Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
CUCUMBER VARIETY NUN 93099 CUL
FIELD OF THE DISCLOSURE
100011 The disclosure relates to the field of plant breeding, specifically to
cucumber variety
NUN 93099 CUL. The disclosure further relates to vegetative reproductions of
cucumber variety
NUN 93099 CUL, methods for tissue culture of cucumber variety NUN 93099 CUL
and
regenerating a plant from such a tissue culture and to phenotypic variants of
cucumber variety
NUN 93099 CUL. The disclosure also relates to progeny of cucumber variety NUN
93099 CUL
and the hybrid varieties obtained by crossing cucumber variety NUN 93099 CUL
as a parent line
with plants of other varieties or parent lines.
BACKGROUND OF THE DISCLOSURE
[0002] The goal of plant breeding is to combine various desirable traits in a
single variety. Such
desirable traits may include greater yield, resistance to diseases, insects or
other pests, tolerance
to heat and drought, better agronomic quality, higher nutritional value,
enhanced growth rate,
and improved fruit properties.
100031 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 genotype. A plant cross-pollinates if
pollen comes to it
from a flower of a different genotype. Plants that have been self-pollinated
and selected for
(uniform) type over many generations become homozygous at almost all gene loci
and produce
a uniform population of true breeding progeny of homozygous plants. A cross
between two such
homozygous plants of different lines produces a uniform population of hybrid
plants that are
heterozygous for many gene loci. The extent of heterozygosity in the hybrid is
a function of the
genetic distance between the parents. Conversely, a cross of two plants each
heterozygous at a
number of loci produces a segregating population of hybrid plants that differ
genetically and are
not uniform. The resulting non-uniformity makes performance unpredictable.
[0004] The development of uniform varieties requires the development of
homozygous inbred
plants, the crossing of these inbred plants to make hybrids, and the
evaluation of the hybrids
resulting from 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
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Date Recue/Date Received 2022-09-09
sources into breeding pools from which new lines are developed by selfing and
selection of
desired phenotypes. The new plants are evaluated to determine which have
commercial potential.
[0005] Cucumber (Cucumis sativus L.) belongs to the Cucurbitaceae family and
is naturally a
diploid (2n=14) outcrossing species, although haploid, doubled-haploid (see,
e.g., US
5,492,827), and triploid (see, e.g., Sarreb et al. (2002), Plant Cell Tissue,
Organ Culture 71: 231-
235) types have been developed. The fruit of cucumber is typically cylindrical
and elongated. It
has thin, dark green skin and mild flavor, which can be eaten without peeling.
[0006] In the United States, the fourth largest cucumber producer, the slicing
(fresh market) and
pickling (processing) are the two main types of cucumber fruit grown
commercially. Varieties
and production methods are typically adapted to the end use. Slicing cucumbers
are often longer,
larger and have darker and thicker skin, whereas pickling/processing cucumbers
have shorter
fruit, thinner skin with interior flesh that make them more amenable to
pickling. Seedless
varieties are generally preferable for both fresh and processing markets as
seeds are not palatable.
[0007] Cucumber plants that set fruit parthenocarpically (without pollination
and fertilization)
have more recently been available. These plants produce seedless fruit unless
pollinated. Growth
of parthenocarpic varieties is beneficial in that setting of fruit on these
cultivars does not produce
an inhibiting effect on plant growth, unlike the case of fertilized, seeded
fruit. The seedless
varieties are usually higher yielding and of higher quality due to the lack of
seeds. However,
growth of these plants requires isolation from seeded cucumbers to avoid
pollination and
subsequent seeded fruit.
[0008] Most of the cucumbers currently grown for processing (for pickles and
pickle products)
in the United States are seeded hybrid varieties. Hybrid varieties offer the
advantages of easy
combination of dominant and recessive traits, such as disease resistance, from
a set of inbred
parents, as well as careful control of parentage. The production of Fl hybrid
cucumber seeds
from a pollen parent bearing only male flowers has been reported (see, e.g.,
US 4,822,949).
[0009] Advances in biotechnology have also resulted in genetically engineered
cucumber plants
with improved traits. For example, cucumbers resistant to Cucumber Mosaic
Virus (CMV) have
been developed by expression of CMV protein coat genes (see, e.g., US
5,349,128). Transgenic
plants exhibiting, for example, other viral resistance traits or high levels
of superoxide dismutase
have also been reported (see, e.g., US 6,084,152).
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Date Recue/Date Received 2024-01-26
[0010] While breeding efforts to date have provided a number of useful
cucumber varieties with
beneficial traits, there remains a great need in the art for new varieties
with further improved
traits. Such plants would benefit farmers and consumers alike by improving
crop yields and/or
quality. Breeding objectives include varying the color, texture, and flavor of
the fruit, minimizing
the occurrence of bitterness, optimizing flesh thickness, solid content (% dry
matter), storage
properties, and sugar content. Breeding programs also focus on developing
plants with earlier
fruit maturity, more restricted vine growth, improved disease resistance or
tolerance, and
improved adaptability to environmental conditions.
SUMMARY OF THE VARIOUS ASPECTS OF THE DISCLOSURE
[0011] The disclosure provides for cucumber variety NUN 93099 CUL, products
thereof, and
methods of using the same. NUN 93099 CUL is a Dutch long cucumber variety of
the Kasj a type
and- is suitable for growing in a greenhouse, e,g., indoor cultivation with
drip irrigation.
[0012] In another aspect, the plant of variety NUN 93099 CUL, or part thereof,
or progeny
thereof comprises resistance to Cladosporium cucumerinum, Cucumber Mosaic
Virus (CMV),
Podosphaera xanthii, Corynespora cassiicola, Cucumber Vein Yellowing Virus
(CVYV), and
Cucumber Green Mottle Mosaic Virus (CGMMV), measured according to TG/61/7.
[0013] In another aspect, the plant of variety NUN 93099 CUL has 22, 23, or
more, or all of the
following distinguishing characteristics when compared to the Reference
Variety as shown in
Table 3:
a. shorter internode length;
b. shorter length of first 15 internodes;
c. medium plant vigor;
d. shorter mature leaf length;
e. thinner mature leaf width;
f. shorter mature leaf petiole length;
g. horizontal mature leaf attitude;
h. thinner mature leaf petiole diameter;
i. larger mature fruit diameter at medial;
j. dark green color at stem end;
k. dark green color at blossom end;
1. necked fruit shape;
m. dull skin luster;
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Date Recue/Date Received 2022-09-09
n. medium ratio of length/diameter;
o. larger core diameter in relation to diameter of fruit;
p. longer length of neck;
q. rounded shape of calyx end;
r. lighter intensity of ground color of skin;
s. very sparse density of vestiture;
t. shorter length of peduncle;
u. intermediately resistant to Cucumber Mosaic Virus (CMV); and
v. highly resistant to Cucumber Vein Yellowing Virus (CVYV), when determined
at 5%
significance level for numerical characteristics and determined by type or
degree for non-
numerical characteristics for plants grown under the same environmental
conditions.
100141 In another aspect, the disclosure provides a plant or a progeny having
all the
physiological and morphological characteristics of variety NUN 93099 CUL when
grown under
the same environmental conditions. In another aspect, the plant or progeny has
all or all but one,
two, or three of the physiological and morphological characteristics of
cucumber variety NUN
93099 CUL when measured under the same environmental conditions and e.g.,
evaluated at
significance levels of 1%, 5%, or 10% significance (which can also be
expressed as a p-value)
for quantitative characteristics and identical (same type or degree) for non-
quantitative
characteristics, wherein a representative sample of seed of variety NUN 93099
CUL has been
deposited under Accession Number NCIMB 43946. In another aspect, the plant or
progeny has
all or all but one, two, or three of the physiological and morphological
characteristics as listed in
Tables 1 and 2 for variety NUN 93099 CUL when measured under the same
environmental
conditions and e.g., evaluated at significance levels of 1%, 5% or 10%
significance (which can
also be expressed as a p-value) for quantitative characteristics and identical
(same type or degree)
for non-quantitative characteristics.
[0015] In another aspect, the disclosure provides a seed of cucumber variety
NUN 93099 CUL,
wherein a representative sample of said seed has been deposited under
Accession Number
NCIMB 43946. The disclosure also provides for a plurality of seeds of cucumber
variety NUN
93099 CUL. The cucumber seed of variety NUN 93099 CUL may be provided as an
essentially
homogeneous population of cucumber seed. The population of seed of cucumber
variety NUN
93099 CUL may be particularly defined as essentially free from other seed. The
seed population
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Date Recue/Date Received 2022-09-09
may be grown into plants to provide an essentially homogeneous population of
cucumber
plants as described herein.
[0016] The disclosure also provides a plant grown from a seed of cucumber
variety NUN 93099
CUL and a plant part thereof.
[0017] In another aspect, the disclosure provides for a hybrid variety of NUN
93099 CUL.
[0018] The disclosure also provides for a progeny of variety NUN 93099 CUL. In
another
aspect, the disclosure provides a plant or a progeny retaining all or all but
one, two, or three of
the "distinguishing characteristics" or all or all but one, two, or three of
the "morphological and
physiological characteristics" of variety NUN 93099 CUL and methods for
producing that plant
or progeny.
[0019] In another aspect, the disclosure provides a seed growing or grown on a
plant of variety
NUN 93099 CUL (i.e., produced after pollination of the flower of cucumber
variety NUN 93099
CUL). The disclosure also provides an Fl progeny of cucumber variety NUN 93099
CUL.
[0020] The disclosure furthermore provides a cucumber fruit produced on a
plant grown from
a seed of cucumber variety NUN 93099 CUL.
[0021] In another aspect, the disclosure provides for a plant part obtained
from variety NUN
93099 CUL, wherein said plant part is: a fruit, a harvested fruit, a part of a
fruit, a leaf, a part of
a leaf, pollen, an ovule, a cell, a petiole, a shoot or a part thereof, a stem
or a part thereof, a root
or a part thereof, a root tip, a cutting, a seed, a part of a seed, seed coat
or another maternal tissue
which is part of a seed grown on said variety, a hypocotyl, a cotyledon, a
scion, a stock, a
rootstock, a pistil, an anther, or a flower or a part thereof. Fruits are
particularly important plant
parts. In another aspect, the plant part obtained from variety NUN 93099 CUL
is a cell, optionally
a cell in a cell or tissue culture. That cell may be grown into a plant of
variety NUN 93099 CUL.
[0022] The disclosure also provides a cell culture of cucumber variety NUN
93099 CUL and a
plant regenerated from cucumber variety NUN 93099 CUL, wherein the plant has
all the
characteristics of cucumber variety NUN 93099 CUL, when grown under the same
environmental
conditions, as well as methods for culturing and regenerating cucumber variety
NUN 93099 CUL.
Alternatively, a regenerated plant may have one characteristic that is
different from cucumber
variety NUN 93099 CUL.
Date Recue/Date Received 2022-09-09
[0023] The disclosure further provides a vegetatively propagated plant of
variety NUN 93099
CUL having all or all but one, two, or three of the morphological and
physiological characteristics
of cucumber variety NUN 93099 CUL when grown under the same environmental
conditions.
[0024] In another aspect, the disclosure provides a method of producing a
cucumber plant
comprising crossing cucumber variety NUN 93099 CUL with itself or with another
cucumber
variety and selecting a progeny cucumber plant from said crossing.
[0025] The disclosure also provides a method of producing a cucumber plant
derived from
cucumber variety NUN 93099 CUL.
[0026] In a further aspect, the disclosure provides a method of producing a
hybrid cucumber
seed comprising crossing a first parent cucumber plant with a second parent
cucumber plant and
harvesting the resultant hybrid cucumber seed, wherein said first parent plant
or second parent
cucumber plant is cucumber variety NUN 93099 CUL. Also provided is a hybrid
cucumber seed
produced from crossing a first parent cucumber plant and second parent
cucumber plant and
harvesting the resultant hybrid cucumber seed, wherein said first parent
cucumber plant or second
parent cucumber plant is cucumber variety NUN 93099 CUL. Moreover, the hybrid
cucumber
plant grown from the hybrid cucumber seed is provided.
[0027] In another aspect, the disclosure provides a method of introducing a
single locus
conversion into the plant of variety NUN 93099 CUL, wherein a representative
sample of seed
of said cucumber variety has been deposited under Accession Number NCIMB
43946, wherein
the plant otherwise retains all of the physiological and morphological
characteristics of cucumber
variety NUN 93099 CUL and further comprises the single locus conversion.
[0028] In yet another aspect, the disclosure provides a method of introducing
a desired trait into
cucumber variety NUN 93099 CUL, said method comprises transforming the plant
of cucumber
variety NUN 93099 CUL with a transgene that confers the desired trait, wherein
the transformed
plant otherwise has all of the physiological and morphological characteristics
of cucumber
variety NUN 93099 CUL and contains the desired trait.
[0029] The disclosure also provides a method of producing a modified cucumber
plant with a
desired trait, wherein the method comprises mutating a cucumber plant or plant
part of cucumber
variety NUN 93099 CUL, wherein a representative sample of seed of said
cucumber variety has
been deposited under Accession Number NCIMB 43946, and wherein the mutated
plant
otherwise retains all of the physiological and morphological characteristics
of cucumber variety
NUN 93099 CUL and contains the desired trait.
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Date Recue/Date Received 2022-09-09
[0030] In one aspect, the single locus conversion or desired trait is yield,
size, compactness, dry
matter content, firmness, flavor, fruit quality, enhanced nutritional quality,
post-harvest quality,
storage properties, color, male sterility, herbicide tolerance, insect
resistance, pest resistance,
disease resistance, Powdery mildew resistance without necrosis, environmental
stress tolerance,
modified carbohydrate metabolism, modified protein metabolism, or ripening.
[0031] In another aspect, the disclosure provides a container comprising the
plant, plant part,
or seed of cucumber variety NUN 93099 CUL.
[0032] Also provided is a food, a feed, or a processed product comprising the
plant part of
cucumber variety NUN 93099 CUL, wherein the plant part is a fruit or part
thereof.
[0033] The invention as claimed relates to:
1. A cell of cucumber plant variety NUN 93099 CUL, or a cell of a part
thereof, wherein the
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited
under Accession Number NCIMB 43946.
2. A cell of a seed of cucumber variety NUN 93099 CUL, wherein a
representative sample
of seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number
NCIMB 43946.
3. A cell of a plant or a cell of a part thereof, produced by growing the seed
of cucumber
variety NUN 93099 CUL, wherein a representative sample of seed of said
cucumber
variety NUN 93099 CUL is deposited under Accession Number NCIMB 43946.
4. A cell of a seed that produces cucumber variety NUN 93099 CUL, wherein a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited
under Accession Number NCIMB 43946.
5. A cell of a plant part of cucumber variety NUN 93099 CUL, wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession
Number NCIMB 43946, wherein the plant part is a leaf, pollen, an ovule, a
fruit, a scion,
a root, a rootstock, a cutting, or a flower.
6. The cell of a plant part of embodiment 5, when the plant part is a fruit or
part thereof.
7. A cell of a cucumber plant, or a part thereof, having all of the
physiological and
morphological characteristics of cucumber variety NUN 93099 CUL listed in
Tables 1
and 2 when grown under the same environmental conditions, wherein a
representative
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Date Recue/Date Received 2022-09-09
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession
Number NCIMB 43946.
8. A tissue culture or cell culture of regenerable cells of cucumber variety
NUN 93099 CUL,
wherein a representative sample of seed of said cucumber variety NUN 93099 CUL
is
deposited under Accession Number NCIMB 43946.
9. The tissue culture or cell culture according to embodiment 8, comprising
cells or
protoplast from a plant part, wherein the plant part is a meristem, a
cotyledon, a hypocotyl,
a pollen, a leaf, an anther, a root, a root tip, a pistil, a petiole, a
flower, a fruit, a stalk, or
a stem.
10. A cell of a cucumber plant regenerated from the tissue culture or cell
culture of
embodiment 8, wherein the plant has all of the physiological and morphological
characteristics of the plant of cucumber variety NUN 93099 CUL listed in
Tables 1 and 2
when grown under the same environmental conditions, and wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession
Number NCIMB 43946.
11. A cell of a cucumber plant produced by vegetatively propagating at least a
part of the
plant of cucumber variety NUN 93099 CUL, wherein a representative sample of
seed of
said cucumber variety NUN 93099 CUL is deposited under Accession Number NCIMB
43946.
12. The cell of embodiment 11, wherein said vegetative propagation comprises
regenerating
a whole plant from said part of the plant of cucumber variety NUN 93099 CUL,
wherein
a representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited
under Accession Number NCIMB 43946.
13. The cell of embodiment 11, wherein said part is a cutting, a cell culture,
or a tissue culture.
