U.S. patent application number 14/932778 was filed with the patent office on 2017-05-04 for melon hybrid sv3394mf and parents thereof.
The applicant listed for this patent is SEMINIS VEGETABLE SEEDS, INC.. Invention is credited to Jeffrey M. Mills.
Application Number | 20170118934 14/932778 |
Document ID | / |
Family ID | 58637228 |
Filed Date | 2017-05-04 |
United States Patent
Application |
20170118934 |
Kind Code |
A1 |
Mills; Jeffrey M. |
May 4, 2017 |
MELON HYBRID SV3394MF AND PARENTS THEREOF
Abstract
The invention provides seed and plants of melon hybrid SV3394MF
and the parent lines thereof. The invention thus relates to the
plants, seeds and tissue cultures of melon hybrid SV3394MF and the
parent lines thereof, and to methods for producing a melon plant
produced by crossing such plants with themselves or with another
melon plant, such as a plant of another genotype. The invention
further relates to seeds and plants produced by such crossing. The
invention further relates to parts of such plants, including the
fruit and gametes of such plants.
Inventors: |
Mills; Jeffrey M.;
(Woodland, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
SEMINIS VEGETABLE SEEDS, INC. |
St. Louis |
MO |
US |
|
|
Family ID: |
58637228 |
Appl. No.: |
14/932778 |
Filed: |
November 4, 2015 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12N 15/8289 20130101;
C12N 15/8274 20130101; C12N 15/8241 20130101; C12N 15/8286
20130101; C12N 15/8271 20130101; C12N 15/8247 20130101; A01H 5/08
20130101; C12N 15/8245 20130101; C12N 15/8279 20130101; C12N
15/8251 20130101; A01H 1/02 20130101 |
International
Class: |
A01H 5/08 20060101
A01H005/08; C12N 15/82 20060101 C12N015/82; A01G 1/00 20060101
A01G001/00; A01H 1/02 20060101 A01H001/02 |
Claims
1. A melon plant comprising at least a first set of the chromosomes
of melon line HARDV10-4029MO, a sample of seed of said line having
been deposited under ATCC Accession Number PTA-122539.
2. A melon seed comprising at least a first set of the chromosomes
of melon line HARDV10-4029MO, a sample of seed of said line having
been deposited under ATCC Accession Number PTA-122539.
3. The plant of claim 1, which is an inbred.
4. The plant of claim 1, which is a hybrid.
5. The seed of claim 2, which is an inbred.
6. The seed of claim 2, which is a hybrid.
7. The plant of claim 4, wherein the hybrid plant is melon hybrid
SV3394MF, a sample of seed of said hybrid SV3394MF having been
deposited under ATCC Accession Number PTA-122538.
8. The seed of claim 6, defined as a seed of melon hybrid SV3394MF,
a sample of seed of said hybrid SV3394MF having been deposited
under ATCC Accession Number PTA-122538.
9. The seed of claim 2, defined as a seed of line
HARDV10-4029MO.
10. A plant part of the plant of claim 1.
11. The plant part of claim 10, further defined as a leaf, an
ovule, pollen, a fruit, or a cell.
12. A melon plant having all the physiological and morphological
characteristics of the melon plant of claim 7.
13. A tissue culture of regenerable cells of the plant of claim
1.
14. The tissue culture according to claim 13, 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.
15. A melon plant regenerated from the tissue culture of claim 13,
wherein said plant comprises all of the morphological and
physiological characteristics of the melon plant comprising at
least a first set of the chromosomes of melon line HARDV10-4029MO,
a sample of seed of said line having been deposited under ATCC
Accession Number PTA-122539.
16. A method of vegetatively propagating the melon plant of claim 1
comprising the steps of: (a) collecting tissue capable of being
propagated from the plant according to claim 1; (b) cultivating
said tissue to obtain proliferated shoots; and (c) rooting said
proliferated shoots to obtain rooted plantlets.
17. The method of claim 16, further comprising growing at least a
first melon plant from said rooted plantlets.
18. A method of introducing a desired trait into a melon line
comprising: (a) utilizing as a recurrent parent a plant of melon
line HARDV10-4029MO, by crossing a plant of melon line
HARDV10-4029MO with a second donor melon plant that comprises a
desired trait to produce F1 progeny, a sample of seed of said line
having been deposited under ATCC Accession Number PTA-122539; (b)
selecting an F1 progeny that comprises the desired trait; (c)
backcrossing the selected F1 progeny with a plant of the same melon
line used as the recurrent parent in step (a), to produce backcross
progeny; (d) selecting backcross progeny comprising the desired
trait; and (e) repeating steps (c) and (d) three or more times to
produce selected fourth or higher backcross progeny plants that
comprise the desired trait, and otherwise comprise essentially all
of the morphological and physiological characteristics of the
recurrent parent melon line used in step (a).
19. A melon plant produced by the method of claim 18.
20. A method of producing a melon plant comprising an added trait,
the method comprising introducing a transgene conferring the trait
into a plant of melon hybrid SV3394MF or melon line HARDV10-4029MO,
a sample of seed of said hybrid and line having been deposited
under ATCC Accession Number PTA-122538 and ATCC Accession Number
PTA-122539, respectively.
21. A melon plant produced by the method of claim 20.
22. The plant of claim 1, further comprising a transgene.
23. The plant of claim 22, wherein the transgene confers a trait
selected from the group consisting of male sterility, herbicide
tolerance, insect resistance, pest resistance, disease resistance,
modified fatty acid metabolism, environmental stress tolerance,
modified carbohydrate metabolism and modified protein
metabolism.
