U.S. patent application number 17/100096 was filed with the patent office on 2021-04-08 for lettuce variety nun 09165 ltl.
The applicant listed for this patent is NUNHEMS B.V.. Invention is credited to Johan VAN ZEE.
Application Number | 20210100188 17/100096 |
Document ID | / |
Family ID | 1000005301775 |
Filed Date | 2021-04-08 |
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United States Patent
Application |
20210100188 |
Kind Code |
A1 |
VAN ZEE; Johan |
April 8, 2021 |
LETTUCE VARIETY NUN 09165 LTL
Abstract
The disclosure relates to lettuce variety NUN 09165 LTL as well
as seeds and plants and heads or leaves thereof. NUN 09165 LTL is a
multileaf green lettuce heavy variety comprising resistance to
Downy Mildew (Bremia lactucae) Isolates Bl:16-36EU and Nasonovia
ribisnigri Biotype Nr:0.
Inventors: |
VAN ZEE; Johan;
(s-Gravenzande, NL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
NUNHEMS B.V. |
Nunhem |
|
NL |
|
|
Family ID: |
1000005301775 |
Appl. No.: |
17/100096 |
Filed: |
November 20, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62939457 |
Nov 22, 2019 |
|
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|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A01H 6/1472 20180501;
A01H 5/10 20130101 |
International
Class: |
A01H 6/14 20060101
A01H006/14; A01H 5/10 20060101 A01H005/10 |
Claims
1. A plant, plant part, or seed of lettuce variety NUN 09165 LTL,
wherein a representative sample of seed of said variety has been
deposited under Accession Number NCIMB ______.
2. The plant part of claim 1, wherein the plant part is a head, a
leaf, pollen, an ovule, a fruit, a scion, a rootstock, a cutting, a
flower, or a cell.
3. A seed that produces the plant of claim 1.
4. A seed grown on the plant of claim 1.
5. A lettuce plant grown from the seed of claim 4.
6. A lettuce plant or a part thereof having all the physiological
and morphological characteristics of the plant of claim 1.
7. A lettuce plant or a part thereof which does not differ from the
plant of claim 1 in all of the characteristics listed in Table 1-3,
when determined at the 5% significance level for numerical
characteristics and determined by type or degree for non-numerical
characteristics, when grown under the same environmental conditions
and wherein a representative sample of seed of lettuce variety NUN
09165 LTL has been deposited under Accession Number NCIMB
______.
8. A tissue or cell culture of regenerable cells of the plant or
plant part of claim 1.
9. The tissue or cell culture according to claim 8, comprising
cells or protoplasts derived from a plant part suitable for
vegetative reproduction, wherein the plant part is 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, a
stem, or a stalk.
10. A lettuce plant regenerated from the tissue or cell culture of
claim 8, wherein the plant has all of the physiological and
morphological characteristics of the plant of lettuce variety NUN
09165 LTL, 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, and wherein a representative sample of
seed of said lettuce variety has been deposited under Accession
Number NCIMB
11. A method of producing of the plant of claim 1, or a part
thereof, said method comprising vegetative propagation of the plant
of lettuce variety NUN 09165 LTL, wherein a representative sample
of seed of said lettuce variety has been deposited under Accession
Number NCIMB ______.
12. The method of claim 11, wherein said vegetative propagation
comprises regenerating a whole plant from a part of lettuce variety
NUN 09165 LTL, wherein a representative sample of seed of said
lettuce variety has been deposited under Accession Number NCIMB
43496.
13. The method of claim 11, wherein said part is a cutting, a cell
culture, or a tissue culture.
14. A vegetatively propagated plant of or part thereof produced by
the method of claim 11, wherein the plant has all of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL, 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.
15. A method of producing a lettuce plant, said method comprising
crossing the plant of claim 1 with itself or a second lettuce plant
at least once, and selecting a progeny lettuce plant from said
crossing and allowing the progeny lettuce plant to form seed.
16. A first generation progeny of the lettuce plant of claim 1
obtained by crossing the plant of claim 1 with itself or another
lettuce plant, wherein the progeny lettuce plant has all of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL, wherein a representative sample of seed of said
lettuce variety has been deposited under Accession Number NCIMB
______.
17. A method of producing a F1 hybrid lettuce seed, said method
comprising crossing a first parent lettuce plant with a second
parent lettuce plant and harvesting the resultant hybrid lettuce
seed, wherein at the first parent lettuce plant or second parent
lettuce plant is the plant of claim 1.
18. A F1 hybrid lettuce seed produced by the method of claim
17.
19. An F1 hybrid lettuce plant or part thereof, produced by growing
the seed of claim 18.
20. A food, a feed, or a processed product comprising the plant
part of claim 2.
21. A container comprising the plant, plant part, or seed of claim
1.
22. A method of introducing a desired trait into the plant of claim
1, said method comprises transforming the plant of claim 1 with a
transgene that confers the desired trait, wherein the desired trait
is yield, storage properties, color, enhanced nutritional quality,
male sterility, herbicide tolerance, insect resistance, pest
resistance, disease resistance, environmental stress tolerance,
modified carbohydrate metabolism, or modified protein metabolism or
the mutation occurs in any of the following genes:
Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL, PPO.
23. A lettuce plant produced by the method of claim 22, wherein the
transformed plant contains the desired trait and otherwise retains
all of the morphological and physiological characteristics of
lettuce variety NUN 09165 LTL.
24. A method of introducing a single locus conversion into the
plant of claim 1, comprising: a. crossing the plant of claim 1 with
a second lettuce plant comprising a desired single locus to produce
F1 progeny plants; b. selecting F1 progeny plants that have the
single locus to produce selected F1 progeny plants; c. crossing the
selected F1 progeny plants with lettuce variety NUN 09165 LTL to
produce backcross progeny plants; d. selecting backcross progeny
plants that have the single locus and otherwise comprise all of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL to produce selected backcross progeny plants; and e.
repeating steps (c) and (d) one or more times in succession to
produce selected second or higher backcross progeny plants that
comprise the single locus and otherwise comprise all of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL, wherein a representative sample of seed of said
variety has been deposited under Accession Number NCIMB ______.
25. The method of claim 24, wherein the single locus confers yield,
storage properties, color, enhanced nutritional quality, male
sterility, herbicide tolerance, insect resistance, pest resistance,
disease resistance, environmental stress tolerance, modified
carbohydrate metabolism, or modified protein metabolism.
26. A lettuce plant produced by the method of claim 24, wherein the
single locus converted plant comprises the single locus conversion
and otherwise has all of the morphological and physiological
characteristics of lettuce variety NUN 09165 LTL.
27. A method of producing a modified lettuce plant, said method
comprising mutating a target gene in lettuce plant or plant part of
lettuce variety NUN 09165 LTL, wherein a representative sample of
seed of said variety has been deposited under Accession Number
NCIMB ______.
28. The method of claim 27, wherein the target gene modifies a
desired trait and wherein the desired trait is yield, storage
properties, color, male sterility, herbicide tolerance, insect
resistance, pest resistance, disease resistance, environmental
stress tolerance, modified carbohydrate metabolism, or modified
protein metabolism or the mutation occurs in any of the following
genes: Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL, PPO.
29. The method of claim 27, wherein the target gene is mutated by
targeted gene editing.
30. A modified lettuce plant produced by the method of claim 27,
wherein the plant contains the desired trait and otherwise all of
the physiological and morphological characteristics of lettuce
variety NUN 09165 LTL.
31. A method of producing doubled haploid cells of the plant of
claim 1, said method comprising making doubled haploid cells from
haploid cells from the plant or plant part of lettuce variety NUN
09165 LTL, wherein a representative sample of seed of said lettuce
variety has been deposited under Accession Number NCIMB ______.
32. A method of producing a lettuce head or a lettuce leaf, said
method comprising growing the plant of claim 1 until it develops at
least a leaf or a head, and collecting the leaf or head.
33. A lettuce head or a lettuce leaf produced by the method of
claim 32.
34. A method for determining the genotype of the plant of claim 1,
said method comprising obtaining a sample of nucleic acids from
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.
35. A method for developing a lettuce plant in a lettuce breeding
program, said method comprising applying plant breeding techniques
comprising recurrent selection, backcrossing, pedigree breeding,
mass selection, mutation breeding, genetic marker enhanced
selection, or genetic transformation to the plant of claim 1 or
part thereof.
36. A method of producing a lettuce plant derived from the plant of
claim 1, comprising: a. preparing a progeny lettuce plant derived
from lettuce variety NUN 09165 LTL by crossing the plant of claim 1
with itself or with a second lettuce plant; b. crossing the progeny
plant with itself or a second lettuce plant to produce seed of a
progeny plant of a subsequent generation; c. growing a progeny
plant of a subsequent generation from said seed and crossing the
progeny plant of a subsequent generation with itself or a second
lettuce plant; and d. repeating step (b) and/or (c) for at least
one more generation to produce a lettuce plant derived from lettuce
variety NUN 09165 LTL.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims priority to U.S. Provisional
Application No. 62/939,457, filed on Nov. 22, 2019, which is hereby
incorporated by reference in its entirety.