14. A cell of a vegetatively propagated cucumber plant, or a cell of a part
thereof, wherein
the plant or part thereof have all of the physiological and morphological
characteristics of
the plant of cucumber variety NUN 93099 CUL listed in Tables 1 and 2 when
grown under
the same environmental conditions, and wherein a representative sample of seed
of said
cucumber variety NUN 93099 CUL is deposited under Accession Number NCIMB
43946.
15. A cell of a cucumber plant produced by a method comprising crossing the
plant of
cucumber variety NUN 93099 CUL with a second cucumber plant at least once, and
8
Date Recue/Date Received 2022-09-09
selecting a progeny cucumber plant from said crossing and allowing the progeny
cucumber plant to form seed, wherein a representative sample of seed of said
cucumber
variety NUN 93099 CUL is deposited under Accession Number NCIMB 43946.
16. A cell of a cucumber seed produced by a method comprising crossing
cucumber plants
and harvesting the resultant seed, wherein at least one cucumber plant is the
plant of
cucumber variety NUN 93099 CUL, wherein a representative sample of seed of
said
cucumber variety NUN 93099 CUL is deposited under Accession Number NCIMB
43946.
17. A cell of a single locus converted plant of cucumber variety NUN 93099
CUL, wherein
the plant has otherwise all of the physiological and morphological
characteristics of the
plant of cucumber variety NUN 93099 CUL listed in Tables 1 and 2 when grown
under
the same environmental conditions, wherein the single locus is introduced by
genetic
transformation, and wherein a representative sample of seed of said cucumber
variety
NUN 93099 CUL is deposited under Accession Number NCIMB 43946.
18. The cell of embodiment 17, wherein the single locus conversion confers
yield, size,
compactness, dry matter content, firmness, flavor, fruit quality, enhanced
nutritional
quality, post-harvest quality, storage properties, color, male sterility,
herbicide tolerance,
insect resistance, pest resistance, disease resistance, Powdery mildew
resistance without
necrosis, environmental stress tolerance, modified carbohydrate metabolism,
modified
protein metabolism, or ripening.
19. A cell of a doubled haploid cucumber plant produced by a method of making
doubled
haploids from haploid cells made from a plant or a plant part of cucumber
variety NUN
93099 CUL, wherein a representative sample of seed of said cucumber variety
NUN
93099 CUL is deposited under Accession Number NUMB 43946.
20. A cell of a cucumber plant produced by a method comprising transforming
the plant of
cucumber variety NUN 93099 CUL with a transgene that confers the desired
trait, wherein
the transformed plant otherwise retains all of the physiological and
morphological
characteristics of cucumber variety NUN 93099 CUL and contains the desired
trait,
wherein a representative sample of seed of said cucumber variety NUN 93099 CUL
is
deposited under Accession Number NCIMB 43946.
21. The cell of embodiment 20, wherein the desired trait is the desired trait
is yield, size,
compactness, dry matter content, firmness, flavor, fruit quality, enhanced
nutritional
quality, post-harvest quality, storage properties, color, male sterility,
herbicide tolerance,
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Date Recue/Date Received 2022-09-09
insect resistance, pest resistance, disease resistance, Powdery mildew
resistance without
necrosis, environmental stress tolerance, modified carbohydrate metabolism,
modified
protein metabolism, or ripening.
22. A cell of a cucumber plant produced by a method comprising growing a plant
of cucumber
variety NUN 93099 CUL until it sets at least one fruit and collecting the
fruit, wherein a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited
under Accession Number NCIMB 43946.
23. A cell of a fruit produced by the method as defined in embodiment 22.
24. A method of determining the genotype of a plant of cucumber variety NUN
93099 CUL,
comprising obtaining a sample of nucleic acids form said plant and detecting
in said
nucleic acids a plurality of polymorphisms, thereby determining the genotype
of the plant,
and storing the results of detecting the plurality of polymorphisms on a
computer readable
medium, wherein a representative sample of seed of said cucumber variety NUN
93099
CUL is deposited under Accession Number NCIMB 43946.
25. A method comprising packaging seeds of cucumber variety NUN 93099 CUL in a
container, wherein a representative sample of seed of said cucumber variety
NUN 93099
CUL is deposited under Accession Number NCIMB 43946.
26. A method comprising packaging a cucumber fruit of cucumber variety NUN
93099 CUL
in a container, wherein a representative sample of seed of said cucumber
variety NUN
93099 CUL is deposited under Accession Number NCIMB 43946.
27. Use of cucumber variety NUN 93099 CUL as a scion or a rootstock, herein a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited
under Accession Number NCIMB 43946.
28. Use of cucumber plant variety NUN 93099 CUL, or a part thereof, for
producing a seed,
a crop, or a food or a processed product, wherein a representative sample of
seed of said
cucumber variety NUN 93099 CUL is deposited under Accession Number NCIMB
43946.
29. Use of a seed of cucumber variety NUN 93099 CUL, for producing cucumber
plants,
wherein a representative sample of seed of said cucumber variety NUN 93099 CUL
is
deposited under Accession Number NCIMB 43946.
Date Recue/Date Received 2022-09-09
30. Use of a plant of cucumber variety NUN 93099 CUL, wherein a representative
sample of
seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number
NCIMB 43946, to cross with a second cucumber plant.
31. Use of a plant of cucumber variety NUN 93099 CUL, wherein a representative
sample of
seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number
NC1MB 43946, and a second cucumber plant to produce a cucumber seed.
32. Use of a plant of cucumber variety NUN 93099 CUL, wherein a representative
sample of
seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number
NCIMB 43946, and a second cucumber plant to produce a descendant plant.
33. Use of a plant of cucumber variety NUN 93099 CUL, wherein a representative
sample of
seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number
NCIMB 43946, to breed a cucumber plant.
34. Use of cucumber variety NUN 93099 CUL, and wherein a representative sample
of seed
of said cucumber variety NUN 93099 CUL is deposited under Accession Number
NCIMB
43946, as a recipient of a single locus conversion.
35. Use of cucumber variety NUN 93099 CUL, and wherein a representative sample
of seed
of said cucumber variety NUN 93099 CUL is deposited under Accession Number
NCIMB
43946, a recipient of a transgene.
BRIEF DESCRIPTION OF THE DRAWINGS
[0034] Fig. 1 shows the fruits at edible maturity of cucumber variety NUN
93099 CUL.
[0035] Fig. 2 shows the fruit comparison at edible maturity of cucumber
variety NUN 93099
CUL and the Reference Variety.
[0036] Fig 3 shows the stem end, medial, and blossom end cross-section of
cucumber variety
NUN 93099 CUL and the Reference Variety.
[0037] Fig. 4 shows the horizontal mature leaf attitude of cucumber variety
NUN 93099 CUL.
[0038] Fig. 5 shows the mature leaf comparison of cucumber variety NUN 93099
CUL and the
Reference Variety.
DEFINITIONS
[0039] "Cucumber" refers herein to plants of the species Cucumis sativus. The
most commonly
eaten part of a cucumber is the fruit or pepo. The fruit comprises a stem and
peduncle or pedicel,
11
Date Recue/Date Received 2022-09-09
receptacle, ectocarp, rind, fruit flesh, exocarp, mesocarp, external phloem,
internal phloem,
xylem, vascular bundle, carpel, placenta and optionally seed. The stem and
peduncle or pedicel,
receptacle, ectocarp, rind, fruit flesh, exocarp, mesocarp, external phloem,
internal phloem,
xylem, vascular bundle, carpel, placenta and seed coat of the seed are
maternal tissues and are
genetically identical to the plant on which they grow.
[0040] "Cultivated cucumber" refers to plants of Cucumis sativus i.e.,
varieties, breeding lines
or cultivars of the species C. sativus, cultivated by humans and having good
agronomic
characteristics.
[0041] The terms "cucumber plant designated NUN 93099 CUL," "NUN 93099 CUL,"
"NUN
93099," "NUN 93099 Fl," "93099 CUL," "cucumber 93099," or "Serein" are used
interchangeably herein and refer to a cucumber plant of variety NUN 93099 CUL,
representative
seed of which having been deposited under Accession Number NCIMB 43946.
[0042] A "seed of NUN 93099 CUL" refers to a cucumber seed which can be grown
into a plant
of variety NUN 93099 CUL, wherein a representative sample of viable seed of
variety NUN
93099 CUL has been deposited under Accession Number NCIMB 43946. A seed can be
in any
stage of maturity, for example, a mature, viable seed, or an immature, non-
viable seed. A seed
comprises an embryo and maternal tissues.
[0043] An "embryo of NUN 93099 CUL" refers to an "Fl hybrid embryo" as present
in a seed
of cucumber variety NUN 93099 CUL, a representative sample of said seed of NUN
93099 CUL
having been deposited under Accession Number NCIMB 43946.
[0044] A "seed grown on NUN 93099 CUL" refers to a seed grown on a mature
plant of variety
NUN 93099 CUL or inside a fruit of cucumber variety NUN 93099 CUL. The "seed
grown on
NUN 93099 CUL" contains tissues and DNA of the maternal parent, cucumber
variety NUN
93099 CUL. When said seed is planted, it grows into a first generation progeny
plant of variety
NUN 93099 CUL.
[0045] A "fruit of NUN 93099 CUL" refers to a fruit containing maternal
tissues of cucumber
variety NUN 93099 CUL as deposited under Accession Number NCIMB 43946. In one
aspect,
the fruit contains seed grown on cucumber variety NUN 93099 CUL. In another
aspect, the fruit
does not contain seed, i.e., the fruit is parthenocarpic. The skilled person
is familiar with methods
for inducing parthenocarpy. Those methods comprise chemically or genetically
inducing
parthenocarpy. Compounds suitable for chemically inducing parthenocarpy
include auxins,
gibberellins and cytokinins. Genetic parthenocarpy can be induced (see, e.g.,
US 2018/0054990
12
Date Recue/Date Received 2022-09-09
and US 2017/0335339 (PIN4)) or can be provided by reduced or eliminated
expression of
PISTILATA (PI) or APETALA3 (AP3). A fruit can be in any stage of maturity, for
example, a
mature fruit in the yellow stage comprising viable seed, or an immature fruit
in the edible green
stage comprising non-viable seed.
[0046] "Plant" includes the whole plant or any part or derivatives thereof
having the same
genetic makeup as the plant from which it is obtained.
[0047] "Plant part" includes any part of a plant, such as a plant organ (e.g.,
harvested or non-
harvested fruits), a plant cell, a plant protoplast, a plant cell tissue
culture or a tissue culture from
which a whole plant can be regenerated, a plant cell that is intact in a
plant, a clone, a
micropropagation, plant callus, a plant cell clump, a plant transplant, a
vegetative propagation, a
seedling, a fruit, a harvested fruit, a part of a fruit, a leaf, a part of a
leaf, pollen, an ovule, an
embryo, a petiole, a shoot or a part thereof, a stem or a part thereof, a root
or a part thereof, a
root tip, a cutting, a seed, a part of a seed, a hypocotyl, a cotyledon, a
scion, a graft, a stock, a
rootstock, a pistil, an anther, and a flower or part thereof. Seed can be
mature or immature. Pollen
or ovules may be viable or non-viable. Also, any developmental stage is
included, such as
seedlings, cuttings prior or after rooting, mature plants or leaves.
Alternatively, a plant part may
also include a plant seed which comprises maternal tissues of cucumber variety
NUN 93099 CUL
and an embryo having one or two sets of chromosomes derived from the parent
plant, e.g., from
cucumber variety NUN 93099 CUL. Such an embryo comprises two sets of
chromosomes derived
from cucumber variety NUN 93099 CUL if it is produced from self-pollination of
cucumber
variety NUN 93099 CUL, while an embryo derived from cross-fertilization of
cucumber
variety NUN 93099 CUL, will comprise only one set of chromosomes from cucumber
variety NUN 93099 CUL, and the other set of chromosomes from the other parent.
[0048] An "essentially homogeneous population of cucumber seed" is a
population of seeds
where at least 97%, 98%, 99% or more of the total population of seeds are
seeds of cucumber
variety NUN 93099 CUL.
[0049] An "essentially homogeneous population of cucumber plants" is a
population of plants
where at least 97%, 98%, 99% or more of the total population of plants are
plants of variety NUN
93099 CUL.
[0050] The phrase "essentially free from other seed" refers to a population of
seed where less
than 3%, 2%, 1% or less of the total population of seed is seed that is not a
cucumber seed or, in
13
Date Recue/Date Received 2022-09-09
another aspect, less than 3%, 2%, 1% or less of the total population of seed
is seed that is not a
seed of cucumber variety NUN 93099 CUL.
[0051] "Harvest maturity" refers to the stage at which a cucumber fruit is
ripe or ready for
harvest or the optimal time to harvest the fruit for the market, for
processing or for consumption.
In one aspect, harvest maturity is the stage which allows proper completion of
the normal
ripening.
[0052] "Flavor" refers to the sensory impression of a food or other substance,
especially a
cucumber fruit or fruit part (fruit flesh) and is determined mainly by the
chemical senses of taste
and smell. Flavor is influenced by texture properties and by volatile and/or
non-volatile chemical
components (organic acids, lipids, carbohydrates, salts etc.).
[0053] "Aroma" refers to smell (or odor) characteristics of cucumber fruits or
fruit parts (fruit
flesh).
[0054] "Yield" means the total weight of all cucumber fruits harvested per
hectare of a
particular line or variety. It is understood that "yield" expressed as weight
of all cucumber fruits
harvested per hectare can be obtained by multiplying the number of plants per
hectare times the
"yield per plant".
[0055] "Marketable yield" means the total weight of all marketable cucumber
fruits, especially
fruit which is not cracked, damaged or diseased, harvested per hectare of a
particular line or
variety. A "marketable fruit" is a fruit that has commercial value.
[0056] "Harvested plant material" refers herein to plant parts (e.g., fruits
detached from the
whole plant), which have been collected for further storage and/or further
use.
[0057] "USDA descriptors" are the plant variety descriptors described for
cucumber in the
"Objective Description of Variety- Cucumber (Cucumis sativus L.)," as
published by the US
Department of Agriculture, Agricultural Marketing Service, Plant Variety
Protection Office,
Beltsville, MD 20705 and which can be downloaded from the world-wide web at
ams.usda.gov/
under servi ces/plant-vari ety-protection/pvpo-c-forms under cucumber. "Non-
USDA descriptors"
are other descriptors suitable for describing cucumber.
[0058] "UPOV descriptors" are the plant variety descriptors described for
cucumber in the
"Guidelines for the Conduct of Tests for Distinctness, Uniformity and
Stability, TG/61/7 (Geneva
2007, revised 2019-03-13), as published by UPOV (International Union for the
Protection of
New Varieties and Plants) and which can be downloaded from the world-wide web
at
14
Date Recue/Date Received 2022-09-09
upov.int/edocs/tgdocs/en/tg061.pdf. Likewise, "UPOV methods" to determine
specific
parameters for the characterization of cucumber are described at upov.int.
[0059] "Calibration Book for Cucumber & Gherkin" refers to the calibration
book for
cucumbers and gherkins which provides guidance for describing a cucumber
variety, as published
by Naktuinbouw (version 1, December 2010). The calibration book is based on
the CPVO
(Community Plant Variety Protection Office) Protocol CPVO-TP/06/12 and UPOV
Guideline
TG/6/1/7.
[0060] "RHS" or "RHS color chart" refers to the color chart of the Royal
Horticultural Society
(UK), which publishes an official botanical color chart quantitatively
identifying colors
according to a defined numbering system. The chart may be purchased from Royal
Horticulture
Society Enterprise Ltd RHS Garden; Wisley, Woking; Surrey GU236QB, UK, e.g.,
the RHS
color chart 2007.
[0061] "Reference Variety for Cucumber Variety NUN 93099 CUL" refers herein to
variety
NUN 83067 CUL, a variety from Nunhems B.V. With commercial name, Sequence,
which has
been planted in a trial together with cucumber variety NUN 93099 CUL. The
characteristics of
cucumber variety NUN 93099 CUL were compared with the characteristics of the
Reference
Variety as shown in Tables 1 and 2. The distinguishing characteristics between
cucumber variety
NUN 93099 CUL and the Reference Variety are shown in Table 3.
100621 A plant having "all the physiological and morphological
characteristics" of a referred-
to-plant means a plant showing the physiological and morphological
characteristics of the
referred-to-plant when grown under the same environmental conditions,
preferably in the same
experiment; the referred-to-plant can be a plant from which it was derived,
e.g., the progenitor
plant, the parent, the recurrent parent, the plant used for tissue- or cell
culture, etc. A
physiological or morphological characteristic can be a numerical
characteristic or a non-
numerical characteristic. In one aspect, a plant has "all but one, two or
three of the physiological
and morphological characteristics" of a referred-to-plant, or "all the
physiological and
morphological characteristics" of Tables 1 and 2 or "all or all but one, two
or three of the
physiological and morphological characteristics" of Tables 1 and 2.
