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| United States Patent Application |
20090133141
|
| Kind Code
|
A1
|
|
Zhang; Xingping
;   et al.
|
May 21, 2009
|
ENHANCED POLLENIZER AND METHOD FOR INCREASING SEEDLESS WATERMELON YIELD
Abstract
An enhanced, diploid pollenizer watermelon plant and method used to
maximize the yield of triploid seedless watermelons per area. The
enhanced pollenizer watermelon plant of the invention is either a hybrid
variety, an open-pollinated variety or a synthetic variety, that exhibits
the characteristics of small leaves and fruit with a brittle rind that
splits when the fruit is overripe or breaks when relatively small
physical forces are applied. In one embodiment, the watermelon plant of
the invention is also characterized by extended flowering duration,
thereby increasing the number of triploid watermelon flowers that are
pollinated and set fruit. The method for producing a seedless watermelon
fruit, includes the steps of providing a pollenizer diploid watermelon
plant, extending the duration of flowering of the pollenizer plant while
reducing the number of such plants needed to pollinate the same number of
triploid watermelon plants, and maximizing dispersal of the pollenizer
watermelon plant throughout the field of triploid watermelon plants.
| Inventors: |
Zhang; Xingping; (Woodland, CA)
; Williams; Thomas Vare; (Naples, FL)
|
| Correspondence Address:
|
SYNGENTA BIOTECHNOLOGY, INC.;PATENT DEPARTMENT
3054 CORNWALLIS ROAD, P.O. BOX 12257
RESEARCH TRIANGLE PARK
NC
27709-2257
US
|
| Assignee: |
Syngenta Participations AG
|
| Serial No.:
|
360991 |
| Series Code:
|
12
|
| Filed:
|
January 28, 2009 |
| Current U.S. Class: |
800/260; 800/308 |
| Class at Publication: |
800/260; 800/308 |
| International Class: |
A01H 5/00 20060101 A01H005/00; A01H 1/02 20060101 A01H001/02 |
Claims
1. A watermelon plant comprising, at maturity:a) a fruit, the rind of
which breaks under a pressure of not more than about 1,800 g when a fruit
tester with a 2 mm tip is used or under a pressure of not more than about
2,300 g when a fruit tester with a 3 mm tip is used, an example of a
plant that produces a fruit with such characteristics being the
commercially available watermelon variety Tiny Orchid; andb) a 5th leaf
from the smallest new leaf on a vine having a surface area of not more
than about 50 cm.sup.2, an example of a plant with such characteristics
being publicly available line XinJiang.
2. A watermelon plant having morphological and physiological
characteristics of the fruit and leaves of NO1F3203B, said
characteristics comprising at maturity:a) a fruit, the rind of which
breaks under a pressure of not more than about 1,800 g when a fruit
tester with a 2 mm tip is used or under a pressure of not more than about
2,300 g when a fruit tester with a 3 mm tip is used; and,b) a 5.sup.th
leaf from the smallest new leaf on a vine having a surface area of not
more than about 50 cm.sup.2;wherein representative seed of NO1F3203B
having been deposited as Accession No. PTA-4856.
3. A watermelon plant comprising, at maturity:a) a fruit sharing the rind
characteristics of the commercially available watermelon variety Tiny
Orchid; andb) leaves sharing the leaf characteristics of the publicly
available line XinJiang.
4. A watermelon plant that produces:a) fruits having the rind
characteristics of the commercially available watermelon variety Tiny
Orchid; andb) leaves having the leaf characteristics of the publicly
available line XinJuiang.
5. The watermelon plant of claim 3, wherein the rind characteristics are
that said rind of said fruit breaks under a pressure of not more than
about 1,800 g when a fruit tester with a 2 mm tip is used or under a
pressure of not more than about 2,300 g when a fruit tester with a 3 mm
tip is used; and wherein the leaf characteristics are that a 5th leaf
from the smallest new leaf on a vine of said watermelon plant has a
surface area of not more than about 50 cm.sup.2.
6. The watermelon plant according to claim 5, wherein the rind of said
fruit breaks under a pressure in the range of about 400 g to about 1,800
g when a fruit tester with a 2 mm tip is used.
7. The watermelon plant according to claim 5, wherein the rind of said
fruit breaks under a pressure in the range of about 400 g to about 1,800
g when a fruit tester with a 2 mm tip is used or under a pressure in the
range of about 1,000 g to about 2,300 g when a fruit tester when a 3 mm
tip is used.
8. The watermelon plant according to claim 5, wherein said 5th leaf has a
surface area in the range of about 15 cm.sup.2 to about 50 cm.sup.2.
9. The watermelon plant according to claim 5, wherein said leaf is
characterized by deep, non-overlapping leaf lobes.
10. The watermelon plant according to claim 5, further comprising heavily
branched vines.
11. The watermelon plant according to claim 5, wherein the weight of said
fruit in the range of about 2 lbs to about 7 lbs.
12. The watermelon plant according to claim 5, wherein said plant is an
inbred, a hybrid or a dihaploid.
13. Pollen of the plant of claim 5.
14. An ovule of the plant of claim 5.
15. A fruit of the plant of claim 5.
16. A seed of the plant of claim 5.
17. A progeny of said watermelon plants claimed in claim 5, wherein said
progeny retain the characteristics of the watermelon plants as set forth
in claim 5.
18. A method of producing seeds of a watermelon plant comprising the steps
of:a) growing the watermelon plant according to claim 5;b) allowing
self-pollination of said plant; andc) harvesting seeds from said plant.
19. The method according to claim 18, further comprises washing and drying
said seed.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001]This application is a continuation of U.S. application Ser. No.
10/933,791, filed Sep. 3, 2004, which is a continuation-in-part of U.S.
application Ser. No. 10/349,509, filed Jan. 22, 2003, now U.S. Pat. No.
7,071,374, which is a continuation-in-part of U.S. application Ser. No.
10/091,154, filed Mar. 5, 2002, now U.S. Pat. No. 6,759,576. The
aforementioned applications are incorporated herein by reference in their
entireties.
FIELD OF THE INVENTION
[0002]This invention is in the field of watermelon breeding, specifically
relating to diploid watermelons used to pollinate triploid watermelon
plants for the commercial production of seedless watermelon fruit, and
includes a novel method for the production of triploid watermelon fruit.
BACKGROUND OF THE INVENTION
[0003]Watermelon is an important horticultural crop that accounts for 2%
of the world area devoted to vegetable crops. There were 6,024,000 acres
of watermelon grown in the world and 187,000 acres of watermelons grown
in the United States in 1997 (FAO Production Yearbook 51, 1998). The
estimated annual world watermelon value exceeded $7.6 billion when using
the United States average price for 1995-1997. The United States
watermelon crop amounted to over 41 million cwt, from over 174,000
harvested acres, and a farm value of over $266 million, accounted for
9.2% of the harvested acres, 10.0% of the production, and 3.5% of the
value of the United States fresh vegetable industry in 1999 (USDA
Agricultural Statistics 2001). California was the leading state in
watermelon farm gate value, exceeded $72 million in 2000, due to high
percentage of triploid seedless watermelon grown in California. Seedless
watermelon receives well above the average price for seeded watermelons
in the market.
[0004]The goal of plant breeding is to combine in a single variety or
hybrid various desirable traits. Desirable traits may include resistance
to diseases and insects, tolerance to heat and drought, reducing the time
to crop maturity, greater yield, and better agronomic quality. With
mechanical harvesting of many crops, uniformity of plant characteristics
such as germination and stand establishment, growth rate, and maturity,
are important. Other desired traits may include particular nutrient
content, color, fruit shape, as well as taste characteristics.
[0005]As with many different plants, watermelon contains a fruit part and
a plant part. Each part contains different traits that are desired by
consumers and/or growers, including such traits as flavor, texture,
disease resistance, and appearance traits such as shape and color. The
seedless trait in the watermelon fruit is highly desired by consumers.
For production of seedless watermelon, optimum pollination
characteristics of the pollinating plant are desired.
[0006]Seedless watermelon plants are triploid and must be pollinated by
the pollen of diploid watermelon plants. To provide adequate pollination
of seedless watermelon plants, it is current practice to plant diploid
pollenizer plants over approximately 25-33% of the field surface. The
remaining portion of the field is planted with the triploid plants. Thus,
to maximize the value of the crop in the field, growers use high yield
marketable diploid watermelon varieties, which ultimately compete with
the triploid seedless varieties for sun, nutrients, and space.
[0007]A pollenizer for seedless watermelon producing small and
unmarketable fruits, which are not harvested, has been disclosed
(WO00/70933). However, when this pollenizer is used, a lower total yield
of marketable fruit is observed when compared to a commercial pollenizer
line. Also, the fruits of the pollenizer described in WO00/70933 that are
not harvested become hosts for diseases in the future, and their seeds
will germinate and grow into weeds, thus reducing future yields.
[0008]The present invention recognizes the need to increase the yield of
the seedless watermelon, preferably without loss in total yields of
marketable fruits. The present invention also recognizes that novel
phenotypic characteristics of the diploid pollenizer plants are needed to
permit these diploids to be planted in close proximity to the triploid
plants and to share the field surface with the triploid plants, thereby
effectively decreasing the surface area of the field required for the
diploid pollenizers of the invention. The present invention also
recognizes the need to minimize the carryover of un-harvested pollenizer
fruits as weeds into the subsequent season. The present invention also
recognizes the need to increase the pollinating capacity of diploid
watermelon plants in order to further decrease the ratio of diploid to
triploid plants in the field, thereby also increasing the yield of the
seedless watermelon. The present invention also further recognizes the
needs to allow farmers to distinguish the seedless fruits from the fruits
of the pollenizer in the field and to provide marketable value to the
pollenizer fruits themselves.
