NZ598733B - Pea line dlsc7v0955 - Google Patents
Pea line dlsc7v0955 Download PDFInfo
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- NZ598733B NZ598733B NZ598733A NZ59873312A NZ598733B NZ 598733 B NZ598733 B NZ 598733B NZ 598733 A NZ598733 A NZ 598733A NZ 59873312 A NZ59873312 A NZ 59873312A NZ 598733 B NZ598733 B NZ 598733B
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- pea
- dlsc7v0955
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- seed
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
- A01H5/10—Seeds
Abstract
Patent 598733 Disclosed is a seed or plant of pea line DLSC7V0955, a sample of seed of said line having been deposited under ATCC Accession Number PTA-11600.
Description
NEW ZEALAND
Patents Act 1953
Patents Form No. 5
COMPLETE SPECIFICATION
Title:
PEA LINE DLSC7V0955
We, Seminis Vegetable Seeds, Inc., of 800 N. Lindbergh Blvd., St. Louis, MO 63167,
United States of America (United States), do hereby declare the invention for which we
pray that a patent may be granted to us and the method by which it is to be performed, to
be particularly described in and by the following statement.
FIELD OF THE INVENTION
The present invention relates to the field of plant breeding and, more specifically,
to the development of pea line DLSC7V0955.
BACKGROUND OF THE INVENTION
The goal of vegetable breeding is to combine various desirable traits in a single
variety/hybrid. Such desirable traits may include greater yield, resistance to insects or pests,
tolerance to heat and drought, better agronomic quality, higher nutritional value, growth rate and
fruit or pod properties.
Breeding techniques take advantage of a plant’s method of pollination. There are
two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred
to the same or another flower of the same plant or plant variety. A plant cross-pollinates if
pollen comes to it from a flower of a different plant variety.
Plants that have been self-pollinated and selected for type over many generations
become homozygous at almost all gene loci and produce a uniform population of true breeding
progeny, a homozygous plant. A cross between two such homozygous plants of different
varieties produces a uniform population of hybrid plants that are heterozygous for many gene
loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a
population of hybrid plants that differ genetically and are not uniform. The resulting non-
uniformity makes performance unpredictable.
The development of uniform varieties requires the development of homozygous
inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree
breeding and recurrent selection are examples of breeding methods that have been used to
develop inbred plants from breeding populations. Those breeding methods combine the genetic
backgrounds from two or more plants or various other broad-based sources into breeding pools
from which new lines are developed by selfing and selection of desired phenotypes. The new
lines are evaluated to determine which of those have commercial potential.
Pea plants are able to reproduce by self-fertilization and cross-fertilization. Thus
far, however, commercial pea varieties have been inbred lines prepared through self fertilization
(McPhee, 2005).
Peas are one of the top vegetables used for processing in the United States; with
approximately 90% of the grown pea acreage used for processed consumption (NASS Census of
Agriculture 2002). The pea is an annual cool season plant, growing best in slightly acidic soil.
Many cultivars reach maturity about 60 days after planting. Pea plants can have both low-
growing and vining cultivars. The vining cultivars grow thin tendrils from the leaves of the plant,
which coil around available supports. The pea pods form at the leaf axils of the plant.
As with other legumes, pea plants are able to obtain fixed nitrogen compounds
from symbiotic soil bacteria. Pea plants therefore have a substantially reduced fertilizer
requirement compared to non-leguminous crops. This advantage adds to their commercial value,
particularly in view of increasing fertilizer costs, and has generated considerable interest in the
creation of new pea plant cultivars.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a pea plant of the line designated
DLSC7V0955. Also provided are pea plants having all the physiological and morphological
characteristics of the pea line designated DLSC7V0955. Parts of the pea plant of the present
invention are also provided, for example, including pollen, an ovule, a seed, a pod, and a cell of
the plant.
The invention also concerns the seed of pea line DLSC7V0955. The pea seed of
the invention may be provided as an essentially homogeneous population of pea seed of the line
designated DLSC7V0955. Essentially homogeneous populations of seed are generally free from
substantial numbers of other seed. Therefore, certain embodiments of the invention concern seed
of line DLSC7V0955 that forms at least about 97% of the total seed, including at least about
98%, 99% or more of the seed. The population of pea seed may be particularly defined as being
essentially free from hybrid seed. The seed population may be separately grown to provide an
essentially homogeneous population of pea plants designated DLSC7V0955.
In another aspect of the invention, a plant of pea line DLSC7V0955 comprising
an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for
example, a dominant or recessive allele. In one embodiment of the invention, a plant of pea line
DLSC7V0955 is defined as comprising a single locus conversion. In specific embodiments of
the invention, an added genetic locus confers one or more traits such as, for example, herbicide
tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In
further embodiments, the trait may be conferred by a naturally occurring gene introduced into the
genome of the line by backcrossing, a natural or induced mutation, or a transgene introduced
through genetic transformation techniques into the plant or a progenitor of any previous
generation thereof. When introduced through transformation, a genetic locus may comprise one
or more genes integrated at a single chromosomal location.
In another aspect of the invention, a tissue culture of regenerable cells of a pea
plant of line DLSC7V0955 is provided. The tissue culture will preferably be capable of
regenerating pea plants capable of expressing all of the physiological and morphological
characteristics of the line, and of regenerating plants having substantially the same genotype as
other plants of the line. Examples of some of the physiological and morphological
characteristics of the line DLSC7V0955 include those traits set forth in the tables herein. The
regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems,
cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks. Still further,
the present invention provides pea plants regenerated from a tissue culture of the invention, the
plants having all the physiological and morphological characteristics of line DLSC7V0955.