[0063] The physiological and/or morphological characteristics mentioned above
are commonly
evaluated at significance levels of 1%, 5% or 10% if they are numerical, or
for having an identical
degree (or type) if not numerical, measured under the same environmental
conditions. For
example, a progeny plant or a Single Locus Converted plant or a mutated plant
of variety NUN
Date Recue/Date Received 2022-09-09
93099 CUL, may have one or more (or all) of the essential physiological and/or
morphological
characteristics of said variety listed in Tables 1 and 2, as determined at the
5% significance level
(i.e., p < 0.05), when grown under the same environmental conditions.
[0064] "Distinguishing characteristics" or "distinguishing morphological
and/or physiological
characteristics" refers herein to the characteristics which distinguish the
new variety from other
cucumber varieties, such as the Reference Variety (i.e., are different), when
grown under the
same environmental conditions. The distinguishing characteristics between
cucumber variety
NUN 93099 CUL and the Reference Variety are described herein and is shown in
Table 3. When
comparing cucumber variety NUN 93099 CUL to other varieties, the
distinguishing
characteristics may be different. In one aspect, the distinguishing
characteristics may include
one, two, three, or more (or all) of the characteristics listed in Tables 1
and 2. Preferably, all
numerical distinguishing characteristics are statistically significantly
different at p < 0.05
between cucumber variety NUN 93099 CUL or the other variety (e.g., the
Reference Variety).
[0065] Cucumber variety NUN 93099 CUL has the following distinguishing
characteristics
when compared to the Reference Variety as shown in Table 3:
a. shorter intemode length;
b. shorter length of first 15 internodes;
c. medium plant vigor;
d. shorter mature leaf length;
e. thinner mature leaf width;
f. shorter mature leaf petiole length;
g. horizontal mature leaf attitude;
h. thinner mature leaf petiole diameter;
i. larger mature fruit diameter at medial;
j. dark green color at stem end;
k. dark green color at blossom end;
1. necked fruit shape;
m. dull skin luster;
n. medium ratio of length/diameter;
o. larger core diameter in relation to diameter of fruit;
p. longer length of neck;
q. rounded shape of calyx end;
16
Date Recue/Date Received 2022-09-09
r. lighter intensity of ground color of skin;
s. very sparse density of vestiture;
t. shorter length of peduncle;
u. intermediately resistant to Cucumber Mosaic Virus (CMV); and
v. highly resistant to Cucumber Vein Yellowing Virus (CVYV), when determined
at 5%
significance level for numerical characteristics and determined by type or
degree for non-
numerical characteristics for plants grown under the same environmental
conditions.
[0066] Thus, a cucumber plant "comprising the distinguishing characteristics
of cucumber
variety NUN 93099 CUL" (such as a progeny plant) refers herein to a plant
which does not differ
significantly from said variety in the distinguishing characteristics above,
when determined at
5% significance level for numerical characteristics and determined by type or
degree for non-
numerical characteristics for plants grown under the same environmental
conditions. Therefore,
in one aspect, a cucumber plant is provided which does not differ
significantly from cucumber
variety NUN 93099 CUL.
[0067] Similarity and differences between two different plant lines or
varieties can be
determined by comparing the number of morphological and/or physiological
characteristics that
are the same (i.e., statistically not significantly different) or that are
different (i.e., statistically
significantly different) between the two plant lines or varieties using plants
grown under the same
environmental conditions. Preferably, a numerical characteristic is considered
to be "the same"
when the value for a numeric characteristic is not significantly different at
the 1% (p <0.01) or
5% (p < 0.05) significance level, using a T-test, a standard method known to
the skilled person.
Preferably, non-numerical or "degree" or "type" characteristics are considered
"the same" when
the values have the same "degree" or "type" when scored using USDA and/ or
UPOV descriptors,
if the plants are grown under the same environmental conditions.
[0068] In one aspect, a statistical analysis of the quantitative
characteristics showing the degree
of significance may be provided. Statistical significance is the likelihood
that a relationship
between two or more variables is caused by something other than chance, i.e.,
that the differences
in the means for quantitative characteristics of the plant of cucumber variety
NUN 93099 CUL
and the Reference Variety are significant due to chance. For the purpose of
proving differences
or distinction between cucumber variety NUN 93099 CUL and the Reference
Variety, a p-value
of 5% or 0.05 or lower is considered statistically significant. This means
that there is only a 5%
17
Date Recue/Date Received 2022-09-09
probability or lower probability that the observed result could have happened
just by chance or
random variation.
[0069] The statistical analysis is drawn from a small sample of at least 15
plants or plant parts
of cucumber variety NUN 93099 CUL and the Reference Variety. Statistical
points or parameters
such as mean, minimum, median, maximum, and standard deviation are collected
from the
sample data to analyze where the average is, how varied the data set is, and
whether the data is
skewed. For the purpose of determining whether the result of the data set is
statistically
significant, a T-Test is used, a statistical tool for proving significance in
the means of the two
groups (e.g., cucumber variety NUN 93099 CUL and the Reference Variety) at 5%
significance
level (a p-value of 5% or 0.05).
100701 "Variety" or "cultivar" means a plant grouping within a single
botanical taxon of the
lowest rank.
[0071] A "plant line" is, for example, a breeding line which can be used to
develop one or more
varieties. A breeding line is typically highly homozygous.
[0072] "Harvested seeds" refer to seeds harvested from a line or variety,
e.g., produced after
self-fertilization or cross-fertilization and collected.
[00731 "Hybrid variety" or "Fl hybrid" refers to the seeds harvested from
crossing two inbred
(nearly homozygous) parental lines. For example, the female parent is
pollinated with pollen of
the male parent to produce hybrid (F1) seeds on the female parent.
[0074] "Locus" (plural loci) refers to the specific location, place or site of
a DNA Sequence on
a chromosome where, for example, a gene or genetic marker is found. A locus
may confer a
specific trait.
100751 "Allele" refers to one or more alternative forms of a gene locus. All
of these loci relate
to one trait. Sometimes, different alleles can result in different observable
phenotypic traits, such
as different pigmentation. However, many variations at the genetic level
result in little or no
observable variation. If a multicellular organism has two sets of chromosomes,
i.e., diploid, these
chromosomes are referred to as homologous chromosomes. Diploid organisms have
one copy
each of gene (and therefore one allele) on each chromosome. If both alleles
are the same, they
are homozygotes. If the alleles are different, they are heterozygotes.
[0076] "Genotype" refers to the genetic composition of a cell or organism.
18
Date Recue/Date Received 2022-09-09
[0077] "Phenotype" refers to the detectable characteristics of a plant, a
cell, or an organism,
which characteristics are the manifestation of gene expression.
[0078] "Haploid" refers to a cell or organism having one set of two sets of
chromosomes in a
diploid.
[0079] "Diploid" refers to a plant, a vegetative plant part(s), or seed having
two sets of
chromosomes, designated herein as 2n.
[0080] "Triploid" refers to a plant, a vegetative plant part (s), or seed
having three sets of
chromosomes, designated herein as 3n.
[0081] "Tetraploid" refers to a plant, vegetative plant part(s), or seed
having four sets of
chromosomes, designated herein as 4n.
[0082] "Rootstock" or "stock" refers to the plant selected for its roots, in
particular for the
resistance of the roots to diseases or stress (e.g., heat, cold, salinity
etc.). Normally, the quality
of the fruit of the plant providing the rootstock is less important.
[0083] "Scion" refers to a part of the plant attached to the rootstock. This
plant is selected for
its stems, leaves, flowers, or fruits. The scion contains the desired genes to
be duplicated in future
production by the stock/scion plant and may produce the desired cucumber
fruit.
[0084] "Stock/scion" or "grafted plant" refers to a cucumber plant comprising
a rootstock from
one plant grafted to a scion from another plant.
[0085] "Grafting" refers to the method of joining of (genetically) different
plant parts,
especially scions and rootstocks, together so that they grow as a single
plant. A grafted seedlings
or a grafted plant is a seedling or plant (produced by grafting) consisting of
such different plant
parts and which grows as one plant.
[0086] "Tissue culture" or "cell culture" refers to a composition comprising
isolated cells of the
same or a different type or a collection of such cells organized into parts of
a plant. Tissue culture
of various tissues of cucumber and regeneration of plants therefrom is well
known and widely
published (see, e.g., Sang-Gu et al. (1988), Plant Cell, Tissue and Organ
Culture 12: 67-74;
Colijn-Hooymans (1994), Plant Cell, Tissue and Organ Culture 39: 211-217).
Similarly, methods
of preparing cell cultures are known in the art.
[0087] "Vegetative propagation," "vegetative reproduction," or "clonal
propagation" are used
interchangeably herein and mean a method of taking a plant part and inducing
or allowing that
plant part to form at least roots, and also refer to the plant or plantlet
obtained by that method.
19
Date Recue/Date Received 2022-09-09
Optionally, the vegetative propagation is grown into a mature plant. The
skilled person is aware
of what plant parts are suitable for use in the method.
[0088] "Regeneration" refers to the development of a plant from cell culture,
or tissue culture,
or vegetative propagation.
[0089] The term "traditional breeding techniques" encompasses herein crossing,
selfing,
selection, doubled haploid production, embryo rescue, protoplast fusion,
marker assisted
selection, mutation breeding etc. as known to the breeder (i.e., methods other
than genetic
modification/transformation/transgenic methods), by which, for example, a
genetically heritable
trait can be transferred from one cucumber line or variety to another.
[0090] "Crossing" refers to the mating of two parent plants. The term
encompasses "cross-
pollination" and "selfing".
[0091] "Selfing" refers to self-pollination of a plant, i.e., the transfer of
pollen from the anther
to the stigma of the same plant.
[0092] "Cross-pollination" refers to the fertilization by the union of two
gametes from different
plants.
[0093] "Backcrossing" is a traditional breeding technique used to introduce a
trait into a plant
line or variety. The plant containing the trait is called the donor plant and
the plant into which
the trait is transferred is called the recurrent parent. An initial cross is
made between the donor
parent and the recurrent parent to produce a progeny plant. Progeny plants
which have the trait
are then crossed to the recurrent parent. After several generations of
backcrossing and/or selfing
the recurrent parent comprises the trait of the donor. The plant generated in
this way may be
referred to as a "single trait converted plant". The technique can also be
used on a parental line
of a hybrid.
[0094] "Progeny" as used herein refers to a plant obtained from a plant
designated NUN 93099
CUL. A progeny may be obtained by regeneration of cell culture or tissue
culture or parts of a
plant of said variety or selfing of a plant of said variety or by producing
seeds of a plant of said
variety. In further aspects, progeny may also encompass plants obtained from
crossing of at least
one plant of said variety with another cucumber plant of the same variety or
another variety or
line, or with wild cucumber plants. A progeny may comprise a mutation or a
transgene. A "first
generation progeny" is the progeny directly derived from, obtained from,
obtainable from or
derivable from the parent plant by, e.g., traditional breeding methods
(selfing and/or cross-
pollinating) or regeneration (optionally combined with transformation or
mutation). Thus, a plant
Date Recue/Date Received 2022-09-09
of variety NUN 93099 CUL is the male parent, the female parent or both of a
first generation
progeny of cucumber variety NUN 93099 CUL. Progeny may have all the
physiological and
morphological characteristics of cucumber variety NUN 93099 CUL, when grown
under the same
environmental conditions. Using methods such as backcrossing, recurrent
selection, mutation or
transformation, one or more specific characteristics may be introduced into
said variety, to
provide or a plant comprising all but 1, 2, or 3 of the morphological and
physiological
characteristics of cucumber variety NUN 93099 CUL.
[0095] The terms "gene converted" or "conversion plant" or "single locus
converted plant" in
this context refer to cucumber plants which are developed by traditional
breeding techniques,
e.g., backcrossing or via genetic engineering or through mutation breeding,
wherein essentially
all of the desired morphological and physiological characteristics of the
parent variety or line are
recovered, in addition to the one or more characteristics introduced into the
parent via e.g., the
backcrossing technique (optionally including reverse breeding or reverse
synthesis of breeding
lines). It is understood that not only the addition of a further
characteristic (e.g., addition of gene
conferring a further characteristic, such as a disease resistance gene), but
also the
replacement/modification of an existing characteristic by a different
characteristic is
encompassed herein (e.g., mutant allele of a gene can modify the phenotype of
a characteristic).
[0096] Likewise, a "Single Locus Converted (Conversion) Plant" refers to
plants developed by
plant breeding techniques comprising or consisting of mutation and/or by
genetic transformation
and/or by traditional breeding techniques, such as backcrossing, wherein
essentially all of the
desired morphological and physiological characteristics of a cucumber variety
are recovered in
addition to the characteristics of the single locus having been transferred
into the variety via the
abovementioned technique, or wherein the morphological and physiological
characteristics of
the variety has been replaced/modified in the variety. In case of a hybrid,
the gene may be
introduced or modified in the male or female parental line.
[0097] "Transgene" or "chimeric gene" refers to a genetic locus comprising a
DNA Sequence
which has been introduced into the genome of the plant by transformation. A
plant comprising a
transgene stably integrated into its genome is referred to as "transgenic
plant"
[0098] As used herein, the terms "resistance" and "tolerance" are used
interchangeably to
describe plants that show no symptoms or significantly reduced symptoms to a
specified biotic
pest, pathogen, abiotic influence or environmental condition compared to a
susceptible plant.
21
Date Recue/Date Received 2022-09-09
These terms are optionally also used to describe plants showing some symptoms
but that are still
able to produce marketable product with an acceptable yield.
[0099] "Average" refers herein to the arithmetic mean.
[0100] The term "mean" refers to the arithmetic mean of several measurements.
The mean, if
not indicated otherwise within this application, refers to the arithmetic mean
of measurements
on at least 15 different, randomly selected plants of a variety or line.
DETAILED DESCRIPTION OF THE VARIOUS ASPECTS OF THE DISCLOSURE
[0101] The disclosure relates to a plant of variety NUN 93099 CUL, wherein a
representative
sample of seeds of said variety has been deposited under the Budapest Treaty,
with Accession
Number NCIMB 43946. NUN 93099 CUL is a Dutch long cucumber variety and- is
suitable for
growing in a greenhouse, e.g., indoor cultivation with drip irrigation.
[0102] In another aspect, the plant of variety NUN 93099 CUL, or part thereof,
or progeny
thereof comprises resistance to Cladosporium cucumerinum, Cucumber Mosaic
Virus (CMV),
Podosphaera xanthii, Corynespora cassiicola, Cucumber Vein Yellowing Virus
(CVYV), and
Cucumber Green Mottle Mosaic Virus (CGMMV), measured according to TG/61/7.
[0103] The disclosure also provides a cucumber plant or part thereof having
all of the
physiological and morphological characteristics of the plant of cucumber
variety NUN 93099
CUL when grown under the same environmental conditions.
[0104] In another aspect, the plant of cucumber variety NUN 93099 CUL, or part
thereof, of
progeny plant thereof, comprises all of the morphological and physiological
characteristics (i.e.,
average values, as indicated on the USDA Objective description of variety ¨
cucumber, unless
indicated otherwise) as shown in Tables 1 and 2, when determined at the 5%
significance level
for numerical characteristics and determined by type or degree for non-
numerical characteristics
for plants grown under the same environmental conditions. A part of this plant
is also provided.
[0105] The disclosure further provides a cucumber plant which does not differ
from the
physiological and morphological characteristics of the plant of cucumber
variety NUN 93099
CUL as determined at the 1%, 2%, 3%, 4% or 5% significance level for numerical
characteristics
and identical (same type or degree) for non-numerical characteristics when
grown under the same
environmental conditions. In a particular aspect, the plants are measured in
the same trial (e.g.,
22
Date Recue/Date Received 2022-09-09
the trial is conducted as recommended by the USDA or UPOV). The disclosure
also comprises a
part of said plant, preferably a fruit or a part thereof.
[0106] The disclosure further relates to a cucumber variety NUN 93099 CUL
which when
compared to the Reference Variety has the following distinguishing
characteristics as shown in
Table 3:
a. shorter internode length;
b. shorter length of first 15 internodes;
c. medium plant vigor;
d. shorter mature leaf length;
e. thinner mature leaf width;
f. shorter mature leaf petiole length;
g. horizontal mature leaf attitude;
h. thinner mature leaf petiole diameter;
i. larger mature fruit diameter at medial;
j. dark green color at stem end;
k. dark green color at blossom end;
1. necked fruit shape;
m. dull skin luster;
n. medium ratio of length/diameter;
o. larger core diameter in relation to diameter of fruit;
p. longer length of neck;
q. rounded shape of calyx end;
r. lighter intensity of ground color of skin;
s. very sparse density of vestiture;
t. shorter length of peduncle;
u. intermediately resistant to Cucumber Mosaic Virus (CMV); and
v. highly resistant to Cucumber Vein Yellowing Virus (CVYV), when determined
at the 5%
significance level for numerical characteristics and determined by type or
degree for non-
numerical characteristics for plants grown under the same environmental
conditions.
