plant transformation are well known in the
transformation arts, and include,
for example, genes that code for enzymes that metabolically detoxify a
selective chemical agent which
may be an antibiotic or an herbicide, or genes that encode an altered target
which is insensitive to the
inhibitor. A few positive selection methods are also known in the art.
One commonly used selectable marker gene for plant transformation is the
neomycin
phosphotransferase II (nptII) gene which, when under the control of plant
regulatory signals, confers
resistance to kanamycin. Another commonly used selectable marker gene is the
hygromycin
phosphotransferase gene which confers resistance to the antibiotic hygromycin.
Additional selectable marker genes of bacterial origin that confer resistance
to antibiotics include
gentamycin acetyl transferase, streptomycin phosphotransferase, aminoglycoside-
3'-adenyl transferase
and the bleomycin resistance determinant. Other selectable marker genes confer
resistance to herbicides
such as glyphosate, glufosinate or bromoxynil. Selectable marker genes for
plant transformation not of
Date Recue/Date Received 2022-09-28
bacterial origin include, for example, mouse dihydrofolate reductase, plant 5 -
enolpyruvylshikimate-3-
phosphate synthase and plant acetolactate synthase.
Another class of marker genes for plant transformation requires screening of
presumptively
transformed plant cells rather than direct genetic selection of transformed
cells for resistance to a toxic
substance such as an antibiotic. These genes are particularly useful to
quantify or visualize the spatial
pattern of expression of a gene in specific tissues and are frequently
referred to as reporter genes because
they can be fused to a gene or gene regulatory sequence for the investigation
of gene expression.
Commonly used genes for screening presumptively transformed cells include P-
glucuronidase (GUS),
0-galactosidase, luciferase and chloramphenicol acetyltransferase. Any of the
above, or other marker
genes, may be utilized in the present invention.
In vivo methods for visualizing GUS activity that do not require destruction
of plant tissue are
available and can be used in embodiments of the invention. Additionally, Green
Fluorescent Protein
(GFP) can be utilized as a marker for gene expression in prokaryotic and
eukaryotic cells. GFP and
mutants of GFP may be used as screenable markers.
EXPRESSION VECTORS FOR CANOLA TRANSFORMATION: PROMOTERS
Genes included in expression vectors must be driven by a nucleotide sequence
comprising a
regulatory element, for example, a promoter. Several types of promoters are
well known in the
transformation arts, as are other regulatory elements that can be used alone
or in combination with
promoters.
As used herein, "promoter" includes reference to a region of DNA upstream from
the start of
transcription and involved in recognition and binding of RNA polymerase and
other proteins to initiate
transcription. A "plant promoter" is a promoter capable of initiating
transcription in plant cells.
Examples of promoters under developmental control include promoters that
preferentially initiate
transcription in certain tissues, such as leaves, roots, seeds, fibers, xylem
vessels, tracheids, or
sclerenchyma. Such promoters are referred to as "tissue-preferred". Promoters
which initiate
transcription only in certain tissues are referred to as "tissue-specific". A
"cell type" specific promoter
primarily drives expression in certain cell types in one or more organs, for
example, vascular cells in
roots or leaves. An "inducible" promoter is a promoter which is under
environmental control. Examples
of environmental conditions that may affect transcription by inducible
promoters include anaerobic
21
Date Recue/Date Received 2022-09-28
conditions or the presence of light. Tissue-specific, tissue-preferred, cell
type specific, and inducible
promoters constitute the class of "non-constitutive" promoters. A
"constitutive" promoter is a promoter
which is active under most environmental conditions.
A. Inducible Promoters - An inducible promoter is operably linked to a gene
for expression in
canola. Optionally, the inducible promoter is operably linked to a nucleotide
sequence encoding a signal
sequence which is operably linked to a gene for expression in canola. With an
inducible promoter the
rate of transcription increases in response to an inducing agent.
Any inducible promoter can be used in the instant invention. Exemplary
inducible promoters
include, but are not limited to, those from the ACEI system which respond to
copper, the In2 gene from
maize which responds to benzenesulfonamide herbicide safeners, or the Tet
repressor from Tn10. A
particularly preferred inducible promoter is a promoter that responds to an
inducing agent to which plants
do not normally respond. An exemplary inducible promoter is the inducible
promoter from a steroid
hormone gene, the transcriptional activity of which is induced by a
glucocorticosteroid hormone.
B. Constitutive Promoters - A constitutive promoter is operably linked to a
gene for expression
in canola or the constitutive promoter is operably linked to a nucleotide
sequence encoding a signal
sequence which is operably linked to a gene for expression in canola.
Many different constitutive promoters can be utilized in the instant
invention. Exemplary
constitutive promoters include, but are not limited to, the promoters from
plant viruses such as the 35S
promoter from CaMV and the promoters from such genes as rice actin, ubiquitin,
pEMU, MAS, and
maize H3 histone. The ALS promoter, Xbal/Ncol fragment 5' to the Brassica
napus ALS3 structural
gene (or a nucleotide sequence similarity to said Xbal/Ncol fragment) could
also be utilized herein.
C. Tissue-specific or Tissue-preferred Promoters - A tissue-specific promoter
is operably linked
to a gene for expression in canola. Optionally, the tissue-specific promoter
is operably linked to a
nucleotide sequence encoding a signal sequence which is operably linked to a
gene for expression in
canola. Plants transformed with a gene of interest operably linked to a tissue-
specific promoter produce
the protein product of the transgene exclusively, or preferentially, in a
specific tissue.
Any tissue-specific or tissue-preferred promoter can be utilized in the
instant invention.
Exemplary tissue-specific or tissue-preferred promoters include, but are not
limited to, a root-preferred
22
Date Recue/Date Received 2022-09-28
promoter such as that from the phaseolin gene, a leaf-specific and light-
induced promoter such as that
from cab or rubisco, an anther-specific promoter such as that from LAT52, a
pollen-specific promoter
such as that from Zm13, or a microspore-preferred promoter such as that from
apg.