FIELD OF THE DISCLOSURE
[0002] The disclosure relates to the field of plant breeding and,
more specifically, to the development of lettuce variety NUN 09165
LTL. The disclosure further relates to vegetative reproductions of
lettuce variety NUN 09165 LTL, methods for tissue culture of
lettuce variety NUN 09165 LTL, and to phenotypic variants of
lettuce variety NUN 09165 LTL. The disclosure also relates to
progeny of lettuce variety NUN 09165 LTL and the hybrid varieties
obtained by crossing lettuce variety NUN 09165 LTL as a parent line
with plants of other varieties or parent lines.
BACKGROUND OF THE DISCLOSURE
[0003] The goal of vegetable 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 shelf
life.
[0004] The development of commercial lettuce cultivars or varieties
requires the crossing of lettuce plants, and the evaluation of the
crosses. Pedigree breeding and recurrent selection are examples of
breeding methods used to develop cultivars from breeding
populations. Breeding programs combine desirable traits from two or
more varieties or various broad-based sources into breeding pools
from which cultivars are developed by selfing and selection of
desired phenotypes. The new cultivars are crossed with other
varieties and the inbred lines or hybrids from these crosses are
evaluated to determine which have commercial potential.
[0005] All cultivated forms of lettuce belong to the highly
polymorphic species Lactuca sativa that is grown for its edible
head and leaves. Lactuca sativa is in the Asteraceae (Compositae)
family. Lettuce is related to chicory, sunflower, aster, dandelion,
artichoke, and chrysanthemum. L. sativa is one of about 300 species
in the genus Lactuca. There are many types of lettuce, and new
types are constantly in development. Types of lettuce include
Cutting/Leaf, Iceberg/Crisphead, Cos or Romaine, Batavian, Salinas
Group, Latin, Butterhead, Great Lakes Group, Eastern (Ithaca)
Group, Bibb, Vanguard Group, Multileaf, or Stem lettuce. Lettuce is
typically consumed fresh and occasionally as a cooked vegetable. It
is popularly used in salads, wraps, and sandwiches.
[0006] Fresh lettuce is available in the United States year-round
although the greatest supply is from May through October. For
planting purposes, the lettuce season is typically divided into
three categories, early, mid and late, with the coastal areas
planting from January to August, and the desert regions planting
from August to December. California and Arizona are the two largest
producers of lettuce in the United States.
[0007] Changes in lifestyle primarily due to increasing health
awareness results to growing demand for healthy convenience food.
Supermarkets, restaurants, catering firms, and convenience stores
are constantly looking for more colorful garnishing for sandwiches,
wraps, and ready-to-eat snacks such as salads. The changing food
and consumer trends present opportunities for breeding companies to
develop new varieties with specific shapes of leaves, specific
average size of leaves, prominent color, glossiness, taste, and a
wide variety of texture. Other breeding objectives include disease
or pest resistance, yield, prolonged shelf life, and suitability to
climatic requirements.
SUMMARY OF VARIOUS ASPECTS OF THE DISCLOSURE
[0008] The disclosure provides for lettuce variety NUN 09165 LTL,
products thereof, and methods of using the same. NUN 09165 LTL is a
multileaf green heavy lettuce variety and is suitable for growing
in the open field.
[0009] In another aspect, the plant of lettuce variety NUN 09165
LTL, a part thereof, or a progeny thereof comprises resistance to
Downy Mildew (Bremia lactucae) Isolates Bl:16-36EU and Nasonovia
ribisnigri Biotype Nr:0, measured according to UPOV standards
described in TG/13/11.
[0010] In another aspect, the disclosure provides for a hybrid
lettuce variety NUN 09165 LTL. The disclosure also provides for a
progeny of lettuce variety NUN 09165 LTL. In another aspect, the
disclosure provides a plant or a progeny retaining all or all but
one, two, or three of the "distinguishing characteristics" of
lettuce variety NUN 09165 LTL, or all but one, two, or three of the
"morphological and physiological characteristics" of lettuce
variety NUN 09165 LTL and methods of producing that plant or
progeny.
[0011] In one aspect, the disclosure provides a plant or a progeny
having all of the physiological and morphological characteristics
of lettuce variety NUN 09165 LTL, 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 lettuce variety NUN 09165 LTL when
grown under the same environmental conditions and e.g., evaluated
at significance levels of 1%, 5%, or 10% significance (which can be
expressed as a p-value) for quantitative characteristics and
determined by type or degree for non-quantitative characteristics,
wherein a representative sample of seed of variety NUN 09165 LTL
has been deposited under Accession Number NCIMB ______. 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
Table 1-3 of lettuce variety NUN 09165 LTL 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 determined by
type or degree for non-quantitative characteristics.
[0012] In another aspect, the plant of lettuce variety NUN 09165
LTL, or part thereof, or a progeny thereof has 14, 15, or more or
all of the following distinguishing characteristics when compared
to Reference Variety as shown in Table 4: 1) black seed color; 2)
longer base of head to apex (core length); 3) wider base of head
(core width); 4) fewer mature leaf divisions; 5) stronger intensity
of green color of leaves; 6) deeper incisions of margin of mature
leaves; 7) less dense incision of margin of mature leaves; 8)
smaller mature leaves; 9) resistant to Bremia lactucae Isolate
Bl:27EU; 10) resistant to Bremia lactucae Isolate Bl:29EU; 11)
resistant to Bremia lactucae Isolate Bl:30EU; 12) resistant to
Bremia lactucae Isolate Bl:33EU; 13) resistant to Bremia lactucae
Isolate Bl:34EU; 14) resistant to Bremia lactucae Isolate Bl:35EU;
and 15) resistant to Bremia lactucae Isolate Bl:36EU, when
determined at the 5% significance level for numerical
characteristics and determined by type or degree for non-numerical
characteristics when grown under the same environmental
conditions.
[0013] In another aspect, the disclosure provides a seed of lettuce
variety NUN 09165 LTL, wherein a representative sample of said seed
has been deposited under Accession Number NCIMB ______. The
disclosure also provides for a plurality of seeds of lettuce
variety NUN 09165 LTL. The lettuce seed of variety NUN 09165 LTL
may be provided as an essentially homogeneous population of lettuce
seed. The population of seed of lettuce variety NUN 09165 LTL may
be particularly defined as an essentially free from other seed. The
seed population may be grown into plants to provide an essentially
homogeneous population of lettuce plants described herein.
[0014] The disclosure also provides a plant grown from a seed of
lettuce variety NUN 09165 LTL and a plant part thereof.
[0015] The disclosure further provides a lettuce head and/or a
lettuce leaf produced on a plant grown from a seed of lettuce
variety NUN 09165 LTL.
[0016] The disclosure furthermore provides a seed growing or grown
on a plant of lettuce variety NUN 09165 LTL (e.g., produced after
pollination of the flower of lettuce variety NUN 09165 LTL).
[0017] In another aspect, the disclosure provides for a plant part
obtained from lettuce variety NUN 09165 LTL, wherein said plant
part is: a leaf, a part of a leaf, a head, a part of a head, a
fruit, a part of a fruit, 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 pistil, an anther, or a flower
or a part thereof. Heads and leaves are particularly important
plant parts. Such plant parts may be suitable for sexual
reproduction, vegetative reproduction, or a tissue culture. In
another aspect, the plant part obtained from variety NUN 09165 LTL
is a cell, optionally a cell in a cell or tissue culture. That cell
may be grown into a plant of lettuce variety NUN 09165 LTL.
[0018] In another aspect, the disclosure provides for an inbred
variety of NUN 09165 LTL.
[0019] In another aspect, the disclosure provides a cell culture of
lettuce variety NUN 09165 LTL and a plant regenerated from lettuce
variety NUN 09165 LTL, wherein the plant has all of the
characteristics of lettuce variety NUN 09165 LTL when grown under
the same environmental conditions, as well as methods for culturing
and regenerating lettuce variety NUN 09165 LTL. Alternatively, a
regenerated plant may have one characteristic that is different
from lettuce variety NUN 09165 LTL.
[0020] The disclosure also provides a vegetatively propagated plant
of variety NUN 09165 LTL having all or all but one, two, or three
of the morphological and physiological characteristics of lettuce
variety NUN 09165 LTL when grown under the same environmental
conditions as well as methods for vegetatively propagating lettuce
variety NUN 09165 LTL.
[0021] In another aspect, the disclosure provides a method of
producing a lettuce plant comprising crossing lettuce variety NUN
09165 LTL with itself or another lettuce variety and selecting a
progeny lettuce plant from said crossing or selfing.
[0022] The disclosure also provides a method of producing a lettuce
plant derived from lettuce variety NUN 09165 LTL.