[0107] The morphological and/or physiological differences between two
different individual
plants described herein (e.g., between cucumber variety NUN 93099 CUL and a
progeny of said
variety) or between a plant of variety NUN 93099 CUL or progeny of said
variety, or a plant
23
Date Recue/Date Received 2022-09-09
having all, or all but 1, 2, or 3, of the physiological and morphological
characteristics of
cucumber variety NUN 93099 CUL (or all, or all but 1, 2, or 3 of the
characteristics as listed in
Tables 1 and 2) and another known variety can easily be established by growing
said variety next
to each other or next to the other variety (in the same field, under the same
environmental
conditions), preferably in several locations which are suitable for said
cucumber cultivation, and
measuring morphological and/or physiological characteristics of a number of
plants (e.g., to
calculate an average value and to determine the variation range/uniformity
within the variety).
Various characteristics, for example, days from seeding to market maturity,
plant habit, plant
growth and sex, stem form, fruit skin color, fruit neck shape, fruit tapering,
skin thickness,
toughness, and luster, flavor, disease resistance, insect resistance, can be
measured and directly
compared for species of cucumber.
[0108] Thus, the disclosure comprises a cucumber plant having one, two, or
three physiological
or morphological characteristics which are different from those of the plant
of variety NUN
93099 CUL and which otherwise has all the physiological and morphological
characteristics of
the plant of variety NUN 93099 CUL, when determined (e.g., at the 5%
significance level for
quantitative characteristics and identical for non-numerical characteristics)
for plants grown
under the same environmental conditions. In another aspect, the different
characteristic is
affected by a mutation, optionally induced mutation, or by transformation.
[0109] The disclosure also relates to a seed of cucumber variety NUN 93099 CUL
wherein a
representative sample of said seed has been deposited under the Budapest
Treaty, with Accession
Number NCIMB 43946.
[0110] In another aspect, a seed of hybrid variety NUN 93099 CUL is obtainable
by crossing
the male parent of cucumber variety NUN 93099 CUL with the female parent of
cucumber variety
NUN 93099 CUL and harvesting the seeds produced on the female parent. The
resultant seeds of
said variety can be grown to produce plants of cucumber variety NUN 93099 CUL.
[0111] The disclosure also provides a plant grown from a seed of cucumber
variety NUN 93099
CUL and plant part thereof, wherein a representative sample of seed has been
deposited under
Accession Number NCIMB 43946.
[0112] The disclosure also provides a cucumber fruit produced on a plant grown
from a seed of
cucumber variety NUN 93099 CUL, wherein a representative sample of seed has
been deposited
under Accession Number NCIMB 43946.
24
Date Recue/Date Received 2022-09-09
[0113] In another aspect, the disclosure provides for a cucumber plant part of
variety NUN
93099 CUL, preferably a fruit or part thereof, a representative sample of seed
from said variety
has been deposited under the Budapest Treaty, with Accession Number NCIMB
43946.
[0114] Also provided is a plant of variety NUN 93099 CUL, or a fruit, or other
plant part
thereof, produced from a seed, wherein a representative sample of said seeds
has been deposited
under the Budapest Treaty, with Accession Number NCI1VIB 43946.
[0115] Also provided is a plant part obtained from variety NUN 93099 CUL,
wherein said plant
part is a fruit, a harvested fruit, a part of a fruit, a leaf, a part of a
leaf, pollen, an ovule, a cell, a
petiole, a shoot or a part thereof, a stem or a part thereof, a root or a part
thereof, a root tip, a
cutting, a seed, a part of a seed, seed coat or another maternal tissue which
is part of a seed grown
on said variety, a hypocotyl, a cotyledon, a scion, a stock, a rootstock, a
pistil, an anther, and a
flower or a part thereof. Such plant parts may be suitable for sexual
reproduction (e.g., a pollen,
a flower, an ovary, an ovule, an embryo, etc.), vegetative reproduction (e.g.,
a cutting, a root, a
stem, a cell, a protoplast, a leaf, a cotyledon, a meristem, etc.) or tissue
culture (e.g., a leaf, a
pollen, an embryo, a cotyledon, a hypocotyl, a cell, a root, a root tip, an
anther, a flower, a seed,
a stem, etc.). Fruits are particularly important plant parts. Fruits may be
parthenocarpic, or
seedless, or contain immature or nonviable seeds, or contain viable seeds.
[0116] In a further aspect, the plant part obtained from variety NUN 93099 CUL
is a cell,
optionally a cell in a cell or tissue culture. That cell may be grown into a
plant of variety NUN
93099 CUL. A part of the plant of variety NUN 93099 CUL (or of a progeny of
that variety or
of a plant having all physiological and/or morphological characteristics but
one, two, or three
which are different from those of cucumber variety NUN 93099 CUL, further
encompasses any
cells, tissues, organs obtainable from the seedlings or plants in any stage of
maturity.
[0117] The disclosure also provides a tissue or cell culture comprising cells
of cucumber variety
NUN 93099 CUL. Such a tissue culture can, for example, be grown on plates or
in liquid culture,
or be frozen for long term storage. The cells of cucumber variety NUN 93099
CUL used to start
the culture can be selected from any plant partsuitable for vegetative
reproduction, or in a
particular aspect can be cells of an embryo, a meristem, a cotyledon, a
hypocotyl, pollen, a leaf,
an anther, a root, a root tip, a pistil, a petiole, a flower, a fruit, a seed
or a stem. In another
particular aspect, the tissue culture does not contain somaclonal variation or
has reduced
somaclonal variation. The skilled person is familiar with methods to reduce or
prevent
somaclonal variation, including regular re-initiation.
Date Recue/Date Received 2022-09-09
[0118] In one aspect, the disclosure provides a cucumber plant regenerated
from the tissue or
cell culture of cucumber variety NUN 93099 CUL, wherein the regenerated plant
is not
significantly different from cucumber variety NUN 93099 CUL, in all, or all
but one, two, or
three, of the physiological and morphological characteristics, e.g.,
determined at the 5%
significance level for numerical characteristics and identical (same type or
degree) for non-
numerical characteristics, when grown under the same environmental conditions.
Optionally, the
plant has one, two, or three the physiological or morphological
characteristics that are affected
by a mutation or by transformation with a transgene.
[0119] In another aspect, the disclosure provides a cucumber plant regenerated
from the tissue
or cell culture of cucumber variety NUN 93099 CUL, wherein the plant has all
or all but one,
two, or three of the physiological and morphological characteristics of said
variety, e.g.,
determined at the 5% significance level for numerical characteristics and
identical (same type or
degree) for non-numerical characteristics, when grown under the same
environmental conditions.
Similarity or difference of a characteristic is determined by measuring that
characteristics on a
representative number of plants grown under the same environmental conditions,
determining
whether the type/degree characteristics are the same and determining whether
numerical
characteristics are different at the 5% significance level.
[01201 Cucumber variety NUN 93099 CUL, or its progeny, or a plant having all
physiological
and/or morphological characteristics but one, two, or three which are
different from those of
cucumber variety NUN 93099 CUL, can also be reproduced using vegetative
reproduction
methods. Therefore, the disclosure provides for a method of producing a plant,
or a plant part, of
cucumber variety NUN 93099 CUL, comprising vegetative propagation of said
variety.
Vegetative propagation comprises regenerating a whole plant from a plant part
of variety NUN
93099 CUL or from a progeny or from or a plant having all physiological and/or
morphological
characteristics of said variety but one, two, or three different
characteristics, such as a cutting, a
cell culture, or a tissue culture.
101211 The disclosure also provides methods of vegetatively propagating a part
of the plant of
variety NUN 93099 CUL. In certain aspects, the method comprises: (a)
cultivating tissue or cells
capable of being propagated from cucumber variety NUN 93099 CUL to obtain
proliferated
shoots; and (b) rooting said proliferated shoots, to obtain rooted plantlets.
Steps (a) and (b) may
also be reversed, i.e., first cultivating said tissue to obtain roots and then
cultivating the tissue to
obtain shoots, thereby obtaining rooted plantlets. The rooted plantlets may
then be further grown,
26
Date Recue/Date Received 2022-09-09
to obtain plants. In one aspect, the method further comprises step (c) growing
plants from said
rooted plantlets. Therefore, the method also comprises regenerating a whole
plant from a part of
the plant of variety NUN 93099 CUL. In a particular aspect, the part of the
plant to be propagated
is is a cutting, a cell culture, or a tissue culture.
[0122] The disclosure also provides for a vegetatively propagated plant of
variety NUN 93099
CUL, or part thereof (or from progeny of said variety or from or a plant
having all but one, two,
or three physiological and/or morphological characteristics of cucumber
variety NUN 93099
CUL), wherein the plant or part thereof has all of the morphological and
physiological
characteristics of cucumber variety NUN 93099 CUL, e.g., determined at the 5%
significance
level for numerical characteristics and identical (same type or degree) for
non-numerical
characteristics, when grown under the same environmental conditions. In
another aspect, the
propagated plant has all but one, two, or three of the morphological and
physiological
characteristics of cucumber variety NUN 93099 CUL, e.g., determined at the 5%
significance
level for numerical characteristics and identical (same type or degree) for
non-numerical
characteristics, when grown under the same environmental conditions.
[0123] In another aspect, the plant and plant parts of cucumber variety NUN
93099 CUL and
progeny of said variety are provided, e.g., grown from seeds, produced by
sexual or vegetative
reproduction, regenerated from the above-described plant parts, or regenerated
from a cell or
tissue culture of cucumber variety NUN 93099 CUL, in which the reproduced
(seed propagated
or vegetatively propagated) plant has all of the physiological and
morphological characteristics
of cucumber variety NUN 93099 CUL. In one aspect, said progeny of cucumber
variety NUN
93099 CUL can be modified in one, two, or three characteristics, in which the
modification is a
result of mutagenesis or transformation with a transgene.
[0124] In another aspect, the disclosure provides a method for producing a
cucumber plant part,
preferably a fruit, comprising growing the plant of variety NUN 93099 CUL
until it sets at least
one fruit, and collecting the fruit. Preferably, the fruit is collected at
harvest maturity. In another
aspect, the fruit is collected when the seed is ripe.
[0125] In another aspect, the plant of variety NUN 93099 CUL can be produced
by seeding
directly in the soil (e.g., the field) or by germinating the seeds in a
controlled environment (e.g.,
a greenhouse) and optionally then transplanting the seedlings into the field.
For example, a seed
is sown into a prepared seed bed in a field where the plant remains for its
entire life. Alternatively,
the cucumber seed may be planted through a black plastic mulch. The dark
plastic will absorb
27
Date Recue/Date Received 2022-09-09
heat from the sun, warming the soil early. It will also help to conserve
moisture during the
growing season, controls weed and makes harvesting easier and cleaner. Tunnel
row covers are
also used for protection against frost and wind, optionally with drip of
irrigation system (see, e.g.,
Schrader, et. al., University of California Agriculture and Natural Resources,
Publication 8050,
2002, 1-8). Cucumber can also be grown on poles or trellises to keep the fruit
suspended or
entirely in the greenhouse (available at world-wide web at wifss.ucdavis.edu
under wp-
c ontent/uploads/2016/05/FDA_WIF SS_-Cucumbers_PDF .pdf). High wire
cultivation system
and the use of artificial lightning have also been introduced for year-round
production of
cucumber (available at world-wide web at nunhems.com under
gb/en/solutions/high-wire.html).
101261 In still another aspect, the disclosure provides a method of producing
a cucumber plant,
comprising crossing the plant of variety NUN 93099 CUL with a second cucumber
plant at least
once, allowing seed to develop and optionally harvesting said progeny seed.
The skilled person
can select progeny from said crossing. Optionally, the progeny (grown from the
progeny seed) is
crossed twice, thrice, or four, five, six or seven times, and allowed to set
seed. In one aspect, the
first "crossing" further comprises planting seeds of a first and a second
parent cucumber 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. After
pollination the plant can produce seed.
101271 The disclosure also provides a method for collecting pollen of cucumber
variety NUN
93099 CUL, comprising collecting pollen from the plant of variety NUN 93099
CUL.
Alternatively, the method comprises growing a plant of variety NUN 93099 CUL
until at least
one flower contains pollen and collecting the pollen. In a particular aspect,
the pollen is collected
when it is mature or ripe. A suitable method for collecting pollen comprises
collecting anthers or
the part of the anther that contains pollen, for example, by cutting the
anther or the part of the
anther off. Pollen can be collected in a container. Optionally, collected
pollen can be used to
pollinate a cucumber flower.
[0128] In still another aspect, the disclosure provides a method of producing
a cucumber plant,
comprising selfing the plant of variety NUN 93099 CUL one or more times, and
selecting a
progeny cucumber plant from said selfing. In one aspect, the progeny plant
retains all or all but
one, two, or three of the physiological and morphological characteristic of
cucumber variety
NUN 93099 CUL, when grown under the same environmental conditions. In a
different aspect,
28
Date Recue/Date Received 2022-09-09
the progeny plant comprises all of the physiological and morphological
characteristic of
cucumber variety NUN 93099 CUL of Tables 1 and 2.
[0129] The disclosure also provides a method for developing a cucumber plant
in a breeding
program, using the plant of cucumber variety NUN 93099 CUL, or its parts as a
source of plant
breeding material. Suitable plant breeding techniques are recurrent selection,
backcrossing,
pedigree breeding, mass selection, mutation breeding and/or genetic marker
enhanced selection.
In one aspect, the method comprises crossing cucumber variety NUN 93099 CUL or
its progeny,
or a plant comprising all but 1, 2, or 3 or more of the morphological and
physiological
characteristics of cucumber variety NUN 93099 CUL (e.g., as listed in Tables 1
and 2), with a
different cucumber plant, and wherein one or more offspring of the crossing
are subject to one
or more plant breeding techniques: recurrent selection, backcrossing, pedigree
breeding, mass
selection, mutation breeding and genetic marker enhanced selection (see, e.g.,
Vidaysky and
Czosnek, (1998) Phytopathology 88(9): 910-4). For breeding methods in general,
see, e.g.,
Principles of Plant Genetics and Breeding, 2007, George Acquaah, Blackwell
Publishing, ISBN-
13: 978-1-4051-3646-4.
[0130] In other aspects, the disclosure provides a progeny plant of variety
NUN 93099 CUL,
such as a progeny plant obtained by further breeding of cucumber variety NUN
93099 CUL.
Further breeding with cucumber variety NUN 93099 CUL includes selfing that
variety and/or
cross-pollinating said variety with another cucumber plant one or more times.
In a particular
aspect, the disclosure provides for a progeny plant that retains all the
morphological and
physiological characteristics of cucumber variety NUN 93099 CUL, or in another
aspect, a
progeny plant that retains all, or all but one, two, or three of the
morphological and physiological
characteristics of cucumber variety NUN 93099 CUL, optionally all or all but
one, two, or three
of the characteristics as listed in Tables 1 and 2, determined at the 5%
significance level for
numerical characteristics and identical for non-numerical characteristics,
when grown under the
same environmental conditions. In another aspect, the progeny is a first
generation progeny, i.e.,
the ovule or the pollen (or both) used in the crossing is an ovule or pollen
of cucumber variety
NUN 93099 CUL, where the pollen comes from an anther of cucumber variety NUN
93099 CUL,
and the ovule comes from an ovary of cucumber variety NUN 93099 CUL. In
another aspect, the
disclosure provides for a vegetative reproduction of the variety and a plant
having all, or all but
1, 2, or 3 of the physiological and morphological characteristics of cucumber
variety NUN 93099
CUL (e.g. as listed in Tables 1 and 2).
29
Date Recue/Date Received 2022-09-09
[0131] In one aspect, pedigree selection is used as a breeding method for
developing a cucumber
variety. Pedigree selection is also known as the "Vilmorin System of
Selection," see, e.g., Allard,
John Wiley & Sons, Inc., 1999, 64-67. In general, selection is first practiced
among F2 plants. In
the next season, the most desirable F3 lines are first identified, then
desirable F3 plants within
each line are selected. The following season and in all subsequent generations
of inbreeding, the
most desirable families are identified first, then desirable lines within the
selected families are
chosen. A family refers to lines that were derived from plants selected from
the same progeny
from the preceding generation.