24. The plant of claim 1, further comprising a single locus
conversion.
25. The plant of claim 24, wherein the single locus conversion
confers a trait selected from the group consisting of male
sterility, herbicide tolerance, insect resistance, pest resistance,
disease resistance, modified fatty acid metabolism, environmental
stress tolerance, modified carbohydrate metabolism and modified
protein metabolism.
26. A method for producing a seed of a melon plant derived from at
least one of melon hybrid SV3394MF or melon line HARDV10-4029MO
comprising the steps of: (a) crossing a melon plant of hybrid
SV3394MF or line HARDV10-4029MO with itself or a second melon
plant; a sample of seed of said hybrid and line having been
deposited under ATCC Accession Number PTA-122538 and ATCC Accession
Number PTA-122539, respectively; and (b) allowing seed of a hybrid
SV3394MF or line HARDV10-4029MO-derived melon plant to form.
27. A method of producing a seed of a hybrid SV3394MF or line
HARDV10-4029MO-derived melon plant comprising the steps of: (a)
producing a hybrid SV3394MF or line HARDV10-4029MO-derived melon
plant from a seed produced by crossing a melon plant of hybrid
SV3394MF or line HARDV10-4029MO with itself or a second melon
plant, a sample of seed of said hybrid and line having been
deposited under ATCC Accession Number PTA-122538 and ATCC Accession
Number PTA-122539, respectively; and (b) crossing the hybrid
SV3394MF or line HARDV10-4029MO-derived melon plant with itself or
a different melon plant to obtain a seed of a further hybrid
SV3394MF or line HARDV10-4029MO-derived melon plant.
28. The method of claim 27, further comprising repeating said
producing and crossing steps of (a) and (b) using a seed from said
step (b) for producing a plant according to step (a) for at least
one generation to produce a seed of an additional hybrid SV3394MF
or line HARDV10-4029MO-derived melon plant.
29. A plant part of the plant of claim 7.
30. The plant part of claim 29, further defined as a leaf, a
flower, a fruit, an ovule, pollen, or a cell.
31. A method of producing a melon seed comprising crossing the
plant of claim 1 with itself or a second melon plant and allowing
seed to form.
32. A method of producing a melon fruit comprising: (a) obtaining
the plant according to claim 1, wherein the plant has been
cultivated to maturity; and (b) collecting a melon from the plant.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to the field of plant breeding
and, more specifically, to the development of melon hybrid SV3394MF
and the inbred melon lines HARDV10-4029MO and HARDV12-4098AN.
BACKGROUND OF THE INVENTION
[0002] The goal of vegetable breeding is to combine various
desirable traits in a single variety/hybrid. Such desirable traits
may include any trait deemed beneficial by a grower and/or
consumer, including greater yield, resistance to insects or
disease, tolerance to environmental stress, and nutritional
value.
[0003] 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.
[0004] 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
genotypes 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.
[0005] 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 and hybrids derived therefrom are developed by
selfing and selection of desired phenotypes. The new lines and
hybrids are evaluated to determine which of those have commercial
potential.
SUMMARY OF THE INVENTION
[0006] In one aspect, the present invention provides a melon plant
of the hybrid designated SV3394MF, the melon line HARDV10-4029MO or
melon line HARDV12-4098AN. Also provided are melon plants having
all the physiological and morphological characteristics of such a
plant. Parts of these melon plants are also provided, for example,
including pollen, an ovule, scion, a rootstock, a fruit, and a cell
of the plant.
[0007] In another aspect of the invention, a plant of melon hybrid
SV3394MF and/or melon lines HARDV10-4029MO and HARDV12-4098AN
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
melon hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN 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 a 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.
[0008] The invention also concerns the seed of melon hybrid
SV3394MF and/or melon lines HARDV10-4029MO and HARDV12-4098AN. The
melon seed of the invention may be provided, in particular
embodiments, as an essentially homogeneous population of melon seed
of melon hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN. Essentially homogeneous populations of seed are
generally free from substantial numbers of other seed. Therefore,
seed of hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN may be provided, in certain embodiments of the
invention, as forming at least about 97% of the total seed,
including at least about 98%, 99% or more of the seed. The seed
population may be separately grown to provide an essentially
homogeneous population of melon plants designated SV3394MF and/or
melon lines HARDV10-4029MO and HARDV12-4098AN.
[0009] In yet another aspect of the invention, a tissue culture of
regenerable cells of a melon plant of hybrid SV3394MF and/or melon
lines HARDV10-4029MO and HARDV12-4098AN is provided. The tissue
culture will preferably be capable of regenerating melon plants
capable of expressing all of the physiological and morphological
characteristics of the starting plant, and of regenerating plants
having substantially the same genotype as the starting plant.
Examples of some of the physiological and morphological
characteristics of the hybrid SV3394MF and/or melon lines
HARDV10-4029MO and HARDV12-4098AN 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, pistils, flowers, seed
and stalks. Still further, the present invention provides melon
plants regenerated from a tissue culture of the invention, the
plants having all the physiological and morphological
characteristics of hybrid SV3394MF and/or melon lines
HARDV10-4029MO and HARDV12-4098AN.
[0010] In still yet another aspect of the invention, processes are
provided for producing melon seeds, plants and fruit, which
processes generally comprise crossing a first parent melon plant
with a second parent melon plant, wherein at least one of the first
or second parent melon plants is a plant of melon line
HARDV10-4029MO or melon line HARDV12-4098AN. These processes may be
further exemplified as processes for preparing hybrid melon seed or
plants, wherein a first melon plant is crossed with a second melon
plant of a different, distinct genotype to provide a hybrid that
has, as one of its parents, a plant of melon line HARDV10-4029MO or
melon line HARDV12-4098AN. In these processes, crossing will result
in the production of seed. The seed production occurs regardless of
whether the seed is collected or not.