SIGNAL SEQUENCES FOR TARGETING PROTEINS TO SUBCELLULAR
COMPARTMENTS
Transport of protein produced by transgenes to a subcellular compartment such
as the chloroplast,
vacuole, peroxisome, glyoxysome, cell wall or mitochondrion or for secretion
into the apoplast, is
accomplished by means of operably linking the nucleotide sequence encoding a
signal sequence to the
5' and/or 3' region of a gene encoding the protein of interest. Targeting
sequences at the 5' and/or 3' end
of the structural gene may determine, during protein synthesis and processing,
where the encoded protein
is ultimately compartmentalized. The presence of a signal sequence directs a
polypeptide to either an
intracellular organelle or subcellular compartment or for secretion to the
apoplast. Many signal
sequences are known in the art and can be utilized in the present invention.
FOREIGN PROTEIN GENES AND AGRONOMIC GENES
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, are within the scope of the invention. In an
embodiment, a foreign protein
then can be extracted from a tissue of interest or from the total biomass by
known methods.
According to a preferred embodiment, the transgenic plant provided for
commercial production
of foreign protein is a canola plant. In another preferred embodiment, the
biomass of interest is seed.
For the relatively small number of transgenic plants that show higher levels
of expression, a genetic map
can be generated, primarily via conventional RFLP, PCR and SSR analysis, which
identifies the
approximate chromosomal location of the integrated DNA molecule. Map
information concerning
chromosomal location is useful for proprietary protection of a subject
transgenic plant. If unauthorized
propagation is undertaken and crosses are made with other germplasm, the map
of the integration region
can be compared to similar maps for suspect plants, to determine if the latter
have a common parentage
with the subject plant. Map comparisons would involve hybridizations, RFLP,
PCR, SSR and
sequencing, all of which are conventional techniques. SNPs may also be used
alone or in combination
with other techniques.
23
Date Recue/Date Received 2022-09-28
Likewise, by means of the present invention, plants can be genetically
engineered to express
various phenotypes of agronomic interest. Through the transformation of
canola, the expression of genes
can be altered to enhance disease resistance, insect resistance, herbicide
resistance, agronomic, grain
quality and other traits. Transformation can also be used to insert DNA
sequences which control, or help
control, male-sterility. DNA sequences native to canola, as well as non-native
DNA sequences, can be
transformed into canola and used to alter levels of native or non-native
proteins. Various promoters,
targeting sequences, enhancing sequences, and other DNA sequences can be
inserted into the genome
for the purpose of altering the expression of proteins. Reduction of the
activity of specific genes (also
known as gene silencing, or gene suppression) is desirable for several aspects
of genetic engineering in
plants.
Many techniques for gene silencing are well known to one of skill in the art,
including but not
limited to knock-outs (such as by insertion of a transposable element such as
Mu or other genetic
elements such as a FRT, Lox or other site specific integration site),
antisense technology, co-suppression,
RNA interference, virus-induced gene silencing, target-RNA-specific ribozymes,
hairpin structures,
MicroRNA, ribozymes, oligonucleotide-mediated targeted modification, Zn-finger
targeted molecules,
and other methods or combinations of the above methods known to those of skill
in the art.
Likewise, by means of the present invention, agronomic genes can be expressed
in transformed
plants. More particularly, plants can be genetically engineered to express
various phenotypes of
agronomic interest. Exemplary genes implicated in this regard include, but are
not limited to, those
categorized below:
1. Genes That Confer Resistance to Pests or Disease and That Encode:
A. Plant disease resistance genes. Plant defences 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 genes to engineer plants
that are resistant to specific pathogen strains.
B. A gene conferring resistance to fungal pathogens, such as oxalate oxidase
or oxalate
decarboxylase.
C. A Bacillus thuringiensis protein, a derivative thereof, or a synthetic
polypeptide modeled
24
Date Recue/Date Received 2022-09-28
thereon, for example, a Bt 6-endotoxin gene.
D. A lectin.
E. A vitamin-binding protein such as avidin or a homolog.
F. An enzyme inhibitor, for example, a protease or proteinase inhibitor or an
amylase inhibitor.
G. An insect-specific hormone or pheromone such as an ecdysteroid or juvenile
hormone, a
variant thereof, a mimetic based thereon, or an antagonist or agonist thereof.
H. An insect-specific peptide or neuropeptide which, upon expression, disrupts
the physiology
of the affected pest.
I. An insect-specific venom produced in nature by a snake, a wasp, etc.
J. An enzyme responsible for a hyperaccumulation of a monoterpene, a
sesquiterpene, a steroid,
hydroxamic acid, a phenylpropanoid derivative or another non-protein molecule
with insecticidal
activity.
K. An enzyme involved in the modification, including the post-translational
modification, of a
biologically active molecule; for example, a glycolytic enzyme, a proteolytic
enzyme, a lipolytic
enzyme, a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, a
phosphatase, a kinase, a
phosphorylase, a polymerase, an elastase, a chitinase and a glucanase, whether
natural or synthetic.
L. A molecule that stimulates signal transduction.
M. A hydrophobic moment peptide.
N. A membrane permease, a channel former or a channel blocker.
0. 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. 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.
P. An insect-specific antibody or an immunotoxin derived therefrom. An
antibody targeted to a
Date Recue/Date Received 2022-09-28
critical metabolic function in the insect gut would inactivate an affected
enzyme, killing the insect.
Q. A virus-specific antibody.
R. A developmental-arrestive protein produced in nature by a pathogen or a
parasite. Thus,
fungal endo-a-1, 4-D-polygalacturonases facilitate fungal colonization and
plant nutrient release by
solubilizing plant cell wall homo-a-1, 4-D-galacturonase.
S. A developmental-arrestive protein produced in nature by a plant.
T. Genes involved in the Systemic Acquired Resistance (SAR) Response and/or
the
pathogenesis-related genes.
U. Antifungal genes.
V. Detoxification genes, such as for fumonisin, beauvericin, moniliformin and
zearalenone and
their structurally related derivatives.
W. Cystatin and cysteine proteinase inhibitors.
X. Defensin genes.
Y. 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.
2. Genes That Confer Resistance to an Herbicide, for Example:
A. An herbicide that inhibits the growing point or meristem, such as an
imidazolinone or a
sulfonylurea.
B. Glyphosate (resistance conferred by mutant 5-enolpyruvylshikimate-3-
phosphate synthase
(EPSP) and aroA genes, respectively) and other phosphono compounds such as
glufosinate
(phosphinothricin acetyl transferase (PAT) and Streptomyces hygroscopicus PAT
bar genes), and
pyridinoxy or phenoxy propionic acids and cyclohexanediones (ACCase inhibitor-
encoding genes).