In yet another aspect of the invention, processes are provided for producing pea
seeds, pods and plants, which processes generally comprise crossing a first parent pea plant with
a second parent pea plant, wherein at least one of the first or second parent pea plants is a plant
of the line designated DLSC7V0955. These processes may be further exemplified as processes
for preparing hybrid pea seed or plants, wherein a first pea plant is crossed with a second pea
plant of a different, distinct line to provide a hybrid that has, as one of its parents, the pea plant
line DLSC7V0955. In these processes, crossing will result in the production of seed. The seed
production occurs regardless of whether the seed is collected or not.
In one embodiment of the invention, the first step in “crossing” comprises
planting seeds of a first and second parent pea plant, often in proximity so that pollination will
occur for example, mediated by insect vectors. Alternatively, pollen can be transferred
manually. Where the plant is self-pollinated, pollination may occur without the need for direct
human intervention other than plant cultivation.
A second step may comprise cultivating or growing the seeds of first and second
parent pea plants into plants that bear flowers. A third step may comprise preventing self-
pollination of the plants, such as by emasculating the male portions of flowers, (i.e., treating or
manipulating the flowers to produce an emasculated parent pea plant). Self-incompatibility
systems may also be used in some hybrid crops for the same purpose. Self-incompatible plants
still shed viable pollen and can pollinate plants of other varieties but are incapable of pollinating
themselves or other plants of the same line.
A fourth step for a hybrid cross may comprise cross-pollination between the first
and second parent pea plants. Yet another step comprises harvesting the seeds from at least one
of the parent pea plants. The harvested seed can be grown to produce a pea plant or hybrid pea
plant.
The present invention also provides the pea seeds and plants produced by a
process that comprises crossing a first parent pea plant with a second parent pea plant, wherein at
least one of the first or second parent pea plants is a plant of the line designated DLSC7V0955.
In one embodiment of the invention, pea seed and plants produced by the process are first
generation (F ) hybrid pea seed and plants produced by crossing a plant in accordance with the
invention with another, distinct plant. The present invention further contemplates plant parts of
such an F hybrid pea plant, and methods of use thereof. Therefore, certain exemplary
embodiments of the invention provide an F hybrid pea plant and seed thereof.
In still yet another aspect, the present invention provides a method of producing a
plant derived from line DLSC7V0955, the method comprising the steps of: (a) preparing a
progeny plant derived from line DLSC7V0955, wherein said preparing comprises crossing a
plant of the line DLSC7V0955 with a second plant; and (b) crossing the progeny plant with
itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In
further embodiments, the method may additionally comprise: (c) growing a progeny plant of a
subsequent generation from said seed of a progeny plant of a subsequent generation and crossing
the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps
for an additional 3-10 generations to produce a plant derived from line DLSC7V0955. The plant
derived from line DLSC7V0955 may be an inbred line, and the aforementioned repeated
crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In
the method, it may be desirable to select particular plants resulting from step (c) for continued
crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a
plant derived from line DLSC7V0955 is obtained which possesses some of the desirable traits of
the line as well as potentially other selected traits.
In certain embodiments, the present invention provides a method of producing
peas comprising: (a) obtaining a plant of pea line DLSC7V0955, wherein the plant has been
cultivated to maturity, and (b) collecting peas from the plant.
In still yet another aspect of the invention, the genetic complement of the pea
plant line designated DLSC7V0955 is provided. The phrase “genetic complement” is used to
refer to the aggregate of nucleotide sequences, the expression of which sequences defines the
phenotype of, in the present case, a pea plant, or a cell or tissue of that plant. A genetic
complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic
complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus
provides pea plant cells that have a genetic complement in accordance with the pea plant cells
disclosed herein, and plants, seeds and plants containing such cells.
Plant genetic complements may be assessed by genetic marker profiles, and by
the expression of phenotypic traits that are characteristic of the expression of the genetic
complement, e.g., isozyme typing profiles. It is understood that line DLSC7V0955 could be
identified by any of the many well known techniques such as, for example, Simple Sequence
Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified Polymorphic
DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified
Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified
Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein by
reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., 1998).
In still yet another aspect, the present invention provides hybrid genetic
complements, as represented by pea plant cells, tissues, plants, and seeds, formed by the
combination of a haploid genetic complement of a pea plant of the invention with a haploid
genetic complement of a second pea plant, preferably, another, distinct pea plant. In another
aspect, the present invention provides a pea plant regenerated from a tissue culture that
comprises a hybrid genetic complement of this invention.
Any embodiment discussed herein with respect to one aspect of the invention
applies to other aspects of the invention as well, unless specifically noted.
The term “about” is used to indicate that a value includes the standard deviation
of error for the device or method being employed to determine the value. The use of the term
“or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only
or the alternatives are mutually exclusive, although the disclosure supports a definition that refers
to only alternatives and to “and/or.” When used in conjunction with the word “comprising” or
other open language in the claims, the words “a” and “an” denote “one or more,” unless
specifically noted. The terms “comprise,” “have” and “include” are open-ended linking verbs.
Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,”
“having,” “includes” and “including,” are also open-ended. For example, any method that
“comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or
more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or
“includes” one or more traits is not limited to possessing only those one or more traits and covers
other unlisted traits.
Other objects, features and advantages of the present invention will become
apparent from the following detailed description. It should be understood, however, that the
detailed description and any specific examples provided, while indicating specific embodiments
of the invention, are given by way of illustration only, since various changes and modifications
within the spirit and scope of the invention will become apparent to those skilled in the art from
this detailed description.
BRIEF DESCRIPTION OF THE DRAWINGS
is a graph showing yield per sieve size of peas of line DLSC7V0955 and
several selected varieties. The graph is a column representation of the data given in Table 2, for
values at 100 tenderometer. The different hatching represent the amount of peas in the different
sieve size, as indicated. The number after the name of the variety stands for Maturity in days at
harvest compared to the reference variety SPRING. The alternate designation “085 7 0955” is
used for line DLSC7V0955 in the figure. These designations have the same meaning.
DETAILED DESCRIPTION OF THE INVENTION
The invention provides methods and compositions relating to plants, seeds and
derivatives of pea line DLSC7V0955. This line shows uniformity and stability within the limits
of environmental influence for the traits described hereinafter. Pea line DLSC7V0955 provides
sufficient seed yield. By crossing with a distinct second plant, uniform F1 hybrid progeny can be
obtained. The development of pea line DLSC7V0955 can be summarized as follows.
A. Origin and Breeding History of Pea Line DLSC7V0955
DLSC7V0955 is a mid-early maturing large sieve dark green pea with a normal
(leafy) foliage. It was selected based on productivity, disease resistance and enhanced
sweetness: DLSC7V0955 caries a dominant allele for resistance to race 0 of the downy mildew
fungus, Peronospora viciae, that traces back to JI85 (a wild accession of the John Innes
collection), the Fw1 and Fw2 alleles for resistance to race 1 and 2 of the wilt fungus, Fusarium
oxysporum fsp pisi. It has a unique earliness in combination with these diseases resistances and
an enhanced sweetness given by the presence of two alleles for wrinkled seed r and rb.
DLSC7V0955 was developed by pedigree selection at Filer, Idaho. The initial
cross that led to the development of DLSC7V0955 was made between the Seminis variety Early
Sweet 414 as seed parent and a Seminis breeding lines of complex parentage: 2513ASKrSpDM
as pollen parent. Early Sweet 414 is an early large sieve pea with normal foliage, which carries
the Fw1 allele for resistance to Race 1 of the wilt fungus Fusarium oxysporum fsp pisi, known
for its high yield potential. 2513ASKr derived from a cross of the variety Alsweet III which
carries the alleles R and rb giving the seed characteristic named “Alsweet” and a Seminis variety
KRITER, selected for plant type improvement and bringing the Fw1 and Fw2 alleles for resistance
to race 1 and 2 of the wilt fungus Fusarium oxysporum fsp pisi in the new variety.
SpDM is derived from a backcross of the variety Spring on to JI85 (a wild
accession of the John Innes collection), selected for resistance to Peronospora viciae. The goal
of the cross between those 2 lines was to put the double recessive alleles r and rb, in the right
Peronospora viciae resistance and plant type background. The cross was followed by 7
generations of self-pollination (in field and greenhouse) until fixation. It was selected based on
productivity, diseases resistance, adaptation to different environment and sweetness of the fresh
product
B. Physiological and Morphological Characteristics of Pea Line DLSC7V0955
In accordance with one aspect of the present invention, there is provided a plant
having the physiological and morphological characteristics of pea line DLSC7V0955. A
description of the physiological and morphological characteristics of pea line DLSC7V0955 is
presented in Table 1.
Table 1: Physiological and Morphological Characteristics of Line DLSC7V0955
CHARACTERISTIC DLSC7V0955
1. Type Garden
2. Maturity
node number of first bloom 11
no. of days processing 89
heat units 1290
9 days earlier than Wando
3. Plant Height
height 60 cm
cm shorter than check cultivar Karina
4. Vine
habit indeterminate
branching 1-2 branches (Little Marvel)
internodes straight
stockiness medium (Thomas Laxton WR)
total number of nodes 16
. Leaflets
color dark green (Alderman)
wax medium
molding marbled (Alaska)
number of leaflet pairs two
leaflet type normal
6. Stipules
present or lacking? present
clasping or not clasping? clasping
marbled or not marbled? marbled
size (compared with leaflets) larger
color (compared with leaflets) same
color dark green
CHARACTERISTIC DLSC7V0955
stipule size medium
Comparative Variety (1):
variety name: Karina
stipule size: large
color: same
7. Flower Color
venation greenish
standard greenish
wing greenish
keel greenish
8. Pods
shape straight
end blunt (Alaska)
color dark green (Alderman)
surface (smooth or rough?) smooth
surface (shiny or dull?) dull
borne single and double
length 7 cm
width (between sutures) 11 mm
number of seeds per pod 7
9. Seeds (95-100 Tenderometer)
color dark green
sieve (% of seeds of indicated size)
1 10
2 15
3 30
4 35
10
average sieve size 3.5
. Seeds (dry-mature)
CHARACTERISTIC DLSC7V0955
shape angular
surface wrinkled
luster dull
color pattern monocolor
primary color cream & green
secondary color cream & green
hilum color white
cotyledon color green
grams per 100 seeds 17
11. Disease
Fusarium Wilt-Race 1 resistant
Fusarium Wilt (Near Wilt)-Race 2 resistant
Ascochyta Blight not tested
Common Mosaic Virus not tested
Bacterial Blight not tested
Pea Enation Mosaic Virus susceptible
Downy Mildew resistant
Seedborne Mosaic Virus not tested
Powdery Mildew susceptible
Yellow Bean Mosaic Virus resistant
Leaf Roll Virus not tested
12. Insect
Aphids not tested
*These are typical values. Values may vary due to environment. Other values that are
substantially equivalent are within the scope of the invention.