[0011] In one embodiment of the invention, the first step in
"crossing" comprises planting seeds of a first and second parent
melon 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.
[0012] A second step may comprise cultivating or growing the seeds
of first and second parent melon plants into plants that bear
flowers. A third step may comprise preventing self-pollination of
the plants, such as by emasculating the flowers (i.e., killing or
removing the pollen).
[0013] A fourth step for a hybrid cross may comprise
cross-pollination between the first and second parent melon plants.
Yet another step comprises harvesting the seeds from at least one
of the parent melon plants. The harvested seed can be grown to
produce a melon plant or hybrid melon plant.
[0014] The present invention also provides the melon seeds and
plants produced by a process that comprises crossing a first parent
melon plant with a second parent melon plant, wherein at least one
of the first or second parent melon plants is a plant of melon
hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN. In one embodiment of the invention, melon seed and
plants produced by the process are first generation (F.sub.1)
hybrid melon 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.sub.1 hybrid melon plant, and methods of use thereof. Therefore,
certain exemplary embodiments of the invention provide an F.sub.1
hybrid melon plant and seed thereof.
[0015] In still yet another aspect, the present invention provides
a method of producing a plant derived from hybrid SV3394MF and/or
melon lines HARDV10-4029MO and HARDV12-4098AN, the method
comprising the steps of: (a) preparing a progeny plant derived from
hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN, wherein said preparing comprises crossing a plant
of the hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN 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 hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN. The plant derived from hybrid SV3394MF and/or melon
lines HARDV10-4029MO and HARDV12-4098AN 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 hybrid SV3394MF and/or melon lines
HARDV10-4029MO and HARDV12-4098AN is obtained which possesses some
of the desirable traits of the line/hybrid as well as potentially
other selected traits.
[0016] In certain embodiments, the present invention provides a
method of producing food or feed comprising: (a) obtaining a plant
of melon hybrid SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN, wherein the plant has been cultivated to maturity,
and (b) collecting at least one melon from the plant.
[0017] In still yet another aspect of the invention, the genetic
complement of melon hybrid SV3394MF and/or melon lines
HARDV10-4029MO and HARDV12-4098AN 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 melon 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 melon plant cells that have a genetic
complement in accordance with the melon plant cells disclosed
herein, and seeds and plants containing such cells.
[0018] 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 hybrid SV3394MF
and/or melon lines HARDV10-4029MO and HARDV12-4098AN could be
identified by any of the many well known techniques such as, for
example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et
al., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly Amplified
Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF),
Sequence Characterized Amplified Regions (SCARs), Arbitrary Primed
Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length
Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein
by reference in its entirety), and Single Nucleotide Polymorphisms
(SNPs) (Wang et al., Science, 280:1077-1082, 1998).
[0019] In still yet another aspect, the present invention provides
hybrid genetic complements, as represented by melon plant cells,
tissues, plants, and seeds, formed by the combination of a haploid
genetic complement of a melon plant of the invention with a haploid
genetic complement of a second melon plant, preferably, another,
distinct melon plant. In another aspect, the present invention
provides a melon plant regenerated from a tissue culture that
comprises a hybrid genetic complement of this invention.
[0020] In still yet another aspect, the invention provides a method
of determining the genotype of a plant of melon hybrid SV3394MF
and/or melon lines HARDV10-4029MO and HARDV12-4098AN comprising
detecting in the genome of the plant at least a first polymorphism.
The method may, in certain embodiments, comprise detecting a
plurality of polymorphisms in the genome of the plant. The method
may further comprise storing the results of the step of detecting
the plurality of polymorphisms on a computer readable medium. The
invention further provides a computer readable medium produced by
such a method.
[0021] Any embodiment discussed herein with respect to one aspect
of the invention applies to other aspects of the invention as well,
unless specifically noted.
[0022] The term "about" is used to indicate that a value includes
the standard deviation of the mean 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. 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 otherwise. 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.
[0023] 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.
DETAILED DESCRIPTION OF THE INVENTION
[0024] The invention provides methods and compositions relating to
plants, seeds and derivatives of melon hybrid SV3394MF, melon line
HARDV10-4029MO and melon line HARDV12-4098AN. The hybrid SV3394MF
was produced by the cross of parent lines HARDV10-4029MO and
HARDV12-4098AN. The parent lines show uniformity and stability
within the limits of environmental influence. By crossing the
parent lines, uniform seed hybrid SV3394MF can be obtained.
[0025] Hybrid SV3394MF, also known as 12-DV-HAR-3394, is a
Harper-type hybrid melon similar in appearance to the hybrid
Caribbean Gold. The fruit of SV3394MF is non-sutured, have light
silver to creamy colored skin, and have a long shelf life. SV3394MF
has a deeper orange flesh color, and a smaller seed cavity than the
variety Caribbean Gold. SV3394MF shows broad adaptation to many
melon growing regions.
A. ORIGIN AND BREEDING HISTORY OF MELON HYBRID SV3394MF
[0026] The parents of hybrid SV3394MF are HARDV10-4029MO and
HARDV12-4098AN. The parent lines are uniform and stable, as is a
hybrid produced therefrom. A small percentage of variants can occur
within commercially acceptable limits for almost any characteristic
during the course of repeated multiplication. However no variants
are expected.
B. PHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF MELON HYBRID
SV3394MF, MELON LINE HARDV10-4029MO AND MELON LINE
HARDV12-4098AN
[0027] In accordance with one aspect of the present invention,
there is provided a plant having the physiological and
morphological characteristics of melon hybrid SV3394MF and the
parent lines thereof. A description of the physiological and
morphological characteristics of such plants is presented in Tables
1-3.
TABLE-US-00001 TABLE 1 Physiological and Morphological
Characteristics of Hybrid SV3394MF Comparison Variety-
CHARACTERISTIC SV3394MF Caribbean Gold 1. Type Other: Harper Other:
Harper 3. Seedling length of hypocotyl (just before short short
development of the first true leaf) size of cotyledon small medium
intensity of green color of medium medium cotyledon 4. Leaf (mature
blade of third leaf) shape reniform reniform lobes not lobed not
lobed color dark green dark green RHS Color Chart value 147A 147A
length 150.6 mm 138.1 mm width 161.3 mm 142.5 mm surface pubescent
pubescent 5. Leaf Blade (fully developed but not old leaves,
preferably between the 5.sup.th and 8.sup.th node when the plant
has at least 11 nodes) size large medium intensity of green color
dark dark development of lobes weak medium length of terminal lobe
short medium dentation of margin weak weak blistering medium medium
6. Petiole attitude erect erect length long long 7. Plant fertility
- sex expression Monoecious monoecious (at full flowering) habit
vine vine 8. Young fruit (green, unripe fruit before color change)
hue of green color of skin green greyish green intensity of green
color of skin light light density of dots absent or absent or very
sparse very sparse conspicuousness of groove absent or absent or
coloring very weak very weak intensity of groove light length of
peduncle medium short thickness of peduncle 1 cm thin thin from
fruit extension of darker area small small around peduncle change
of skin color from late in late in young fruit to maturity fruit
fruit development development 9. Fruit length long medium length
(at edible maturity) 17.4 cm 15.1 cm diameter medium medium
diameter (at edible maturity) 15.6 cm 12.8 cm ratio length/diameter
small small weight (at edible maturity) 2216.3 gm 1320.1 gm
position of maximum diameter at middle at middle shape round round
shape in longitudinal section circular circular surface (at edible
maturity) netted netted blossom scar (at edible obscure obscure
maturity) rib presence (at edible absent absent maturity) shipping
quality (at edible excellent excellent maturity) abscission (at
edible when when maturity) overripe overripe maturity (number of
days 102.sup. 102.sup. from seeding to harvest) Rind Net net
presence abundant abundant distribution covers covers entire fruit
entire fruit coarseness very coarse very coarse interlacing
complete complete interstices medium deep medium deep Rind texture:
hard hard thickness at medial 4.1 mm 1.6 mm Rind Color Primary
Color (at edible gray/green gray/green maturity) RHS Color Chart
value 194B 194B Net Color (at edible gray/yellow orange/white
maturity) RHS Color Chart value 161D 159C Primary Color (at full
yellow/green yellow/green maturity) RHS Color Chart value N144A
148B Net Color (at full maturity cream white/green RHS Color Chart
value 162D 157C Fruit: ground color of skin yellow yellow intensity
of ground color medium light of skin hue of ground color of skin
yellowish yellowish density of dots absent or absent or very sparse
very sparse density of patches absent or absent or very sparse very
sparse warts absent absent strength of attachment of strong medium
peduncle at maturity shape of base rounded rounded shape of apex
rounded rounded size of pistil scar medium small grooves absent or
absent or very weakly very weakly expressed expressed creasing of
surface absent or absent or very weak very weak cork formation
absent absent rate of change of skin color absent or absent or from
maturity to over maturity very slow very slow width of flesh in
longitudinal thick medium section (at position of maximum fruit
diameter) main color of flesh orange orange intensity of orange
color dark medium of flesh (only varieties with main color of flesh
orange) firmness of flesh firm firm hue of color of skin (at over
creamish creamish maturity): only varieties with change of skin
color from maturity to over maturity intensity of yellow color
medium medium of skin (at over maturity) only varieties with change
of skin color from maturity to over maturity and with yellow or
orangish yellow color of skin 10. Flesh Color Near Cavity (at
edible orange orange maturity) RHS Color Chart value 24B 24C Color
in Center (at edible orange orange maturity) RHS Color Chart value
24B 24C Color Near Rind (at edible orange orange maturity) RHS
Color Chart value 24B 24C Flesh: refractometer % 16.40% 12.80%
soluable solids (center of flesh) aroma (at edible maturity) strong
faint flavor (at edible maturity) very somewhat spicy spicy 12.
Seed length of the seed cavity 119.7 mm 101.6 mm width of seed
cavity 69.7 mm 56.7 mm shape in cross section circular circular
length (made on fully short short developed and dry seeds, after
washing and drying in the shade) width medium medium shape pine-nut
pine-nut shape shape color cream cream yellow yellow intensity of
color (only medium light varieties with cream yellow seed color)
Time of male flowering early early Time of female flowering early
medium Time of ripening medium medium Shelf life of fruit very long
long Seeds: measurements 868.1 621.3 (number of seeds per fruit)
Seeds: measurements 24.0 gm 21.0 gm (grams per 1,000 seeds) *These
are typical values. Values may vary due to environment. Other
values that are substantially equivalent are also within the scope
of the invention.