Glyphosate resistance is also imparted to plants that express a gene that
encodes a glyphosate oxido-
reductase enzyme. In addition glyphosate resistance can be imparted to plants
by the over expression of
genes encoding glyphosate N-acetyltransferase. Nucleotide sequences of
glutamine synthetase genes
which confer resistance to herbicides such as L-phosphinothricin are known and
can be used herein. The
26
Date Recue/Date Received 2022-09-28
nucleotide sequence of a PAT gene is also known and can be used. Exemplary of
genes conferring
resistance to phenoxy propionic acids and cyclohexanediones, such as
sethoxydim and haloxyfop are the
Accl -S1, Accl -S2 and Accl -S3 genes.
C. An herbicide that inhibits photosynthesis, such as a triazine (psbA and gs+
genes) and a
benzonitrile (nitrilase gene). The transformation of Chlamydomonas with
plasmids encoding mutant
psbA genes are known and can be used.
D. Acetohydroxy acid synthase, which has been found to make plants that
express this enzyme
resistant to multiple types of herbicides. Other genes that confer tolerance
to herbicides include a gene
encoding a chimeric protein of rat cytochrome P4507A1 and yeast NADPH-
cytochrome P450
oxidoreductase, genes for glutathione reductase and superoxide dismutase, and
genes for various
phosphotransferases.
E. Protoporphyrinogen oxidase (protox), which is necessary for the production
of chlorophyll.
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.
3. Genes That Confer or Contribute to a Value-Added Trait, Such as:
A. Modified fatty acid metabolism, for example, by transforming a plant with
an antisense gene
of stearyl-ACP desaturase to increase stearic acid content of the plant.
B. Decreased phytate content. Introduction of a phytase-encoding gene, such as
Aspergillus niger
phytase gene, may enhance breakdown of phytate, adding more free phosphate to
the transformed plant.
Alternatively, a gene could be introduced that reduces phytate content. In
maize for example, this could
be accomplished by cloning and then reintroducing DNA associated with the
single allele which is
responsible for maize mutants characterized by low levels of phytic acid.
C. Modified carbohydrate composition effected, for example, by transforming
plants with a gene
coding for an enzyme that alters the branching pattern of starch, or, a gene
altering thioredoxin such as
NTR and/or TRX and/or a gamma zein knock out or mutant such as cs27 or TUSC27
or en27. Any
known fatty acid modification genes may also be used to affect starch content
and/or composition
through the interrelationship of the starch and oil pathways.
D. Elevated oleic acid via FAD-2 gene modification and/or decreased linolenic
acid via FAD-3
27
Date Recue/Date Received 2022-09-28
gene modification.
E. Altering conjugated linolenic or linoleic acid content. Altering LEC 1,
AGP, Dekl, Superal 1,
milps, various Ipa genes such as Ipal, Ipa3, hpt or hggt.
F. Altered antioxidant content or composition, such as alteration of
tocopherol or tocotrienols.
In an embodiment, antioxidant levels may be manipulated through alteration of
a phytl prenyl transferase
(ppt) or through alteration of a homogentisate geranyl geranyl transferase
(hggt).
G. Altered essential seed amino acids.
4. Genes that Control Male Sterility
There are several methods of conferring genetic male sterility available and
within the scope of
the invention. As one example, nuclear male sterility may be accomplished by
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. Other possible examples include the
introduction of a deacetylase gene
under the control of a tapetum-specific promoter and with the application of
the chemical N-Ac-PPT,
the introduction of various stamen-specific promoters, or the introduction of
the barnase and the barstar
genes.
5. Genes that create a site for site specific DNA integration.
This may include 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. Other systems that may be
used include the Gin
recombinase of phage Mu, the Pin recombinase of E. coli, and the R/RS system
of the pSR1 plasmid.
6. Genes that affect abiotic stress resistance (including but not limited to
flowering, pod and seed
development, enhancement of nitrogen utilization efficiency, altered nitrogen
responsiveness,
drought resistance or tolerance, cold resistance or tolerance, and salt
resistance or tolerance) and
increased yield under stress.
28
Date Recue/Date Received 2022-09-28
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.
METHODS FOR CANOLA TRANSFORMATION
Numerous methods for plant transformation have been developed, including
biological and
physical plant transformation protocols. In addition, expression vectors and
in vitro culture methods for
plant cell or tissue transformation and regeneration of plants are available.
A. Agrobacterium-mediated Transformation - One method for introducing an
expression vector
into plants is based on the natural transformation system of Agrobacterium. A.
tumefaciens and A.
rhizogenes are plant pathogenic soil bacteria which genetically transform
plant cells. The Ti and Ri
plasmids of A. tumefaciens and A. rhizogenes, respectively, carry genes
responsible for genetic
transformation of the plant. Agrobacterium vector systems and methods for
Agrobacterium-mediated
gene transfer can be used in the present invention.
B. Direct Gene Transfer - Several methods of plant transformation,
collectively referred to as
direct gene transfer, have been developed as an alternative to Agrobacterium-
mediated transformation.
A generally applicable method of plant transformation is microprojectile-
mediated transformation
wherein DNA is carried on the surface of microprojectiles measuring 1 to 4
!um. The expression vector
is introduced into plant tissues with a ballistic device that accelerates the
microprojectiles to speeds of
300 to 600 m/s which is sufficient to penetrate plant cell walls and
membranes. Another method for
physical delivery of DNA to plants is sonicati on of target cells, which may
be used herein. Alternatively,
liposome and spheroplast fusion may be used to introduce expression vectors
into plants. Direct uptake
of DNA into protoplasts using CaCl2 precipitation, polyvinyl alcohol or poly-L-
ornithine may also be
useful. Electroporation of protoplasts and whole cells and tissues may also be
utilized.
Following transformation of canola target tissues, expression of the above-
described selectable
marker genes allows for preferential selection of transformed cells, tissues
and/or plants, using
regeneration and selection methods well known in the art.