SUMMARY OF THE INVENTION
[0009]The present invention uses a novel diploid watermelon to improve
current methods of commercial production of seedless watermelon and to
increase seedless watermelon yield. According to the invention, there is
provided a novel enhanced, pollenizer diploid watermelon (hereinafter
referred to as "enhanced pollenizer") and method for pollinating seedless
watermelon plants. The present invention includes an enhanced pollenizer
comprising, at maturity, small leaves. The present invention further
includes an enhanced pollenizer comprising, at maturity, brittle fruits.
The present invention includes an enhanced pollenizer comprising, at
maturity, small leaves and bearing brittle fruits. The small leaves allow
the enhanced pollenizer to be grown in close proximity to the triploid
watermelon plants without competing with them, thereby increasing yields
of seedless fruits. The brittleness of the fruit offers the advantage
that un-harvested fruits of the pollenizer can be easily destroyed
through conventional field preparation for minimizing carry over as weeds
in future plantings.
[0010]In one embodiment, the enhanced pollenizer of the present invention
further comprises heavily branching lacy vines (also referred to as
heavily branched open vines) and therefore preferably comprises a high
number of open (lacy) branches. The heavily branching vines allow an
enhanced pollenizer of this invention to produce very large amount of
male flowers to pollinate the female flowers of the triploid plant, to
produce the seedless fruit. In one embodiment, the leaves of the enhanced
pollenizer also comprise non-overlapping, deep lobes. The openness of the
branched or lacy vine results, in part, from the distinct small and
non-overlapping, deep lobed leaves. The lacy branches and the small
leaves, in one embodiment with non-overlapping, deep lobes, of the
invention have the additional advantage to provide more access of bees to
the flowers of both the pollenizing and the triploid plant, thereby
enhancing transfer of the pollen from enhanced pollenizer watermelon to
the female flowers of the triploid watermelon. Easier access by bees to
the male flowers of the enhanced pollenizer and coupled with a greater
frequency of male flowers provides a greater pollen source for triploid
fruit production.
[0011]A second advantage of small leaves, in one embodiment characterized
by deep, non-overlapping lobes, is that more sunlight is able to
penetrate to adjacent triploid plants. A third advantage of small leaves,
in one embodiment characterized by deep, non-overlapping lobes, is that
these leaves take up less field area than the substantially larger leaves
of the diploid pollenizers currently used in the production of seedless
watermelon. Thus, as it is less competitive for light, water and
fertilizers, the enhanced pollenizer of the present invention can also be
grown closer to the triploid plants, and it does not need dedicated space
to grow. In one embodiment, when the enhanced pollenizer and method of
the present invention are used, the triploid seedless watermelon are
grown in solid rows at a standard spacing, the enhanced pollenizer being
then inter-planted between the plants within the rows. This results in
significantly higher numbers of triploid plants per acre compared to the
number of triploid watermelon plants that has traditionally been planted,
and higher yields of seedless fruits.
[0012]In one embodiment, the fruit of the enhanced pollenizer of the
present invention are small and therefore easier to distinguish from the
seedless fruits in the field. Therefore, also according to the present
invention, there is provided a novel enhanced pollenizer comprising small
fruits with brittle rind. The small fruits with brittle rind also reduce
the load to the plant and allow the plant to continue flowering for
extended periods of time, significantly greater than pollenizer
watermelons that are currently used in the production of seedless
watermelon. The longer flowering duration of the enhanced pollenizer,
compared to traditional pollenizer diploid watermelons, results in
increased fruit set and yield of seedless watermelon. The brittle rind
also offers the advantage that un-harvested fruits of the pollenizer
quickly decompose in the fields, and can be easily eliminated from
further re-production through conventional crop disposal (discing and
plowing). The brittle rind also offers the advantage of differentiating
the fruit of pollenizer from the fruit of triploid seedless watermelon,
even when they are similar in appearance.
[0013]An additional advantage of the enhanced pollenizer of the present
invention is also that its fruits contain very large amounts of seeds,
which can be harvested and sold as edible watermelon seeds for food or
feed uses, or for use in medicines. This provides additional value to the
grower who can harvest and market the fruits of the enhanced pollenizer
as such or its seeds.
[0014]The present invention also includes an enhanced pollenizer fruit
that weighs approximately in the range of about 2 to 7 lbs, in one
embodiment about 2 to about 6 lbs, in one embodiment about 2 to about 5
lbs. In one embodiment, the average weight for the fruits of the enhanced
pollenizer is about 3.2 lbs.
[0015]In one embodiment, the present invention further includes an
enhanced pollenizer fruit rind that is brittle, breaking under a pressure
approximately in the range of about 7 to about 11 lbs/in.sup.2. In
another embodiment, an enhanced pollenizer fruit rind breaks under a
pressure approximately in the range of about 90 to about 150 g/mm.sup.2,
in one embodiment about 100 to about 148 g/mm.sup.2, in one embodiment
about 110 to about 145 g/mm.sup.2, in one embodiment about 120 to about
140 g/mm.sup.2.
[0016]In one embodiment, the rind of a fruit an enhanced pollenizer of the
instant invention breaks under a pressure of not more than about 1,800 g
when a fruit tester with a 2 mm tip is used. In one embodiment, the fruit
rind of an enhanced pollenizer of the instant invention breaks under a
pressure approximately in the range of about 400 g to about 1,800 g when
a fruit tester with a 2 mm tip is used.
[0017]In one embodiment, the fruit rind of an enhanced pollenizer of the
instant invention breaks under a pressure of not more than about 2,300 g
when a fruit tester with a 3 mm tip is used. In one embodiment, the fruit
rind of an enhanced pollenizer of the instant invention breaks under a
pressure approximately in the range of about 1,000 g to about 2,300 g
when a fruit tester with a 3 mm tip is used.
[0018]In one embodiment, the average pressure to break the rind of a fruit
of an enhanced pollenizer of the present invention is not more than about
1,300 g when a fruit tester with a 2 mm tip is used. In one embodiment,
the average pressure to break the rind of a fruit of an enhanced
pollenizer of the present invention is approximately in the range of
about 700 g to about 1,300 g when a fruit tester with a 2 mm tip is used.
[0019]In one embodiment, the average pressure to break the rind of a fruit
of an enhanced pollenizer of the present invention is not more than about
2,000 g when a fruit tester with a 3 mm tip is used. In one embodiment,
the average pressure to break the rind of a fruit of an enhanced
pollenizer of the present invention is approximately in the range of
about 1,400 g to about 2,000 g when a fruit tester with a 3 mm tip is
used.
[0020]In one embodiment, the present invention includes an enhanced
pollenizer having leaves with a surface area approximately in the range
of about 20 to about 70 cm.sup.2, in one embodiment about 22.5 to about
50 cm.sup.2, in one embodiment about 25 to about 40 cm.sup.2. In one
embodiment, the average leaf surface area of leaves of the enhanced
pollenizer is approximately about 25 to about 40 cm.sup.2, in one
embodiment about 27.5 to about 37.5 cm.sup.2, in one embodiment about 30
to about 35 cm.sup.2.
[0021]In one embodiment, the surface area of the 5.sup.th leaf on a vine
of an enhanced pollenizer of the present invention counted from the
smallest new leaf at the tip of the vine towards the crown of the plant
is not more than about 50 cm.sup.2. In one embodiment, the surface area
of such 5.sup.th leaf from the smallest new leaf on a vine of an enhanced
pollenizer of the instant invention is approximately in the range of
about 15 cm.sup.2 to about 50 cm.sup.2. In one embodiment, the average
leaf surface area of such 5.sup.th leaves from the smallest new leaf on a
vine of an enhanced pollenizer of the instant invention is not more than
about 40 cm.sup.2. In one embodiment, the average leaf surface area of
the 5.sup.th leaves from the smallest new leaf on a vine of an enhanced
pollenizer is approximately in the range of about 20 cm.sup.2 to about 40
cm.sup.2.
[0022]Also included in the invention is an enhanced pollenizer plant for
pollinating triploid plants producing seedless watermelon fruit,
comprising, at maturity, the characteristics of smaller leaf size
compared to the watermelon variety Sangria.TM., wherein the fruit rind is
more brittle than the rind of the variety Sangria.TM. (a commercial
variety of Syngenta Seeds, Inc.). In one embodiment, the average leaf
surface area of leaves of an enhanced pollenizer of the present invention
is about 3 to about 14 times smaller than that of watermelon variety
Sangria.TM.. In one embodiment, the fruit rind of an enhanced pollenizer
of the present invention is about 2 to about 4 times more brittle than
the fruit rind of watermelon variety Sangria.TM.. In one embodiment, the
enhanced pollenizer further comprises small fruits. In one embodiment,
the leaves of the enhanced pollenizer comprises deep, non-overlapping
lobes.
[0023]The pollenizer diploid watermelon of the invention is further
enhanced by including resistance to various pests and herbicides via
conventional plant breeding methods or genetic transformation.
[0024]The present invention also provides a method for inter-planting
enhanced pollenizer plants amongst the triploid watermelon plants in a
field in a pattern that decreases the ratio of pollenizer plants to
triploid plants and increases the field surface for triploid plants. This
allows for a higher population of triploid plants, than conventional
practices, and results in 25-33% higher yield of seedless fruits.