TABLE-US-00002 TABLE 2 Physiological and Morphological
Characteristics of Line HARDV10-4029MO Comparison HARDV10- Variety-
CHARACTERISTIC 4029MO Caribbean Gold 1. Type other: Harper other:
Harper 3. Seedling length of hypocotyl (just before medium medium
development of the first true leaf) size of cotyledon medium medium
intensity of green color of medium medium cotyledon 4. Leaf (mature
blade of third leaf) shape reniform reniform lobes shallowly
shallowly lobed lobed color dark green dark green RHS Color Chart
value 147A 147A length 90.3 mm 114.5 mm width 128.1 mm 154.6 mm
surface glabrous pubescent 5. Leaf Blade (fully developed but not
old leaves, preferably between the 5.sup.th and 8.sup.th node when
the plant has at least 11 nodes) size medium large intensity of
green color dark dark development of lobes medium medium length of
terminal lobe medium medium dentation of margin medium weak
blistering medium weak 6. Petiole attitude erect erect length
medium long 7. Plant fertility - sex expression monoecious
monoecious (at full flowering) habit vine vine 8. Young fruit
(green, unripe fruit before color change) hue of green color of
skin whitish green green intensity of green color of skin light
medium density of dots medium sparse size of dots small small
contrast of dot color/ground medium medium color conspicuousness of
groove absent or absent or coloring very weak very weak length of
peduncle medium long thickness of peduncle 1 cm medium thick from
fruit extension of darker area small medium around peduncle change
of skin color from very late very late young fruit to maturity in
fruit in fruit development development or no change or no change 9.
Fruit length medium medium length (at edible maturity) 16.2 cm 17.2
cm diameter medium medium diameter (at edible maturity) 13.9 cm
14.7 cm ratio length/diameter small small weight (at edible
maturity) 1552.3 gm 1815.8 gm position of maximum diameter at
middle at middle shape oval round shape in longitudinal section
broad broad elliptic elliptic surface (at edible maturity) netted
netted blossom scar (at edible conspicuous conspicuous maturity)
rib presence (at edible absent absent maturity) shipping quality
(at edible excellent excellent maturity) abscission (at edible do
not do not maturity) abscise abscise maturity (number of days
101.sup. 101.sup. from seeding to harvest) Rind Net net presence
abundant abundant distribution covers covers entire fruit entire
fruit coarseness very coarse very coarse interlacing complete
complete interstices medium deep medium deep Rind texture: hard
hard thickness at medial 5.9 mm 2.1 mm Rind Color Primary Color (at
edible green/gray gray/green maturity) RHS Color Chart value 191A
194A Net Color (at edible green/yellow gray/yellow maturity) RHS
Color Chart value 160D 161D Primary Color (at full green/gray
yellow/green maturity) RHS Color Chart value 191A 154A Net Color
(at full maturity) white/green orange/white RHS Color Chart value
157A 159C Fruit: ground color of skin yellow yellow intensity of
ground color medium medium of skin hue of ground color of skin
yellowish yellowish density of dots absent or absent or very sparse
very sparse density of patches absent or absent or very sparse very
sparse warts absent absent strength of attachment of very strong
peduncle at maturity strong shape of base rounded rounded shape of
apex rounded rounded size of pistil scar medium medium grooves
absent or absent or very weakly very weakly expressed expressed
creasing of surface absent or absent or very weak very weak cork
formation absent present rate of change of skin color absent or
absent or from maturity to over maturity very slow very slow width
of flesh in longitudinal medium medium section (at position of
maximum fruit diameter) main color of flesh orange orange intensity
of orange color medium medium of flesh (only varieties with main
color of flesh orange) firmness of flesh firm firm 10. Flesh Color
Near Cavity (at edible orange orange maturity) RHS Color Chart
value 26B 26B Color in Center (at edible orange orange maturity)
RHS Color Chart value 26B 26B Color Near Rind (at edible orange
orange maturity) RHS Color Chart value 26B 26B Flesh: refractometer
% 14.40% 16.20% soluable solids (center of flesh) aroma (at edible
maturity) faint faint flavor (at edible maturity) very very spicy
spicy 12. Seed length of the seed cavity 106.3 mm 107.6 mm width of
seed cavity 57.2 mm 57.4 mm shape in cross section triangular
circular length (made on fully short medium developed and dry
seeds, after washing and drying in the shade) width medium medium
shape pine-nut pine-nut shape shape color cream cream yellow yellow
intensity of color (only light medium varieties with cream yellow
seed color) Time of male flowering early early Time of female
flowering early early Time of ripening late late Shelf life of
fruit long medium Seeds: measurements 651.4 722.8 (number of seeds
per fruit) Seeds: measurements 19.5 gm 27.5 gm (grams per 1,000
seeds) *These are typical values. Values may vary due to
environment. Other values that are substantially equivalent are
also within the scope of the invention.