The foregoing methods for transformation would typically be used for producing
a transgenic
variety. The transgenic variety could then be crossed with another (non-
transformed or transformed)
variety in order to produce a new transgenic variety. Alternatively, a genetic
trait which has been
29
Date Recue/Date Received 2022-09-28
engineered into a particular canola variety using the foregoing transformation
techniques could be moved
into another variety using traditional backcrossing techniques that are well
known in the plant breeding
arts. For example, a backcrossing approach could be used to move an engineered
trait from a public,
non-elite variety into an elite variety, or from a variety containing a
foreign gene in its genome into a
variety or varieties which do not contain that gene. As used herein,
"crossing" can refer to a simple X
by Y cross, or the process of backcrossing, depending on the context.
GENETIC MARKER PROFILE THROUGH SSR AND FIRST GENERATION PROGENY
In addition to phenotypic observations, a plant can also be identified by its
genotype. The
genotype of a plant can be characterized through a genetic marker profile
which can identify plants of
the same variety or a related variety or be used to determine or validate a
pedigree. Genetic marker
profiles can be obtained by techniques such as Restriction Fragment Length
Polymorphisms (RFLPs),
Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase
Chain Reaction
(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized
Amplified Regions
(SCARs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence
Repeats (SSRs)
which are also referred to as Microsatellites, and Single Nucleotide
Polymorphisms (SNPs).
Particular markers used for these purposes are not limited to any particular
set of markers, but
are envisioned to include any type of marker and marker profile which provides
a means of
distinguishing varieties. One method of comparison is to use only homozygous
loci for 5CV686590.
In addition to being used for identification of canola variety 5CV686590 and
plant parts and plant
cells of variety 5CV686590, the genetic profile may be used to identify a
canola plant produced through
the use of 5CV686590 or to verify a pedigree for progeny plants produced
through the use of
5CV686590. The genetic marker profile is also useful in breeding and
developing backcross
conversions.
The present invention comprises a canola plant characterized by molecular and
physiological
data obtained from the representative sample of said variety deposited with
the Provasoli-Guillar
National Center for Marine Algae and Microbiota (NCMA) at Bigelow Laboratory
for Ocean Sciences,
60 Bigelow Drive, East Boothbay, ME 04544 USA. Further provided by the
invention is a canola plant
formed by the combination of the disclosed canola plant or plant cell with
another canola plant or cell
and comprising the homozygous alleles of the variety.
Date Recue/Date Received 2022-09-28
Means of performing genetic marker profiles using SSR polymorphisms are well
known in the
art. SSRs are genetic markers based on polymorphisms in repeated nucleotide
sequences, such as
microsatellites. A marker system based on SSRs can be highly informative in
linkage analysis relative
to other marker systems in that multiple alleles may be present. Another
advantage of this type of marker
is that, through use of flanking primers, detection of SSRs can be achieved,
for example, by the
polymerase chain reaction (PCR), thereby eliminating the need for labor-
intensive Southern
hybridization. The PCR detection is done by use of two oligonucleotide primers
flanking the
polymorphic segment of repetitive DNA. Repeated cycles of heat denaturation of
the DNA followed by
annealing of the primers to their complementary sequences at low temperatures,
and extension of the
annealed primers with DNA polymerase, comprise the major part of the
methodology.
Following amplification, markers can be scored by electrophoresis of the
amplification products.
Scoring of marker genotype is based on the size of the amplified fragment,
which may be measured by
the number of base pairs of the fragment. While variation in the primer used
or in laboratory procedures
can affect the reported fragment size, relative values should remain constant
regardless of the specific
primer or laboratory used. When comparing varieties it is preferable if all
SSR profiles are performed in
the same lab.
The SSR profile of canola plant 5CV686590 can be used to identify plants
comprising
5CV686590 as a parent, since such plants will comprise the same homozygous
alleles as 5CV686590.
Because the canola variety is essentially homozygous at all relevant loci,
most loci should have only one
type of allele present. In contrast, a genetic marker profile of an Fi progeny
should be the sum of those
parents, e.g., if one parent was homozygous for allele x at a particular
locus, and the other parent
homozygous for allele y at that locus, then the F 1 progeny will be xy
(heterozygous) at that locus.
Subsequent generations of progeny produced by selection and breeding are
expected to be of genotype
x (homozygous), y (homozygous), or xy (heterozygous) for that locus position.
When the F 1 plant is
selfed or sibbed for successive filial generations, the locus should be either
x or y for that position.
In addition, plants and plant parts substantially benefiting from the use of
5CV686590 in their
development, such as 5CV686590 comprising a backcross conversion, transgene,
or genetic sterility
factor, may be identified by having a molecular marker profile with a high
percent identity to
5CV686590. Such a percent identity might be 95%, 96%, 97%, 98%, 99%, 99.5% or
99.9% identical to
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Date Recue/Date Received 2022-09-28
SCV686590.
The SSR profile of SCV686590 also can be used to identify essentially derived
varieties and
other progeny varieties developed from the use of SCV686590, as well as cells
and other plant parts
thereof. Progeny plants and plant parts produced using SCV686590 may be
identified by having a
molecular marker profile of at least 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%,
65%, 70%, 75%, 76%,
77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,
92%, 93%, 94%,
95%, 96%, 97%, 98%, 99% or 99.5% genetic contribution from canola variety, as
measured by either
percent identity or percent similarity. Such progeny may be further
characterized as being within a
pedigree distance of 5CV686590, such as within 1 ,2, 3 ,4 or 5 or less cross-
pollinations to a canola plant
other than 5CV686590 or a plant that has 5CV686590 as a progenitor. Unique
molecular profiles may
be identified with other molecular tools such as SNPs and RFLPs.
While determining the SSR genetic marker profile of the plants described
supra, several unique
SSR profiles may also be identified which did not appear in either parent of
such plant. Such unique
SSR profiles may arise during the breeding process from recombination or
mutation. A combination of
several unique alleles provides a means of identifying a plant variety, an Fi
progeny produced from such
variety, and progeny produced from such variety.