[0023] In further aspect, the disclosure provides a method of
producing hybrid lettuce seed comprising crossing a first parent
lettuce plant with a second parent lettuce plant and harvesting the
resultant hybrid lettuce seed, wherein said first parent lettuce
plant or second parent lettuce plant is lettuce variety NUN 09165
LTL. Also provided is a hybrid lettuce seed produced from crossing
a first parent lettuce plant with a second parent lettuce plant and
harvesting the resultant hybrid lettuce seed, wherein said first
parent lettuce plant or second parent lettuce plant is lettuce
variety NUN 09165 LTL. Moreover, a hybrid lettuce plant grown from
the hybrid lettuce seed is provided.
[0024] In another aspect, the disclosure provides a method of
introducing a single locus conversion into a plant of variety NUN
09165 LTL, wherein a representative sample of said seed has been
deposited under Accession Number NCIMB ______, wherein the plant
otherwise retains all of the physiological and morphological
characteristics of lettuce variety NUN 09165 LTL and further
comprises the single locus conversion.
[0025] In yet another aspect, the disclosure provides a method of
introducing a desired trait into lettuce variety NUN 09165 LTL,
said method comprises transforming the plant of variety NUN 09165
LTL with a transgene that confers the desired trait, wherein the
transformed plant otherwise has all of the physiological and
morphological characteristics of lettuce variety NUN 09165 LTL and
contains the desired trait.
[0026] The disclosure also provides a method of producing a
modified lettuce plant with a desired trait, wherein the method
comprises mutating a lettuce plant or plant part of lettuce variety
NUN 09165 LTL, wherein a representative sample of said seed has
been deposited under Accession Number NCIMB ______, and wherein the
mutated plant otherwise retains all of the physiological and
morphological characteristics of lettuce variety NUN 09165 LTL and
contains the desired trait.
[0027] In one aspect, the single locus conversion or desired trait
is yield, size, storage properties, color, taste, enhanced
nutritional quality, post-harvest quality, male sterility,
herbicide tolerance, insect resistance, pest resistance, disease
resistance, environmental stress tolerance, modified carbohydrate
metabolism, or modified protein metabolism, or the mutation occurs
in any of the following genes: Ferulate-5-hydrxylase, dmr1, dmr6,
NCED4, PAL, PPO.
[0028] In another aspect, the disclosure provides a container
comprising the plant, plant part, or seed of lettuce variety NUN
09165 LTL.
[0029] [29] Also provided is a food, a feed, or a processed product
comprising the plant part of lettuce variety NUN 09165 LTL, wherein
the plant part is a leaf or part thereof.
BRIEF DESCRIPTION OF THE DRAWINGS
[0030] FIG. 1 shows the mature plants of lettuce variety NUN 09165
LTL.
[0031] FIG. 2 shows the mature plant comparison of lettuce variety
NUN 09165 LTL and the Reference Variety.
[0032] FIG. 3 shows the mature leaves of lettuce variety NUN 09165
LTL.
[0033] FIG. 4 shows the leaf division of lettuce variety NUN 09165
LTL.
[0034] FIG. 5 shows the leaf division of the Reference Variety.
[0035] FIG. 6 shows the mature leaf comparison of lettuce variety
NUN 09165 LTL and the Reference Variety.
DEFINITIONS
[0036] "Lettuce" refers herein to plants of the species Lactuca
sativa L. The most commonly eaten parts of a lettuce plant are the
head or a leaf. The head comprises a core and leaves, which may be
divided in inner and outer leaves.
[0037] "Cultivated lettuce" refers to plants of Lactuca sativa
(e.g., varieties, breeding lines or cultivars of the species L.
sativa as well as crossbreds thereof, or crossbreds with other
Lactuca sativa species, or even with other Lactuca species),
cultivated by humans and having good agronomic characteristics.
[0038] "Leaf lettuce" or "cutting lettuce" refers to a type of
lettuce having very loose leaves that does not form a head.
[0039] The terms "lettuce plant designated NUN 09165 LTL," "NUN
09165 LTL," "inbred NUN 09165," "09165 LTL," or "lettuce 09165" are
used interchangeably herein and refer to a lettuce plant of variety
NUN 09165 LTL, representative sample of said seed has been
deposited under Accession Number NCIMB ______.
[0040] "Plant" includes the whole plant or any parts or derivatives
thereof, preferably having the same genetic makeup as the plant
from which it is obtained.
[0041] "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
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, or a
flower. 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 one or two sets of chromosomes derived from
the parent plant (e.g., from lettuce variety NUN 09165 LTL). An F1
progeny produced from self-pollination of the inbred variety NUN
09165 LTL will thus comprise two sets of chromosomes derived from
lettuce variety NUN 09165 LTL, while an F1 progeny derived from
cross-fertilization of NUN 09165 LTL will comprise only one set of
chromosomes from lettuce variety NUN 09165 LTL and the other set of
chromosomes from the other parent.
[0042] A "seed of lettuce variety NUN 09165 LTL" refers to a
lettuce seed which can be grown into a plant of lettuce variety NUN
09165 LTL, wherein a representative sample of viable seeds of
lettuce variety NUN 09165 LTL has been deposited under Accession
Number NCIMB ______. 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 lettuce variety NUN 09165 LTL" refers to an
embryo as present in a seed of lettuce variety NUN 09165 LTL, a
representative sample of said seed of NUN 09165 LTL having been
deposited under Accession Number NCIMB ______.
[0044] A "seed grown on lettuce variety NUN 09165 LTL" refers to a
seed grown on a mature plant of variety NUN 09165 LTL or inside a
fruit of lettuce variety NUN 09165 LTL. The "seed grown on lettuce
variety NUN 09165 LTL" contains tissues and DNA of the maternal
parent, lettuce variety NUN 09165 LTL. The "seed grown on lettuce
variety NUN 09165 LTL" contains an F1 embryo. When said seed is
planted, it grows into a first generation progeny plant of variety
NUN 09165 LTL. Since NUN 09165 LTL is an inbred variety and thus
highly homozygous, the set of chromosomes inherited by the first
generation is predictable.
[0045] An "essentially homogeneous population of lettuce seed" is a
population of seeds where at least 77%, 98%, 99% or more of the
total population of seed are seed of lettuce variety NUN 09165
LTL.
[0046] An "essentially homogeneous population of lettuce plants" is
a population of plants where at least 97%, 97%, 99% or more of the
total population of plants are plants of lettuce variety NUN 09165
LTL.
[0047] The phrase "essentially free from other seed" refers to a
population of seeds where less than 3%, 2%, 1%, or even less, of
the total population of seed is seed that is not a lettuce seed or,
in another option, less than 3%, 2%, 1%, or less, of the total
population of seed is seed that is not seed of lettuce variety NUN
09165 LTL.
[0048] "Cotyledon" refers to one of the first leaves of the embryo
of a seed plant.
[0049] "Head" as used herein refers to lettuce heads, i.e., the
plant without the root system, for example, substantially all
harvested leaves. Encompassed are immature leaves (e.g., "baby
leaf") and mature leaves.
[0050] The "base" of a plant is the part of a lettuce plant where
the leaves are attached to the root system of the plant.
[0051] "Core length" of the internal lettuce stem is measured from
the base of the cut and trimmed head to the tip of the stem.
[0052] "Core Length to Head Diameter Ratio (CLHD Ratio)" refers to
the mean core length/head diameter ratio. It is calculated by
dividing the mean core length with the mean head diameter. This is
an indication of the head shape and of the ability of a lettuce
plant to reduce the amount of surface which is on or close to the
ground.
[0053] "Head weight" refers to the mean weight of saleable lettuce
head, cut, and trimmed to market specifications.
[0054] "Head diameter" refers to the mean diameter of the cut and
trimmed head, sliced vertically, and measured at the widest point
perpendicular to the stem.
[0055] "Head height" refers to the mean height of the cut and
trimmed head, sliced vertically, and measured from the base of the
cut stem to the leaf tip.
[0056] "Harvested plant material" refers herein to plant parts
(e.g., leaves or heads detached from the whole plant) which have
been collected for further storage and/or further use.
[0057] "Harvested seeds" refers to seeds harvested from a line or
variety, e.g., produced after self-fertilization or
cross-fertilization and collected.
[0058] "First water date" refers to the date the seed first
receives adequate moisture to germinate. This can and often does
equal the planting date.
[0059] "Maturity date" refers to the stage when the plants are of
full size or optimum weight, in marketable form or shape to be of
commercial or economic value. This is also the time when measuring
parameters of "mature" leaves.
[0060] "Yield" means the total weight of all lettuce heads or
leaves harvested per hectare of a particular line or variety. It is
understood that "yield" expressed as weight of all lettuce heads or
leaves harvested per hectare can be obtained by multiplying the
number of plants per hectare times the "yield per plant."
[0061] "Marketable yield" means the total weight of all marketable
lettuce heads or leaves harvested per hectare of a particular line
or variety, e.g., lettuce heads or leaves suitable for being sold
for fresh consumption, having good color, glossiness size and
texture and no or very low levels of deficiencies. A "marketable
lettuce head or leaf" is a head or leaf that has commercial
value.