C. Breeding Pea Line DLSC7V0955
One aspect of the current invention concerns methods for crossing the pea line
DLSC7V0955 with itself or a second plant and the seeds and plants produced by such methods.
These methods can be used for propagation of line DLSC7V0955, or can be used to produce
hybrid pea seeds and the plants grown therefrom. Hybrid seeds are produced by crossing line
DLSC7V0955 with second pea parent line.
The development of new varieties using one or more starting varieties is well
known in the art. In accordance with the invention, novel varieties may be created by crossing
line DLSC7V0955 followed by multiple generations of breeding according to such well known
methods. New varieties may be created by crossing with any second plant. In selecting such a
second plant to cross for the purpose of developing novel lines, it may be desired to choose those
plants which either themselves exhibit one or more selected desirable characteristics or which
exhibit the desired characteristic(s) when in hybrid combination. Once initial crosses have been
made, inbreeding and selection take place to produce new varieties. For development of a
uniform line, often five or more generations of selfing and selection are involved.
Uniform lines of new varieties may also be developed by way of double-haploids.
This technique allows the creation of true breeding lines without the need for multiple
generations of selfing and selection. In this manner true breeding lines can be produced in as
little as one generation. Haploid embryos may be produced from microspores, pollen, anther
cultures, or ovary cultures. The haploid embryos may then be doubled autonomously, or by
chemical treatments (e.g. colchicine treatment). Alternatively, haploid embryos may be grown
into haploid plants and treated to induce chromosome doubling. In either case, fertile
homozygous plants are obtained. In accordance with the invention, any of such techniques may
be used in connection with line DLSC7V0955 and progeny thereof to achieve a homozygous
line.
Backcrossing can also be used to improve an inbred plant. Backcrossing
transfers a specific desirable trait from one inbred or non-inbred source to an inbred that lacks
that trait. This can be accomplished, for example, by first crossing a superior inbred (A)
(recurrent parent) to a donor inbred (non-recurrent parent), which carries the appropriate locus or
loci for the trait in question. The progeny of this cross are then mated back to the superior
recurrent parent (A) followed by selection in the resultant progeny for the desired trait to be
transferred from the non-recurrent parent. After five or more backcross generations with
selection for the desired trait, the progeny are heterozygous for loci controlling the characteristic
being transferred, but are like the superior parent for most or almost all other loci. The last
backcross generation would be selfed to give pure breeding progeny for the trait being
transferred.
The line of the present invention is particularly well suited for the development of
new lines based on the elite nature of the genetic background of the line. In selecting a second
plant to cross with DLSC7V0955 for the purpose of developing novel pea lines, it will typically
be preferred to choose those plants which either themselves exhibit one or more selected
desirable characteristics or which exhibit the desired characteristic(s) when in hybrid
combination. Examples of potentially desirable traits include, but are not necessarily limited to,
improved resistance to viral, fungal, and bacterial pathogens, improved insect resistance, pod
morphology, herbicide tolerance, environmental tolerance (e.g. tolerance to temperature,
drought, and soil conditions, such as acidity, alkalinity, and salinity), growth characteristics,
nutritional content, taste, and texture. Improved taste and texture applies not only to the peas
themselves, but also to the pods of those varieties yielding edible pods. In peas, as in other
legumes, taste and nutritional content are particularly affected by the sucrose and starch content.
Among fungal diseases of particular concern in peas are Ascochyla pisi,
Cladosporium pisicola (leaf spot or scab), Erysiphe polygoni (powdery mildew), Fusarium
oxysporum (wilt), Fusarium solani (Fusarium root rot), Mycosphaerella pinodes (Mycospharella
blight), Peronospora viciae (downy mildew), Phythium sp. (pre emergence damping-off),
Botrytis cinerea (grey mold), Aphanomyces euteiches (common root rot), Thielaviopsis basicola
(black root rot), and Sclerotina sclerotiorum (sclerotina white mold). Pea plant viral diseases
include: Bean yellow mosaic virus (BYMV), Bean leaf roll virus (BLRV), Pea Early Browning
Virus (PEBV), Pea Enation Mosaic virus (PEMV), Pea Mosaic Virus (PMV), Pea seed-borne
Mosaic Virus (PSbMV) and Pea Streak Virus (PSV). An important bacterial disease affecting
pea plants is caused by Pseudomonas pisi (bacterial blight), (Muehlbauer et al., 1983; Davies et
al. 1985; van Emden et al., 1988).