SINGLE-GENE CONVERSIONS
When the term "canola plant" is used in the context of the present invention,
this also includes
any single gene conversions of that variety. The term single gene converted
plant as used herein refers
to those canola plants which are developed by backcrossing, wherein
essentially all of the desired
morphological and physiological characteristics of a variety are recovered in
addition to the single gene
transferred into the variety via the backcrossing technique. Backcrossing
methods can be used with the
present invention to improve or introduce a characteristic into the variety. A
hybrid progeny may be
backcrossed to the recurrent parent 1, 2, 3, 4, 5, 6, 7, 8 or more times as
part of this invention. The
parental canola plant that contributes the gene for the desired characteristic
is termed the nonrecurrent
or donor parent. This terminology refers to the fact that the nonrecurrent
parent is used one time in the
backcross protocol and therefore does not recur. The parental canola plant to
which the gene or genes
from the nonrecurrent parent are transferred is known as the recurrent parent
as it is used for several
rounds in the backcrossing protocol. In a typical backcross protocol, the
original variety of interest
32
Date Recue/Date Received 2022-09-28
(recurrent parent) is crossed to a second variety (nonrecurrent parent) that
carries the single gene of
interest to be transferred. The resulting progeny from this cross are then
crossed again to the recurrent
parent and the process is repeated until a canola plant is obtained wherein
essentially all of the desired
morphological and physiological characteristics of the recurrent parent are
recovered in the converted
plant, in addition to the single transferred gene from the nonrecurrent
parent.
The selection of a suitable recurrent parent is an important step for a
successful backcrossing
procedure. The goal of a backcross protocol is to alter or substitute a single
trait or characteristic in the
original variety. To accomplish this, a single gene of the recurrent variety
is modified or substituted
with the desired gene from the nonrecurrent parent, while retaining
essentially all of the rest of the
desired genetic, and therefore the desired morphological and physiological,
constitution of the original
variety. The choice of the particular nonrecurrent parent will depend on the
purpose of the backcross;
one of the major purposes is to add some agronomically important trait to the
plant. The exact
backcrossing protocol will depend on the characteristic or trait being altered
to determine an appropriate
testing protocol. Although backcrossing methods are simplified when the
characteristic being
transferred is a dominant allele, a recessive allele may also be transferred.
In this instance it may be
necessary to introduce a test of the progeny to determine if the desired
characteristic has been
successfully transferred.
Many single gene traits have been identified that are not regularly selected
for in the development
of a new variety but that can be improved by backcrossing techniques. Single
gene traits may or may not
be transgenic; examples of these traits include but are not limited to, male
sterility, waxy starch, herbicide
resistance, resistance for bacterial, fungal, or viral disease, insect
resistance, male fertility, enhanced
nutritional quality, industrial usage, yield stability and yield enhancement.
These genes are generally
inherited through the nucleus.
INTRODUCTION OF A NEW TRAIT OR LOCUS INTO 5CV686590
Variety 5CV686590 represents a new base genetic variety into which a new locus
or trait may
be introgressed. Direct transformation and backcrossing represent two methods
that can be used to
accomplish such an introgression. The term backcross conversion and single
locus conversion are used
interchangeably to designate the product of a backcrossing program.
33
Date Recue/Date Received 2022-09-28
BACKCROSS CONVERSIONS OF SCV686590
A backcross conversion of SCV686590 may occur when DNA sequences are
introduced through
backcrossing with SCV686590 utilized as the recurrent parent. Both naturally
occurring and transgenic
DNA sequences may be introduced through backcrossing techniques. Molecular
marker assisted
breeding or selection may be utilized to reduce the number of backcrosses
necessary to achieve the
backcross conversion.
The complexity of the backcross conversion method depends on the type of trait
being transferred
(single genes or closely linked genes as vs. unlinked genes), the level of
expression of the trait, the type
of inheritance (cytoplasmic or nuclear) and the types of parents included in
the cross. It is understood by
those of ordinary skill in the art that for single gene traits that are
relatively easy to classify, the backcross
method is effective and relatively easy to manage. Desired traits that may be
transferred through
backcross conversion include, but are not limited to, sterility (nuclear and
cytoplasmic), fertility
restoration, nutritional enhancements, drought tolerance, nitrogen
utilization, altered fatty acid profile,
altered seed amino acid levels, altered seed oil levels, low phytate,
industrial enhancements, disease
resistance (bacterial, fungal or viral), insect resistance and herbicide
resistance. In addition, an
introgression site itself, such as an FRT site, Lox site or other site
specific integration site, may be
inserted by backcrossing and utilized for direct insertion of one or more
genes of interest into a specific
plant variety. In some embodiments of the invention, the number of loci that
may be backcrossed into
SCV686590 is at least 1, 2, 3, 4, or 5 and/or no more than 6, 5, 4, 3, or 2. A
single locus may contain
several transgenes, such as a transgene for disease resistance that, in the
same expression vector, also
contains a transgene for herbicide resistance. The gene for herbicide
resistance may be used as a
selectable marker and/or as a phenotypic trait. A single locus conversion of
site specific integration
system allows for the integration of multiple genes at the converted loci.
The backcross conversion may result from either the transfer of a dominant
allele or a recessive
allele. Selection of progeny containing the trait of interest is accomplished
by direct selection for a trait
associated with a dominant allele. Transgenes transferred via backcrossing
typically function as a
dominant single gene trait and are relatively easy to classify. Selection of
progeny for a trait that is
transferred via a recessive allele requires growing and selfing the first
backcross generation to determine
which plants carry the recessive alleles. Recessive traits may require
additional progeny testing in
34
Date Recue/Date Received 2022-09-28
successive backcross generations to determine the presence of the locus of
interest. The last backcross
generation is usually selfed to give pure breeding progeny for the gene(s)
being transferred, although a
backcross conversion with a stably introgressed trait may also be maintained
by further backcrossing to
the recurrent parent with selection for the converted trait.
Along with selection for the trait of interest, progeny are selected for the
phenotype of the
recurrent parent. The backcross is a form of inbreeding, and the features of
the recurrent parent are
automatically recovered after successive backcrosses. As noted above, the
number of backcrosses
necessary can be reduced with the use of molecular markers. Other factors,
such as a genetically similar
donor parent, may also reduce the number of backcrosses necessary. As noted,
backcrossing is easiest
for simply inherited, dominant and easily recognized traits.
One process for adding or modifying a trait or locus in canola variety
SCV686590 comprises
crossing SCV686590 plants grown from SCV686590 seed with plants of another
canola variety that
comprise the desired trait or locus, selecting Fi progeny plants that comprise
the desired trait or locus to
produce selected Fi progeny plants, crossing the selected progeny plants with
the SCV686590 plants to
produce backcross progeny plants, selecting for backcross progeny plants that
have the desired trait or
locus and the morphological characteristics of canola variety SCV686590 to
produce selected backcross
progeny plants; and backcrossing to SCV686590 three or more times in
succession to produce selected
fourth or higher backcross progeny plants that comprise said trait or locus.