[0025]Also included in the present invention is a method of increasing the
yield of triploid, seedless watermelon comprising the steps of reducing
fruit load of said enhanced pollenizer watermelon, increasing the
flowering duration of said pollenizer watermelon, planting said enhanced
pollenizer watermelon in a field of triploid watermelon; and harvesting
said triploid watermelon.
[0026]The invention also provides a method of increasing the yield of
triploid seedless watermelon plants by using enhanced pollenizer
watermelon plants, in one embodiment with small fruits, wherein the fruit
as such are not harvested for human consumption. In one embodiment, the
seeds of the fruits of the enhanced pollenizer are used as food or feed,
or in medicines.
[0027]The present invention also provides a method for producing an
enhanced pollenizer according to the present invention comprising
crossing a first watermelon plant having small leaves with a second
watermelon plant producing fruit with brittle rind that splits easily and
selecting for a watermelon plant having the characteristics of the
enhanced pollenizer disclosed herein. In one embodiment, the first
watermelon plant further comprises the characteristic of a heavily
branching lacy vine. In one embodiment, the leaves of the enhanced
pollenizer comprises deep, non-overlapping lobes. In one embodiment, the
first watermelon plant has the characteristics of OW824 disclosed herein.
In one embodiment, the second watermelon plant bears small fruit. In one
embodiment, the second watermelon plant has the characteristics of OW823
disclosed herein. In one embodiment, the first watermelon plant is OW824.
In one embodiment, the second watermelon plant is OW823. In another
embodiment, the first watermelon plant is OW824 and the second watermelon
plant is OW823. In one embodiment, the method further comprises fixing
the traits of the enhanced pollenizer.
[0028]The present invention also discloses a watermelon enhanced
pollenizer obtainable by a method comprising the steps of a) crossing a
watermelon plant with a plant of NO1F3203B (now called SP-1) deposited
under Accession No. PTA-4856, b) obtaining a progeny, c) selecting said
progeny for the characteristics of the enhanced pollenizer, preferably
small leaves and brittle fruit. In one embodiment, it is further selected
for heavily branching lacy vines, in one embodiment for small fruit. In
one embodiment, the method further comprises crossing said progeny either
with itself or with a plant of NO1F3203B, or with another enhanced
pollenizer, and selecting for the said characteristics of the enhanced
pollenizer. In one embodiment, the method further comprises fixing the
traits of the enhanced pollenizer.
[0029]In one embodiment, an enhanced pollenizer of the instant invention
is an inbred or a hybrid. In one embodiment, an enhanced pollenizer of
the instant invention is a dihaploid. In one embodiment, the present
invention discloses a method of producing seeds of an enhanced pollenizer
comprising: a) growing a plant of an enhanced pollenizer according to the
present invention; b) allowing self-pollination of said plant; c)
harvesting seeds from said plant. In one embodiment, the method further
comprises washing and drying said seed.
[0030]In one embodiment, the present invention discloses a method of
vegetative propagating an enhanced pollenizer of the present invention
comprising: a) collecting shoot tissue of a plant of an enhanced
pollenizer; b) cultivating said tissue to obtain proliferated shoots; c)
rooting said proliferated shoots to obtain rooted plantlets. In one
embodiment, the method further comprises growing plants from said rooted
plantlets. In one embodiment, the method further comprises harvesting
seeds from said plants. In one embodiment, the method further comprises
washing and drying said seed.
[0031]In one embodiment, the present invention discloses a method for
producing triploid, seedless watermelon fruit comprising the steps of: a)
inter-planting a seed or a plant of an enhanced pollenizer according to
the present invention and seed or plants of triploid watermelon plants in
a field; and b) allowing pollination of said triploid watermelon plants
by pollen of said diploid watermelon plant to obtain triploid, seedless
watermelon fruit. In one embodiment, the method further comprises
harvesting seeds from said plants. In one embodiment, the method further
comprises washing and drying said seed.
DESCRIPTION OF THE DRAWINGS
[0032]FIG. 1 is a p
hotographic depiction of a leaf of the enhanced
pollenizer plant of the invention.
[0033]FIG. 2 is a p
hotographic depiction of a leaf of the pollenizer
referred to as Sangria.TM. that is currently used in commerce.
DETAILED DESCRIPTION OF THE INVENTION
Development of Seedless Watermelons
[0034]Triploid watermelons are created by crossing a tetraploid (4.times.)
female parent line with diploid (2.times.) male parent line. The
resulting triploid (3.times.) watermelon seeds or plants are planted in a
field with diploid watermelon pollenizers. The resulting fruit of the
triploid watermelon are seedless.
[0035]To create a tetraploid female watermelon line, it is known in the
art to use chemicals that alter mitosis of a diploid inbred line so that
unusual numbers of chromosomes are obtained. For example, colchicine is a
chemical that alters the mitotic spindle fibers of diploid cells
resulting in a number of cells that are tetraploid. The diploid line used
to create a tetraploid is selected based on the traits desired for the
tetraploid line. Traits that are desired for a tetraploid line may
therefore first be introgressed into the diploid inbred lines that will
be used to develop the tetraploid lines by breeding methods well known to
those skilled in the art. Thus, the diploid and tetraploid parent lines
are bred separately for the desired traits.
[0036]It usually requires at least two generations of self-pollination and
selection to "fix" the 4.times. condition, after the colchicine treatment
generation because, often, chromosomal aberrations are encountered that
affect seed fertility, and must be eliminated. Once the stable tetraploid
containing the desired characteristics is verified, it then can be used
as a stable female parent for the production of the triploid hybrid. A
stable diploid inbred is selected for use as the male parent. Methods for
developing tetraploid plants are described in Kihara, H., 1951, Triploid
Watermelons, Proceedings of American Society for Horticultural Science
58:217-230; and Eigsti, O. J., 1971, Seedless Triploids, HortScience 6,
pgs. 1-2.
[0037]The tetraploid female parent line and diploid male parent line are
planted in a seed production field. The pollen of the diploid male parent
is transferred to the female tetraploid flower by methods well known to
those skilled in the art. The triploid seed that is produced is present
in the resulting fruit and is planted to produce the triploid plants. The
breeding of watermelon is further described in Mark Bassett (Editor),
1986, Breeding Vegetable Crops, AVI Publishing, ISBN 0-87055-499-9.
[0038]A triploid seedless watermelon is a true F1 hybrid between a
tetraploid watermelon, as the female parent, and a diploid watermelon, as
the male parent (Kihara, H. 1951. Triploid Watermelons. Proceedings of
American Society for Horticultural Science 58:217-230). The seedless
condition in triploid watermelon is the result of the presence of three
homologous sets of chromosomes per somatic cell rather than the usual
two. Cells with three sets of homologous chromosomes are said to be
triploid and are designated as 3.times.. The triploid seedless
watermelons have 33 chromosomes (2N=3.times.=33) in their somatic cells.
The inability of the triploid zygote to produce normal viable gametes
(pollen and egg cells) causes the absence of seeds in triploid fruits.
Typically, seedless watermelons contain small edible white ovules,
similar to those in immature cucumbers.
[0039]Adequate viable pollen supply from the diploid pollenizer watermelon
is essential for the triploid female flowers to set and develop into
regular seedless fruit. The female flowers of triploid watermelon will
not set if they are not pollinated by viable pollen of diploid
watermelon. (Maynard, D. N. (editor), 2001, Watermelons: Characteristics,
Production and Marketing, ASHS Press, ISBN 0-9707546-1-2). The diploid
watermelon grown in a field of triploid plants is referred to herein as
the "pollenizer." In current commercial triploid watermelon production
fields, the triploid watermelon and diploid pollenizer are inter-planted,
either within row or between rows, in a ratio of approximately 1 diploid
to 2 or 3 triploids. Although research has indicated a 1:4 ratio is
acceptable, it is rarely used in commercial plots. (NeSmith, D. S.,
Duval, J. R. Fruit Set of Triploid Watermelons as a Function of Distance
from a Diploid Pollenizer, HortScience 36(1): 60-61, 2001)
Development of Enhanced Pollenizer Diploid Watermelon
[0040]According to the present invention, a watermelon (OW824) is selected
having the characteristics of a heavily branching lacy vine, early and
prolific male flowers, and small leaves with deep, non-overlapping leaf
lobes. In this example, the fruit of OW824 is relatively large, the rind
and flesh are very firm, the seed size is very big and the flesh is
white. OW824 is a publicly available edible seed watermelon variety
generally referred to as XinJiang edible seed watermelon.
[0041]Also according to the invention, a hybrid watermelon (OW823) is
selected for its small fruit (2-3 kg) with brittle rind that splits
easily. OW823 also includes the characteristics of mid-sized seeds with
yellow flesh and has relatively large leaves. OW823 is a commercially
available variety, Tiny Orchid, from Known-You Seeds, Ltd. of Taiwan.
[0042]Crossing OW824 X OW823 generated progeny having the characteristics
of the enhanced pollenizer diploid watermelon of the present invention as
described in more detail below.