Insect pests that may be of particular concern in peas include Aphis cracivora
(Groundnut aphid), Acyrthosiphon pisum (Pea aphid), Kakothrips robustus (Pea thrips), Bruchis
pisorum (Pea seed beetle), Callosobruchus chinensis (Adzuki bean seed beetle), Apion sp. (Seed
weevil), Sitona lineatus (Bean weevil), Contarina pisi (Pea midge), Helicoverpa armigera
(African bollworm), Diachrysia obliqua (Pod borer), Agriotis sp. (Cut worms), Cydia nigricana
(Pea moth), Phytomuza horticola (Leaf minor), Heliothis Zea (American bollworm), Etiella
Zinckenella (Lima bean pod borer), Ophiomyia phaseoli (Bean fly), Delia platura (Bean seed
fly), Tetranychus sp. (Spider mites), Pratylenchus penetrants (Root lesion nematodes),
Ditylenchus dipsaci (Stem nematode), Heterodera goettingiana (Pea cyst nematode), and
Meloidogyne javanica (Root knot nematode), (van Emden et al., 1988; Muehlbauer et al., 1983).
D. Performance Characteristics
Performance characteristics of the line DLSC7V0955 were the subject of an
objective analysis of the performance traits of the line relative to other lines. The results of the
analysis are presented below.
Table 2: Performance Data For Line DLSC7V0955 and Selected Varieties
* The alternate designation “085 7 0955” is used to refer to line DLSC7V0955 in the table. These designations have the same meaning.
Nber Years = Number of years of field trials from which data was collected and pooled.;
Fol = type of foliage (A=Afila, Afa=Afila faciated);
FullFl = days to full flowering;
HU = Heat Units to harvest;
Mat = Maturity in days at harvest compared to the reference variety SPRING;
Yield (qx/ha): in quintals (100kg), quantity of fresh peas harvested on one Hectar;
TdrTV = Tenderometer value at harvest;
<7,5 to >10,2 = in mm, the percentage of berries with the mentioned caliber;
AV SS = coefficient calculated from the amount of peas per caliber;
Thresh : percentage of non full pods in the total harvest;
Col. Fresh = Fresh color of the berries;
Col.af.Blanch = Color of the berries after blanching;
AIS = percentage of Alcohol Insoluble Solids in the berries at the given Tenderometer value;
Tdr at AIS 12% : Tenderometer value at which you should harvest the peas in order to have an AIS % of 12.
E. Further Embodiments of the Invention
In certain aspects, the invention provides plants modified to include at least a first
desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding
technique called backcrossing, wherein essentially all of the morphological and physiological
characteristics of a variety are recovered in addition to a genetic locus transferred into the plant
via the backcrossing technique. The term single locus converted plant as used herein refers to
those pea plants which are developed by a plant breeding technique called backcrossing, wherein
essentially all of the desired morphological and physiological characteristics of a variety are
recovered in addition to the single locus 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 present variety. The parental pea plant which contributes the
locus 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 pea plant to which the locus or loci 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 (recurrent parent) is
crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be
transferred. The resulting progeny from this cross are then crossed again to the recurrent parent
and the process is repeated until a pea 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 locus 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 locus of the recurrent variety is
modified or substituted with the desired locus from the nonrecurrent parent, while retaining
essentially all of the rest of the desired genetic, and therefore the desired physiological and
morphological constitution of the original variety. The choice of the particular nonrecurrent
parent will depend on the purpose of the backcross; one of the major purposes is to add some
commercially desirable trait to the plant. The exact backcrossing protocol will depend on the
characteristic or trait being altered and the genetic distance between the recurrent and
nonrecurrent parents. Although backcrossing methods are simplified when the characteristic
being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive
and dominant), 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.
In one embodiment, progeny pea plants of a backcross in which DLSC7V0955 is
the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the
physiological and morphological characteristics of pea line DLSC7V0955 as determined at the
% significance level when grown in the same environmental conditions.
Pea varieties can also be developed from more than two parents. The technique,
known as modified backcrossing, uses different recurrent parents during the backcrossing.
Modified backcrossing may be used to replace the original recurrent parent with a variety having
certain more desirable characteristics or multiple parents may be used to obtain different
desirable characteristics from each.
Many single locus traits have been identified that are not regularly selected for in
the development of a new inbred but that can be improved by backcrossing techniques. Single
locus traits may or may not be transgenic; examples of these traits include, but are not limited to,
male sterility, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect
resistance, restoration of male fertility, modified fatty acid or carbohydrate metabolism, and
enhanced nutritional quality. These comprise genes generally inherited through the nucleus.
Direct selection may be applied where the single locus acts as a dominant trait.
An example of a dominant trait is the downy mildew resistance trait. For this selection process,
the progeny of the initial cross are sprayed with downy mildew spores prior to the backcrossing.
The spraying eliminates any plants which do not have the desired downy mildew resistance
characteristic, and only those plants which have the downy mildew resistance gene are used in
the subsequent backcross. This process is then repeated for all additional backcross generations.
Selection of pea plants for breeding is not necessarily dependent on the phenotype
of a plant and instead can be based on genetic investigations. For example, one can utilize a
suitable genetic marker which is closely genetically linked to a trait of interest. One of these
markers can be used to identify the presence or absence of a trait in the offspring of a particular
cross, and can be used in selection of progeny for continued breeding. This technique is
commonly referred to as marker assisted selection. Any other type of genetic marker or other
assay which is able to identify the relative presence or absence of a trait of interest in a plant can
also be useful for breeding purposes. Procedures for marker assisted selection applicable to the
breeding of pea are well known in the art. Such methods will be of particular utility in the case
of recessive traits and variable phenotypes, or where conventional assays may be more
expensive, time consuming or otherwise disadvantageous. Types of genetic markers which
could be used in accordance with the invention include, but are not necessarily limited to, Simple
Sequence Length Polymorphisms (SSLPs) (Williams et al., 1990), Randomly Amplified
Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence
Characterized Amplified Regions (SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-
PCR), Amplified Fragment Length Polymorphisms (AFLPs) (EP 534 858, specifically
incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs)
(Wang et al., 1998).