[0062] "USDA descriptors" are the plant variety descriptors
described for lettuce in the "Objective description of
Variety--Lettuce (Lactuca sativa L.)," as published by U.S.
Department of Agriculture, Agricultural Marketing Service, Science
and Technology, Plant Variety Protection Office (June 2015, revised
December 2018), and which can be downloaded from the world-wide web
at ams.usda.gov/ under
services/plant-variety-protection/pvpo-c-forms under lettuce.
"Non-USDA descriptors" are other descriptors suitable for
describing lettuce.
[0063] "UPOV descriptors" are the plant variety descriptors
described for lettuce in the "Guidelines for the Conduct of Tests
for Distinctness, Uniformity and Stability," TG/13/11 (Geneva 2006,
last updated Jun. 14, 2019), 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 upov.int/ under
edocs/tgdocs/en/tg013.pdf, which is hereby incorporated by
reference in its entirety. Likewise, "UPOV methods" to determine
specific parameters for the characterization of lettuce are
described at upov.int.
[0064] "RHS" or "RHS color" refers to the color chart of the Royal
Horticultural Society (UK), which publishes a 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.
[0065] "Calibration Manual: DUS Test for Lettuce" refers to the
calibration book for lettuce which provides guidance for describing
a lettuce variety, as published by Naktuinbow (Netherlands) and
National Agriculture and Food Research Organization (NARO Japan)
(Dec. 26, 2016) and based on the UPOV Guideline TG/13/10 Rev. 2
[0066] "Reference Variety" refers herein to variety NUN 09074 LTL,
a variety from Nunhems B.V. with commercial name Skrunch Green 74,
which has been planted in a trial together with lettuce variety NUN
09165 LTL. The characteristics of lettuce variety NUN 09165 LTL
were compared with the characteristics of the Reference Variety as
shown in Tables 2 and 3. The distinguishing characteristics between
lettuce variety NUN 09165 LTL and the Reference Variety are shown
in Table 4.
[0067] 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 progenitor 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 Table 1-3 or "all or all but
one, two or three of the physiological and morphological
characteristics" of Table 1-3.
[0068] The physiological and/or morphological characteristics
mentioned above are commonly evaluated at significance levels of
1%, 5%, or 10% if they are numerical (quantitative), or for having
an identical degree (or type) if not numerical (not quantitative),
if measured under the same environmental conditions. For example, a
progeny plant or a single locus converted plant or a mutated plant
of lettuce variety NUN 09165 LTL may have one or more (or all) of
the essential physiological and/or morphological characteristics of
said variety listed in Table 1-3, as determined at the 5%
significance level (i.e., p<0.05) for quantitative
characteristics and determined by type or degree for
non-quantitative characteristics, when grown under the same
environmental conditions.
[0069] "Distinguishing characteristics" or "distinguishing
morphological and/or physiological characteristics" refers herein
to the characteristics which distinguish a new variety from other
lettuce varieties (i.e., are different), when grown under the same
environmental conditions. The distinguishing characteristics
between lettuce variety NUN 09165 LTL and the Reference Variety are
described Table 4. When comparing lettuce variety NUN 09165 LTL
with different varieties, the distinguishing characteristics will
be different. In one aspect, the distinguishing characteristics may
therefore include at least one, two, three or more (or all) of the
characteristics listed in Table 1-3. All numerical distinguishing
characteristics are statistically significantly different at
p<0.05 between lettuce variety NUN 09165 LTL and the other
variety (e.g., the Reference Variety).
[0070] Lettuce Variety NUN 09165 LTL has the following
distinguishing characteristics when compared to the Reference
Variety as shown in Table 4: 1) black seed color; 2) longer base of
head to apex (core length); 3) wider base of head (core width); 4)
fewer mature leaf divisions; 5) stronger intensity of green color
of leaves; 6) deeper incisions of margin of mature leaves; 7) less
dense incision of margin of mature leaves; 8) smaller mature
leaves; 9) resistant to Bremia lactucae Isolate Bl:27EU; 10)
resistant to Bremia lactucae Isolate Bl:29EU; 11) resistant to
Bremia lactucae Isolate Bl:30EU; 12) resistant to Bremia lactucae
Isolate Bl:33EU; 13) resistant to Bremia lactucae Isolate Bl:34EU;
14) resistant to Bremia lactucae Isolate Bl:35EU; and 15) resistant
to Bremia lactucae Isolate Bl:36EU, when determined at the 5%
significance level for numerical characteristics and determined by
type or degree for non-numerical characteristics when grown under
the same environmental conditions.
[0071] Thus, a lettuce plant "comprising the distinguishing
characteristics of lettuce variety NUN 09165 LTL" (such as a
progeny plant) refers herein to a plant which does not differ from
said variety in the distinguishing characteristics above.
Therefore, in one aspect, the disclosure provides a plant that does
not differ from lettuce variety NUN 09165 LTL in the distinguishing
characteristics above.
[0072] 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 when grown under the same
environmental conditions. 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 T-test, a standard method known to the
skilled person. A non-numerical characteristic is considered "the
same" when the values have the same "degree" or "type" when scored
using USDA and/or UPOV descriptors, for plants grown under the same
environmental conditions.
[0073] 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 lettuce variety NUN 09165 LTL and the Reference
Variety are significant or due to chance. For the purpose of
proving differences or distinction between lettuce variety NUN
09165 LTL 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% probability that the observed result could have
happened just by chance or random variation.
[0074] The statistical analysis is drawn from a small sample of at
least 15 plants or plant parts of lettuce variety NUN 09165 LTL 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 a data set is
statistically significant, a T-test is used, a statistical tool for
proving significance in the means of the two groups at 5%
significance level (p-value of 5% or 0.05).
[0075] 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 (e.g., methods other
than genetic modification/transformation/transgenic methods, e.g.,
gene editing), by which, for example, a genetically heritable trait
can be transferred from one lettuce line or variety to another.
[0076] "Variety" or "cultivar" means a plant grouping within a
single botanical taxon of the lowest known rank.
[0077] "Plant line" is, for example, an inbred variety or a
breeding line which can be used to develop one or more varieties.
Both are typically highly homozygous. Progeny obtained by selfing
such a plant line has the same phenotype as its parents.
[0078] "Inbred variety" refers to an inbred (nearly homozygous)
line or seeds thereof. For example, the (nearly homozygous) plant
is self-pollinated or the (nearly homozygous) female parent is
pollinated with pollen of the same plant line to produce inbred
seeds on the female parent.
[0079] "Progeny" as used herein refers to a plant obtained from a
plant designated NUN 09165 LTL. 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 lettuce plant of the
same or another variety or line, or wild lettuce plants. A progeny
may comprise a mutation or a transgene. A "first generation
progeny" is the progeny is 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 of lettuce variety NUN 09165 LTL is the male parent,
the female parent, or both of a first generation progeny of lettuce
variety NUN 09165 LTL. Progeny may have all of the physiological
and morphological characteristics of lettuce variety NUN 09165 LTL
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, or to a plant comprising all but one,
two, or three of the morphological and physiological
characteristics of lettuce variety NUN 09165 LTL.
[0080] "Tissue 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 lettuce and regeneration of plants therefrom is well known and
widely published (see, e.g., Teng et al., HortScience. 1992, 27(9):
1030-1032; Teng et al., HortScience. 1993, 28(6): 669-1671; Zhang
et al., Journal of Genetics and Breeding. 1992, 46(3): 287-290).
Similarly, methods of preparing cell cultures are known in the
art.
[0081] "Vegetative propagation," "vegetative reproduction," or
"clonal propagation" are used interchangeably herein and mean the
method of taking part of a plant and allowing that plant part to
form at least roots, and also refer to the plant or plantlet
obtained by that method. Optionally, the vegetative propagation is
grown into a mature plant.
[0082] "Regeneration" refers to the development of a plant from
cell culture or tissue culture or vegetative propagation.
[0083] "Crossing" refers to the mating of two parent plants. The
term encompasses "cross-pollination" and "selfing."
[0084] "Selfing" refers to self-pollination of a plant, i.e., the
transfer of pollen from the anther to the stigma of the same
plant.
[0085] "Cross-pollination" refers to the fertilization by the union
of two gametes from different plants.
[0086] "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.
[0087] The terms "gene converted," "conversion plant," or "single
locus converted plant" in this context refer to lettuce plants
which are developed by traditional backcrossing techniques, e.g.,
backcrossing, or via genetic engineering (e.g., gene editing) or
through mutation breeding, wherein essentially all of the desired
morphological and physiological characteristics of parent variety
or line are recovered, in addition to the one or more
characteristics introduced into the parent via e.g., backcrossing
technique (optionally including reverse synthesis of breeding
line). It is understood that 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., a mutant
allele of a gene can modify the phenotype of a characteristic).
[0088] Likewise, a "Single Locus Converted (Conversion) Plant"
refers to plants developed by plant breeding techniques comprising
or consisting of mutation breeding and/or by genetic transformation
(e.g., gene editing) and/or by traditional breeding techniques,
such as backcrossing, wherein essentially all of the desired
morphological and physiological characteristics of a lettuce
variety are recovered in addition to the characteristics of the
single locus having been transferred into the variety via the
above-mentioned techniques, or wherein a morphological and
physiological characteristic 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.