[0132] Thus, progeny in connection with pedigree selection are either the
generation (seeds)
produced from the first cross (F1) or selfing (Si), or any further generation
produced by crossing
and/or selfing (F2, F3, etc.) and/or backcrossing (BC1, BC2, etc.) one or more
selected plants of
the Fl and/or S1 and/or BC1 generation (or plants of any further generation,
e.g., F2) with another
cucumber plant (and/or with a wild relative of cucumber). Progeny may have all
the physiological
and morphological characteristics of cucumber variety NUN 93099 CUL, when
grown under the
same environmental conditions and/or progeny may have (be selected for having)
one or more of
the distinguishing characteristics of cucumber variety NUN 93099 CUL.
[0133] In yet a further aspect, the disclosure provides for a method of
producing a new
cucumber plant. The method comprises crossing cucumber variety NUN 93099 CUL,
or a plant
comprising all but 1, 2, or 3 of the morphological and physiological
characteristics of cucumber
variety NUN 93099 CUL, (as listed in Tables 1 and 2), or a progeny thereof,
either as male or as
female parent, with a second cucumber plant (or a wild relative of cucumber)
one or more times,
and/or selfing a cucumber plant of variety NUN 93099 CUL or a progeny plant
thereof, one or
more times, and selecting progeny from said crossing and/or selfing. The
second cucumber plant
may, for example, be a line or variety of the species C. sativus L., Cucumis
hystrix, Cucumis
ritchiei (syn. Dicaelospermum ritchiei) or Cucumis maderaspatana (syn. Mukia
maderaspatana).
[0134] In a further aspect, cucumber variety NUN 93099 CUL is used in crosses
with other
different cucumber varieties to produce first generation (F1) cucumber hybrid
seeds and plants
with superior characteristics. In a particular aspect, the disclosure provides
a cucumber seed and
a cucumber plant produced by crossing a first parent cucumber plant with a
second parent
cucumber plant, wherein at least one of the first or second parent cucumber
plant is cucumber
variety NUN 93099 CUL. In another aspect, the cucumber seed and cucumber plant
produced
Date Recue/Date Received 2022-09-09
are the first filial generation (F1) cucumber seed and plant produced by
crossing the plant of
cucumber variety NUN 93099 CUL with another cucumber plant.
[0135] The morphological and physiological characteristics of cucumber variety
NUN 93099
CUL are provided in Tables 1 and 2, as collected in a trial according to USDA
and/or UPOV
standards. Encompassed herein is also a plant obtainable from cucumber variety
NUN 93099
CUL (e.g., by selfing and/or crossing and/or backcrossing with said variety
and/or progeny of
said variety) comprising all or all but one, two, or three of the
physiological and morphological
characteristics of cucumber variety NUN 93099 CUL as listed in Tables 1 and 2
(e.g., as
determined at the 5% significance level for numerical characteristics and
identical for non-
numerical characteristics), when grown under the same environmental conditions
and/or
comprising one or more (or all; or all except one, two, or three)
characteristics when grown under
the same environmental conditions. The morphological and/or physiological
characteristics may
vary somewhat with variation in the environment (such as temperature, light
intensity, day
length, humidity, soil, fertilizer use), which is why a comparison under the
same environmental
conditions is preferred. Colors can best be measured using the Royal
Horticultural Society (RHS)
Chart.
101361 In another aspect, the disclosure provides a method of producing a
plant derived from a
cucumber variety NUN 93099 CUL, comprising crossing the plant of variety NUN
93099 CUL
either as a male or female parent with a second cucumber plant or selfing
cucumber variety NUN
93099 CUL or vegetatively propagating cucumber NUN 93099 CUL and collecting
seeds from
said crossing or selfing or regenerating a whole plant from the vegetable cell-
or tissue culture.
Also provided are seeds and/or plants obtained by this method. All plants
produced using
cucumber variety NUN 93099 CUL as a parent are within the scope of the
disclosure including
plant parts derived from cucumber variety NUN 93099 CUL.
[0137] In further aspects, the method comprises growing 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 additional 3-10 generations to produce a plant
derived from cucumber
variety NUN 93099 CUL. The plant derived from cucumber variety NUN 93099 CUL
may be an
inbred line and the aforementioned repeating crossing steps may be defined as
comprising
sufficient inbreeding to produce the inbred line. By selecting plants having
one or more desirable
traits of the line as well as potentially other selected traits.
31
Date Recue/Date Received 2022-09-09
[0138] The disclosure provides for methods of producing plants which retain
all the
morphological and physiological characteristics of the plant described herein.
The disclosure also
provides for methods of producing a plant comprising all but 1, 2, or 3 or
more of the
morphological and physiological characteristics of cucumber variety NUN 93099
CUL (e.g., as
listed in Tables 1 and 2), but which are still genetically closely related to
said variety. The
relatedness can, for example, be determined by fingerprinting techniques
(e.g., making use of
isozyme markers and/or molecular markers such as Single-nucleotide
polymorphism (SNP)
markers, amplified fragment length polymorphism (AFLP) markers,
microsatellites,
mini satellites, Random Amplified Polymorphic DNA (RAPD) markers, restriction
fragment
length polymorphism (RFLP) markers and others). A plant is "closely related"
to cucumber
variety NUN 93099 CUL if its DNA fingerprint is at least 80%, 90%, 95% or 98%
identical to
the fingerprint of cucumber variety NUN 93099 CUL. In a particular aspect,
AFLP markers are
used for DNA fingerprinting (see, e.g., Vos et al. 1995, Nucleic Acid Research
23: 4407-4414).
A closely related plant may have a Jaccard's Similarity index of at least
about 0.8, preferably at
least about 0.9, 0.95, 0.98 or more (see, e.g., Parvathaneni et al., J. Crop
Sci. Biotech. 2011
(March) 14 (1) : 39-43). The disclosure also provides a plant and a variety
obtained or selected
by applying these methods on cucumber variety NUN 93099 CUL. Such a plant may
be produced
by crossing and/or selfing, or alternatively, a plant may simply be identified
and selected amongst
plants of said variety, or progeny of said variety, e.g., by identifying a
variant within cucumber
variety NUN 93099 CUL or within progeny of said variety (e.g., produced by
selfing) which
variant differs from cucumber variety NUN 93099 CUL in one, two, or three of
the morphological
and/or physiological characteristics (e.g., in one, two, or three
distinguishing characteristics),
e.g. those listed in Tables 1 and 2. In another aspect, the disclosure
provides a cucumber plant
having a Jaccard's Similarity index with cucumber variety NUN 93099 CUL of at
least 0.8, e.g.,
at least 0.85, 0.9, 0.95, 0.98 or even at least 0.99.
[0139] In some aspects, the disclosure provides a cucumber plant comprising
genomic DNA
having at least 95%, 96%, 97%, 98% or 99% Sequence identity compared to the
genomic DNA
Sequence of a plant of variety NUN 93099 CUL as deposited under Accession
Number NCIMB
43946. In some aspects, the cucumber plant further comprises all or all but 1,
2, or 3 of the
morphological and physiological characteristics of cucumber variety NUN 93099
CUL (e.g. as
listed in Tables 1 and 2). In other aspects, the cucumber plant is a hybrid
derived from a seed or
32
Date Recue/Date Received 2022-09-09
plant of variety NUN 93099 CUL. In another aspects the cucumber plant
comprises the
physiological and morphological characteristics of cucumber variety NUN 93099
CUL.
[0140] For the purpose of this disclosure, the "Sequence identity" of
nucleotide Sequences,
expressed as a percentage, refers to the number of positions in the two
optimally aligned
Sequences which have identical residues (x100) divided by the number of
positions compared.
A gap, i.e., a position in the pairwise alignment where a residue is present
in one Sequence but
not in the other, is regarded as a position with non-identical residues. A
pairwise global Sequence
alignment of two nucleotide Sequences is found by aligning the two Sequences
over the entire
length according to the Needleman and Wunsch global alignment algorithm
described in
Needleman and Wunsch, 1970, J. Mol, Biol. 48(3):443-53). A full implementation
of the
Needleman-Wunsch global alignment algorithm is found in the needle program in
The European
Molecular Biology Open Software Suite (see, e.g., EMBOSS, Rice et al., Trends
in Genetics June
2000, vol. 16, No. 6. pp.276-277).
[0141] In one aspect, the plant of cucumber variety NUN 93099 CUL may also be
mutated (by
e.g., irradiation, chemical mutagenesis, heat treatment, etc.) and mutated
seeds or plants may be
selected in order to change one or more characteristics of said variety.
Methods such as TILLING
(Targeting Induced Local Lesions in Genomes) may be applied to cucumber
populations in order
to identify mutants.
[0142] Similarly, cucumber variety NUN 93099 CUL may be transformed and
regenerated,
whereby one or more chimeric genes are introduced into the variety or into a
plant comprising
all but 1, 2, 3, or more of the morphological and physiological
characteristics (e.g., as listed in
Tables 1 and 2). Many useful traits can be introduced into cucumber variety
93099 CUL by e.g.,
crossing cucumber variety NUN 93099 CUL with a transgenic cucumber plant
comprising
desired transgene, as well as by directly introducing a transgene into
cucumber variety NUN
93099 CUL by genetic transformation techniques.
[0143] Transformation can be carried out using standard methods, such as
Agrabacterium
tumefaciens mediated transformation or biolistics, followed by selection of
the transformed cells
and regeneration into plants. A desired trait (e.g., gene(s) conferring pest
or disease resistance,
herbicide, fungicide or insecticide tolerance, etc.) can be introduced into
cucumber variety NUN
93099 CUL, or progeny of said variety, by transforming said variety or progeny
of said variety
with a transgene that confers the desired trait, wherein the transformed plant
retains all or all but
one, two or three of the phenotypic and/or morphological and/or physiological
characteristics of
33
Date Recue/Date Received 2022-09-09
cucumber variety NUN 93099 CUL or the progeny of said variety and contains the
desired trait.
In another aspect, the transformation or mutation confers a trait wherein the
trait is the desired
trait is yield, size, compactness, dry matter content, fruit quality, enhanced
nutritional quality,
post-harvest quality, storage properties, color, male sterility, herbicide
tolerance, insect
resistance, pest resistance, disease resistance, Powdery mildew resistance
without necrosis,
environmental stress tolerance, modified carbohydrate metabolism, modified
protein
metabolism, or ripening.
[0144] Any pest or disease resistance genes may be introduced into the plant
of variety NUN
93099 CUL, progeny of said variety or into a plant comprising all but 1, 2, or
3 or more of the
morphological and physiological characteristics of cucumber variety NUN 93099
CUL (e.g., as
listed in Tables 1 and 2). Resistance to one or more of the following diseases
or pests may be
introduced into plants described herein: Angular Leaf Spot (Pseudomonas
lachrymans),
Anthracnose Race 1 (Colletotrichum lagenaria), Anthracnose Race 2, Bacterial
Wilt (Erwinia
tracheiphilus), Cucumber Scab (Gummosis) (Cladosporium cucumerinum), Downy
Mildew
(Pseudoperonospora cubensis), Powdery Mildew (Erysiphe chicoracearum),
Alternaria Leaf
Blight (Alternaria cucumerina), Target Spot (Corynespora cassiicola), Cucumber
Rust, Root
Rot, Crown Blight, Verticillum Wilt, Sulphur Burn, Fusarium oxysporum fsp.
cucumerinum,
Fusarium Wilt Race2, Root Knot (Nematode), Anthracnose, and/or Squash Mosaic.
Other
resistances, against pathogenic viruses (e.g., Cucumber Mosaic Virus (CMV),
Cucumber Yellow
Mottle Mosaic Virus (CYMMV), Cucumber Green Mottle Mosaic Virus (CGMMV),
Cucumber
Aucuba Mosaic Virus (CAMV), Muskmelon Mosaic Virus (MMV), Cucumber Vein
Yellowing
Virus (CVYV), Cucurbit Yellow Stunting Disorder Virus (CYSDV), Watermelon
Mosaic Virus
(WMV), Papaya Ring Spot Virus (PRSV), Zucchini Mosaic Virus (ZMV)), fungi,
bacteria,
nematodes, insects, or other pests may also be introduced.
[0145] Genetic transformation may, therefore, be used to insert a selected
transgene into the
cucumber plants of the disclosure described herein or may, alternatively, be
used for the
preparation of transgenic cucumber plants which can be used as a source of the
transgene(s),
which can be introduced into cucumber variety NUN 93099 CUL by e.g.,
backcrossing. A genetic
trait which has been engineered into the genome of a particular cucumber plant
may then be
moved into the genome of another cucumber plant (e.g., another variety) using
traditional
breeding techniques which are well-known in the art. For example, backcrossing
is commonly
used to move a transgene from a transformed cucumber variety into an already
developed
34
Date Recue/Date Received 2022-09-09
cucumber variety and the resulting backcross conversion plant will then
comprise the
transgene(s).Any DNA Sequences, whether from a different species or from the
same species,
which are inserted into the genome using transformation, are referred to
herein collectively as
"transgenes." A "transgene" also encompasses antisense, or sense and antisense
Sequences
capable of gene silencing. Thus, the disclosure also related to transgenic
plants of cucumber
variety NUN 93099 CUL. In some aspects, a transgenic plant of cucumber variety
NUN 93099
CUL may contain at least one transgene but could also contain at least 1, 2,
3, 4, or more
transgenes.
[0146] Plant transformation involves the construction of an expression vector
which will
function in plant cells. Such a vector comprises DNA comprising a gene under
control of, or
operatively linked to a regulatory element active in plant cells (e.g.,
promoter). The expression
vector may contain one or more such operably linked gene/regulatory element
combinations. The
vector may be in the form of a plasmid and can be used alone or in combination
with other
plasmids to provide transformed cucumber plants using transformation methods
to incorporate
transgenes into the genetic material of the cucumber plant(s). transformation
can be carried out
using standard methods, such as Agrobacterium tumefaciens mediated
transformation,
electroporation, biolistics particle delivery stem, or microprojectile
bombardment, followed by
selection of the transformed cells and regeneration into plants.
[0147] Plants can also be genetically engineered, modified, or manipulated to
express various
phenotypes of horticultural interest. Through the transformation of cucumber,
the expression of
genes can be altered to enhance disease resistance, insect resistance,
herbicide resistance, stress
tolerance, horticultural quality, and other traits. Transformation can also be
used to insert DNA
Sequences which control or help control male sterility or fertility
restoration. DNA Sequences
native to cucumber as well as non-native DNA Sequences can be transformed into
cucumber and
used to alter levels of native or non-native proteins. Reduction of the
specific activity of specific
genes (also known as gene silencing or gene suppression) is desirable for
several aspects of
genetic engineering in plants.
[0148] Genome editing is another method recently developed to genetically
engineer plants.
Specific modification of chromosomal loci or targeted mutation can be done
through Sequence-
specific nucleases (SSNs) by introducing a targeted DNA double strand break in
the locus to be
altered. Examples of SSNs that been applied to plants are: finger nucleases
(ZFNs), transcription
activator-like effector nucleases (TALENs), engineered homing endonucleases or
Date Recue/Date Received 2022-09-09
meganucleases, and clustered regularly interspaced short palindromic repeats
(CRISPR)/(CRISPR associated protein 9 (Cas9)), see, e.g., Songtad, et. al.,
Critical Reviews in
Plant Sciences, 2017, 36:1, 1-23.
[0149] Thus, the disclosure also provides a method of producing a cucumber
plant having a
desired trait comprising mutating the plant or plant part of variety NUN 93099
CUL and selecting
a plant with the desired trait, wherein the mutated plant retains all or all
but one, two, or three of
the physiological and morphological characteristics of cucumber variety NUN
93099 CUL,
optionally as described Tables 1 and 2, and contains the desired trait and
wherein a representative
sample of seed of variety NUN 93099 CUL has been deposited under Accession
Number NCIMB
43946. In a further aspect, the desired trait is yield, size, compactness, dry
matter content, fruit
quality, enhanced nutritional quality, post-harvest quality, storage
properties, color, male
sterility, herbicide tolerance, insect resistance, pest resistance, disease
resistance, Powdery
mildew resistance without necrosis, environmental stress tolerance, modified
carbohydrate
metabolism, modified protein metabolism, or ripening.
[0150] In one aspect, the disclosure provides a method for inducing a mutation
in cucumber
variety NUN 93099 CUL comprising:
a. exposing the seed, plant, plant part, or cell of cucumber variety NUN 93099
CUL to a
mutagenic compound or to radiation, wherein a representative sample of seed of
said
cucumber variety is deposited under Accession Number NCIMB 43946;
b. selecting the seed, plant, plant part, or a cell of cucumber variety NUN
93099 CUL,
having a mutation; and
c. optionally growing and/or multiplying the seed, plant, plant part, or cell
of cucumber
variety NUN 93099 CUL, having the mutation.
[0151] The disclosure also provides a method of producing a cucumber plant
having a desired
trait, wherein the method comprises transforming the cucumber plant with a
transgene that
confers the desired trait, wherein the transformed plant otherwise retains all
of the physiological
and morphological characteristics of the cucumber variety NUN 93099 CUL and
contains the
desired trait. Thus, a transgenic cucumber plant is provided which is produced
by the method
described above, wherein the plant otherwise has all of the physiological and
morphological
characteristics of cucumber variety NUN 93099 CUL and the desired trait.