TABLE-US-00003 TABLE 3 Physiological and Morphological
Characteristics of Line HARDV12-4098AN Comparison HARDV12- Variety-
CHARACTERISTIC 4098AN Caribbean Gold 1. Type other: Harper other:
Harper 3. Seedling length of hypocotyl (just before short short
development of the first true leaf) size of cotyledon medium medium
intensity of green color of medium medium cotyledon 4. Leaf (mature
blade of third leaf) shape reniform reniform lobes not lobed not
lobed color dark green dark green RHS Color Chart value 147A 147A
length 165.5 mm 138.1 mm width 173.7 mm 142.5 mm surface pubescent
pubescent 5. Leaf Blade (fully developed but not old leaves,
preferably between the 5.sup.th and 8.sup.th node when the plant
has at least 11 nodes) size large medium intensity of green color
dark dark development of lobes weak medium length of terminal lobe
short medium dentation of margin weak weak blistering medium medium
6. Petiole attitude erect erect length long long 7. Plant fertility
- sex expression andromon- monoecious (at full flowering) oecious
habit vine vine 8. Young fruit (green, unripe fruit before color
change) hue of green color of skin green greyish green intensity of
green color of skin light light density of dots absent or absent or
very sparse very sparse contrast of dot color/ground weak color
conspicuousness of groove medium absent or coloring very weak
intensity of groove coloring medium length of peduncle short short
thickness of peduncle 1 cm medium thin from fruit extension of
darker area absent or small around peduncle very small change of
skin color from late in late in young fruit to maturity fruit fruit
development development 9. Fruit length short medium length (at
edible maturity) 10.0 cm 15.1 cm diameter narrow narrow diameter
(at edible maturity) 11.1 cm 12.8 cm ratio length/diameter very
small small to small weight (at edible maturity) 699.2 gm 1320.1 gm
position of maximum diameter at middle at middle shape oblate round
shape in longitudinal section oblate circular surface (at edible
maturity) netted netted blossom scar (at edible obscure obscure
maturity) rib presence (at edible present absent maturity) number
of ribs per fruit 9.5 (at edible maturity) rib width at medial 34.4
mm (at edible maturity) ribs surface (at edible netted maturity)
suture depth (at edible shallow maturity) suture surface (at edible
netted maturity) shipping quality (at edible excellent excellent
maturity) abscission (at edible when ripe when overripe maturity)
maturity (number of days 91 102.sup. from seeding to harvest) Rind
Net net presence abundant abundant distribution covers covers
entire fruit entire fruit coarseness very coarse very coarse
interlacing complete complete interstices medium deep medium deep
Rind texture: hard hard thickness at medial 3.7 mm 1.6 mm Rind
Color Primary Color (at edible yellow/gray gray/green maturity) RHS
Color Chart value 160A 194B Net Color (at edible green/gray
orange/white maturity) RHS Color Chart value 161C 159C Furrow
(Suture) Color green/gray (at edible maturity) RHS Color Chart
value 194B Primary Color (at full maturity) yellow/green
yellow/green RHS Color Chart value 160B 148B Net Color (at full
maturity yellow/green white/green RHS Color Chart value 165D 157C
Furrow (Suture) Color yellow/green (at full maturity) RHS Color
Chart value 160B Fruit: ground color of skin yellow yellow
intensity of ground color medium light of skin hue of ground color
of skin yellowish yellowish density of dots absent or absent or
very sparse very sparse density of patches absent or absent or very
sparse very sparse warts absent absent strength of attachment of
very weak medium peduncle at maturity shape of base truncate
rounded shape of apex truncate rounded size of pistil scar large
small grooves weakly absent or expressed very weakly expressed
width of grooves narrow depth of grooves very shallow color of
grooves green creasing of surface absent or absent or very weak
very weak cork formation absent absent rate of change of skin slow
absent or color from maturity to over very slow maturity width of
flesh in longitudinal medium medium section (at position of maximum
fruit diameter) main color of flesh orange orange intensity of
orange color dark medium of flesh (only varieties with main color
of flesh orange) firmness of flesh firm firm at over maturity: hue
of color yellow creamish of skin (only varieties with change of
skin color from maturity to over maturity) at over maturity:
intensity medium medium of yellow color of skin (only varieties
with change of skin color from maturity to over maturity and with
yellow or orangish yellow color of skin) 10. Flesh Color Near
Cavity (at edible orange orange maturity) RHS Color Chart value 26A
24C Color in Center (at edible orange orange maturity) RHS Color
Chart value 26A 24C Color Near Rind (at edible orange orange
maturity) RHS Color Chart value 26A 24C Flesh: refractometer %
16.00% 12.80% soluable solids (center of flesh) aroma (at edible
maturity) strong faint flavor (at edible maturity) very somewhat
spicy spicy 12. Seed length of the seed cavity 68.4 mm 101.6 mm
width of seed cavity 50.7 mm 56.7 mm shape in cross section
circular circular length (made on fully medium short developed and
dry seeds, after washing and drying in the shade) width medium
medium shape pine-nut pine-nut shape shape color cream cream yellow
yellow intensity of color (only medium light varieties with cream
yellow seed color) Time of male flowering medium early Time of
female flowering late medium Time of ripening early medium Shelf
life of fruit long long Seeds: measurements 381.9 621.3 (number of
seeds per fruit) Seeds: measurements 24.0 gm 21.0 gm (grams per
1,000 seeds) *These are typical values. Values may vary due to
environment. Other values that are substantially equivalent are
also within the scope of the invention.
C. BREEDING MELON PLANTS
[0028] One aspect of the current invention concerns methods for
producing seed of melon hybrid SV3394MF involving crossing melon
lines HARDV10-4029MO and HARDV12-4098AN. Alternatively, in other
embodiments of the invention, hybrid SV3394MF, line HARDV10-4029MO,
or line HARDV12-4098AN may be crossed with itself or with any
second plant. Such methods can be used for propagation of hybrid
SV3394MF and/or the melon lines HARDV10-4029MO and HARDV12-4098AN,
or can be used to produce plants that are derived from hybrid
SV3394MF and/or the melon lines HARDV10-4029MO and HARDV12-4098AN.
Plants derived from hybrid SV3394MF and/or the melon lines
HARDV10-4029MO and HARDV12-4098AN may be used, in certain
embodiments, for the development of new melon varieties.
[0029] 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 hybrid
SV3394MF 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.
[0030] 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 a plant of the invention and progeny
thereof to achieve a homozygous line.
[0031] 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 have 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.