[0089] "Transgene" refers to a genetic locus comprising a DNA
sequence which has been introduced into the genome of a lettuce
plant by transformation. A plant comprising a transgene stably
integrated into its genome is referred to as "transgenic
plant."
[0090] "Locus" (plural loci) refers to the specific location of a
gene or DNA sequence on a chromosome. A locus may confer a specific
trait.
[0091] "Genotype" refers to the genetic composition of a cell or
organism.
[0092] "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.
[0093] 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. 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.
[0094] "Average" refers herein to the arithmetic mean.
[0095] 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 VARIOUS ASPECTS OF THE DISCLOSURE
[0096] The disclosure relates to the plant of lettuce variety NUN
09165 LTL, wherein a representative sample of seeds of said variety
has been deposited under Budapest Treaty, with Accession Number
NCIMB ______. NUN 09165 LTL is a multileaf green heavy lettuce
variety and is suitable for growing in the open field.
[0097] In another aspect, the plant of lettuce variety NUN 09165
LTL, or part thereof, or a progeny thereof comprises resistance
Downy Mildew (Bremia lactucae) Isolates Bl:16-36EU and Nasonovia
ribisnigri Biotype Nr:0, measured according to UPOV standards
described in TG/13/11.
[0098] The disclosure also provides a lettuce plant or part thereof
having all of the physiological and morphological characteristics
of lettuce variety NUN 09165 LTL, when grown under the same
environmental conditions.
[0099] The disclosure also provides a plant of lettuce variety NUN
09165 LTL or a progeny plant thereof comprises all of the
morphological and/or physiological characteristics (i.e., average
values, as indicated on the USDA Objective Description of
variety--lettuce (unless indicated otherwise)) as shown in Table
1-3, when the characteristics are determined at the 5% significance
level for plants grown under the same environmental conditions. A
part of this plant is also provided.
[0100] The disclosure further provides a lettuce plant which does
not differ from the physiological and morphological characteristics
of the plant of lettuce variety NUN 09165 LTL, as determined at the
1%, 2%, 3%, 4%, or 5% significance level, when grown under the same
environmental conditions. In a particular aspect, the plants are
measured in the same trial (e.g., the trial is conducted as
recommended by the USDA or UPOV). The disclosure also comprises
part of said plant, preferably a leaf or a part thereof.
[0101] The morphological and/or physiological differences between
two different individual plants described herein (e.g., between
lettuce variety NUN 09165 LTL and a progeny of lettuce variety NUN
09165 LTL) or between a plant of lettuce variety NUN 09165 LTL or
progeny of said variety, or a plant having all, or all but 1, 2, or
3 of the physiological and morphological characteristics of lettuce
variety NUN 09165 LTL (or all, or all but 1, 2, or 3 of the
characteristics as listed in Table 1-3) and another known variety
can easily be established by growing said variety next to the each
other or next to the other variety (e.g., in the same field, under
the same environmental conditions), preferably in several locations
which are suitable for said lettuce 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). For example, trials
can be carried out in Acampo Calif., USA (N 38 degrees 07'261''/W
121 degrees 18' 807'', USA), whereby various characteristics, for
example, maturity, leaf shape, size and texture, leaf color and
glossiness, bolt shape, surface and length, flower size and color,
head weight, disease resistance, insect resistance, and resistance
to physiological stress can be measured and directly compared for
species of lettuce.
[0102] Thus, the disclosure comprises a lettuce plant having one,
two, or three of the physiological and/or morphological
characteristics which are different from those of the plant of
lettuce variety NUN 09165 LTL, and which otherwise has all of the
physiological and morphological characteristics of the plant of
lettuce variety NUN 09165 LTL, when determined (e.g., at the 5%
significance level for plants for quantitative characteristics or
determined by type for non-quantitative characteristics) grown
under the same environmental conditions. In one aspect, the
different characteristic(s) is/are a result of breeding with
lettuce variety NUN 09165 LTL and selection of a progeny plant
comprising one, two, or three characteristics which are different
than in lettuce variety NUN 09165 LTL. In another aspect, the
different characteristic is the result of a mutation (e.g.,
spontaneous mutation or a human induced mutation through e.g.,
targeted mutagenesis or traditional mutagenesis such as chemically
or radiation induced mutagenesis), or it is the result of
transformation.
[0103] The disclosure further relates to lettuce variety NUN 09165
LTL, which when compared to Reference Variety has the following
distinguishing characteristics as shown in Table 4: 1) black seed
color; 2) longer base of head to apex (core length); 3) wider base
of head (core width); 4) fewer mature leaf divisions; 5) stronger
intensity of green color of leaves; 6) deeper incisions of margin
of mature leaves; 7) less dense incision of margin of mature
leaves; 8) smaller mature leaves; 9) resistant to Bremia lactucae
Isolate Bl:27EU; 10) resistant to Bremia lactucae Isolate Bl:29EU;
11) resistant to Bremia lactucae Isolate Bl:30EU; 12) resistant to
Bremia lactucae Isolate Bl:33EU; 13) resistant to Bremia lactucae
Isolate Bl:34EU; 14) resistant to Bremia lactucae Isolate Bl:35EU;
and 15) resistant to Bremia lactucae Isolate Bl:36EU, when
determined at the 5% significance level for numerical
characteristics and determined by type or degree for non-numerical
characteristics when grown under the same environmental
conditions.
[0104] The disclosure also relates to a seed of lettuce variety NUN
09165 LTL, wherein a representative sample of said seed has been
deposited under the Budapest Treaty with Accession Number NCIMB
______.
[0105] In another aspect, a seed of hybrid variety NUN 09165 LTL is
obtainable by crossing the male parent of lettuce variety NUN 09165
LTL with the female parent of lettuce variety NUN 09165 LTL and
harvesting the seeds produced on the female parent. The resultant
seed of said variety can be grown to produce plants of said
variety.
[0106] In another aspect, the disclosure provides a plant grown
from a seed of lettuce variety NUN 09165 LTL and a plant part
thereof.
[0107] The disclosure also provides a lettuce head and/or a lettuce
leaf produced on a plant grown from a seed of lettuce variety NUN
09165 LTL.
[0108] In another aspect, the disclosure provides for a plant part
of lettuce variety NUN 09165 LTL, preferably a head or a leaf, a
representative sample of seed from said variety has been deposited
under Accession Number NCIMB ______.
[0109] Also provided is a plant of lettuce variety NUN 09165 LTL,
or a head or a leaf 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 NCIMB
______.
[0110] Also provided is a plant part obtained from variety NUN
09165 LTL, wherein said plant part is a leaf, a part of a leaf, a
head, a part of a head, a fruit, a part of a fruit, 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 a said variety, a hypocotyl, a
cotyledon, a pistil, an anther, or a flower or a part thereof. Such
plant parts may be suitable for sexual reproduction (e.g., 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.). Heads
and leaves are particularly important plant parts.
[0111] In a further aspect, the plant part obtained from variety
NUN 09165 LTL is a cell, optionally a cell in a cell or tissue
culture. The cell may be grown into a plant of lettuce variety NUN
09165 LTL. A part of lettuce variety NUN 09165 LTL (or of a progeny
of that variety or of a plant having all of the physiological and
morphological characteristics but one, two, or three of lettuce
variety NUN 09165 LTL) further encompasses any cells, tissues,
organs obtainable from the seedlings or plants in any stage of
maturity.
[0112] The disclosure also provides a tissue or cell culture
comprising regenerable cells of lettuce variety NUN 09165 LTL. Such
tissue culture can, for example, be grown on plates or in liquid
culture or be frozen for long term storage. The cells of lettuce
variety NUN 09165 LTL used to start the culture can be any plant
part suitable for vegetative reproduction, or, in a particular
aspect, can be one or more of: an embryo, a meristem, a cotyledon,
a hypocotyl, a pollen, a leaf, an anther, a root, a root tip, 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 reinitiation.
[0113] In another aspect, the disclosure provides a lettuce plant
regenerated from the tissue or cell culture of lettuce variety NUN
09165 LTL, wherein the regenerated plant is not different from
lettuce variety NUN 09165 LTL in all, or all but one, two or three,
of the physiological and morphological characteristics (determined
at the 5% significance level when grown under the same
environmental conditions. Optionally, the plant has one, two, or
three physiological or morphological characteristic that is
different from lettuce variety NUN 09165 LTL, wherein the
difference or modification is effected by mutation or
transformation with a transgene.
[0114] In another aspect, the disclosure provides a lettuce plant
regenerated from the tissue or cell culture of variety NUN 09165
LTL, wherein the plant has all or all but one, two, or three of the
physiological and morphological characteristics of said variety
determined at the 5% significance level when grown under the same
environmental conditions. Similarity or difference of a
characteristic is determined by measuring the characteristics of a
representative number of plants grown under the same environmental
conditions, determining whether type/degree characteristics are the
same and determining whether numerical characteristics are
different at the 5% significance level.