[0152] In another aspect, the disclosure provides a method of producing a
progeny of cucumber
variety NUN 93099 CUL, further comprising a desired trait, said method
comprising
36
Date Recue/Date Received 2022-09-09
transforming the plant of cucumber variety NUN 93099 CUL with a transgene that
confers the
desired trait and/or crossing the plant of cucumber variety NUN 93099 CUL with
a transgenic
cucumber plant comprising a desired transgene so that the genetic material of
the progeny that
resulted from the cross contains the desired transgene(s). Also encompassed is
the progeny
produced by this method.
101531 A desired trait (e.g., gene(s)) conferring pest or disease resistance,
or tolerance for
protection, etc. can be introduced into cucumber plant variety NUN 93099 CUL,
or progeny of
said variety, by transforming said variety or progeny of said variety with a
transgene that confers
the desired trait, wherein the transformed plant otherwise retains all or all
but one, two, or three
of the physiological and morphological characteristics of cucumber variety NUN
93099 CUL,
and contains the desired trait. In another aspect, the transformation or
mutation confers a trait
wherein the trait is yield, size, compactness, dry matter content, fruit
quality, enhanced
nutritional quality, post-harvest quality, storage properties, color, male
sterility, herbicide
tolerance, insect resistance, pest resistance, disease resistance, Powdery
mildew resistance
without necrosis, environmental stress tolerance, modified carbohydrate
metabolism, modified
protein metabolism, or ripening. In a particular aspect, the specific
transgene may be any known
in the art or listed herein, including a polynucleotide Sequence conferring
resistance to
imidazolinone, sulfonylurea, glyphosate, glufosinate, triazine, benzonitrile,
cy clohexanedi one,
phenoxy proprionic acid and L-phosphinothricin or a polynucleotide conferring
resistance to
Angular Leaf Spot (Pseudomonas lachtymans), Anthracnose Race 1 (Colletotrichum
lagenaria),
Anthracnose Race 2, Bacterial Wilt (Erwinia tracheiphilus), Cucumber Scab
(Gummosis)
(Cladosporium cucumerinum), Downy Mildew (Pseudoperonospora cubensis), Powdery
Mildew
(Erysiphe chicoracearum), Altemaria Leaf Blight (Alternaria cucumerina),
Target Spot
(Corynespora cassiicola), Cucumber Rust, Root Rot, Crown Blight, Verticillum
Wilt, Sulphur
Burn, Fusarium oxyspo rum fsp. cucumerinum, Fusarium Wilt Race2, Root Knot
(Nematode),
Anthracnose, and/or Squash Mosaic. Other resistances, against pathogenic
viruses (e.g.,
Cucumber Mosaic Virus (CMV), Cucumber Yellow Mottle Mosaic Virus (CYMMV),
Cucumber
Green Mottle Mosaic Virus (CGMMV), Cucumber Aucuba Mosaic Virus (CAMV),
Muskmelon
Mosaic Virus (MMV), Cucumber Vein Yellowing Virus (CVYV), Cucurbit Yellow
Stunting
Disorder Virus (CYSDV), Watermelon Mosaic Virus (WMV), Papaya Ring Spot Virus
(PRSV),
Zucchini Mosaic Virus (ZMV)), fungi, bacteria, nematodes, insects, or other
pests may also be
introduced.
37
Date Recue/Date Received 2022-09-09
[0154] By crossing and/or selfing, (one or more) single traits may be
introduced into cucumber
variety NUN 93099 CUL (e.g., using backcrossing breeding schemes), while
retaining the
morphological and physiological characteristics of said variety and/or while
retaining one or
more or all distinguishing characteristics. A single trait converted plant may
thereby be produced.
For example, disease resistance genes may be introduced, genes responsible for
one or more
quality traits, yield, etc. Both single genes (e.g., dominant or recessive)
and one or more QTLs
(quantitative trait loci) may be transferred into cucumber variety NUN 93099
CUL by breeding
with said variety.
[0155] Alternatively, a single trait converted plant or single locus converted
plant of variety
NUN 93099 CUL may be produced by (i) genetically transforming or mutating
cells of NUN
93099 CUL; (ii) growing the cells into a plant; and (iii) optionally selecting
a plant that contains
the desired single locus conversion. The skilled person is familiar with
various techniques for
genetically transforming a single locus in a plant cell, or mutating said
cells.
[0156] In another aspect the disclosure provides a method of introducing a
single locus
conversion, a single trait conversion, or a desired trait into cucumber
variety NUN 93099 CUL,
comprising introducing a single locus conversion, a single trait conversion,
or a desired trait in
at least one of the parents of cucumber variety NUN 93099 CUL, and crossing
the converted
parent with the other parent of cucumber variety NUN 93099 CUL to obtain seed
of said variety.
[0157] In another aspect, the step of introducing a single locus conversion, a
single trait
conversion, or a desired trait in at least one of the parents comprises:
a. crossing the parental line of cucumber variety NUN 93099 CUL with a second
cucumber
plant comprising the single locus conversion, the single trait conversion, or
the desired trait;
b. selecting F 1progeny plants that contain the single locus conversion, the
single trait
conversion, or the desired trait;
c. crossing said selected progeny plants of step b) with the parental line of
step a) to produce
a backcross progeny plant;
d. selecting backcross progeny plants comprising the single locus conversion,
the single trait
conversion, or the desired trait and otherwise all or all but one, two or
three of the
morphological and physiological characteristics the parental line of step a)
to produce
selected backcross progeny plants; and
e. optionally repeating steps c) and d) one or more times in succession to
produce selected
second, third or fourth or higher backcross progeny plants comprising the
single locus
38
Date Recue/Date Received 2022-09-09
conversion, the single trait conversion or the desired trait and otherwise all
or all but one, two
or three of the morphological and physiological characteristics the parental
line of step a) to
produce selected backcross progeny plants, when grown in the same
environmental
conditions.
[0158] The disclosure further relates to plants obtained by this method.
[0159] In another aspect, the step of introducing a single locus conversion, a
single trait
conversion, or a desired trait in at least one of the parents comprises:
a. obtaining a cell or tissue culture of cells of the parental line of
cucumber variety NUN
93099 CUL;
b. genetically transforming or mutating said cells;
c. growing the cells into a plant; and
d. optionally selecting plants that contain the single locus conversion, the
single trait
conversion or the desired trait.
[0160] In another aspect, the disclosure provides a method of introducing a
single locus
conversion, a single trait conversion or a desired trait into cucumber variety
NUN 93099 CUL
comprising:
a. obtaining a combination of a parental lines of cucumber variety NUN 93099
CUL,
optionally through reverse synthesis of breeding lines;
b. introducing a single locus conversion, a single trait conversion, or a
desired trait in at least
one of the parents of step a); and
c. crossing the converted parent with the other parent of step a to obtain
seed of cucumber
variety NUN 93099 CUL.
[0161] In another method, the step of introducing a single locus conversion,
single trait
conversion, or a desired trait in at least one of the parents comprises
genetically transforming or
mutating cells the parental line of cucumber variety NUN 93099 CUL, growing
the cells into a
plant, and optionally selecting plants that contain the single locus
conversion, the single trait
conversion, or the desired trait.
[0162] In any of the above methods, where the single locus conversion concerns
a trait, the trait
may be yield or pest resistance or disease resistance. In one aspect, the
trait is disease resistance
and the resistance is conferred to Angular Leaf Spot (Pseudomonas lachrymans),
Anthracnose
Race 1 (Colletotrichum lagenaria), Anthracnose Race 2, Bacterial Wilt (Erwinia
tracheiphilus),
Cucumber Scab (Gummosis) (Cladosporium cucumerinum), Downy Mildew
39
Date Recue/Date Received 2022-09-09
(Pseudoperonospora cubensis), Powdery Mildew (Erysiphe chicoracearum),
Alternari a Leaf
Blight (Alternaria cucumerina), Target Spot (Cwynespora cassiicola), Cucumber
Rust, Root
Rot, Crown Blight, Verticillum Wilt, Sulphur Burn, Fusarium oxysporum !sp.
cucurnerinum,
Fusarium Wilt Race 2, Root Knot (Nematode), Anthracnose, and/or Squash Mosaic.
Other
resistances, against pathogenic viruses, e.g., Cucumber Mosaic Virus (CMV),
Cucumber Yellow
Mottle Mosaic Virus (CYMMV), Cucumber Green Mottle Mosaic Virus (CGMMV),
Cucumber
Aucuba Mosaic Virus (CAMV), Muskmelon Mosaic Virus (MMV), Cucumber Vein
Yellowing
Virus (CVYV), Cucurbit Yellow Stunting Disorder Virus (CYSDV), Watermelon
Mosaic Virus
(WMV), Papaya Ring Spot Virus (PRSV), Zucchini Mosaic Virus (ZMV), fungi,
bacteria,
nematodes, insects, or other pests may also be introduced.
101631 The disclosure also provides a plant having one, two, or three
physiological or
morphological characteristics which are different from those of cucumber
variety NUN 93099
CUL, and which otherwise has all the physiological and morphological
characteristics of said
variety, wherein a representative sample of seed of variety has been deposited
under Accession
Number NCIMB 43946. In particular, variants which differ from cucumber variety
NUN 93099
CUL in none, one, two, or three of the characteristics mentioned in Tables 1
and 2 are
encompassed.
[0164] The disclosure also provides a cucumber plant comprising at least a
first set of the
chromosomes of cucumber variety NUN 93099 CUL, a sample of seed of said
variety having
been deposited under Accession Number NCIMB 43946; optionally further
comprising a single
locus conversion, wherein said plant has essentially all of the morphological
and physiological
characteristics of the plant comprising at least a first set of the
chromosomes of said variety. In
another aspect, this single locus conversion confers a trait wherein the trait
yield, size,
compactness, dry matter content, fruit quality, enhanced nutritional quality,
post-harvest quality,
storage properties, color, male sterility, herbicide tolerance, insect
resistance, pest resistance,
disease resistance, Powdery mildew resistance without necrosis, environmental
stress tolerance,
modified carbohydrate metabolism, modified protein metabolism, or ripening.
[0165] In one aspect, the disclosure provides for a haploid plant and/or a
doubled haploid plant
of variety NUN 93099 CUL, or of a plant having all but one, two, or three
physiological or
morphological characteristics of cucumber variety NUN 93099 CUL, or progeny of
said variety.
Haploid and doubled haploid (DH) plants can, for example, be produced by cell
or tissue culture
and chromosome doubling agents and regeneration into a whole plant. In a
method for DH
Date Recue/Date Received 2022-09-09
production, chromosome doubling may be induced using known methods, such as
colchicine
treatment or the like. In one aspect, the method comprises inducing a cell or
tissue culture with
a chromosome doubling agent and regenerating the cells or tissues into a whole
plant.
101661 In another aspect, the disclosure comprises a method for making doubled
haploid cells
of cucumber variety NUN 93099 CUL, comprising making doubled haploid cells
from haploid
cells from the plant or plant part of cucumber variety NUN 93099 CUL with a
chromosome
doubling agent such as colchicine treatement (see, e.g., Nikolova and
Niemirowicz-Szczytt
(1996) Acta Soc Bot Pol 65:311-317).
101671 In yet another aspect, the disclosure provides for haploid plants
and/or doubled haploid
plants derived from cucumber variety NUN 93099 CUL that, when combined, make a
set of
parents of cucumber variety NUN 93099 CUL. The haploid plant and/or the
doubled haploid
plant of variety NUN 93099 CUL can be used in a method for generating parental
lines of
cucumber variety NUN 93099 CUL.
101681 In one aspect, the disclosure relates to a method of producing a
combination of parental
lines of the plant of variety NUN 93099 CUL, comprising making doubled haploid
cells from
haploid cells from said plant or seed of that plant; and optionally crossing
these parental lines to
produce and collecting seeds. In another aspect, the disclosure relates to a
combination of
parental lines produced by this method. In still another aspect, the
combination of parental lines
can be used to produce a seed or plant of variety NUN 93099 CUL when these
parental lines are
crossed. In still another aspect, the disclosure relates to a combination of
parental lines from
which a seed or plant having all physiological and/or morphological
characteristics of cucumber
variety NUN 93099 CUL, e.g., when the numerical characteristics are determined
at the 5%
significance level and identical (same type or degree) for non-numerical
characteristics for plants
grown under the same environmental conditions.
[0169] The disclosure also provides a method for producing parental lines for
hybrid variety
NUN 93099 CUL comprising: genetically characterizing a doubled haploid line
from cucumber
variety NUN 93099 CUL to determine whether one or more genetic markers are
present in a first
homozygous form or in a second homozygous form in said line, wherein one or
more genetic
markers are present in a heterozygous form in cucumber variety NUN 93099 CUL;
and selecting
at least one pair of doubled haploid lines that have complementary alleles for
one or more genetic
markers, wherein each member of the pair is suitable as a parental line for a
hybrid organism,
optionally this method further comprises defining a set of genetic markers
present in a
41
Date Recue/Date Received 2022-09-09
heterozygous form in cucumber variety NUN 93099 CUL; and producing doubled
haploid lines
from cucumber variety NUN 93099 CUL. Doubled haploid lines generated as
described herein
can be used in such method.
[0170] A combination of a male and a female parental line of cucumber variety
NUN 93099
CUL can also be generated, for example, through reverse synthesis of breeding
lines.
[0171] Using methods known in the art such as "reverse synthesis of breeding
lines" or "reverse
breeding," it is possible to produce parental lines for a hybrid plant such as
cucumber variety
NUN 93099 CUL. A skilled person can take any individual heterozygous plant
(called
a "phenotypically superior plant" inExample 2 of US 2015/0245570; cucumber
variety NUN
93099 CUL is such a plant) and generate a combination of parental lines
(reverse breeding
parental lines) that, when crossed, produce the variety NUN 93099 CUL. It is
not necessary that
the reverse breeding parental lines are identical to the original parental
lines. Such new breeding
methods are based on the segregation of individual alleles in the spores
produced by a desired
plant and/or in the progeny derived from the self-pollination of that desired
plant, and on the
subsequent identification of suitable progeny plants in one generation, or in
a limited number of
inbred cycles. Such a method is known from US 2015/0245570 or from Wijnker et
al., Nature
Protocols Volume: 9, Pages: 761-772 (2014) DOI: doi:10.1038/nprot.2014.049.
Thus, the
disclosure provides a method for producing parental lines for a hybrid
organism (e.g., cucumber
variety NUN 93099 CUL), comprising in one aspect: a) defining a set of genetic
markers present
in a heterozygous form (H) in a partially heterozygous starting organism; b)
producing doubled
haploid lines from spores of the starting organism; c) genetically
characterizing the doubled
haploid lines thus obtained for the said set of genetic markers to determine
whether they are
present in a first homozygous form (A) or in a second homozygous form (B); and
d) selecting at
least one pair of doubled haploid lines that have complementary alleles for at
least a subset of
the genetic markers, wherein each member of the pair is suitable as a parental
line for the hybrid
organism.
[0172] In another aspect, the method for producing parental lines for hybrid
organisms, e.g., of
cucumber variety NUN 93099 CUL, which when crossed reconstitute the genome of
cucumber
variety NUN 93099 CUL, comprising:
a. defining a set of genetic markers that are present in a heterozygous form
(H) in a partially
heterozygous starting organism;
42
Date Recue/Date Received 2022-09-09
b. producing at least one further generation from the starting organism by
self-pollination
(e.g., F2 or F3 generation);
c. selecting at least one pair of progeny organisms in which at least one
genetic marker from
the set is present in a complementary homozygous forms (B vs. A, or A vs. B);
and
d. optionally repeating steps b) and c) until at least one pair of progeny
organisms that have
complementary alleles for at least a subset of the genetic markers has been
selected as parental
lines for a hybrid.
[0173] The disclosure also provides methods for determining the identity of
parental lines of
plants described herein, in particular the identity of the female line. US
2015/0126380, relates to
a non-destructive method for analyzing maternal DNA of a seed. In this method,
the DNA is
dislodged from the seed coat surface and can be used to collect information on
the genome of the
maternal parent of the seed. This method for analyzing maternal DNA of a seed
comprises
contacting a seed with a fluid to dislodge DNA from the seed coat surface, and
analyzing the
DNA thus dislodged from the seed coat surface using methods known in the art.