The modified SCV686590
may be further characterized as having essentially all of the morphological
and physiological
characteristics of canola variety SCV686590 listed in Table 1 and/or may be
characterized by percent
similarity or identity to SCV686590 as determined by SSR markers. The above
method may be utilized
with fewer backcrosses in appropriate situations, such as when the donor
parent is highly related or
markers are used in the selection step. Desired traits that may be used
include those nucleic acids known
in the art, some of which are listed herein, that will affect traits through
nucleic acid expression or
inhibition. Desired loci include the introgression of FRT, Lox and other sites
for site specific integration,
which may also affect a desired trait if a functional nucleic acid is inserted
at the integration site.
In addition, the above process and other similar processes described herein
may be used to
produce first generation progeny canola seed by adding a step at the end of
the process that comprises
crossing SCV686590 with the introgressed trait or locus with a different
canola plant and harvesting the
Date Recue/Date Received 2022-09-28
resultant first generation progeny canola seed.
TISSUE CULTURE OF CANOLA
Further production of the SCV686590 variety can occur by tissue culture and
regeneration.
Culture of various tissues of canola and regeneration of plants therefrom is
known and widely published.
Thus, another aspect of this invention is to provide cells which upon growth
and differentiation produce
canola plants having the morphological and physiological characteristics of
canola variety SCV686590.
As used herein, the term "tissue culture" indicates a composition comprising
isolated cells of the
same or a different type or a collection of such cells organized into parts of
a plant. Exemplary types of
tissue cultures are protoplasts, calli, plant clumps, and plant cells that can
generate tissue culture that are
intact in plants or parts of plants, such as embryos, pollen, flowers, seeds,
pods, leaves, stems, roots, root
tips, anthers, pistils and the like. Means for preparing and maintaining plant
tissue culture are well
known in the art. Tissue culture comprising organs can be used in the present
invention to produce
regenerated plants.
USING 5CV686590 TO DEVELOP OTHER CANOLA VARIETIES
Canola varieties such as SCV686590 are typically developed for use in seed and
grain
production. However, canola varieties such as SCV686590 also provide a source
of breeding material
that may be used to develop new canola varieties. Plant breeding techniques
known in the art and used
in a canola plant breeding program include, but are not limited to, recurrent
selection, mass selection,
bulk selection, mass selection, backcrossing, pedigree breeding, open
pollination breeding, restriction
fragment length polymorphism enhanced selection, genetic marker enhanced
selection, making double
haploids, and transformation. Often combinations of these techniques are used.
The development of
canola varieties in a plant breeding program requires, in general, the
development and evaluation of
homozygous varieties.
ADDITIONAL BREEDING METHODS
This invention is 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 variety
SCV686590. The other parent may be any other canola plant, such as a canola
plant that is part of a
synthetic or natural population. Any such methods using canola variety
SCV686590 are part of this
36
Date Recue/Date Received 2022-09-28
invention: selfing, sibbing, backcrosses, mass selection, pedigree breeding,
bulk selection, hybrid
production, crosses to populations, and the like. These methods are well known
in the art and some of
the more commonly used breeding methods are described below.
The following describes breeding methods that may be used with canola variety
SCV686590 in
the development of further canola plants. One such embodiment is a method for
developing a variety
SCV686590 progeny canola plant in a canola plant breeding program comprising:
obtaining the canola
plant, or a part thereof, of variety SCV686590 utilizing said plant or plant
part as a source of breeding
material and selecting a canola variety SCV686590 progeny plant with molecular
markers in common
with variety SCV686590 and/or with morphological and/or physiological
characteristics selected from
the characteristics listed in Table 1. Breeding steps that may be used in the
canola plant breeding program
include pedigree breeding, backcrossing, mutation breeding, and recurrent
selection. In conjunction with
these steps, techniques such as RFLP-enhanced selection, genetic marker
enhanced selection (for
example SSR markers) and the making of double haploids may be utilized.
Another method involves producing a population of canola variety SCV686590
progeny canola
plants, comprising crossing variety SCV686590 with another canola plant,
thereby producing a
population of canola plants, which, on average, derive 50% of their alleles
from canola variety
SCV686590. A plant of this population may be selected and repeatedly selfed or
sibbed with a canola
variety resulting from these successive filial generations. One embodiment of
this invention is the canola
variety produced by this method and that has obtained at least 50% of its
alleles from canola variety
SCV686590.
One of ordinary skill in the art of plant breeding would know how to evaluate
the traits of two
plant varieties to determine if there is no significant difference between the
two traits expressed by those
varieties. Thus the invention includes canola variety SCV686590 progeny canola
plants comprising a
combination of at least two variety SCV686590 traits selected from the group
consisting of those listed
in Table 1 or the variety SCV686590 combination of traits listed in the
Summary of the Invention, so
that said progeny canola plant is not significantly different for said traits
than canola variety SCV686590
as determined at the 5% significance level when grown in the same
environmental conditions. Using
techniques described herein, molecular markers may be used to identify said
progeny plant as a canola
variety SCV686590 progeny plant. Mean trait values may be used to determine
whether trait differences
37
Date Recue/Date Received 2022-09-28
are significant, and preferably the traits are measured on plants grown under
the same environmental
conditions. Once such a variety is developed its value is substantial since it
is important to advance the
germplasm base as a whole in order to maintain or improve traits such as
yield, disease resistance, pest
resistance, and plant performance in extreme environmental conditions.
Progeny of canola variety SCV686590 may also be characterized through their
filial relationship
with canola variety SCV686590, as for example, being within a certain number
of breeding crosses of
canola variety SCV686590. A breeding cross is a cross made to introduce new
genetics into the progeny,
and is distinguished from a cross, such as a self or a sib cross, made to
select among existing genetic
alleles. The lower the number of breeding crosses in the pedigree, the closer
the relationship between
canola variety SCV686590 and its progeny. For example, progeny produced by the
methods described
herein may be within 1, 2, 3, 4 or 5 breeding crosses of canola variety
SCV686590.