[0115] Lettuce variety NUN 09165 LTL, or its progeny, or a plant
having all physiological and/or morphological characteristics or
all but one, two, or three which are different from those of
lettuce variety NUN 09165 LTL, can also be reproduced using
vegetative reproduction methods. Therefore, the disclosure provides
for a method of producing a plant or plant part of lettuce variety
NUN 09165 LTL, comprising vegetative reproduction of lettuce
variety NUN 09165 LTL. Vegetative propagation comprises
regenerating a whole plant from a plant part of lettuce variety NUN
09165 LTL or from a progeny or from a plant having all of the
physiological and morphological characteristics of said variety or
all but one, two, or three different characteristics, such as a
cutting, a cell culture, or a tissue culture.
[0116] The disclosure also provides methods of vegetatively
propagating a part of the plant of variety NUN 09165 LTL. In
certain aspects, the method comprises: (a) cultivating tissue or
cells capable of being propagated from lettuce variety NUN 09165
LTL 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, to obtain plant. In one aspect, the method further
comprises (c) growing plants from said rooted plantlets. Therefore,
the method also comprises regenerating a whole plant from a part of
lettuce variety NUN 09165 LTL. In a particular aspect, the part to
be propagated is a cutting, a cell culture, or a tissue
culture.
[0117] The disclosure also provides for a vegetatively propagated
plant of variety NUN 09165 LTL (or from progeny of lettuce variety
NUN 09165 LTL or from a plant having all but one, two, or three of
the physiological and/or morphological characteristics of lettuce
variety NUN 09165 LTL), wherein the plant has all of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL when the characteristics are determined at the 5%
significance level for plants grown under the same environmental
conditions. In another aspect, the propagated plant has all but
one, two, or three of the physiological and morphological
characteristics of lettuce variety NUN 09165 LTL when the
characteristics are determined at the 5% significance level for
plants grown under the same environmental conditions. A part of
said propagated plant or said propagated plant with one, two, or
three differences is also provided. In another aspect, the
propagated plant has all or all but one, two, or three of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL (e.g., as listed in Table 1-3).
[0118] In another aspect, the disclosure provides a method for
producing a plant part, preferably a head or a leaf, comprising
growing a plant of lettuce variety NUN 09165 LTL until it develops
at least one leaf or develops a head, and optionally collecting the
head or leaf. Preferably, the head or leaf is collected at harvest
maturity. In another aspect, the leaf is collected at baby leaf
stage.
[0119] In another aspect, a plant of lettuce variety NUN 09165 LTL
can be produced by seeding directly in the soil (e.g., field) or by
germinating the seeds in controlled environment conditions (e.g.,
greenhouses, hydroponic cultures, etc.) and optionally
transplanting the seedlings into the field (see, e.g., Gonai et
al., J. of Exp. Bot., 55(394): 111, 2004; Turini, et. al.,
University of California Agricultural and Natural Resources, 2011,
Publication 7215, 1-6; Smith, et. al., University of California
Agricultural and Natural Resources, 2011, Publication 7216, 1-6).
Lettuce may also be grown in tunnels. Moreover, said variety can be
grown in hydroponic cultures as described in, e.g., US
2008/0222949,which is herein incorporated by reference in its
entirety, and the skilled person is familiar with various types of
hydroponic cultures. Alternatively, seed of lettuce variety NUN
09165 LTL may be grown on peat block for use as root ball lettuce.
Furthermore, said variety may be combined with 1, 2 or 3 different
lettuce varieties to be grown as "composite lettuce" (see, e.g., EP
1197137, which is herein incorporated by reference in its
entirety).
[0120] In another aspect, the plant and plant parts of lettuce
variety NUN 09165 LTL 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 cell or tissue culture of the lettuce variety NUN
09165 LTL, in which the reproduced (seed propagated or vegetatively
propagated) plant has all of the physiological and morphological
characteristics of lettuce variety NUN 09165 LTL, e.g., as listed
in Table 1-3. In one aspect, said progeny of lettuce variety NUN
09165 LTL can be modified in one, two, or three characteristics, in
which the modification is a result of mutagenesis or transformation
with a transgene.
[0121] In other aspects, the disclosure provides for a progeny
plant of variety NUN 09165 LTL such as a progeny plant obtained by
further breeding of lettuce variety NUN 09165 LTL. Further breeding
with said variety includes selfing and/or cross-pollinating lettuce
variety NUN 09165 LTL with another lettuce plant or variety one or
more times. In a particular aspect, the disclosure provides for a
progeny plant that retains all of the morphological and
physiological characteristics of lettuce variety NUN 09165 LTL,
optionally all or all but one, two, or three characteristics as
listed in Table 1-3, determined at the 5% significance level for
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 lettuce variety NUN 09165 LTL,
where the pollen comes from an anther of lettuce variety NUN 09165
LTL and the ovule comes from an ovary of lettuce variety NUN 09165
LTL.
[0122] In still another aspect, the disclosure provides a method of
producing a lettuce plant, comprising crossing a plant of lettuce
variety NUN 09165 LTL with a second lettuce plant at least once,
allowing the seed to develop and optionally harvesting said progeny
seed. The skilled person can select a progeny lettuce plant from
said crossing. Optionally, the progeny (grown from the progeny
seed) is crossed twice, thrice, or four, 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 lettuce
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.
[0123] The disclosure also provides a method for collecting pollen
of lettuce variety NUN 09165 LTL, comprising collecting pollen from
a plant of variety NUN 09165 LTL. Alternatively, the method
comprises growing a plant of lettuce variety NUN 09165 LTL 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 lettuce flower.
[0124] In yet another aspect, the disclosure provides a method of
producing a lettuce plant, comprising selfing a plant of variety
NUN 09165 LTL one or more times, and selecting a progeny plant from
said selfing. In one aspect, the progeny plant retains all or all
but one, two, or three of the physiological and morphological
characteristics of lettuce variety NUN 09165 LTL when grown under
the same environmental conditions. In a different aspect, the
progeny plant comprises all (or all but one, two or three) of the
physiological and morphological characteristic of lettuce variety
NUN 09165 LTL of Table 1-3.
[0125] The disclosure also provides a method for developing a
lettuce plant in a lettuce breeding program, using lettuce variety
NUN 09165 LTL, or its parts as a source of plant breeding material.
Suitable plant breeding techniques are recurrent selection,
backcrossing, pedigree breeding, mass selection, mutation breeding,
genetic marker enhanced selection, and/or genetic transformation.
In one aspect, the method comprises crossing lettuce variety NUN
09165 LTL or progeny of said variety, or a plant comprising all but
1, 2, or 3 or more of the morphological and physiological
characteristics of lettuce variety NUN 09165 LTL (e.g., as listed
in Table 1-3) with a different lettuce 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, genetic marker
enhanced selection, genetic transformation (see, e.g., Brotman et
al., Theor Appl Genet (2002) 104:1055-1063). For breeding methods
in general, see, e.g., Acquaah, Principles of Plant Genetics and
Breeding, 2007, Blackwell Publishing, ISBN-13:
978-1-4051-3646-4.
[0126] In one aspect, pedigree selection is used as a breeding
method for developing a lettuce variety. Pedigree selection is also
known as the "Vilmorin System of Selection," see, e.g., Allard,
John Wiley & Sons, Inc., 1999, pp. 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.
[0127] Using pedigree method, two parents may be crossed using an
emasculated female and a pollen donor (male) to produce F1
offspring. Lettuce is an obligate self-pollination species, which
means that pollen is shed before stigma emergence, assuring 100%
self-fertilization. Therefore, in order to optimize crossing, a
method of misting may be used to wash the pollen off prior to
fertilization to assure crossing or hybridization. The F 1 may be
self-pollinated to produce segregating F2 generation. Individual
plants may then be selected which represent the desired phenotype
in each generation (F3, F4, F5, etc.) until the traits are
homozygous or fixed within a breeding population.
[0128] Thus, progeny in connection with pedigree selection are
either the generation (seeds) produced from the first cross (F1) or
selfing (S1), or any further generation produced by crossing and/or
selfing (F2, F3, F4, F5, F6, F7, etc.) and/or backcrossing (BC1,
BC2, BC3, BC4, BC5, BC6, BC7, etc.) one or more selected plants of
the F 1 and/or S1 and/or BC1 generation (or plants of any further
generation, e.g., the F2) with another lettuce plant (an/or with
wild relative of lettuce).
[0129] In yet another aspect, the disclosure provides for a method
of producing a new lettuce plant. The method comprises crossing
lettuce variety NUN 09165 LTL, or a plant of comprising all but 1,
2, or 3 of the morphological and physiological characteristics of
said lettuce variety (as listed in Table 1-3), or a progeny plant
thereof, either as male or as female parent, with a second lettuce
plant (or a wild relative of lettuce) one or more times, and/or
selfing a lettuce plant of variety NUN 09165 LTL, or a progeny
plant thereof, one or more times, and selecting progeny from said
crossing and/or selfing. The second lettuce plant may, for example,
be a line or variety of the species Lactuca sativa, or other
Lactuca species.