F. Plants Derived From Pea Line DLSC7V0955 by Genetic Engineering
Many useful traits that can be introduced by backcrossing, as well as directly into
a plant, can also be introduced by genetic transformation techniques. Genetic transformation
may therefore be used to insert a selected transgene into the pea line of the invention or may,
alternatively, be used for the preparation of transgenes which can be introduced by backcrossing.
Methods for the transformation of plants, including pea plants, are well known to those of skill in
the art (see, e.g., Schroeder et al., 1993). Techniques which may be employed for the genetic
transformation of pea plants include, but are not limited to, electroporation, microprojectile
bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.
To effect transformation by electroporation, one may employ either friable
tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may
transform immature embryos or other organized tissue directly. In this technique, one would
partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading
enzymes (pectolyases) or mechanically wound tissues in a controlled manner.
A particularly efficient method for delivering transforming DNA segments to
plant cells is microprojectile bombardment. In this method, particles are coated with nucleic
acids and delivered into cells by a propelling force. Exemplary particles include those comprised
of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are
concentrated on filters or solid culture medium. Alternatively, immature embryos or other target
cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an
appropriate distance below the macroprojectile stopping plate.
Microprojectile bombardment techniques are widely applicable, and may be used
to transform virtually any plant species. An illustrative embodiment of a method for delivering
DNA into plant cells by bombardment is the Biolistics Particle Delivery System, which can be
used to propel particles coated with DNA or cells through a screen, such as a stainless steel or
Nytex screen, onto a surface covered with target pea cells. The screen disperses the particles so
that they are not delivered to the recipient cells in large aggregates.
Agrobacterium-mediated transfer is another widely applicable system for
introducing gene loci into plant cells. An advantage of the technique is that DNA can be
introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant
from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E.
coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., 1985).
Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer
have improved the arrangement of genes and restriction sites in the vectors to facilitate the
construction of vectors capable of expressing various polypeptide coding genes. The vectors
described have convenient multi-linker regions flanked by a promoter and a polyadenylation site
for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium
containing both armed and disarmed Ti genes can be used for transformation.
In those plant strains where Agrobacterium-mediated transformation is efficient, it
is the method of choice because of the facile and defined nature of the gene locus transfer. The
use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is
well known in the art (Fraley et al., 1985; U.S. Patent No. 5,563,055). Agrobacterium-mediated
transformation is a particularly beneficial method for producing recombinant pea-plants.
Transformed pea plants may be obtained by incubating pea explant material with Agrobacterium
containing the DNA sequence of interest (U.S. Patent No. 5,286,635; U.S. Patent No. 5,773,693).
Transformation of plant protoplasts also can be achieved using methods based on
calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and
combinations of these treatments (see, e.g., Potrykus et al., 1985; Omirulleh et al., 1993; Fromm
et al., 1986; Uchimiya et al., 1986; Marcotte et al., 1988). Transformation of plants and
expression of foreign genetic elements is exemplified in Choi et al. (1994), and Ellul et al.
(2003).
A number of promoters have utility for plant gene expression for any gene of
interest including but not limited to selectable markers, scoreable markers, genes for pest
tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest.
Examples of constitutive promoters useful for pea plant gene expression include, but are not
limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive,
high-level expression in most plant tissues (see, e.g., Odel et al., 1985), including monocots (see,
e.g., Dekeyser et al., 1990; Terada and Shimamoto, 1990); a tandemly duplicated version of the
CaMV 35S promoter, the enhanced 35S promoter (P-e35S) the nopaline synthase promoter (An
et al., 1988), the octopine synthase promoter (Fromm et al., 1989); and the figwort mosaic virus
(P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV
promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem, the
cauliflower mosaic virus 19S promoter, a sugarcane bacilliform virus promoter, a commelina
yellow mottle virus promoter, and other plant DNA virus promoters known to express in plant
cells.
A variety of plant gene promoters that are regulated in response to environmental,
hormonal, chemical, and/or developmental signals can be used for expression of an operably
linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., 1988), (2) light
(e.g., pea rbcS-3A promoter, Kuhlemeier et al., 1989; maize rbcS promoter, Schaffner and
Sheen, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., 1985), (3) hormones,
such as abscisic acid (Marcotte et al., 1989), (4) wounding (e.g., wunl, Siebertz et al., 1989); or
(5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous
to employ organ-specific promoters (e.g., Roshal et al., 1987; Schernthaner et al., 1988; Bustos
et al., 1989).
Exemplary nucleic acids which may be introduced to the pea lines of this
invention include, for example, DNA sequences or genes from another species, or even genes or
sequences which originate with or are present in the same species, but are incorporated into
recipient cells by genetic engineering methods rather than classical reproduction or breeding
techniques. However, the term “exogenous” is also intended to refer to genes that are not
normally present in the cell being transformed, or perhaps simply not present in the form,
structure, etc., as found in the transforming DNA segment or gene, or genes which are normally
present and that one desires to express in a manner that differs from the natural expression
pattern, e.g., to over-express. Thus, the term "exogenous" gene or DNA is intended to refer to
any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar
gene may already be present in such a cell. The type of DNA included in the exogenous DNA
can include DNA which is already present in the plant cell, DNA from another plant, DNA from
a different organism, or a DNA generated externally, such as a DNA sequence containing an
antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a
gene.