[0043]The initial cross of OW824 X OW823 was made during the summer of
2000 in California. The F.sub.1 generation was grown in the greenhouse in
the fall of 2000. The F.sub.2 population was grown Florida in the spring,
and in California in the summer of 2001. Individuals with the set of
traits required for the enhanced pollenizer were successfully identified
and self-pollinated in F.sub.2 populations grown in both locations. A
total 7 selections were made. The 7 F.sub.3 lines were grown in the field
in Florida and the greenhouse in California in the fall of 2001 for
further selection and evaluation. Three F.sub.3 lines were identified to
best meet our breeding goals and advanced to F.sub.4 generation. They all
have the set of the traits required by the enhanced pollenizer. One line,
NO1F3203B, now called SP-1, is fixed for every trait concerned. NO1F3203B
contains the traits that are illustrative of the traits of the enhanced
pollenizer of the invention. Other enhanced pollenizer lines with similar
characteristics were for example SP-2 with slightly larger leaves than
SP-1, and SP-3 with slightly larger fruits than SP-1 and a different
fruit skin color.
[0044]Leaf: The leaves of the enhanced pollenizer are significantly
smaller and are more numerous than that of the commonly used pollenizers
such as the variety Sangria.TM. (See FIGS. 1 and 2). The size of a leaf
is determined by measuring its surface area. The surface area of
different types of leaves of a watermelon plant can be measured. In one
embodiment, the surface area of the 5.sup.th leaf on a vine, counted from
the smallest new leaf at the tip of the vine towards the crown of the
plant, is measured. The smallest new leaf at the tip of a vine is
typically about 5 cm in length and width, and is counted as the first
leaf. Such 5.sup.th leaf is generally referred to herein as the 5.sup.th
leaf from the smallest new leaf on a vine. In another embodiment, the
surface area of an average mature and fully developed leaf is measured.
An average mature and fully developed leave is for example a leaf at the
fifth node from the crown of a plant. In another embodiment, the surface
area of the leaves, which appear to be the largest on a plant, is
measured.
[0045]In one embodiment, the leaves of an enhanced pollenizer of the
present invention have a surface area approximately in the range of about
20 to about 70 cm.sup.2, in one embodiment about 22.5 to about 50
cm.sup.2, in one embodiment about 25 to about 40 cm.sup.2. In another
embodiment, the average leaf surface area of the leaves of the enhanced
pollenizer is approximately about 25 to about 40 cm.sup.2, in one
embodiment about 27.5 to about 37.5 cm.sup.2, in one embodiment about 30
to about 35 cm.sup.2.
[0046]In one embodiment, the surface area of the 5.sup.th leaf from the
smallest new leaf on a vine of an enhanced pollenizer of the present
invention is measured. In one embodiment, the surface area of the
5.sup.th leaf from the smallest new leaf on a vine of an enhanced
pollenizer of the present invention is not more than about 50 cm.sup.2.
In one embodiment, the surface area of the 5.sup.th leaf from the
smallest new leaf on a vine of an enhanced pollenizer of the instant
invention is approximately in the range of about 15 cm.sup.2 to about 50
cm.sup.2. In one embodiment, the average leaf surface area of 5.sup.th
leaves from the smallest new leaf on a vine of an enhanced pollenizer of
the present invention is not more than about 40 cm.sup.2. In one
embodiment, the average leaf surface area of 5.sup.th leaves from the
smallest new leaf on a vine of an enhanced pollenizer of the present
invention is approximately in the range of about 20 to about 40 cm.sup.2.
For example, Tables 1A and 1D below describe measurements of the surface
area of 5.sup.th leaves from the smallest new leaf on a vine of
NO1F3203B/SP-1.
[0047]In one embodiment, the surface area of average mature and fully
developed leaves of an enhanced pollenizer of the present invention is
measured. In one embodiment, the surface area of such average mature and
fully developed leaves of an enhanced pollenizer of the present invention
is not more about 90 cm.sup.2. In one embodiment, the surface area of an
average mature and fully developed leaf of an enhanced pollenizer of the
present invention is approximately in the range of about 40 cm.sup.2 to
about 90 cm.sup.2. For example, Tables 1C and 1D below describe
measurements of the surface area of average mature and fully developed
leaves of NO1F3203B/SP-1.
[0048]In one embodiment, the surface area of largest leaves of plants of
the enhanced pollenizer of the present invention is determined. In one
embodiment, such largest leaves have a surface area not more than about
120 cm.sup.2, in one embodiment approximately in the range of about 60
cm.sup.2 to about 120 cm.sup.2. For example, Table 1B below describes
measurements of the surface area of such large leaves of NO1F3203B/SP-1.
[0049]Clearly, due to various environmental and physiological conditions,
the size of the leaves of a watermelon plant may vary. Accordingly, in
one embodiment, at least about 80% of the leaves of an enhanced
pollenizer of the instant invention in a field show the above
characteristics of surface area. In one embodiment, at least about 90% of
the leaves of an enhanced pollenizer of the instant invention in a field
show the above characteristics of surface area.
[0050]In one embodiment, the leaves of the enhanced pollenizer have deep,
non-overlapping leaf lobes.
[0051]The leaf surface areas of the enhanced pollenizer NO1F3203B and the
Sangria.TM., a pollenizer favored by growers, are shown for comparison
purposes in Table 1A-D.
[0052]In Table 1A the leaves for both NO1F3203B and Sangria.TM. were taken
from mature plants sowed on Aug. 20, 2001 and harvested on Nov. 8, 2001.
In Table A, 5.sup.th leaves from the smallest new leaf on a vine were
harvested.
[0053]In Table 1B the leaves of both NO1F3203B and Sangria.TM. were taken
from mature plants sowed on Aug. 7, 2003, transplanted to open fields on
Sep. 9, 2003 and harvested on Oct. 31, 2003. In Table 1B, leaves, which
appeared to be the largest leaves on the plants, were harvested.
[0054]In Table 1C leaves of both NO1F3203B and Sangria.TM. were taken from
mature plants sowed on Aug. 7, 2003, transplanted to open fields on Sep.
9, 2003 and harvested on Dec. 4, 2003. The leaves used in Table 1C were
average mature and fully expanded leaves at the fifth node from the crown
of the plant.
[0055]In Table 1D the leaves for both NO1F3203B (SP-1) and Sangria.TM.
were taken from mature plants sowed on Jan. 16, 2004 and harvested on May
16, 2004. Average mature and fully developed leaves (M) and 5.sup.th
leaves from the smallest new leaf on a vine (Y) were harvested. The data
in the Tables are given in cm.sup.2 (also given in square inches (sq.
in.) in Table 1B and 1C).
[0056]Plants were grown at the Naples station in Florida. Leaf samples
were collected and the leaves p
hotocopied. In Table 1A, the surface area
of the leaves was determined using graph paper. In Tables 1B-D, the
p
hotocopies of leaves were scanned and the surface area determined using
a computer. In particular, in Table 1D, a WinRhizo STD 1600+scanner and
the program WinRhizo Pro 2003 were used to analyze the leaf images.
TABLE-US-00001
TABLE 1A
SANGRIA
NO1F3203B LEAF cm.sup.2 LEAF cm.sup.2
A 38.75 A 232.00
B 26.25 B 447.25
C 39.75 C 241.50
D 28.75 D 238.00
E 38.25 E 211.00
F 26.27
Average (.+-.Std Dev) 33.08 (.+-.6.46) 273.95 (.+-.97.60)
TABLE-US-00002
TABLE 1B
NO1F3203B Sangria
Sample cm.sup.2 (sq.in.) cm.sup.2 (sq.in.)
1 63.42 (9.83) 301.81 (46.78)
2 100.19 (15.53) 285.81 (44.30)
3 103.87 (16.10) 212.00 (32.86)
4 78.00 (12.09) 334.77 (51.89)
5 114.58 (17.76) 330.90 (51.29)
Average 92.00 (14.26) 293.03 (45.42)
Std Dev 20.77 (3.22) 49.67 (7.70)
TABLE-US-00003
TABLE 1C
NO1F3203B LEAF SANGRIA LEAF
Sample cm.sup.2 (sq.in.) cm.sup.2 (sq.in.)
1 59.16 (9.17) 213.03 (33.02)
2 51.35 (7.96) 242.37 (37.57)
3 51.48 (7.98) 265.10 (41.09)
4 43.74 (6.78) 245.16 (38.00)
5 57.94 (8.98) 274.25 (42.51)
Average 52.71 (8.17) 248.00 (38.44)
Sdt Dev 5.53 (0.86) 21.19 (3.28)
TABLE-US-00004
TABLE 1D
Leaf Stage Leaf ID Area (cm2) Average Std Dev
SP-1 Y sp6-1 18.79
SP-1 Y sp6-2 40.31
SP-1 Y sp6-3 19.00
SP-1 Y sp6-4 32.34
SP-1 Y sp6-5 22.47 26.58 7.79
Sangria Y sg6-1 120.35
Sangria Y sg6-2 119.00
Sangria Y sg6-3 103.16
Sangria Y sg6-4 152.82
Sangria Y sg6-5 118.09 122.68 12.06
SP-1 M sp-1 60.99
SP-1 M sp-2 59.01
SP-1 M sp-3 44.05
SP-1 M sp-4 80.91
SP-1 M sp-5 81.56
SP-1 M sp-6 64.49
SP-1 M sp-7 60.61
SP-1 M sp-8 51.06
SP-1 M sp-9 51.88
SP-1 M sp-10 41.55 59.61 10.10
Sangria M sg-1 218.92
Sangria M sg-2 317.81
Sangria M sg-3 261.77
Sangria M sg-4 244.82
Sangria M sg-5 235.53
Sangria M sg-6 234.35
Sangria M sg-7 255.25
Sangria M sg-8 261.73
Sangria M sg-9 237.72
Sangria M sg-10 240.58 250.85 18.63
[0057]In one embodiment, the surface area of the enhanced pollenizer leaf
of the invention is approximately 3 to 14 times less than the surface
area of the typical diploid pollenizer, Sangria.TM. plant. In one
embodiment, the surface area of the enhanced pollenizer leaf of the
invention is approximately 5 to 12 times less than the surface area of
the typical diploid pollenizer, Sangria.TM. plant.