As used herein, the term "plant" includes plant cells, 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, flowers, shoots,
pods, leaves, roots, root tips,
anthers, cotyledons, hypocotyls, meristematic cells, stems, pistils, petiole,
seeds, and the like.
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Date Recue/Date Received 2022-09-28
PEDIGREE BREEDING
Pedigree breeding starts with the crossing of two genotypes, such as SCV686590
and another
canola variety having one or more desirable characteristics that is lacking or
which complements
SCV686590. If the two original parents do not provide all the desired
characteristics, other sources can
be included in the breeding population. In the pedigree method, superior
plants are selfed and selected
in successive filial generations. In the succeeding filial generations the
heterozygous condition gives
way to homogeneous varieties as a result of self-pollination and selection.
Typically, in the pedigree
method of breeding, five or more successive filial generations of selfing and
selection is practiced: Fi to
F2; F2 to F3; F3 to F4; F4 to F5, etc. After a sufficient amount of
inbreeding, successive filial generations
will serve to increase seed of the developed variety. Preferably, the
developed variety comprises
homozygous alleles at about 95% or more of its loci.
In addition to being used to create a backcross conversion, backcrossing can
also be used in
combination with pedigree breeding. As discussed previously, backcrossing can
be used to transfer one
or more specifically desirable traits from one variety, the donor parent, to a
developed variety called the
recurrent parent, which has overall good agronomic characteristics yet lacks
that desirable trait or traits.
However, the same procedure can be used to move the progeny toward the
genotype of the recurrent
parent but at the same time retain many components of the non-recurrent parent
by stopping the
backcrossing at an early stage and proceeding with selfing and selection. For
example, a canola variety
may be crossed with another variety to produce a first generation progeny
plant. The first generation
progeny plant may then be backcrossed to one of its parent varieties to create
a BC1 or BC2. Progeny
are selfed and selected so that the newly developed variety has many of the
attributes of the recurrent
parent and yet several of the desired attributes of the non-recurrent parent.
This approach leverages the
value and strengths of the recurrent parent for use in new canola varieties.
Therefore, an embodiment of this invention is a method of making a backcross
conversion of
canola variety SCV686590, comprising the steps of crossing a plant of canola
variety SCV686590 with
a donor plant comprising a desired trait, selecting an Fi progeny plant
comprising the desired trait, and
backcrossing the selected Fi progeny plant to a plant of canola variety
SCV686590. This method may
further comprise the step of obtaining a molecular marker profile of canola
variety SCV686590 and
using the molecular marker profile to select for a progeny plant with the
desired trait and the molecular
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Date Recue/Date Received 2022-09-28
marker profile of SCV686590. In one embodiment the desired trait is a mutant
gene or transgene present
in the donor parent.
RECURRENT SELECTION AND MASS SELECTION
Recurrent selection is a method used in a plant breeding program to improve a
population of
plants. SCV686590 is suitable for use in a recurrent selection program. The
method entails individual
plants cross pollinating with each other to form progeny. The progeny are
grown and the superior
progeny selected by any number of selection methods, which include individual
plant, half-sib progeny,
full-sib progeny and selfed progeny. The selected progeny are cross pollinated
with each other to form
progeny for another population. This population is planted and again superior
plants are selected to cross
pollinate with each other. Recurrent selection is a cyclical process and
therefore can be repeated as many
times as desired. The objective of recurrent selection is to improve the
traits of a population. The
improved population can then be used as a source of breeding material to
obtain new varieties for
commercial or breeding use, including the production of a synthetic variety. A
synthetic variety is the
resultant progeny formed by the intercrossing of several selected varieties.
Mass selection is a useful technique when used in conjunction with molecular
marker enhanced
selection. In mass selection seeds from individuals are selected based on
phenotype or genotype. These
selected seeds are then bulked and used to grow the next generation. Bulk
selection requires growing a
population of plants in a bulk plot, allowing the plants to self-pollinate,
harvesting the seed in bulk and
then using a sample of the seed harvested in bulk to plant the next
generation. Also, instead of self
pollination, directed pollination could be used as part of the breeding
program.
MUTATION BREEDING
Mutation breeding is another method of introducing new traits into canola
variety SCV686590.
Mutations that occur spontaneously or are artificially induced can be useful
sources of variability for a
plant breeder. The goal of artificial mutagenesis is to increase the rate of
mutation for a desired
characteristic. Mutation rates can be increased by many different means
including temperature, long-
term seed storage, tissue culture conditions, radiation; such as X-rays, Gamma
rays (e.g. cobalt 60 or
cesium 137), neutrons, (product of nuclear fission by uranium 235 in an atomic
reactor), Beta radiation
(emitted from radioisotopes such as phosphorus 32 or carbon 14), or
ultraviolet radiation (preferably
from 2500 to 2900 nm), or chemical mutagens (such as base analogues (5-bromo-
uracil), related
Date Recue/Date Received 2022-09-28
compounds (8-ethoxy caffeine), antibiotics (streptonigrin), alkylating agents
(sulfur mustards, nitrogen
mustards, epoxides, ethylenamines, sulfates, sulfonates, sulfones, lactones),
azide, hydroxylamine,
nitrous acid, or acridines. Once a desired trait is observed through
mutagenesis the trait may then be
incorporated into existing germplasm by traditional breeding techniques. In
addition, mutations created
in other canola plants may be used to produce a backcross conversion of canola
variety SCV686590 that
comprises such mutation.
BREEDING WITH MOLECULAR MARKERS
Molecular markers, which include markers identified through the use of
techniques such as
Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs),
Randomly Amplified
Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-
PCR), DNA
Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions
(SCARs), Amplified
Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs) and
Single Nucleotide
Polymorphisms (SNPs), may be used in plant breeding methods utilizing canola
variety 5CV686590.
One use of molecular markers is Quantitative Trait Loci (QTL) mapping. Q
________ IL mapping is the use of
markers, which are known to be closely linked to alleles that have measurable
effects on a quantitative
trait. Selection in the breeding process is based upon the accumulation of
markers linked to the positive
effecting alleles and/or the elimination of the markers linked to the negative
effecting alleles from the
plant's genome.