The skilled
person is thus able to determine whether a seed has grown on a plant of
variety NUN 93099 CUL
or is a progeny of said variety, because the seed coat of the seed is a
maternal tissue genetically
identical to cucumber variety NUN 93099 CUL. In one aspect, the disclosure
relates to a seed
coat comprising maternal tissue of cucumber variety NUN 93099 CUL. In another
aspect, the
disclosure relates to a cucumber seed comprising a maternal tissue of cucumber
variety NUN
93099 CUL. In another particular aspect, the disclosure provides for a method
of identifying the
female parental line of cucumber variety NUN 93099 CUL by analyzing the seed
coat of a seed
of that variety. In another aspect, the skilled person can determine whether a
seed is grown on
cucumber variety NUN 93099 CUL by analysing the seed coat or another maternal
tissue of said
seed.
101741 In another aspect, the disclosure provides a method of determining the
genotype of a
plant described herein comprising detecting in the genome (e.g., a sample of
nucleic acids) of
the plant at least a first polymorphism or an allele. The skilled person is
familiar with many
suitable methods of genotyping, detecting a polymorphism or detecting an
allele including SNP
(single Nucleotide Polymorphism) genotyping, restriction fragment length
polymorphism
identification (RFLP) of genomic DNA, random amplified polymorphic detection
(RAPD) of
genomic DNA, amplified fragment length polymorphism detection (AFLP),
polymerase chain
reaction (PCR), DNA sequencing, allele specific oligonucleotide (ASO) probes,
and
43
Date Recue/Date Received 2022-09-09
hybridization to DNA microarrays or beads. Alternatively, the entire genome
could be sequenced.
The method may, in certain aspects, comprise detecting a plurality of
polymorphisms in the
genome of the plant, for example, by obtaining a sample of nucleic acid from a
plant and detecting
in said nucleic acids a plurality of polymorphisms. The method may further
comprise storing the
results of the step of detecting the plurality of polymorphisms on a computer
readable medium.
[0175] Also provided is a plant part obtainable from variety NUN 93099 CUL or
from progeny
of said variety or from a plant having all but one, two or three physiological
and/or morphological
characteristics which are different from those of cucumber variety NUN 93099
CUL, or from a
vegetatively propagated plant of variety NUN 93099 CUL (or from its progeny or
from a plant
having all or all but one, two or three physiological and/or morphological
characteristics which
are different from those of cucumber variety NUN 93099 CUL, wherein the plant
part is a fruit,
a harvested fruit, a part of a fruit, a leaf, a part of a leaf, pollen, an
ovule, a cell, a petiole, a shoot
or a part thereof, a stem or a part thereof, a root or a part thereof, a root
tip, a cutting, a seed, a
part of a seed, seed-coat or another maternal tissue which is part of a seed
grown on cucumber
variety NUN 93099 CUL or a hypocotyl, a cotyledon, a scion, a stock, a
rootstock, a pistil, an
anther, or a flower or a part thereof.
[0176] Such a plant part of variety NUN 93099 CUL can be stored and/or
processed further.
The disclosure thus, provides for a food or a feed product comprising one or
more of such parts,
such as canned, chopped, cooked, roasted, in a sauce, in a sandwich, pasted,
puréed or
concentrated, juiced, frozen, dried, pickled, or powdered cucumber fruit from
cucumber variety
NUN 93099 CUL or from progeny of said variety, or from a derived variety, such
as a plant
having all but one, two, or three physiological and/or morphological
characteristics of cucumber
variety NUN 93099 CUL. Preferably, the plant part is a cucumber fruit or part
thereof and/or an
extract from a fruit of the plant described herein comprising at least one
cell of cucumber variety
NUN 93099 CUL. The food or feed product may be fresh or processed, e.g.,
dried, grinded,
powdered, pickled, chopped, cooked, roasted, in a sauce, in a sandwich,
pasted, puréed or
concentrated, juiced, pickled, canned, steamed, boiled, fried, blanched and/or
frozen, etc.
[0177] In another aspect, the disclosure provides for a cucumber fruit of
variety NUN 93099
CUL, or a part of a fruit of said variety. The fruit can be in any stage of
maturity, for example,
immature or mature. In another aspect, the disclosure provides for a container
comprising or
consisting of a plurality of harvested cucumber fruits or parts of fruits of
said variety, or fruits
44
Date Recue/Date Received 2022-09-09
of progeny thereof, or fruits of a derived variety. Marketable fruits are
generally sorted by size
and quality after harvest.
[0178] In another aspect, the plant, plant part, or seed of cucumber variety
NUN 93099 CUL is
inside one or more containers. For example, the disclosure provides containers
such as cans,
boxes, crates, bags, cartons, Modified Atmosphere Packaging, films (e.g.,
biodegradable films),
etc. comprising a plant or a plant part (fresh and/or processed) or a seed of
cucumber variety
NUN 93099 CUL. In a particular aspect, the container comprises a plurality of
seeds of cucumber
variety NUN 93099 CUL, or a plurality of plant parts of cucumber variety NUN
93099 CUL.
The seeds may be disinfected, primed and/or treated with various compounds,
such as seed
coatings or crop protection compounds. The seed produces a plant of variety
NUN 93099 CUL.
[0179] Cucumbers may also be grown for use as rootstocks (stocks) or scions.
Typically,
different types of cucumbers are grafted to enhance disease resistance, which
is usually conferred
by the rootstock, while retaining the horticultural qualities usually
conferred by the scion. It is
not uncommon for grafting to occur between cultivated cucumber varieties and
related cucumber
species. Methods of grafting and vegetative propagation are well-known in the
art.
[0180] In another aspect, the disclosure provides to a plant comprising a
rootstock or scion of
cucumber variety NUN 93099 CUL.
[0181] References:
Naktuinbouw, "Calibration Book for Cucumber & Gherkin," Netherlands, December
2010.
UPOV, "Guidelines for the Conduct of Tests for Distinctness, Uniformity and
Stability", TG/61/7, March 2019.
US Department of Agriculture, "Objective Description of Variety Cucumber
(Cucumis
sativus L.)", June 2015.
Colijn-Hooymans, J.C., et. al., "Competence for Regeneration of Cucumber
Cotyledons is
Restricted to Specific Developmental Stages", Plant Cell, Tissue and Organ
Culture, 1994, vol.
39, pp. 211-217.
Needleman, S.B., et. al., "A General Method Applicable to the Search for
Similarities in the
Amino Acid Sequence of Two Proteins", Journal of Molecular Biology, 1970, vol.
48(3), pp.
443-53.
Date Recue/Date Received 2022-09-09
Nikolova, V., et. al., "Diploidization of Cucumber (Cucumis sativus L.)
Haploids by Colchini
Treatment", Acta Societas Botanicorum Poloniae, 1996, vol. 65, pp. 311-317.
Parvathaneni, R.K., et al., "Fingerprinting in Cucumber and Melon (Cucumis
spp.) genotypes
Using Morphological and ISSR Markers", Journal of Crop Science and
Biotechnology, 2011,
vol. 14, no. 1, pp. 39-43. DOI No. 10.1007/s12892-010-0080-1.
Rice, P., et al., "EMBOSS: The European Molecular Biology Open Software
Suite", Trends in
Genetics, 2000, vol. 16, Issue 6. pp. 276-277.
Sang-Gu, K., et. al., "Callus growth and Plant Regeneration in Diverse
Cultivars of Cucumber
(Cucumis sativus L.), Plant Cell, Tissue and Organ Culture, 1998, vol. 12, pp.
67-74.
Sarreb, D.A., et. al., "Comparison of Triploid and Diploid Cucumber in Long-
term Liquid
Cultures," Plant Cell, Tissue and Organ Culture, 2002, vol. 71-3, pp. 231-235.
Schrader, et. al., University of California Agriculture and Natural Resources,
Publication 8050,
2002,1-8
Vidaysky, F., et. al., "Tomato Breeding Lines Resistant and Tolerant to Tomato
Yellow Leaf
Curl Virus Issued from Lycopersicum hirsulum", The American Phytopathology
Society, 1998,
vol. 88, no. 9, pp. 910-914.
Vos, P., et al., "AFLP: A New Technique for DNA Fingerprinting", Nucleic Acids
Research,
1995, vol. 23(21), pp. 4407-4414.
Wijnker, E., et al., "Hybrid Recreation by Reverse breeding in Arabidopsis
thaliana", Nature
Protocols, 2014, vol. 9, pp. 761-772. DOT: doi: 10.1038/nprot.2014.049.
US 4,822,949
US 5,349,128
US 5,492,827
US 6,084,152
US 2015/0126380
46
Date Recue/Date Received 2022-09-09
US 2015/0245570
US 2018/0054990
US 2017/0335339
Development of Cucumber Variety NUN 93099 CUL
101821 The hybrid variety NUN 93099 CUL was developed from a male and female
proprietary
inbred line of Nunhems. The female and male parents were crossed to produce
hybrid (F1) seeds
of cucumber variety NUN 93099 CUL. The seeds of cucumber variety NUN 93099 CUL
can be
grown to produce hybrid plants and parts thereof (e.g., cucumber fruit). The
hybrid variety NUN
93099 CUL can be propagated by seeds or vegetatively.
101831 The hybrid variety is uniform and genetically stable. This has been
established through
evaluation of horticultural characteristics. Several hybrid seed production
events resulted in no
observable deviation in genetic stability. Coupled with the confirmation of
genetic stability of
the female and male parents the Applicant has concluded that cucumber variety
NUN 93099 CUL
is uniform and stable.
Deposit Information
101841 A total of 625 seeds of the hybrid variety NUN 93099 CUL was made and
accepted
according to the Budapest Treaty by Nunhems B.V. on March 3, 2022 at the NCIMB
Ltd.,
Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen AB21 9YA, United
Kingdom
(NUMB). The deposit has been assigned Accession Number NCIMB 43946. A
statement
indicating the viability of the sample has been provided. A deposit of
cucumber variety NUN
93099 CUL and of the male and female parent line is also maintained at Nunhems
B.V. The seed
lot number for cucumber variety NUN 93099 CUL is 29597902004.
[0185] The deposit will be maintained for a period of 30 years, or 5 years
after the most recent
request, or for the enforceable life of the patent whichever is longer and
will be replaced if it ever
becomes nonviable during that period. Access to the deposits will be available
during the
pendency of this application to persons determined by the Director of the U.S.
Patent Office to
be entitled thereto upon request. Subject to 37 C.F.R. 1.808(b), all
restrictions imposed by the
depositor on the availability to the public of the deposited material will be
irrevocably removed
upon the granting of the patent. Applicant does not waive any rights granted
under this patent on
47
Date Recue/Date Received 2022-09-09
this application or under the Plant Variety Protection Act (7 U.S.C. 2321 et
seq.). Accordingly,
the requirements of 37 CFR 1.801-1.809 have been satisfied.
Characteristics of Cucumber Variety NUN 93099 CUL
[0186] The most similar variety to NUN 93099 CUL refers to variety NUN 83067
CUL, a
variety from Nunhems B.V., with commercial name, Sequence.
[0187] In Tables 1 and 2, a comparison between cucumber variety NUN 93099 CUL
and the
Reference Variety are shown based on a trial in Acampo, California, USA under
high tech
conditions (i.e., greenhouse). Seeding date: May 12, 2022; Transplanting date:
May 31, 2022;
Harvesting date: July 7, 2022. In Table 3, the distinguishing characteristics
between cucumber
variety NUN 93099 CUL and the Reference Variety are presented.
[0188] One replication of 30 plants per variety, from which at least 15 plants
or plant parts were
randomly selected and were used to measure the characteristics. For numerical
characteristics,
averages were calculated. For non-numerical characteristics, the type or
degree were determined.
Similarity and differences between two different plant lines or varieties can
be determined by
comparing the number of morphological and/or physiological characteristics
that are the same
(i.e., statistically not significantly different) or that are different (i.e.,
statistically significantly
different) between the two plant lines or varieties using plants grown under
the same
environmental conditions. Preferably, a numerical characteristic is considered
to be "the same"
when the value for a numeric characteristic is not significantly different at
the 1% (p <0.01) or
5% (p < 0.05) significance level, using a T-test, a standard method known to
the skilled person.
Preferably, non-numerical or "degree" or "type" characteristics are considered
"the same" when
the values have the same "degree" or "type" when scored using USDA and/ or
UPOV descriptors,
if the plants are grown under the same environmental conditions. In one
aspect, a statistical
analysis using the T-test at 5% significance level is provided (see, Tables 4-
14).
[0189] In another aspect, the disclosure provides a cucumber plant having all
of the
physiological and morphological characteristics of cucumber variety NUN 93099
CUL as
presented in Tables 1 and 2 when grown under the same environmental
conditions, wherein a
representative sample of seed of said cucumber variety NUN 93099 CUL has been
deposited
under Accession Number NCIMB 43946.