[0058]FIG. 1 illustrates the non-overlapping characteristic of the deep,
non-overlapping lobed leaves of the enhanced pollenizer. Clearly, due to
various environmental and physical forces, some of the leaves in this
population may have some overlapping lobes, but overlapping lobes are not
characteristic thereof. In contrast, the Sangria.TM. leaf shown in FIG. 2
is characterized as having leaf lobes that habitually overlap each other.
The small, deeply lobed and non-overlapping leaves of the invention allow
more sunlight through to adjacent triploid watermelon plants.
[0059]Branching: In one embodiment, an enhanced pollenizer of the
invention is also heavily branched (also referred to as "lacy vines" or
"open vines"), having significantly more branches (average of 25.9) than
the variety referred to as Sangria.TM. M, (average of 13). In one
embodiment, an enhanced pollenizer of the present invention produces
secondary and tertiary branches on the main branch, thus allowing for
very large amounts of male flowers. Moreover, in one embodiment, an
enhanced pollenizer of the present invention, for example, a plant of
SP-1, also develops tertiary branches late in the season, a
characteristic rarely observed on regular watermelon plant like
Sangria.TM.. The lacy vine characteristic enables the enhanced pollenizer
to produce more accessible male flowers than current diploid pollenizers,
thereby enhancing exposure of the flowers to bees. The open or lacy vines
also permit the inter-planting of the enhanced pollenizer between
triploid plants thereby allowing for higher triploid populations and
greater seedless fruit production.
[0060]Fruit: The fruit rind of the enhanced pollenizer is very brittle and
is easily broken. The brittle fruit rind splits easily, due to natural
maturation or by breaking or splitting of the fruit during harvest of the
seedless triploid watermelon (for example from foot traffic). Splitting
of fruit signals the plant that it hasn't completed its reproductive
process inducing the plant to continue flowering for a longer period of
time. Brittleness is conferred by a gene e (explosive rind, thin, and
tender rind, bursting when cut (Rhodes & Dane, 1999, Gene List for
Watermelon, Cucurbit Genetics Cooperative Report 22:71-77; Nihat Guner &
Todd C. Wehner, 2003, Gene List for Watermelon, Cucurbit Genetics
Cooperative Report 26:76-92; Porter D. R. (1937) Inheritance of certain
fruit and see characters in watermelon Hilgardia 10: 489-509; Poole C. F.
(1944) Genetics of cultivated cucurbits J. Hered. 35: 122-128).
Accordingly, in one embodiment, the brittleness of the fruit of an
enhanced pollenizer of the present invention is conferred by a gene e,
and the present invention includes the use of a watermelon plant
comprising a gene e as pollenizer for triploid watermelon plants.
[0061]When measured by a penetrometer, a fruit of NO1F3203B breaks at
about 7-11 lbs/in.sup.2, whereas a fruit of a typical watermelon such as
Sangria.TM. breaks at about 21-27 lbs/in.sup.2. Accordingly, in one
embodiment, the fruit rind of an enhanced pollenizer of the present
invention is about 2 to about 4 times more brittle than the fruit rind of
watermelon variety Sangria.TM.. In one embodiment, using a Tester FT02 of
Wagner Instruments, Greenwich, Conn. 06836, the fruit of the enhanced
pollenizer breaks under a pressure approximately in the range of about 90
to about 150 g/mm.sup.2, in one embodiment about 100 to about 148
g/mm.sup.2, in one embodiment about 110 to about 145 g/mm.sup.2, in one
embodiment about 120 to about 140 g/mm.sup.2. By comparison, the fruit of
Sangria.TM. breaks under a pressure of approximately about 300
g/mm.sup.2.
[0062]In one embodiment, measurements of the brittleness of a watermelon
fruit are carried out with a Tester FT327, a Tester FT011 or a Tester
FT02 from Wagner Instruments, Greenwich, Conn. 06836, with a 2 mm or a 3
mm tip.
[0063]In one embodiment, the rind of a fruit of an enhanced an enhanced
pollenizer of the instant invention breaks under a pressure under about
1,800 g when a fruit tester with a 2 mm tip is used. In one embodiment,
the fruit rind of an enhanced an enhanced pollenizer of the instant
invention breaks under a pressure approximately in the range of about 400
g to about 1,800 g when a fruit tester with a 2 mm tip is used.
[0064]In one embodiment, the fruit rind of an enhanced an enhanced
pollenizer of the instant invention breaks under a pressure under about
2,300 g when a fruit tester with a 3 mm tip is used. In one embodiment,
the fruit rind of an enhanced an enhanced pollenizer of the instant
invention breaks under a pressure approximately in the range of about
1,000 g to about 2,300 g when a fruit tester with a 3 mm tip is used.
[0065]In one embodiment, the average pressure to break the rind of a fruit
of an enhanced pollenizer of the present invention is under about 1,300 g
when a fruit tester with a 2 mm tip is used. In one embodiment, the
average pressure to break the rind of a fruit of an enhanced pollenizer
of the present invention is approximately in the range of about 700 g to
about 1,300 g when a fruit tester with a 2 mm tip is used.
[0066]In one embodiment, the average pressure to break the rind of a fruit
of an enhanced pollenizer of the present invention is under about 2,000 g
when a fruit tester with a 3 mm tip is used. In one embodiment, the
average pressure to break the rind of a fruit of an enhanced pollenizer
of the present invention is approximately in the range of about 1,400 g
to about 2,000 g when a fruit tester with a 3 mm tip is used.
[0067]In one embodiment, at least about 80% of the fruits of a plant of
the instant invention in a field show the above characteristics of rind
brittleness. In one embodiment, at least about 90% of the fruits of a
plant of the instant invention in a field show the above characteristics
of rind brittleness.
[0068]The environmental conditions generally influence the brittleness of
the rind of a fruit of the present invention. For example, the rind of
fruits grown under warm and sunny conditions tends to be more brittle
than that of fruits grown under cooler and shadier conditions. This is
for example reflected in the measurements of Example 12, where a Spring
crop was tested compared to a Fall crop in Examples 10 and 11.
[0069]In one embodiment, the fruit size of the enhanced pollenizer is
approximately in the range of about 5 to about 7 inches long.times.about
6 to about 8 inches wide. In one embodiment, the fruit size of the
enhanced pollenizer is approximately about 6 inches long.times.about 7
inches wide, whereas the typical pollenizer is about 10 inches
long.times.20 inches wide. Small fruit size, as well its brittleness was
selected to decrease the load on the plant, thereby extending the
duration of plant growth and flower production. Another advantage of the
small fruit size is that it enables the harvester to easily distinguish
the seedless fruit from seeded fruit, is often difficult with currently
used pollenizers, which are selected based on their overall similarity to
the seedless triploid plants. The fruit of the enhanced pollenizer weighs
approximately in the range of about 2 to about 7 lbs, in one embodiment
about 2 to about 6 lbs, in one embodiment about 2 to about 5 lbs. In one
embodiment, the average weight for the fruits of the enhanced pollenizer
is about 3.2 lbs.
[0070]In one embodiment, the rind color of the enhanced pollenizer is
light green with very thin dark green lines. The fruit of the enhanced
pollenizer of the invention can be distinguished from the fruit of most
(about 99%) of the commercially available seedless watermelon varieties.
[0071]Flowering: The plants of the enhanced pollenizer, e.g. of NO1F3203B,
also flower approximately 7 to 10 days earlier than diploid pollenizer
plants currently used for the production of seedless watermelon, and
continue flowering during fruit harvest time of the seedless watermelon,
2 to 3 weeks longer than standard diploid pollenizer plants. Thus, the
pollenizer plant of the invention has a flowering duration that is
approximately 3 to 5 weeks longer than pollenizers currently used.
[0072]Other Traits: The enhanced pollenizer, e.g. NO1F3203B, can be used
either as donor of the set of traits disclosed above, or as the recurrent
parent to develop additional enhanced pollenizer lines. In accordance
with the invention, the enhanced pollenizer watermelon contains traits of
disease resistance (e.g. Fusarium wilt, Anthracnose, Gummy Stem Blight,
Powdery Mildew, and Bacterial Fruit Blotch), insect resistance (e.g.
cucumber beetle, aphids, white flies and mites), salt tolerance, cold
tolerance and/or herbicide resistance added. These traits can be added to
existing lines by using either conventional backcrossing method, pedigree
breeding method or genetic transformation. The methods of conventional
watermelon breeding are taught in several reference books, e.g. Maynard,
D. N. (editor), 2001, WATERMELONS Characteristics, Production and
Marketing, ASHS Press; Mohr, H. C., Watermelon Breeding, in Mark J.
Bassett (editor), 1986, Breeding Vegetable Crops, AVI Publishing Company,
Inc. General methods of genetic transformation can be learned from
publish references, e.g. Glich et al., (Eds), 1993, Methods in Plant
Molecular Biology & Biotechnology, CRC Press, and more specifically for
watermelon in WO02/14523.