Molecular markers can also be used during the breeding process for the
selection of qualitative
traits. For example, markers closely linked to alleles or markers containing
sequences within the actual
alleles of interest can be used to select plants that contain the alleles of
interest during a backcrossing
breeding program. The markers can also be used to select for the genome of the
recurrent parent and
against the genome of the donor parent. Using this procedure can minimize the
amount of genome from
the donor parent that remains in the selected plants. It can also be used to
reduce the number of crosses
back to the recurrent parent needed in a backcrossing program. The use of
molecular markers in the
selection process is often called genetic marker enhanced selection. Molecular
markers may also be
used to identify and exclude certain sources of germplasm as parental
varieties or ancestors of a plant by
providing a means of tracking genetic profiles through crosses.
Genome-wide selection (GWS)/genomic selection (GS) can also be used as an
alternative to, or in
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Date Recue/Date Received 2022-09-28
combination to, marker assisted selection and phenotype selection. GS utilizes
quantitative models over
a large number of markers distributed across the genome to predict the genomic
estimated breeding
values (GEBVs) of individual plants that has been genotyped but not
phenotyped. GS can improve
complex traits or combination of multiple traits without the need to identify
markers associated with the
traits. GS can replace phenotyping in a few selection cycles, thus reducing
the cost and the time required
for variety development (Crossa et al., Trends in Plant Science, November
2017, Vol. 22, No.11).
PRODUCTION OF DOUBLE HAPLOIDS
The production of double haploids can also be used for the development of
plants with a
homozygous phenotype in the breeding program. For example, a canola plant for
which canola variety
SCV686590 is a parent can be used to produce double haploid plants. Double
haploids are produced by
the doubling of a set of chromosomes (1 N) from a heterozygous plant to
produce a completely
homozygous individual. This can be advantageous because the process omits the
generations of selfing
needed to obtain a homozygous plant from a heterozygous source.
Haploid induction systems have been developed for various plants to produce
haploid tissues,
plants and seeds. Thus, an embodiment of this invention is a process for
making a substantially
homozygous SCV686590 progeny plant by producing or obtaining a seed from the
cross of SCV686590
and another canola plant and applying double haploid methods to the F 1 seed
or F 1 plant or to any
successive filial generation. Based on studies in maize and currently being
conducted in canola, such
methods would decrease the number of generations required to produce a variety
with similar genetics
or characteristics to 5CV686590.
In particular, a process of making seed retaining the molecular marker profile
of canola variety
SCV686590 is contemplated, such process comprising obtaining or producing Fi
seed for which canola
variety 5CV686590 is a parent, inducing doubled haploids to create progeny
without the occurrence of
meiotic segregation, obtaining the molecular marker profile of canola variety
5CV686590, and selecting
progeny that retain the molecular marker profile of 5CV686590.
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
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Date Recue/Date Received 2022-09-28
hybrid production. It provides stable expression of the male sterility trait
and an effective nuclear restorer
gene.
In developing improved new Brassica hybrid varieties, breeders use self-
incompatible (SI),
cytoplasmic male sterile (CMS) and 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 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 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.
In one instance, production of F 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 Fi
hybrid must have a fertility restorer gene (Rf gene). The presence of an Rf
gene means that the F 1
generation will not be completely or partially sterile, so that either self-
pollination or cross pollination
may occur. Self pollination of the F 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.
An example of a Brassica plant which is cytoplasmic male sterile and used for
breeding is ogura
(OGU) cytoplasmic male sterile. A fertility restorer for ogura cytoplasmic
male sterile plants has been
transferred from Raphanus sativus (radish) to Brassica. The restorer gene is
Rf 1, originating from radish.
Improved versions of this restorer have been developed as well. Other sources
and refinements of CMS
sterility in canola include the Polima cytoplasmic male sterile plant.
Further, as a result of the advances in sterility systems, varieties are
developed that can be used
as an open pollinated line (i.e. a pure line sold to the grower for planting)
and/or as a sterile inbred
(female) used in the production of F 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.
The development of a canola hybrid in a canola plant breeding program involves
three steps: (1)
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Date Recue/Date Received 2022-09-28
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.
Combining ability of a line, as well as the performance of the line per se, is
a factor in the selection
of improved canola varieties 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 is adjusted for known genetic
relationships among the
lines.
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, either through visual or molecular methods.
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
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Date Recue/Date Received 2022-09-28
insect pollinators at a given growing site are greater. Thus open pollination
is often used in commercial
canola production.
INDUSTRIAL USES
Currently Brassica napus canola is 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.
Canola oil has the lowest level of saturated fatty acids of all vegetable
oils. "Canola" refers to
rapeseed (Brassica) which has an erucic acid (C22 1) content of at most 2
percent by weight based on the
total fatty acid content of a seed, and which produces, after crushing, an air-
dried meal containing less
than 30 micromoles ( mol) 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.
Canola variety SCV686590 can be used in the production of an edible vegetable
oil or other food
products in accordance with known techniques. The solid meal component derived
from seeds can be
used as a nutritious livestock feed. Parts of the plant not used for human or
animal food can be used for
biofuel.
Date Recue/Date Received 2022-09-28
DEPOSIT INFORMATION
A deposit of the canola variety SCV686590, which is disclosed herein above and
referenced in the claims, was made with the Provasoli-Guillar National Center
for Marine Algae
and Microbiota (NCMA) at Bigelow Laboratory for Ocean Sciences, 60 Bigelow
Drive, East
Boothbay, ME 04544 USA. The date of deposit was June 1, 2022 and the accession
number
for those deposited seeds of canola variety 5CV686590 is NCMA Accession No.
202206003.
This deposit will be maintained under the terms of the Budapest Treaty on the
International
Recognition of the Deposit of Microorganisms for the Purposes of Patent
Procedure. These
deposits are not an admission that a deposit is required under Section 27(3)
and 38.1(1) of the
Patent Act.
While a number of exemplary aspects and embodiments have been discussed above,
those of skill in the art will recognize certain modifications, permutations,
additions and sub-
combinations thereof. It is therefore intended that the following appended
claims and claims
hereafter introduced are interpreted to include all such modifications,
permutations, additions
and sub-combinations as are within their true spirit and scope.
The scope of the claims should not be limited by the preferred embodiments set
forth
herein, but should be given the broadest interpretation consistent with the
description as a
whole.
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Date Recue/Date Received 2022-09-28