[0032] The plants of the present invention are particularly well
suited for the development of new lines based on the elite nature
of the genetic background of the plants. In selecting a second
plant to cross with SV3394MF and/or melon lines HARDV10-4029MO and
HARDV12-4098AN for the purpose of developing novel melon 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 desirable traits may include, in specific
embodiments, high seed yield, high seed germination, seedling
vigor, high fruit yield, disease tolerance or resistance, and
adaptability for soil and climate conditions. Consumer-driven
traits, such as a fruit shape, color, texture, and taste are other
examples of traits that may be incorporated into new lines of melon
plants developed by this invention.
D. PERFORMANCE CHARACTERISTICS
[0033] As described above, hybrid SV3394MF exhibits desirable
agronomic traits. The performance characteristics of hybrid
SV3394MF were the subject of an objective analysis of the
performance traits relative to other varieties. The results of the
analysis are presented in Tables 4 and 5.
TABLE-US-00004 TABLE 4 Performance data for Hybrid SV3394MF and
Comparative Variety Int. Firmn Plant Diseases Net Cavity Color Brix
(Kg) Overall Comments C. Gold 5 5 2 5 5 12.3 4.2 4 Low brix and no
taste. SV3394MF 4 4 2 3 3 12.5 4.1 3 Good fruit set and net. Good
taste.
TABLE-US-00005 TABLE 5 Performance data for Hybrid SV3394MF and
Comparative Variety Boxes/ha Fruits/m 9j 9 12 15 18 Caribbean 2,055
4.2 10 44 31 14 Gold SV3394MF 2,048 4.8 19 33 41 7
E. FURTHER EMBODIMENTS OF THE INVENTION
[0034] In certain aspects of the invention, plants described herein
are provided 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 melon plants which
are 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 the
single locus transferred into the variety via the backcrossing
technique. By essentially all of the morphological and
physiological characteristics, it is meant that the characteristics
of a plant are recovered that are otherwise present when compared
in the same environment, other than an occasional variant trait
that might arise during backcrossing or direct introduction of a
transgene.
[0035] Backcrossing methods can be used with the present invention
to improve or introduce a characteristic into the present variety.
The parental melon 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 melon 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.
[0036] 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 melon plant is obtained wherein essentially all of the
morphological and physiological characteristics of the recurrent
parent are recovered in the converted plant, in addition to the
single transferred locus from the nonrecurrent parent.
[0037] 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.
[0038] In one embodiment, progeny melon plants of a backcross in
which a plant described herein is the recurrent parent comprise (i)
the desired trait from the non-recurrent parent and (ii) all of the
physiological and morphological characteristics of melon the
recurrent parent as determined at the 5% significance level when
grown in the same environmental conditions.
[0039] New 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.
[0040] With the development of molecular markers associated with
particular traits, it is possible to add additional traits into an
established germ line, such as represented here, with the end
result being substantially the same base germplasm with the
addition of a new trait or traits. Molecular breeding, as described
in Moose and Mumm, 2008 (Plant Physiology, 147: 969-977), for
example, and elsewhere, provides a mechanism for integrating single
or multiple traits or QTL into an elite line. This molecular
breeding-facilitated movement of a trait or traits into an elite
line may encompass incorporation of a particular genomic fragment
associated with a particular trait of interest into the elite line
by the mechanism of identification of the integrated genomic
fragment with the use of flanking or associated marker assays. In
the embodiment represented here, one, two, three or four genomic
loci, for example, may be integrated into an elite line via this
methodology. When this elite line containing the additional loci is
further crossed with another parental elite line to produce hybrid
offspring, it is possible to then incorporate at least eight
separate additional loci into the hybrid. These additional loci may
confer, for example, such traits as a disease resistance or a fruit
quality trait. In one embodiment, each locus may confer a separate
trait. In another embodiment, loci may need to be homozygous and
exist in each parent line to confer a trait in the hybrid. In yet
another embodiment, multiple loci may be combined to confer a
single robust phenotype of a desired trait.
[0041] 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, herbicide resistance, resistance to bacterial,
fungal, or viral disease, insect resistance, modified fatty acid or
carbohydrate metabolism, and altered nutritional quality. These
comprise genes generally inherited through the nucleus.
[0042] Direct selection may be applied where the single locus acts
as a dominant trait. For this selection process, the progeny of the
initial cross are assayed for viral resistance and/or the presence
of the corresponding gene prior to the backcrossing. Selection
eliminates any plants that do not have the desired gene and
resistance trait, and only those plants that have the trait are
used in the subsequent backcross. This process is then repeated for
all additional backcross generations.
[0043] Selection of melon 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 are well known in the art. Such methods
will be of particular utility in the case of recessive traits and
variable phenotypes, or where conventional assays may be more
expensive, time consuming or otherwise disadvantageous. Types of
genetic markers which could be used in accordance with the
invention include, but are not necessarily limited to, Simple
Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic
Acids Res., 1 8:6531-6535, 1990), Randomly Amplified Polymorphic
DNAs (RAPDs), DNA Amplification Fingerprinting (DAF), Sequence
Characterized Amplified Regions (SCARs), Arbitrary Primed
Polymerase Chain Reaction (AP-PCR), Amplified Fragment Length
Polymorphisms (AFLPs) (EP 534 858, specifically incorporated herein
by reference in its entirety), and Single Nucleotide Polymorphisms
(SNPs) (Wang et al., Science, 280:1077-1082, 1998).
F. PLANTS DERIVED BY GENETIC ENGINEERING
[0044] Many useful traits that can be introduced by backcrossing,
as well as directly into a plant, are those which are introduced by
genetic transformation techniques. Genetic transformation may
therefore be used to insert a selected transgene into a plant 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 that are well known to those of skill in
the art and applicable to many crop species include, but are not
limited to, electroporation, microprojectile bombardment,
Agrobacterium-mediated transformation and direct DNA uptake by
protoplasts.