Many hundreds if not thousands of different genes are known and could
potentially be introduced into a pea plant according to the invention. Non-limiting examples of
particular genes and corresponding phenotypes one may choose to introduce into a pea plant
include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest
tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring
glyphosate tolerance, and genes for quality improvements such as yield, nutritional
enhancements, environmental or stress tolerances, or any desirable changes in plant physiology,
growth, development, morphology or plant product(s). For example, structural genes would
include any gene that confers insect tolerance including but not limited to a Bacillus insect
control protein gene as described in WO 99/31248, herein incorporated by reference in its
entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos.
,500,365 and 5,880,275, herein incorporated by reference it their entirety. In another
embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by
genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene
(aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its
entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175,
herein incorporated by reference in its entirety.
Alternatively, the DNA coding sequences can affect these phenotypes by
encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an
endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for
example, Bird et al., 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme)
engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito,
1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or
morphology change of interest is useful for the practice of the present invention.
G. Definitions
In the description and tables herein, 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:
Allele: Any of one or more alternative forms of a gene locus, all of which alleles
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.
Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny,
for example a first generation hybrid (F ), 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.
Crossing: The mating of two parent plants.
Cross-pollination: Fertilization by the union of two gametes from different
plants.
Diploid: A cell or organism having two sets of chromosomes.
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.
Enzymes: Molecules which can act as catalysts in biological reactions.
F Hybrid: The first generation progeny of the cross of two nonisogenic plants.
Genotype: The genetic constitution of a cell or organism.
Haploid: A cell or organism having one set of the two sets of chromosomes in a
diploid.
Linkage: A phenomenon wherein alleles on the same chromosome tend to
segregate together more often than expected by chance if their transmission was independent.
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.
Phenotype: The detectable characteristics of a cell or organism, which
characteristics are the manifestation of gene expression.
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.
Resistance: As used herein, the terms “resistance” and “tolerance” are used
interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen,
abiotic influence or environmental condition. These terms are also used to describe plants
showing some symptoms but that are still able to produce marketable product with an acceptable
yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they
may still produce a crop, even though the plants are stunted and the yield is reduced.
Regeneration: The development of a plant from tissue culture.
Royal Horticultural Society (RHS) color chart value: The RHS color chart is a
standardized reference which allows accurate identification of any color. A color’s designation
on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS
color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group
19A or Red Group 41B.
Self-pollination: The transfer of pollen from the anther to the stigma of the same
plant.
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 an inbred are recovered in addition to the characteristics
conferred by the single locus transferred into the inbred via the backcrossing technique. By
“essentially all,” it is meant that all of the characteristics of a plant are recovered that are
otherwise present when compared in the same environment and save for the converted locus,
other than an occasional variant trait that might arise during backcrossing or direct introduction
of a transgene. A single locus may comprise one gene, or in the case of transgenic plants, one or
more transgenes integrated into the host genome at a single site (locus).
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.
Transgene: A genetic locus comprising a sequence which has been introduced
into the genome of a pea plant by transformation.
H. Deposit Information
A deposit of pea line DLSC7V0955, disclosed above and recited in the claims,
has been made with the American Type Culture Collection (ATCC), 10801 University Blvd.,
Manassas, VA 20110-2209. The date of deposit was January 21, 2011. The accession number
for those deposited seeds of pea line DLSC7V0955 is ATCC Accession Number PTA-11600.
Upon issuance of a patent, all restrictions upon the deposit will be removed, and the deposit is
intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposit will be
maintained in the depository for a period of 30 years, or 5 years after the last request, or for the
effective life of the patent, whichever is longer, and will be replaced if necessary during that
period.
Although the foregoing invention has been described in some detail by way of
illustration and example for purposes of clarity and understanding, it will be obvious that certain
changes and modifications may be practiced within the scope of the invention, as limited only by
the scope of the appended claims.
All references cited herein are hereby expressly incorporated herein by reference.
REFERENCES
The following references, to the extent that they provide exemplary procedural or other
details supplementary to those set forth herein, are specifically incorporated herein by reference:
U.S. Patent 5,286,635
U.S. Patent 5,378,619
U.S. Patent 5,463,175
U.S. Patent 5,500,365
U.S. Patent 5,563,055
U.S. Patent 5,633,435
U.S. Patent 5,689,052
U.S. Patent 5,773,693
U.S. Patent 5,880,275
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Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991.
Bustos et al., Plant Cell, 1:839, 1989.
Callis et al., Plant Physiol., 88:965, 1988.
Choi et al., Plant Cell Rep., 13: 344–348, 1994.
Davies et al., In: Pea (Pisum sativum L.), Summerfield and Roberts (Eds.), Williams Collins
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Dekeyser et al., Plant Cell, 2:591, 1990.
Ellul et al., Theor. Appl. Genet., 107:462–469, 2003.
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Fromm et al., Nature, 312:791-793, 1986.
Fromm et al., Plant Cell, 1:977, 1989.