[0073]Forms of the Enhanced Diploid Pollenizer: Once the enhanced
pollenizer lines are developed, several forms of enhanced pollenizer
varieties can be used in commercial seedless watermelon production.
Specifically, these forms of enhanced pollenizer varieties include: Forms
of Enhanced Pollenizer: (1) Open Pollinated Variety: The stable, enhanced
lines of the enhanced pollenizer are grown in isolated fields, at least
2,000 meters from other watermelon varieties. Pollination is conducted in
the open fields by bees. Seeds are harvested from the seed production
field when the fruit and seeds are fully developed. The seeds are dried
and processed according to the regular watermelon seed handling
procedures. (2) Synthetic Variety: The seed of different enhanced
pollenizer lines are individually produced in isolated fields. Bee
pollination is used in each isolation. The seed of different enhanced
pollenizer are separately harvested and processed. Mixing several
enhanced pollenizer lines in various ratios forms the synthetic
varieties. The synthetic variety can provide a broader pollenizer
population for the triploid watermelons. (3) Open-Pollinated Hybrid
Variety: Two or several enhanced pollenizer lines are planted in the same
seed production field with bee pollination. The harvested seed lot,
therefore, contains both hybrid and inbred seed. (4) Hybrid Variety: Two
enhanced pollenizer lines, the male and female parents, are planted in
the same field. Hand pollination is conducted. Only the seed from female
parent line is harvested and sold to the commercial grower to use as
pollenizer. Table 3 in Example 7 shows the results obtained using various
combinations of inbred and hybrid enhanced pollenizers.
[0074]In one embodiment, an enhanced pollenizer of the present invention
is a dihaploid. A dihaploid is for example produced by gamma-ray
irradiation of the anthers followed by pollination of female flowers with
irradiated pollen and embryo rescue. In one embodiment, an enhanced
pollenizer is grafted on rootstock according to methods standard in the
art.
[0075]Method of Seedless Watermelon Production: Most current commercial
seedless watermelon growers in NAFTA use elongated diploid varieties with
an Allsweet stripe pattern: light green skin with wide green stripes, as
the pollenizer. The variety referred to as Sangria.TM. is the most
preferred Allsweet type pollenizer and is available as a commercial
product from Syngenta Seeds, Inc., Boise Id. Typically, the pollenizer is
inter-planted with the triploid watermelon either between rows or within
row. The current method of planting diploid pollenizers include planting
the diploid plants at a distance from adjacent triploid such that they
have the same field area available per plant as the field area that is
available to the triploid watermelon plants. For example, currently
watermelon growers inter-plant the diploids within a row, whereby the
space between all adjacent plants within the row are approximately
equidistant.
[0076]Alternatively, diploid pollenizer plants are planted in separate
rows between rows of triploid watermelon plants. All rows of diploid and
triploid plants in such a field are planted approximately equidistant
from each other. In other words, under current methods for producing
seedless watermelon, the width of all diploid and triploid rows is the
same.
[0077]In one embodiment, a method of the present invention includes
planting the enhanced pollenizer watermelon plants in rows that are
narrower than the triploid rows, thereby saving field area for production
of triploid seedless watermelon. In one embodiment, a method of the
present invention includes planting an enhanced pollenizer watermelon
plant within a row of triploid watermelon plants. In one embodiment, a
method of the present invention includes planting an enhanced pollenizer
watermelon plant and a triploid watermelon plant in the same hole. In one
embodiment, enhanced pollenizer watermelon plants and triploid watermelon
plants are planted in a ratio of 3-4:1, i.e. in every 3.sup.rd or
4.sup.th hole both an enhanced pollenizer plant and a triploid watermelon
plant are planted in the same hole. In one embodiment, an enhanced
pollenizer watermelon plant of the present invention is planted within
pollinating distance of a triploid watermelon plant.
[0078]In one embodiment, a seed or a plant (e.g. a young plant about 2-4
weeks after sowing) of an enhanced pollenized of the present invention is
planted in a field.
[0079]Table 2 below shows examples of different planting alternatives for
watermelon pollenizer, including a preferred inter-planting according to
the present invention (right column).
TABLE-US-00005
TABLE 2
seeded seedless seedless seeded seedless seedless seeded
Conventional 2:1 pollenizer ratio using the row method
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
.largecircle. X X .largecircle. X X .largecircle.
Conventional 2:1 pollenizer ratio using the within row method
X .largecircle. X X .largecircle. X X
.largecircle. X X .largecircle. X X .largecircle.
X X .largecircle. X X .largecircle. X
X .largecircle. X X .largecircle. X X
.largecircle. X X .largecircle. X X .largecircle.
X X .largecircle. X X .largecircle. X
X .largecircle. X X .largecircle. X X
.largecircle. X X .largecircle. X X .largecircle.
Pollenizer inter-planted at a 3:1 pollenizer ratio
X X X X X X X
.diamond-solid. .diamond-solid.
X X X X X X X
.diamond-solid. .diamond-solid. .diamond-solid.
X X X X X X X
.diamond-solid. .diamond-solid.
X X X X X X X
.diamond-solid. .diamond-solid.
X X X X X X X
.diamond-solid. .diamond-solid. .diamond-solid.
X X X X X X X
.diamond-solid. .diamond-solid.
X X X X X X X
.diamond-solid. .diamond-solid.
X X X X X X X
Seeded = .largecircle.
Seedless = X
Pollenizer = .diamond-solid.
EXAMPLES
[0080]The following Examples are provided to illustrate the present
invention, and should not be construed as limiting thereof.
Example 1
[0081]Triploid watermelon plants are planted in parallel rows 7 feet apart
and 3 feet apart within each row. However, the enhanced diploid
watermelon plants are planted in a narrow row 3.5.degree. wide (1/2 the
width of the triploid rows) between every second and third triploid row.
For example, rows A and B are two consecutive rows of triploids, each
7-foot wide. Row C is a diploid row that is 3.5 feet wide. Row D and E
are the following two 7 foot wide rows of triploids, followed by the
3.5-foot wide row F of diploid plants. This pattern is repeated across
the width of the field. Because the diploid row is narrower according to
the method of the invention, the distance between rows B and D is 10.5
feet instead of the traditional distance of 14 feet. Using this ratio of
1 pollenizer row for every 2 triploid rows (1:2), 33.3% of the field
would normally be used for the pollenizer plants. Reducing the width of
the pollenizer row according to the method of the invention by one-half,
the gain of space for planting additional triploid plants would be
33.3%/2 or approximately 17%.
Example 2
[0082]Triploid watermelon plants are again planted in parallel rows 7 feet
apart and 3 feet apart within each row. As in Example 1, the enhanced
diploid watermelon plants are planted in a narrow row 3.5' wide, but are
planted between every third and fourth triploid row. For example, rows A,
B, and C, are three consecutive rows of triploids, each row being 7'
wide. The following row D is a diploid row that is 3.5 feet wide. Row E,
F, and G are the following three rows of triploids, all 7 feet wide,
followed by a 3.5 foot wide row of enhanced pollenizer plants. This
pattern is repeated across the width of the field. Because the diploid
row is narrower according to the method of the invention, the distance
between rows B and D is again 10.5 feet instead of the traditional
distance of 14 feet. Using this ratio of 1 pollenizer row for every 3
triploid rows (1:3), 25% of the field would normally be used for the
pollenizer plants. Reducing the width of the pollenizer row according to
the method of the invention by one-half, the gain of space for planting
additional triploid plants would be 25%/2 or approximately 12%.
Example 3
[0083]Triploid watermelons are planted in parallel rows 8 feet apart and 3
feet apart within each row. The enhanced diploid watermelon plants are
planted in a narrow row 4.0 feet wide (1/2 the width of the triploid
rows) between every second and third triploid row. For example, rows A
and B are two consecutive rows of triploids, each 8 foot wide. Row C is a
diploid row that is 4.0 feet wide. Row D and E are the following two 8
foot wide rows of triploids, followed by the 4.0 feet wide row F of
diploid plants. This pattern is repeated across the width of the field.
Because the diploid row is narrower according to the method of the
invention, the distance between rows B and D is 12.0 feet instead of the
traditional distance of 16 feet. Using this ratio of 1 pollenizer row for
every 2 triploid rows (1:2), 33.3% of the field would normally be used
for the pollenizer plants. Reducing the width of the pollenizer row
according to the method of the invention by one-half, the gain of space
for planting additional triploid plants would be 33.3%/2 or approximately
17%.
Example 4
[0084]Referring to the above three examples, when triploids are planted in
rows 8 feet apart, and the ratio of diploid to triploid is 1:3, it is now
clear that the reduction of the pollenizer row width by one-half will
gain space for planting additional 12%.
Example 5
[0085]It is also within the scope of the invention to reduce the
pollenizer row width to approximately 1/3 that of the triploid row width.
Thus, according to the present invention, at any row width, when the
ratio of diploid rows to triploid rows is:
[0086](a.) 1:2, the savings of field area for additional triploid plants
is (33%.times.2/3) or 22%.
[0087](b) 1:3, the savings of field area for additional triploid plants is
(25%.times.2/3) or 16.5%.
[0088](c) 1:4, the savings of field area for additional triploid plants is
(20%.times.2/3) or 13.2%.
[0089]It is also within the scope of the invention to reduce the
pollenizer row width to approximately 2/3 that of the triploid row width.