[0045] 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.
[0046] An 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.
[0047] An illustrative embodiment of a method for delivering DNA
into plant cells by acceleration is the Biolistics Particle
Delivery System, which can be used to propel particles coated with
DNA or cells through a screen, such as a stainless steel or Nytex
screen, onto a surface covered with target cells. The screen
disperses the particles so that they are not delivered to the
recipient cells in large aggregates. Microprojectile bombardment
techniques are widely applicable, and may be used to transform
virtually any plant species.
[0048] Agrobacterium-mediated transfer is another widely applicable
system for introducing gene loci into plant cells. An advantage of
the technique is that DNA can be introduced into whole plant
tissues, thereby bypassing the need for regeneration of an intact
plant from a protoplast. Modern Agrobacterium transformation
vectors are capable of replication in E. coli as well as
Agrobacterium, allowing for convenient manipulations (Klee et al.,
Bio-Technology, 3(7):637-642, 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.
[0049] In those plant strains where Agrobacterium-mediated
transformation is efficient, it is the method of choice because of
the facile and defined nature of the gene locus transfer. The use
of Agrobacterium-mediated plant integrating vectors to introduce
DNA into plant cells is well known in the art (Fraley et al.,
Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).
[0050] Transformation of plant protoplasts also can be achieved
using methods based on calcium phosphate precipitation,
polyethylene glycol treatment, electroporation, and combinations of
these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet.,
199:183-188, 1985; Omirulleh et al., Plant Mol. Biol.,
21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986;
Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al.,
Nature, 335:454, 1988). Transformation of plants and expression of
foreign genetic elements is exemplified in Choi et al. (Plant Cell
Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet.,
107:462-469, 2003).
[0051] 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
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., Nature, 313:810, 1985), including in monocots
(see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and
Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated
version of the CaMV 35S promoter, the enhanced 35S promoter
(P-e35S);1 the nopaline synthase promoter (An et al., Plant
Physiol., 88:547, 1988); the octopine synthase promoter (Fromm et
al., Plant Cell, 1:977, 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.
[0052] A variety of plant gene promoters that are regulated in
response to environmental, hormonal, chemical, and/or developmental
signals can also be used for expression of an operably linked gene
in plant cells, including promoters regulated by (1) heat (Callis
et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A
promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS
promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or
chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J.,
4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et
al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl, Siebertz
et al., Plant Cell, 1:961, 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., EMBO J.,
6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et
al., Plant Cell, 1:839, 1989).
[0053] Exemplary nucleic acids which may be introduced to plants 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.
[0054] Many hundreds if not thousands of different genes are known
and could potentially be introduced into a melon plant according to
the invention. Non-limiting examples of particular genes and
corresponding phenotypes one may choose to introduce into a melon
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. 5,500,365
and 5,880,275, herein incorporated by reference in 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.
[0055] 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., Biotech. Gen. Engin. Rev., 9:207, 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, Mol. Biotech., 7:125, 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
[0056] 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:
[0057] 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.
[0058] Backcrossing: A process in which a breeder repeatedly
crosses hybrid progeny, for example a first generation hybrid
(F.sub.1), back to one of the parents of the hybrid progeny.
Backcrossing can be used to introduce one or more single locus
conversions from one genetic background into another.
[0059] Crossing: The mating of two parent plants.
[0060] Cross-pollination: Fertilization by the union of two gametes
from different plants.
[0061] Diploid: A cell or organism having two sets of
chromosomes.
[0062] 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.
[0063] Enzymes: Molecules which can act as catalysts in biological
reactions.
[0064] F.sub.1 Hybrid: The first generation progeny of the cross of
two nonisogenic plants.
[0065] Genotype: The genetic constitution of a cell or
organism.
[0066] Haploid: A cell or organism having one set of the two sets
of chromosomes in a diploid.
[0067] Linkage: A phenomenon wherein alleles on the same chromosome
tend to segregate together more often than expected by chance if
their transmission was independent.
[0068] 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.
[0069] Phenotype: The detectable characteristics of a cell or
organism, which characteristics are the manifestation of gene
expression.
[0070] 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.
[0071] 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.
[0072] Regeneration: The development of a plant from tissue
culture.
[0073] Self-pollination: The transfer of pollen from the anther to
the stigma of the same plant.
[0074] 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 melon 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.
[0075] Substantially Equivalent: A characteristic that, when
compared, does not show a statistically significant difference
(e.g., p=0.05) from the mean.
[0076] 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.
[0077] Transgene: A genetic locus comprising a sequence which has
been introduced into the genome of a melon plant by
transformation.
H. DEPOSIT INFORMATION
[0078] A deposit of melon hybrid SV3394MF and inbred parent lines
HARDV10-4029MO and HARDV12-4098AN, 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 the deposit was Sep. 30, 2015. The accession numbers for those
deposited seeds of melon hybrid SV3394MF and inbred parent lines
HARDV10-4029MO and HARDV12-4098AN are ATCC Accession Number
PTA-122538, ATCC Accession Number PTA-122539, and ATCC Accession
Number PTA-122540, respectively. Upon issuance of a patent, all
restrictions upon the deposits will be removed, and the deposits
are intended to meet all of the requirements of 37 C.F.R.
.sctn.1.801-1.809. The deposits 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.
[0079] 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.
[0080] All references cited herein are hereby expressly
incorporated herein by reference.
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