[0130] In a further aspect, lettuce variety NUN 09165 LTL is used
in crosses with other, different, lettuce varieties to produce
first generation (F1) lettuce hybrid seeds and plants with superior
characteristics. In a particular aspect, the disclosure provides a
method a producing a hybrid lettuce seed comprising crossing a
first parent lettuce plant with a second parent lettuce plant and
harvesting the resultant seed, in which the first parent lettuce
plant or second parent lettuce plant is lettuce variety NUN 09165
LTL. Also provided is a hybrid lettuce seed produced from crossing
a first parent lettuce plant with a second parent lettuce plant and
harvesting the resultant hybrid lettuce seed, wherein said first
parent lettuce plant or second parent lettuce plant is lettuce
variety NUN 09165 LTL. In a further aspect, the hybrid lettuce
plant produce from the hybrid lettuce seed is provided.
[0131] The morphological and physiological characteristics (and the
distinguishing characteristics) of lettuce variety NUN 09165 LTL
are provided, for example, in Table 1-3. Encompassed herein is also
a plant obtainable from lettuce variety NUN 09165 LTL (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
lettuce variety NUN 09165 LTL listed in Table 1-3 as determined at
the 5% significance level for numerical characteristics or
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) when grown under the same
environmental conditions. The morphological and/or physiological
characteristics may vary somewhat with variation in the environment
(e.g., 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
against the Royal Horticultural Society (RHS) Chart. Also,
at-harvest and/or post-harvest characteristics of heads or leaves
can be compared, such as cold storage holding quality, post-harvest
leaf crispness and leaf browning or pinking after cutting can be
measured using known methods.
[0132] In another aspect, a seed of inbred variety NUN 09165 LTL is
obtainable by selfing the variety and harvesting the seeds
produced. The resultant seeds can be grown to produce plants of
said variety.
[0133] In still another aspect, the disclosure provides a method of
producing a plant derived from lettuce variety NUN 09165 LTL, the
method comprising: (a) preparing a progeny plant derived from
lettuce variety NUN 09165 LTL by crossing a plant of variety NUN
09165 LTL either as a male or female parent with a second plant or
selfing lettuce variety NUN 09165 LTL or vegetative reproduction of
lettuce variety NUN 09165 LTL, and (b) collecting seeds from said
crossing or selfing or regenerating a whole plant from the
vegetative cell-or tissue culture. Also provided are seeds and/or
plants obtained by this method. All plants produced using lettuce
variety NUN 09165 LTL as a parent are within the scope of the
disclosure, including plant parts derived from lettuce variety NUN
09165 LTL.
[0134] In further aspects, the method comprises growing a progeny
plant of a subsequent generation from said seed of a progeny plant
of a subsequent generation and crossing the progeny plant of a
subsequent generation with itself or a second plant and repeating
the steps for an additional 3-10 generations to produce a plant
derived from lettuce variety NUN 09165 LTL. The plant derived from
lettuce variety NUN 09165 LTL 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, a plant
derived from lettuce variety NUN 09165 LTL is obtained which has
some of the desirable traits of the line as well as potentially
other selected traits.
[0135] The disclosure provides for methods of producing plants
which retain all the morphological and physiological
characteristics of a 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 lettuce variety NUN 09165 LTL (e.g., as listed in Table 1-3),
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, minisatellites, Random Amplified Polymorphic DNA
(RAPD) markers, restriction fragment length polymorphism (RFLP)
markers and others). A plant is "closely related" to lettuce
variety NUN 09165 LTL if its DNA fingerprint is at least 80%, 90%,
95% or 98% identical to the fingerprint of lettuce variety NUN
09165 LTL. 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 0.95 or 0.96 or more (see, e.g.,
"Guidelines for the Handling of a Dispute on Essential Derivation
in Lettuce" at worldseed.org/wp-content/uploads/2015/10/ Guidelines
EDV_Lettuce_2004.pdf). the disclosure also provides a plant and a
variety obtained or selected by applying these methods on lettuce
variety NUN 09165 LTL. 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 lettuce variety NUN
09165 LTL or progeny of said variety (e.g., produced by selfing)
which variant differs from lettuce variety NUN 09165 LTL in one,
two or three of the morphological and/or physiological
characteristics (e.g., characteristics listed in Table 1-3). In one
aspect, the disclosure provides a plant of lettuce variety NUN
09165 LTL having a Jaccard's Similarity index with said variety of
at least 0.95, 0.96, 0.97, 0.98 or even at least 0.99.
[0136] In some aspects, the disclosure provides a lettuce 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 lettuce variety NUN 09165 LTL as deposited under Accession
Number NCIMB ______. In some aspects, the lettuce plant further
comprises all or all but one, two, or three of the physiological
and morphological characteristics of lettuce variety NUN 09165 LTL
(e.g., as listed in Table 1-3). In other aspects, the lettuce plant
comprises the distinguishing characteristics of lettuce variety NUN
09165 LTL.
[0137] For the purpose of this disclosure, the "sequence identity"
or 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 length 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 consequences over the entire length according to
the Needleman and Wunsch global alignment algorithm described in
Needleman and Wunsch, 1970, J. Mol. Bio. 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 (see, EMBOSS, Rice, et. al., Trends in Genetics, June
2000, 16(6):276-77).
[0138] In one aspect, a plant of lettuce variety NUN 09165 LTL 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 lettuce populations in order to identify
mutants.
[0139] Similarly, lettuce variety NUN 09165 LTL may be transformed
and regenerated, whereby one or more chimeric genes are introduced
into the variety or into a plant comprising all but one, two, or
three of the physiological and morphological characteristics (e.g.,
as listed in Table 1-3). Many useful traits can be introduced into
lettuce variety NUN 09165 LTL by e.g., crossing lettuce variety NUN
09165 LTL with a transgenic lettuce plant comprising a desired
transgene, as well as by directly introducing a transgene into
lettuce variety NUN 09165 LTL by genetic transformation
techniques.
[0140] Any pest or disease resistance genes may be introduced into
lettuce variety NUN 09165 LTL, progeny of said variety, or into a
plant comprising all but 1, 2, or 3 or more of the morphological
and physiological characteristics of lettuce variety NUN 09165 LTL
(e.g., as listed in Table 1-3). Resistance to one or more of the
following diseases or pests may be introduced into the plant
described herein: Rhizomonas suberifaciens (Corky root rot), Bremia
lactucae (Downy mildew), Erysiphe cichoracearum f. sp. lactucae
(Powdery mildew), Sclerotinia minor and Sclerotinia sclerotiorum
(Lettuce Drop), Pseudomonas spp. (Bacterial Soft Rot), Botrytis
cinerea (Grey Mold), Verticillium dahlia (Verticillium Wilt),
Xanthomonas spp. (Bacterial Leaf Spot), Microdochium panattonianum
(Anthracnose), Fusarium oxysporum f sp. lactucae, Rhizoctonia
solani (Bottom Rot), Cabbage Loopers, Lettuce Root Aphid, Myzus
persicae (Green Peach Aphid), Liriomyza langei (Pea Leafminer),
Liriomyza trifolii (Serpentine Leafminer), Liriomyza sativae
(Vegetable Leafminer), Foxglove Aphid, Potato Aphid, Beet Armyworm,
Bemisia argentifolii (Silver Whitefly), and/or Aster Yellows. Other
resistance genes, against pathogenic viruses (e.g., Mirafiori
Lettuce Big Vein Virus (LMBVV), Lettuce Infectious Yellows Virus
(LIYV), Lettuce Mosaic Virus (LMV), Lettuce Necrotic Stunt Virus
(LNSV), Cucumber Mosaic Virus (CMV), Tomato Bushy Stunt Virus
(Dieback), Tomato Spotted Wilt Virus (TSWV), Turnip Mosaic Virus,
Beet Western Yellows Virus (BWYV), Alfalfa mosaic virus (AMV)),
fungi, bacteria, nematodes, insects or other pests may also be
introduced. In one aspect, resistance against Nasonovia ribisnigri
biotype Nr:0 and/or Nr:1 maybe introduced into the plant disclosed
herein. Also, any resistances to physiological stresses may be
introduced into the plant described herein, or progeny thereof or
into a plant comprising all but 1, 2, or 3 or more of the
morphological and physiological characteristics of said plant
(e.g., as listed in Table 1-3). Resistance against one or more of
the following may be introduced into the plant described herein:
Tip burn, Heat, Drought, Cold, Salt and/or Brown rob (Rib
Discoloration/Rib Blight).
[0141] Genetic transformation may, therefore, be used to insert a
selected transgene into the lettuce plants of the disclosure
described herein or may, alternatively, be used for the preparation
of transgenic lettuce plants which can be used as a source of the
transgene(s), which can be introduced into lettuce variety NUN
09165 LTL by e.g., backcrossing. A genetic trait which has been
engineered into the genome of a particular lettuce plant may then
be moved into the genome of another lettuce 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 lettuce variety into an already
developed lettuce variety and the resulting backcross conversion
plant will then comprise the transgene(s).