48
Date Recue/Date Received 2022-09-09
Table 1. Objective Description of Cucumber Variety NUN 93099 CUL and the
Reference Variety
(USDA Descriptors)
Characteristics Application Variety
Reference Variety I
(NUN 93099 CUL) (NUN 83067 CUL)
Type:
Predominant usage: Fresh market Fresh market
Slicing (Fresh Market), Pickling
Predominant Culture: Greenhouse; indoor
Greenhouse; indoor
Outdoor; Greenhouse cultivation with drip
cultivation with drip
irrigation irrigation
Growth: Indeterminate Indeterminate
Determinate, Indeterminate
Sex: Gynoecious Gynoecious
Andromonoecious, Monoecious,
Primarily gynoecious
Flower color: Yellow Yellow
Yellow, Orange, Green, Other
Main Stem:
3rd internode length, cm: 82.12 cm 91.43 cm
Stem form: Grooved, ridged Grooved; ridged
Grooved, ridged; Smooth, round
Leaf (Mature blade of third leaf):
Length, mm: 24.81 cm 29.04 cm
Width, mm: 23.19 cm 26.83 cm
Petiole length, cm: 15.99 cm 18.75 cm
Fruit at edible maturity:
Length, cm: 30.93 cm 29.92 cm
Diameter at medial, cm: 4.39 cm 4.02 cm
Weight, gram: 421.60 g 387.20 g
Skin color: Not mottled Not mottled
Not mottled, Mottled or speckled with
yellow
Yellowish blossom end stripes: Absent Absent
Absent, Extend less than 1/3 of the fruit
length, Extend more than 1/3 of the fruit
length
Predominant color at stem end: Dark green Dark green
White, Light Green, Medium Green, RHS N137B RHS 139A
Dark Green
Predominant color at blossom end: Dark green Dark green
White, Light Green, Medium Green, RHS N137B RHS 139A
Dark Green
Fruit neck shape: Necked Not necked
Not necked, necked
Fruit tapering: Stem end tapered Stem end tapered
49
Date Recue/Date Received 2022-09-09
Characteristics Application Variety
Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Both ends tapered, Stem end tapered,
Blossom end tapered, Ends blunt or
rounded
Stem end cross section: Circular Circular
Circular, Triangular, Square
Medial cross section: Circular Circular
Circular, Triangular, Square
Blossom end cross section: Triangular Triangular
Circular, Triangular, Square
Skin ribs: Ribbed Ribbed
Not ribbed, Ribbed
Skin luster: Dull Glossy
Dull, Glossy
Spine color: White White
White, Black
Spine quality: Fine Fine
Coarse, Fine
Spine density Few Few
Few, Many
Tubercles (Warts): Absent Absent
Few, obscure; Many, obscure; Few,
prominent; Many, prominent
Disease resistances:
Cladosporium cucumerinum(Ccu) Highly resistant Highly
resistant
Cucumber Mosaic Virus (CMV) Intermediately Absent
resistant
Podosphaera xanthii (Powdery mildew) Highly resistant Highly
resistant
Cognespora cassiicola (Corynespora Highly resistant Highly
resistant
blight and target leaf spot)
Cucumber Vein Yellowing Virus Highly resistant Absent
(CVYV)
Cucumber Green Monte Mosaic Virus Highly resistant Highly
resistant
(CGMIVIV)
Table 2. Objective Description of Cucumber Variety NUN 93099 CUL and the
Reference Variety
(Non-USDA Descriptors)
Characteristics Application Variety
Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Cotyledon (bitterness): Absent Absent
absent, present
Plant:
Stem diameter, mm: 9.34 mm 9.32 mm
Total length of first 15 intemodes: Short Medium
very short, very short to short,
Date Recue/Date Received 2022-09-09
Characteristics Application Variety
Reference Variety
(NUN 93099 CUL) _ (NUN 83067 CUL)
short, short to medium, medium,
medium to long, long to very long,
very long
Vigor: Medium Very strong
very weak, weak, medium, strong,
very strong
Number of flowers per node: Predominantly one or two Predominantly one or
predominantly one, predominantly two
one or two, predominantly two,
predominantly two or three,
predominantly three or four,
predominantly four or five,
predominantly more than five
Leaf (mature blade):
Attitude: Horizontal Drooping
erect, horizontal, drooping
Length: Short Long
short, medium, long
Ratio length/width of terminal lobe: Medium Medium
very small, small, medium, large,
very large
Shape of apex of terminal lobe: Right angled Right angled
acute, right-angled, obtuse, rounded
Intensity of green color: Medium Medium
very light, very light to light, light,
light to medium, medium, medium
to dark, dark, dark to very dark,
very dark
Blistering: Medium Medium
absent or very weak, weak,
medium, strong, very strong
Undulation of margin: Moderate Moderate
absent or very weak, moderate,
strong
Dentati on of margin: Weak Weak
very weak, weak, medium, strong,
very strong
Leaf color: Green Green
RHS N137A RHS 137A
Petiole diameter, mm: 6.91 mm 7.78 mm
Ovary:
Color of vestiture: White White
Parthenocarpy: Present Present
Fruit at edible maturity:
Length: Long Long
51
Date Recue/Date Received 2022-09-09
Characteristics Application Variety
Reference Variety
(NUN 93099 CUL) _ (NUN 83067 CUL)
very short, very short to short,
short, short to medium, medium,
medium to long, long, long to very
long, very long
Diameter: Medium Medium
small, medium, large
Ratio length/diameter: Medium Very large
very small, small, medium, large,
very large
Core diameter in relation to Large Medium
diameter of fruit:
Round, Round to angular, Angular
Shape in transverse section: Round to angular Round to angular
round, round to angular, angular
Shape of stem end: Necked Acute
necked, acute, obtuse
Length of neck: Medium Short
very short, short, medium, long,
very long
Shape of calyx end Rounded Obtuse
acute, obtuse, rounded, truncate
Ground color of skin at market Green Green
stage:
white, yellow, green
Intensity of ground color of skin: Medium Dark
very light, light, medium, dark,
very dark
Ribs: Strong Strong
absent or weak, medium, strong
Sutures: Absent Absent
absent, present
Creasing: Present Present
absent, present
Degree of creasing: Weak Weak
very weak, weak, medium, strong,
very strong
Type of vestiture: Prickles only Prickles only
hairs only, hairs and prickles,
prickles only
Density of vestiture: Very sparse Sparse
very sparse, sparse, medium, dense,
very dense
Stripes: Absent Absent
absent, present
Length of stripes: Absent Absent
52
Date Recue/Date Received 2022-09-09
Characteristics Application Variety Reference Variety
(NUN 93099 CUL) _ (NUN 83067 CUL)
absent or very short, short, medium,
long, very long
Mottling; Absent Absent
absent, present
Dots: Absent Absent
absent, present
Glaucosity: Weak Weak
absent or very weak, weak,
medium, strong, very strong
Length of peduncle: Medium Long
short, medium, long
Ground color of skin at Yellow Yellow
physiological ripeness:
white, yellow, green, orange, brown
Peduncle length, mm: 43.95 mm 44.33 mm
Peduncle width, mm: 4.07 mm 3.80 mm
Table 3. Distinguishing Characteristics between Cucumber Variety NUN 93099 CUL
and the
Reference Variety
Characteristics Application Variety Reference Variety I
(NUN 93099 CUL) (NUN 83067 CUL)
Plant:
3rd intemode length, cm: 82.12 cm 91.43 cm
Total length of first 15 internodes: Short Medium
very short, very short to short, short,
short to medium, medium, medium
to long, long to very long, very long
Vigor: Medium Very strong
very weak, weak, medium, strong,
very strong
Leaf (Mature blade of third leaf):
Length, mm: 24.81 cm 29.04 cm
Width, /um: 23.19 cm 26.83 cm
Petiole length, cm: 15.99 cm 18.75 cm
Attitude: Horizontal Drooping
erect, horizontal, drooping
Length: Short Long
short, medium, long
Petiole diameter, mm: 6.91 mm 7.78 mm
Fruit at edible maturity:
Diameter at medial, cm: 4.39 cm 4.02 cm
Predominant color at stem end: Dark green Dark green
White, Light Green, Medium Green, RHS N137B RHS 139A
Dark Green
53
Date Recue/Date Received 2022-09-09
Predominant color at blossom end: Dark green Dark green
White, Light Green, Medium Green, RHS N137B RHS 139A
Dark Green
Fruit neck shape: Necked Not necked
Not necked, necked
Skin luster: Dull Glossy
Dull, Glossy
Ratio length/diameter: Medium Very large
very small, small, medium, large,
very large
Core diameter in relation to Large Medium
diameter of fruit:
Round, Round to angular, Angular
Shape of stem end: Necked Acute
necked, acute, obtuse
Length of neck: Medium Short
very short, short, medium, long,
very long
Shape of calyx end Rounded Obtuse
acute, obtuse, rounded, truncate
Intensity of ground color of skin: Medium Dark
very light, light, medium, dark,
very dark
Density of vestiture: Very sparse Sparse
very sparse, sparse, medium, dense,
very dense
Length of peduncle: Medium Long
short, medium, long
Disease resistances:
Cucumber Mosaic Virus (CMV) Intermediately resistant Absent
Cucumber Vein Yellowing Virus Highly resistant Absent
(CVYV)
[0190] The results of the T-Test show significant difference at 5%
significance level between
cucumber variety NUN 93099 CUL and the Reference Variety for 3r1 intemode
length, mature
leaf length, mature leaf width, and mature leaf petiole length, mature leaf
petiole diameter, and
mature fruit diameter at medial as shown in Tables 4-9.
[0191] Table 4 shows a significant difference at 5% significance level between
cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.003) for 3rd internode
length (cm) based
on a trial conducted in the US in 2022.
Table 4.
54
Date Recue/Date Received 2022-09-09
Statistical Parameter Application Variety Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 69.57 80.32
Maximum 94.51 106.25
Median 81.49 89.12
Mean 82.12 91.43
Standard deviation 7.94 7.94
[0192] Table 5 shows a significant difference at 5% significance level between
cucumber
variety NUN 93099 CUL and the Reference Variety (p<0.001) for mature leaf
length (cm) based
on a trial conducted in the US in 2022.
Table 5.
Statistical Parameter Application Variety Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 22.10 26.50
Maximum 27.0 34.9
Median 25.20 28.60
Mean 24.81 29.04
Standard deviation 1.56 2.14
[0193] Table 6 shows a significant difference at 5% significance level between
cucumber
variety NUN 93099 CUL and the Reference Variety (p<0.001) for mature leaf
width (cm) based
on a trial conducted in the US in 2022.
Table 6.
Statistical Parameter Application Variety Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 21.0 24.40
Maximum 27.40 30.40
Median 22.50 26.50
Mean 23.19 26.83
Standard deviation 1.63 1.97
[0194] Table 7 shows a significant difference at 5% significance level between
cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.001) for mature leaf
petiole length (cm)
based on a trial conducted in the US in 2022.
Table 7.
Date Recue/Date Received 2022-09-09
Statistical Parameter Application Variety Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 1140 16.20
Maximum 19.70 22.50
Median 16.0 19.20
Mean 15.99 18.75
Standard deviation 2.07 2.08
[0195] Table 8 shows a significant difference at 5% significance level between
cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.007) for mature leaf
petiole diameter
(mm) based on a trial conducted in the US during the trial season 2021.
Table 8.
Statistical Parameter Application Variety Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 5.72 6.45
Maximum 8.28 9.72
Median 6.87 7.47
Mean 6.91 7.78
Standard deviation 0.61 0.99
[0196] Table 9 shows a significant difference at 5% significance level between
cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.009) for mature fruit
diameter at medial
(cm) based on a trial conducted in the US in 2022.
Table 9.
Statistical Parameter Application Variety Reference Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 3.80 3.40
Maximum 5.0 4.60
Median 4.50 4.0
Mean 4.39 4.02
Standard deviation 0.35 0.37
[0197] The results of the T-Test show no significant difference at 5%
significance level between
cucumber variety NUN 93099 CUL and the Reference Variety for stem diameter,
mature fruit
weight, mature fruit length, peduncle length, and peduncle width as shown in
Tables 10-14.
56
Date Recue/Date Received 2022-09-09
[0198] Table 10 shows no significant difference at 5% significance level
between cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.974) for stem diameter
(mm) based on
a trial conducted in the US in 2022.
Table 10.
Statistical Parameter Application Variety Reference
Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 7.26 8.09
Maximum 13.98 12.02
Median 9.11 9.05
Mean 9.34 9.32
Standard deviation 1.55 1.10
[0199] Table 11 shows no significant difference at 5% significance level
between cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.196) for mature fruit
weight (g) based
on a trial conducted in the US in 2022.
Table 11.
Statistical Parameter Application Variety Reference
Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 314.0 274.0
Maximum 654.0 480.0
Median 412.0 390.0
Mean 421.60 387.20
Standard deviation 80.28 60.66
102001 Table 12 shows no significant difference at 5% significance level
between cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.100) for mature fruit
length (cm) based
on a trial conducted in the US in 2022.
Table 12.
Statistical Parameter Application Variety Reference
Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 28.0 27.10
Maximum 36.0 32.20
Median 30.60 30.30
Mean 30.93 29.92
Standard deviation 1.80 1.43
57
Date Recue/Date Received 2022-09-09
102011 Table 13 shows no significant difference at 5% significance level
between cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.735) for peduncle length
(mm) based
on a trial conducted in the US in 2022.
Table 13.
Statistical Parameter Application Variety .. Reference
Variety
_ (NUN 93099 CUL) (NUN 83067
CUL)
Number of samples 15 15
Minimum 34.21 35.84
Maximum 52.22 53.68
Median 44.0 42.94
Mean 43.95 43.33
Standard deviation 5.65 4.31
102021 Table 14 shows no significant difference at 5% significance level
between cucumber
variety NUN 93099 CUL and the Reference Variety (p=0.059) for peduncle widtth
(mm) based
on a trial conducted in the US in 2022.
Table 14.
Statistical Parameter Application Variety .. Reference
Variety
(NUN 93099 CUL) (NUN 83067 CUL)
Number of samples 15 15
Minimum 3.25 3.33
Maximum 4.80 4.31
Median 4.19 3.81
Mean 4.07 3.80
Standard deviation 0.44 0.28
102031 In some aspects, embodiments of the present invention as described
herein include the
following items:
Item 1. A cell of cucumber plant variety NUN 93099 CUL, or a cell of a part
thereof,
wherein the representative sample of seed of said cucumber variety NUN 93099
CUL is deposited
under Accession Number NCIMB 43946.
Item 2. A cell of a seed of cucumber variety NUN 93099 CUL, wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession Number
NCIMB 43946.
58
Date Recue/Date Received 2024-01-26
Item 3. A cell of a plant part of cucumber variety NUN 93099 CUL, wherein a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited under
Accession Number NCIMB 43946, wherein the plant part is a leaf, pollen, an
ovule, a fruit, a
scion, a root, a rootstock, a cutting, or a flower.
Item 4. The cell of the plant part of item 3, wherein the plant part is a
fruit or part
thereof.
Item 5. A tissue culture or cell culture of regenerable cells of cucumber
variety NUN
93099 CUL, wherein a representative sample of seed of said cucumber variety
NUN 93099 CUL
is deposited under Accession Number NCIMB 43946.
Item 6. The tissue culture or cell culture according to item 5, comprising
cells or
protoplast from a plant part, wherein the plant part is a meristem, a
cotyledon, a hypocotyl, a
pollen, a leaf, an anther, a root, a root tip, a pistil, a petiole, a flower,
a fruit, a stalk, or a stem.
Item 7. A cell of a cucumber plant regenerated from the tissue culture or cell
culture of
cucumber variety NUN 93099 CUL, wherein a representative sample of seed of
said cucumber
variety NUN 93099 CUL is deposited under Accession Number NCIMB 43946.
Item 8. A cell of a cucumber plant produced by vegetatively propagating a part
of the
plant of cucumber variety NUN 93099 CUL, wherein a representative sample of
seed of said
cucumber variety NUN 93099 CUL is deposited under Accession Number NCIMB
43946.
Item 9. The cell of item 8, wherein said part is a cutting, a cell culture, or
a tissue
culture.
Item 10.A cell of a vegetatively propagated cucumber plant, or a cell of a
part thereof of
cucumber variety NUN 93099 CUL, wherein a representative sample of seed of
said cucumber
variety NUN 93099 CUL is deposited under Accession Number NCIMB 43946.
Item 11.A cell of a cucumber plant produced by a method comprising
transforming the
plant of cucumber variety NUN 93099 CUL with a transgene that confers a
desired trait, wherein
the transformed plant is the same as cucumber variety NUN 93099 CUL except for
the transgene
and otherwise retains all of the physiological and morphological
characteristics of cucumber
variety NUN 93099 CUL and contains the desired trait, wherein a representative
sample of seed
59
Date Recue/Date Received 2024-01-26
of said cucumber variety NUN 93099 CUL is deposited under Accession Number
NCIMB 43946,
wherein the desired trait is the desired trait is yield, size, compactness,
dry matter content,
firmness, flavor, fruit quality, enhanced nutritional quality, post-harvest
quality, storage
properties, color, male sterility, herbicide tolerance, insect resistance,
pest resistance, disease
resistance, Powdery mildew resistance without necrosis, environmental stress
tolerance,
modified carbohydrate metabolism, modified protein metabolism, or ripening.
Item 12.A cell of a cucumber fruit of cucumber variety NUN 93099 CUL, wherein
a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited under
Accession Number NCIMB 43946.
Item 13.A method of determining the genotype of a plant of cucumber variety
NUN
93099 CUL, comprising obtaining a sample of nucleic acids from said plant,
detecting in said
nucleic acids a plurality of polymorphisms, thereby determining the genotype
of the plant, and
storing the results of detecting the plurality of polymorphisms on a computer
readable medium,
wherein a representative sample of seed of said cucumber variety NUN 93099 CUL
is deposited
under Accession Number NCIMB 43946.
Item 14.A container comprising seeds of cucumber variety NUN 93099 CUL,
wherein a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited under
Accession Number NCIMB 43946.
Item 15.A container comprising a cucumber fruit of cucumber variety NUN 93099
CUL,
wherein a representative sample of seed of said cucumber variety NUN 93099 CUL
is deposited
under Accession Number NCIMB 43946.
Item 16.Use of cucumber variety NUN 93099 CUL as a scion or a rootstock,
wherein a
representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited under
Accession Number NCIMB 43946.
Item 17.Use of cucumber plant variety NUN 93099 CUL, or a part thereof, for
producing, a crop, or a food, or a processed product, wherein a representative
sample of seed of
said cucumber variety NUN 93099 CUL is deposited under Accession Number NCIMB
43946.
Date Recue/Date Received 2024-01-26
Item 18.Use of a seed of cucumber variety NUN 93099 CUL, for producing
cucumber
plants, wherein a representative sample of seed of said cucumber variety NUN
93099 CUL is
deposited under Accession Number NCIMB 43946.
Item 19.Use of a plant of cucumber variety NUN 93099 CUL, wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession Number
NCIMB 43946, to cross with another cucumber plant.
Item 20.Use of a plant of cucumber variety NUN 93099 CUL, wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession Number
NCIMB 43946, and a second cucumber plant to produce a cucumber seed.
Item 21.Use of a plant of cucumber variety NUN 93099 CUL, wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession Number
NCIMB 43946, and a second cucumber plant to produce a descendant plant.
Item 22.Use of a plant of cucumber variety NUN 93099 CUL, wherein a
representative
sample of seed of said cucumber variety NUN 93099 CUL is deposited under
Accession Number
NCIMB 43946, to breed a cucumber plant.
Item 23.Use of cucumber variety NUN 93099 CUL as a recipient of a transgene,
wherein
a representative sample of seed of said cucumber variety NUN 93099 CUL is
deposited under
Accession Number NCIMB 43946.
Item 24.Use of cucumber variety NUN 93099 CUL, wherein a representative sample
of
seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number NCIMB
43946, to produce a genetic marker profile.
Item 25.Use of cucumber variety NUN 93099 CUL, wherein a representative sample
of
seed of said cucumber variety NUN 93099 CUL is deposited under Accession
Number NCIMB
43946, to produce a vegetative propagation of cucumber variety NUN 93099 CUL.
Item 26.Use of a plant of cucumber variety NUN 93099 CUL as a recipient or
donor of
a heritable trait, wherein a representative sample of seed of said cucumber
variety NUN 93099
CUL is deposited under Accession Number NCIMB 43946.
61
Date Recue/Date Received 2024-01-26
Item 27.A plant cell from i) a cucumber plant or ii) a cucumber seed, wherein
the plant
or seed is a descendant of cucumber variety NUN 93099 CUL, wherein a
representative sample
of seed of said cucumber variety NUN 93099 CUL has been deposited under
Accession Number
NCIMB 43946, and wherein the descendant comprises a transgene and otherwise
has all the
physiological and morphological characteristics of cucumber variety NUN 93099
CUL.
62
Date Recue/Date Received 2024-01-26