Example 6
[0090]It is also within the scope of the present invention to inter-plant
the diploid plants within the rows of triploid plants. According to the
invention, the triploid plants are first planted by machine or by hand in
regularly spaced rows. The triploid plants within each row are planted,
for example, 3 feet apart. After the triploid plants are in the field as
described, the diploid pollenizer watermelon plants of the invention are
inter-planted, by hand, within each row approximately midway between the
triploid plants, i.e. the diploid pollenizer watermelon plants of the
invention are inserted between the triploid plants. Thus, in this
example, the diploid plants are planted approximately 1.5 feet from the
flanking triploid plants within the row. Due to the characteristics of
the enhanced pollenizer of the invention, the diploid plants can be
inter-planted within each row after every 2, 3, 4, 5, 6, 7, 8, 9, or 10
consecutive triploid plants. It is currently preferred in the industry to
plant the diploid plants after every 2 (1:2) or 3 (1:3) triploid plants
within the row. A 1:4 ratio has been reported, but is not normally used
in commercial fields due to inadequate pollenization of the triploid
plants. The field area saved under this example, when compared with both
the current methods of planting diploids in separate rows or within a row
at the ratios (diploid:triploid) of:
[0091](a) 1:2, is 33.3%,
[0092](b) 1:3, is 25%,
[0093](c) 1:4, is 20%.
[0094]The enhanced pollenizer and method of the present invention
comprises planting the enhanced pollenizer watermelons in rows that are
narrower than the rows containing the triploid plants. Although the
narrower diploid row will encourage diploid plant growth into the
triploid plant row, the novel characteristics of the enhanced pollenizer
watermelon allow it maintain its ability to sufficiently pollinate the
triploid plants in the field. Thus, the enhanced pollenizer watermelon
and method of the present invention increase the yield of seedless
watermelon in a field.
Example 7
[0095]A split-plot design is used for this experiment to test three inbred
enhanced pollenizers and three hybrid enhanced pollenizers against the
commercial checks Sangria 2:1 and Sangria 3:1. All 6 enhanced pollenizers
are inserted between regularly spaced (80''.times.24'') triploid plants
in the ratio of 3:1. For Sangria 2:1 ratio, every third space is a
Sangria plant. For Sangria 3:1 ratio, every 4.sup.th space is a Sangria
plant. A 5:1 ratio is also included in this trial using the mixed
enhanced pollenizers. In this treatment, the enhanced pollenizers plant
is inserted between 5.sup.th and 6.sup.th regularly spaced triploid
plants. So there are total 9 main plots, the 9 main
treatments/pollinators, in this experiment. The 9 main plots are
separated by cantaloupe plants. 3 different triploids, the sub-plots,
with 2 replications are used to test different pollinators (see table 3).
Plants are well grown except the leaf-miner damage. This damage results
in smaller fruit size for Palomar and Tri-X-313. The trials are evaluated
after about two months. The number of triploid fruit in each sub-plot is
counted. The first 15 fruits in each sub-plot are non-selectively
harvested and weighted. 10 fruits are also harvested from each pollinator
and measured for rind firmness. Data are analyzed using S-Plus 6.1. The
enhanced pollenizers varieties are also evaluated for fruit size and
other fruit characteristics.
[0096]As shown in table 3, very similar fruit set per plant is achieved
for all the pollenizer used. Smaller triploid seedless melons are
produced when Sangria is used as pollinator in the ratio of 2:1 in this
experiment. This could be due to Sangria's strong competition to the
triploid plant for space, water and nutrient. A lot more seedless melons
per acre, 25% (compared to the 3:1 ratio) to 33% (compared to the
standard 2:1 ratio), are produced when enhanced pollenizers varieties are
used as pollenizer.
[0097]The rind of enhanced pollenizer varieties of the present invention
is much less durable compared to diploid pollenizer Sangria, as indicated
by the force used to penetrate the rind using a fruit firmness tester
(Fruit Firmness Tester FT02 of Wagner Instruments, Greenwich, Conn.
06836). Should the pollenizer not be harvested for its commercial value,
its brittle rind allows the pollinator fruit to be destroyed during fruit
harvest or soon thereafter. This is helpful for unloading the pollenizer
plant and maintaining the flowering ability of the pollenizer plants for
longer period of time. The brittle rind of the enhanced pollenizer also
reduces the risk of carry-over into the next season, as a weed, since the
fruit, and plant debris can be easily destroyed, after harvest of the
triploid fruit.
[0098]Enhanced pollenizer plants flower about 7 days earlier than diploid
Sangria. Enhanced pollenizer plants produce more than twice many of
branches compared to Sangria. This allows enhanced pollenizer plants to
produce more male flowers, thereby reducing the number of pollenizer
plants needed. The vine of enhanced pollenizer plant is much thinner than
regular diploid plants. The leaf size and leaf-lobe size of enhanced
pollenizer are much smaller than those of Sangria. All these make
enhanced pollenizer much less competitive for light, water and
fertilizer, compared to regular diploid watermelon.
[0099]Enhanced pollenizer plants are producing male flowers after the
harvest of triploid seedless fruits. This gives the potential of having a
second fruit set and multiple har
vests of triploid seedless fruit with
single planting. The male flowers open earlier in the morning compared to
regular watermelons, especially in the cooler days.
TABLE-US-00006
TABLE 3
Seedless Watermelon Fruit Yields Produced by Using Different Pollenizer
and Rind Firmness of Different Pollenizer
Rind
Fruit/Plant Fruit/Acre Frt Wt (lbs) Firmness
Pollinator Palomar RWT8124 TriX313 Mean Palomar RWT8124 TriX313 Mean
Palomar RWT8124 TriX313 Mean (g/mm.sup.2)
SP Hyb 5:1 2.00 3.60 2.15 2.58 6534 11652 6957 8381 13.6 6.0 15.4 11.6 NA
SP1 2.05 3.55 1.95 2.53 6719 11661 6413 8265 12.2 5.7 14.6 10.8 121
SP1 .times. SP3 2.00 3.60 2.15 2.58 6579 11752 7001 8444 13.2 6.0 14.9
11.3 139
SP2 1.90 3.50 1.90 2.43 6258 11479 6137 7958 12.1 6.0 13.3 10.5 123
SP2 .times. SP1 1.85 3.30 2.20 2.45 6004 10728 7106 7946 13.1 5.8 14.0
10.9 129
SP3 1.90 3.40 1.55 2.28 6210 11170 5116 7499 12.8 6.0 14.1 11.0 133
SP3 .times. SP2 1.90 3.60 2.05 2.52 6219 11649 6577 8149 12.5 5.8 13.9
10.7 129
Sangria 2:1 1.90 3.50 2.00 2.47 4086 7596 4375 5352 10.5 5.7 12.5 9.6 302
Sangria 3:1 1.95 3.35 1.95 2.42 4737 8248 4863 5949 12.4 5.6 12.9 10.3
Mean 1.95 3.52 2.02 2.49 5770 10405 5946 7374 12.5 5.8 13.8 10.7 154
Factor P-value P-value P-value P-value
Pollinator 0.0239 0.0000 0.0000 0.0000
Triploid 0.0000 0.0000 0.0000
Pollinator * 0.4121 0.0061 0.0029
Triploid
Replication 0.9372 0.8580 0.6310
Example 8
[0100]Eight triploid varieties (see table 4) are transplanted on two 80''
beds and spaced 24'' apart. These two beds are located in the center of
our regular hybrid evaluation block. A diploid hybrid bed is placed in
each side of the two trial beds to eliminate the pollination factor.
About 90 plants are transplanted for each variety. Two days later, each
triploid plot is divided into 2 sub-plots and the enhanced pollenizer
SP-1 plants of the present invention are inserted in one of the 2
sub-plots in the ratio of 3:1, for each of the 8 triploid varieties. This
planting pattern allows 3260 triploid plants per acre. The 8 triploid
varieties differ in fruit shape, size and maturity. About 10 weeks later,
the first 30 fruits are non-selectively harvested from each sub-plot and
are weighted using a digital scale. Data are analyzed using S-Plus 6.1.
[0101]As shown in table 4, the fruit size differences are solely due to
triploid variety differences. Inserting of enhanced pollenizer SP-1
between regularly spaced triploid plants in the ratio of 3:1 does not
reduce the fruit size of triploid seedless fruit, regardless of the type
of triploid variety. The triploid varieties used in this trial represent
a very broad spectrum of triploids used in commercial production. They
differ in fruit size, fruit shape, and maturity. Thus, inserting enhanced
pollenizer plants of the present invention between regularly spaced
triploid plants does not reduce the fruit size of the triploid seedless
melons. Therefore, a seedless grower can plant his or her fields solid
with triploid plants and then insert the enhanced pollenizer plants in a
ratio of 3:1 or less. This planting pattern and ratio allows growers to
produce significant higher (25 to 33%) yields of seedless fruit per acre.