Gibson and Shillito, Mol. Biotech., 7:125,1997
Kevin McPhee, In: Journal of New Seeds: Innovations in production, biotechnology, quality,
and marketing; ISSN: 1522-886X, 6:2/3, 2005.
Klee et al., Bio-Technology, 3(7):637-642, 1985.
Kuhlemeier et al., Plant Cell, 1:471, 1989.
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Marcotte et al., Plant Cell, 1:969, 1989.
Muehlbauer et al., In: Description and culture of dry peas, USAD-ARS Agricultural Reviews
and Manuals, Western Region, California, 37:92, 1983.
NASS Census of Agriculture, 2002.
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Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993.
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Roshal et al., EMBO J., 6:1155, 1987.
Schaffner and Sheen, Plant Cell, 3:997, 1991.
Schernthaner et al., EMBO J., 7:1249, 1988.
Schroeder et al., Plant Physiol. 101(3): 751–757, 1993.
Siebertz et al., Plant Cell, 1:961, 1989.
Simpson et al., EMBO J., 4:2723, 1985.
Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990.
Uchimiya et al., Mol. Gen. Genet., 204:204, 1986.
van Emden et al., In: Pest, disease and weed problems in pea lentil faba bean and chickpea. p.,
Summerfield (Ed.), Kluwer Academic Publishers, Dordrecht, The Netherlands, 519-534,
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Claims (23)
1. A seed of pea line DLSC7V0955, a sample of seed of said line having been deposited under ATCC Accession Number PTA-11600.
2. A plant of pea line DLSC7V0955, a sample of seed of said line having been deposited under ATCC Accession Number PTA-11600.
3. A plant part of the plant of claim 2.
4. The plant part of claim 3, wherein said part is selected from the group consisting of a pod, pollen, an ovule and a cell.
5. A tissue culture of regenerable cells of pea line DLSC7V0955, a sample of seed of said line having been deposited under ATCC Accession Number PTA-11600.
6. The tissue culture according to claim 5, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.
7. A pea plant regenerated from the tissue culture of claim 5, wherein the regenerated plant expresses all of the physiological and morphological characteristics of pea line DLSC7V0955, a sample of seed of said line having been deposited under ATCC Accession Number PTA-11600.
8. A method of producing seed, comprising crossing the plant of claim 2 with itself or a second plant.
9. The method of claim 8, wherein the plant of pea line DLSC7V0955 is the female parent.
10. The method of claim 8, wherein the plant of pea line DLSC7V0955 is the male parent.
11. An F1 hybrid seed produced by the method of claim 8.
12. An F1 hybrid plant produced by growing the seed of claim 11.
13. A method for producing a seed of a line DLSC7V0955-derived pea plant comprising the steps of: (a) crossing a pea plant of line DLSC7V0955 with a second pea plant, a sample of seed of said line having been deposited under ATCC Accession Number PTA- 11600; and (b) allowing seed of a DLSC7V0955-derived pea plant to form.
14. The method of claim 13, further comprising the steps of: (c) crossing a plant grown from said DLSC7V0955-derived pea seed with itself or a second pea plant to yield additional DLSC7V0955-derived pea seed; (d) growing said additional DLSC7V0955-derived pea seed of step (c) to yield additional DLSC7V0955-derived pea plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate further DLSC7V0955-derived pea plants.
15. A method of vegetatively propagating a plant of pea line DLSC7V0955 comprising the steps of: (a) collecting tissue capable of being propagated from a plant of pea line DLSC7V0955, a sample of seed of said line having been deposited under ATCC Accession Number PTA-11600; (b) cultivating said tissue to obtain proliferated shoots; and (c) rooting said proliferated shoots to obtain rooted plantlets.
16. The method of claim 15, further comprising growing plants from said rooted plantlets.
17. A method of introducing a desired trait into pea line DLSC7V0955 comprising: (a) crossing a plant of line DLSC7V0955 with a second pea plant that comprises a desired trait to produce F1 progeny, a sample of seed of said line DLSC7V0955 having been deposited under ATCC Accession Number PTA-11600; (b) selecting an F1 progeny that comprises the desired trait; (c) crossing the selected F1 progeny with a plant of line DLSC7V0955 to produce backcross progeny; (d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristic of pea line DLSC7V0955; and (e) repeating steps (c) and (d) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait and essentially all of the physiological and morphological characteristics of pea line DLSC7V0955 when grown in the same environmental conditions.
18. A pea plant produced by the method of claim 17, or a selfed progeny thereof.
19. A seed that produces the plant of claim 18.
20. A method of producing a plant of pea line DLSC7V0955 comprising an added desired trait, the method comprising introducing a transgene conferring the desired trait into a plant of pea line DLSC7V0955, a sample of seed of said line DLSC7V0955 having been deposited under ATCC Accession Number PTA-11600.
21. A pea plant produced by the method of claim 20, or a selfed progeny thereof.
22. A seed that produces the plant of claim 21.
23. A method of producing peas comprising: (a) obtaining the plant of claim 2, wherein the plant has been cultivated to maturity, (b) collecting peas from the plant.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US13/048,044 US8415529B2 (en) | 2011-03-15 | 2011-03-15 | Pea line DLSC7V0955 |
US13/048,044 | 2011-03-15 |
Publications (2)
Publication Number | Publication Date |
---|---|
NZ598733A NZ598733A (en) | 2013-08-30 |
NZ598733B true NZ598733B (en) | 2013-12-03 |
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