TABLE-US-00007
TABLE 4
Effect of Inserting Super-Pollenizer Between Regularly Spaced
(80'' .times. 24'') Triploid Plants in the Ratio of 3:1 to the Fruit Size
of Eight Different Triploid Watermelon Varieties
Super-Pollenizer Insertion
Triploid Variety No Yes Mean
3X Sangria 18.05 18.51 18.28
Palomar 14.23 16.21 15.22
RWT 8126 16.97 17.15 17.06
RWT8124 6.26 6.03 6.15
RWT8139 15.46 14.43 14.94
RWT8140 15.31 15.73 15.52
Shadow 15.97 14.73 15.35
Tri-X-313 15.77 15.60 15.69
Mean 14.75 14.80 14.77
Factor P-Value
Triploid Variety 0.0000
Super-Pollenizer 0.8829
Variety*Super- 0.2451
Pollenizer
Example 9
Production of Dihaploid Watermelon Plants
[0102]Anthers of SP-1 plants were gamma-ray irradiated with cobalt 60 for
a dose of 0.4 KGy. Irradiated pollen was gently transferred from the
anthers to the receptive stigma on or before anthesis. Each ovary of the
pollinated female received an application of 50 ppm CPPU (a plant
cytokinin hormone) to stimulate fruit development. Plants were monitored
for pollination take and fruit development. Fruit was harvested 14 days
or 21 days post-pollination.
[0103]Harvested immature fruit were carefully cut open under sterile
conditions and the seeds were meticulously removed from the flesh. The
distal portion of each seed was cut off before plating about 40 seeds to
each plate of culture medium. Sealed plates with seeds were cultured at
25.degree. C. with a 16-hour p
hotoperiod in a culture room on a Murashige
and Skoog Basal Medium, 30 g/L sucrose, 10 g/L agar supplemented either
with 10 .mu.M BA (2.25 mg/L) or 22.2 .mu.M BA (5 mg/L) and 2.85 .mu.M IAA
(0.5 mg/L), pH 5.8 and dispensed into 100.times.15 petri dishes after
autoclaving.
[0104]After 30 days, seeds were screened for greenish immature embryos for
embryo rescue. Those with embryos were moved to fresh medium. As the
embryos germinated and elongated, they were transferred to small culture
jars with the same medium. When sufficient leaf tissue was present on the
plantlet, a leaf was sampled and ploidy analysis was carried out by flow
cytometry.
[0105]Once the plantlets had been confirmed haploid, cuttings/clones are
made and rooted in vitro. The medium consisted of half strength MS basal
salts, 20 g/L sucrose, 1.0 .mu.M IBA (0.2 mg/L), 4 g/L agar and 1 g/L
Phytagel, pH 5.8. Once a good root system had developed, plantlets are
moved into the greenhouse and planted in trays. The chromosome doubling
occurred in the greenhouse by applying 58 .mu.M Surflan (oryzalin) to all
apical and axillary nodes. Once plants were established and new flowers
exhibited the presence of pollen confirming restored fertility, they were
self-pollinated and seed was harvested. Further increase of the dihaploid
SP-1 can be done in a field isolated from any other watermelon plant, or
physically isolated in a net cage.
Example 10
Measurements of Brittleness of Fruits
[0106]Watermelon plants of SP-1 and Sangria.TM. were sown at the Naples,
Fla. research station on Aug. 7, 2003 and transplanted to open field on
Sep. 9, 2003. Fruits were harvested from the plants on Nov. 20, 2003.
Mature fruits were tested using a Tester FT327 of Wagner Instruments,
Greenwich, Conn. 06836 and a 3.0 mm tip. Ten fruits of SP-1 and
Sangria.TM. were tested. The average pressure to puncture the rind of
fruits of SP-1 was 3 lb 8 oz (1,587 g). The average pressure to puncture
the rind of fruits of Sangria.TM. was 11 lb 6 oz (5,159 g).
Example 11
Measurements of Brittleness of Fruits
[0107]Watermelon plants of SP-1 and Sangria.TM. were grown at the Naples,
Fla. research station. Fruits were harvested on Dec. 4, 2003 from plants
about 17 weeks after sowing. Mature fruits (about 35-40 days after
anthesis) were tested. The fruits were tested in the morning at about 9
to 10 am. Different models of Wagner penetrometers (Tester FT327, a
Tester FT011 or a Tester FT02 of Wagner Instruments, Greenwich, Conn.
06836) were used in combination with a 2 mm or a 3 mm tip. The tip of the
penetrometer was placed vertically on the top surface of the fruit in the
middle portion of the fruit. For SP-1, 5 fruits (1 to 5) were tested in
three independent measurements (A, B, C). For Sangria.TM., one fruit was
tested in three independent measurements (A, B, C). The results are shown
in Table 5A (2 mm tip) and Table 5B (3 mm tip).
TABLE-US-00008
TABLE 5A
FT02/2 mm (force in g) FT327/2 mm (force in g) FT011/2 mm (force in g)
Average Average Average
SP-1 A B C per fruit A B C per fruit A B C per fruit
Fruit 1 1450 1460 1520 1477 1700 1300 1350 1450 1650 1250 1300 1400
2 1120 1000 1220 1113 1100 1100 1200 1133 1700 1200 1150 1350
3 970 990 1370 1110 1750 1000 1200 1317 1100 1100 1150 1117
4 1150 1140 1100 1130 1400 1100 1200 1233 1000 1150 1100 1083
5 1150 1190 1270 1203 1200 1200 1100 1167 1200 1150 950 1100
Average all fruits 1207 1260 1210
(std dev all fruits) (175) (216) (208)
Sangria 1 not able to break through 3500 2900 3500 3300 3350 3750 3150
3417
(std dev) rind (346) (306)
TABLE-US-00009
TABLE 5B
FT02/3 mm (force in g) FT327/3 mm (force in g) FT011/3 mm (force in g)
Average Average Average
SP-1 A B C per fruit A B C per fruit A B C per fruit
Fruit 1 2240 1950 2250 2147 1900 2100 2100 2033 1850 2000 2200 2017
2 1720 1690 1720 1710 1600 1700 1800 1700 1800 1800 1700 1767
3 1910 2000 2100 2003 1700 1600 1800 1700 1750 1550 1650 1650
4 1910 1970 1900 1927 1900 1800 1800 1833 1550 1650 1750 1650
5 1930 1840 1880 1883 1900 1900 1800 1867 1600 1700 1700 1667
Average all fruits 1934 1827 1750
(std dev all fruits) (167) (149) (171)
Sangria 1 not able to break through 4200 5400 4800 4800 5000 5000 5200
5067
(std dev) rind (600) (115)
Example 12
Measurements of Brittleness of Fruits
[0108]Watermelon plants of SP-1 and SANGRIA.TM. were sown on Jan. 16, 2004
at the Naples, Fla. station. Fruits were harvested on May 17, 2004 and
tested using Tester FT 327 (Wagner Instruments, Greenwich, Conn. 06836)
in combination with a 2 mm or a 3 mm tip. Ten mature fruit of SP-1 and
Sangria.TM. were tested, and the results are shown in Table 6.
TABLE-US-00010
TABLE 6
Force in Grams (Pounds) Force in Grams (Pounds)
3 mm Tip (FT 30M) 2 mm Tip (FT 20M)
SP-1 1452 (3.2) 680 (1.5)
1270 (2.8) 726 (1.6)
1452 (3.2) 680 (1.5)
1633 (3.6) 1043 (2.3)
1043 (2.3) 590 (1.3)
1043 (2.3) 771 (1.7)
1588 (3.5) 816 (1.8)
1542 (3.4) 998 (2.2)
1633 (3.6) 907 (2.0)
1588 (3.5) 454 (1.0)
1678 (3.7) 816 (1.8)
AVE 1447 (3.2) 771 (1.7)
STD 179 (0.4) 132 (0.3)
Sangria 4763 (10.5) 2903 (6.4)
5670 (12.5) 2903 (6.4)
6804 (15.0) 3992 (8.8)
4717 (10.4) 3175 (7.0)
7439 (16.4) 4082 (9.0)
5171 (11.4) 2631 (5.8)
5352 (11.8) 2722 (6.0)
5534 (12.2) 2722 (6.0)
5897 (13.0) 3175 (7.0)
6350 (14.0) 3266 (7.2)
AVE 5770 (12.7) 3157 (7.0)
STD 682 (1.5) 381 (0.8)
Deposit
[0109]Applicants have made a deposit of at least 2500 seeds of enhanced
watermelon pollenizer line NO1F3203B (now called SP-1) with the American
Type Culture Collection (ATCC), Manassas, Va., 20110-2209 U.S.A., ATCC
Deposit No: PTA-4856. This deposit of the enhanced watermelon pollenizer
line NO1F3203B/SP-1 will be maintained in the ATCC depository, which is a
public depository, for a period of 30 years, or 5 years after the most
recent request, or for the effective life of the patent, whichever is
longer, and will be replaced if it becomes nonviable during that period.
Additionally, Applicants have satisfied all the requirements of 37 C.F.R.
.sctn..sctn.1.801-1.809, including providing an indication of the
viability of the sample. Applicants impose no restrictions on the
availability of the deposited material from the ATCC; however, Applicants
have no authority to waive any restrictions imposed by law on the
transfer of biological material or its transportation in commerce.
Applicants do not waive any infringement of its rights granted under this
patent or under the Plant Variety Protection Act (7 USC 2321 et seq.).
[0110]The foregoing invention has been described in detail by way of
illustration and example for purposes of clarity and understanding.
However, it will be obvious that certain changes and modifications such
as single gene modifications and mutations, somaclonal variants, variant
individuals selected from large populations of the plants of the instant
inbred and the like may be practiced within the scope of the invention,
as limited only by the scope of the appended claims. Thus, although the
foregoing invention has been described in some detail in this document,
it will be obvious that changes and modification may be practiced within
the scope of the invention, as limited only by the scope of the appended
claims.
[0111]All references cited herein are incorporated by reference in the
application in their entireties.
* * * * *