[0142] 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 relates to transgenic plants of lettuce variety NUN
09165 LTL. In some aspects, a transgenic plant of lettuce variety
NUN 09165 LTL may contain at least one transgene but could also
contain at least 1, 2, 3, 4, or more transgenes.
[0143] 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 lettuce
plants using transformation methods to incorporate transgenes into
the genetic material of the lettuce plant(s). Transformation can be
carried out using standard methods, such as Agrobacterium
tumefaciens mediated transformation or biolistic, followed by
selection of the transformed cells and regeneration into
plants.
[0144] Plants can also be genetically engineered, modified, or
manipulated to express various phenotypes of horticultural
interest. Through the transformation of lettuce, 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 lettuce as well
as non-native DNA sequences can be transformed into lettuce and
used to alter levels of native or non-native proteins. Various
promoters, targeting sequences, enhancing sequences, and other DNA
sequences can be inserted into the genome for the purpose of
altering the expression of proteins. Reduction of the activity of
specific genes (also known as gene silencing, or gene suppression)
is desirable for several aspects of genetic engineering in
plants.
[0145] 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 have been applied
to plants are: finger nucleases (ZFNs), transcription
activator-like effector nucleases (TALENs), engineered homing
endonucleases or meganucleases, and clustered regularly interspaced
short palindromic repeats (CRISPR)/CRISPR-associated protein 9
(Cas9), see, e.g., Songstad, et. al., Critical Reviews in Plant
Sciences, 2017, 36:1, 1-23.
[0146] Thus, the disclosure provides a method of producing a plant
of lettuce variety NUN 09165 LTL having a desired trait, comprising
mutating a plant or plant part of variety NUN 09165 LTL, optionally
with a target gene, and selecting a plant the desired trait,
wherein the mutated plant retains all or all but one of the
physiological and morphological characteristics of said variety,
optionally as described Table 1-3, and contains the desired trait,
and wherein a representative sample of seed of variety NUN 09165
LTL has been deposited under Accession Number NCIMB ______. In a
further aspect, the desired trait is yield, nutritional value,
taste, color, crunchiness, male sterility, herbicide tolerance,
insect resistance, pest resistance, disease resistance,
environmental stress tolerance, modified carbohydrate metabolism,
or modified protein metabolism, or the mutation occurs in any of
the following genes: Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL,
PPO.
[0147] The disclosure also provides a method for inducing a
mutation in lettuce variety NUN 09165 LTL comprising: [0148] a.
exposing the seed, plant, plant part, or cell of lettuce variety
NUN 09165 LTL to a mutagenic compound or to radiation, wherein a
representative sample of seed of said variety has been deposited
under Accession Number NCIMB ______; [0149] b. selecting the seed,
plant, or plant part or cell of lettuce variety NUN 09165 LTL
having a mutation; and [0150] c. optionally growing and/or
multiplying the seed, plant, plant part, or cell of lettuce variety
NUN 09165 LTL having the mutation.
[0151] The disclosure also provides a method of producing a lettuce
plant having a desired trait, wherein the method comprises
transforming the lettuce plant with a transgene that confers the
desired trait, wherein the transformed plant otherwise retains all
of the physiological and morphological characteristics of the plant
of variety NUN 09165 LTL and contains the desired trait. Thus, a
transgenic lettuce plant is provided which is produced by the
method described above, wherein the plant comprises the desired
trait and all of the physiological and morphological
characteristics of the plant of variety NUN 09165 LTL.
[0152] In another aspect, the disclosure provides a method of
producing a progeny of plant of variety NUN 09165 LTL further
comprising a desired trait, said method comprising transforming the
plant of lettuce variety NUN 09165 LTL with at least one transgene
that confers the desired trait and/or crossing the plant of lettuce
variety NUN 09165 LTL with a transgenic lettuce 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.
[0153] A desired trait (e.g., gene(s) conferring pest or disease
resistance, herbicide, fungicide or insecticide tolerance, etc.)
can be introduced into lettuce variety NUN 09165 LTL, 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 lettuce variety NUN 09165 LTL or the progeny of
said variety and contains the desired trait, wherein the desired
trait is yield, nutritional value, taste, color, crunchiness, male
sterility, herbicide tolerance, insect resistance, pest resistance,
disease resistance, environmental stress tolerance, modified
carbohydrate metabolism, or modified protein metabolism or the
mutation occurs in any of the following genes:
Ferulate-5-hydrxylase, dmr1, dmr6, NCED4, PAL, PPO. 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, cyclohexanedione, phenoxy proprionic acid
and L-phosphinothricin or a polynucleotide conferring resistance to
Rhizomonas suberifaciens (Corky root rot), Bremia lactucae (Downy
mildew), Erysiphe cichoracearum f sp. lactucae (Powdery mildew),
Sclerotinia minor and Sclerotinia sclerotiorum (Lettuce Drop),
Pseudomonas spp. (Bacterial Soft Rot), Botrytis cinerea (Grey
Mold), Verticillium dahlia (Verticillium Wilt), Xanthomonas spp.
(Bacterial Leaf Spot), Microdochium panattonianum (Anthracnose),
Fusarium oxysporum f sp. lactucae, Rhizoctonia solani (Bottom Rot),
Cabbage Loopers, Lettuce Root Aphid, Myzus persicae (Green Peach
Aphid), Liriomyza langei (Pea Leafminer), Liriomyza trifolii
(Serpentine Leafminer), Liriomyza sativae (Vegetable Leafminer),
Foxglove Aphid, Potato Aphid, Beet Armyworm, Bemisia argentifolii
(Silver Whitefly), and/or Aster Yellows. Other resistance genes,
against pathogenic viruses (e.g., Mirafiori Lettuce Big Vein Virus
(LMBVV), Lettuce Infectious Yellows Virus (LIYV), Lettuce Mosaic
Virus (LMV), Lettuce Necrotic Stunt Virus (LNSV), Cucumber Mosaic
Virus (CMV), Tomato Bushy Stunt Virus (Dieback), Tomato Spotted
Wilt Virus (TSWV), Turnip Mosaic Virus, Beet Western Yellows Virus
(BWYV), Alfalfa mosaic virus (AMV)), fungi, bacteria, nematodes,
insects or other pests may also be introduced.
[0154] By crossing and/or selfing also (one or more), single traits
may be introduced into, or modified in, lettuce variety NUN 09165
LTL (e.g., using backcrossing breeding schemes), while retaining
the remaining 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
lettuce variety NUN 09165 LTL by breeding with said variety.
[0155] In another aspect, the disclosure provides a method of
introducing a single locus conversion, a single trait conversion,
or a desired trait into lettuce variety NUN 09165 LTL, comprising
introducing a single locus conversion, a single trait conversion,
or a desired trait in at least one of the parents of lettuce
variety NUN 09165 LTL, and crossing the converted parent with the
other parent of lettuce variety NUN 09165 LTL, to obtain seed of
said variety.
[0156] 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 parent plants comprises: [0157] a. crossing the
parental line of lettuce variety NUN 09165 LTL, with a second
lettuce plant comprising the single locus conversion, the single
trait conversion, or the desired trait; [0158] b. selecting F1
progeny plants that contain the single locus conversion, the single
trait conversion, or the desired trait. [0159] c. crossing said
selected progeny plants of step b) with the parental line of step
a) to produce a backcross progeny plant; [0160] 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 physiological and
morphological characteristics of the parental line of step a) to
produce selected backcross progeny plants; and [0161] 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 conversion, the single
trait conversion, or the desired trait and otherwise all or all but
one, two, or three of the physiological and morphological
characteristics of the parental line of step a) to produce selected
backcross progeny plants, when grown in the same environmental
conditions. The disclosure further relates to plants obtained by
this method.
[0162] Alternatively, a single trait converted plant or single
locus converted plant may be produced by: [0163] a. obtaining a
cell or tissue culture of cells of the parental line of lettuce
variety NUN 09165 LTL; [0164] b. genetically transforming or
mutating said cells; [0165] c. growing the cells into a plant; and
[0166] d. optionally selecting a plant that contains the desired
single locus conversion, single trait conversion, or the desired
trait.
[0167] In another aspect, the disclosure provides a method of
introducing a single locus conversion, a single trait conversion,
or a desired trait into lettuce variety NUN 09165 LTL, comprising:
[0168] a. obtaining a combination of parental lines of lettuce
variety NUN 09165 LTL, optionally through reverse synthesis of
breeding lines; [0169] 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 [0170] c. crossing the converted parent
with the other parent of step a) to obtain seed of lettuce variety
NUN 09165 LTL.
[0171] In any of the above methods, wherein 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 Rhizomonas
suberifaciens (Corky root rot), Bremia lactucae (Downy mildew),
Erysiphe cichoracearum f. sp. lactucae (Powdery mildew),
Sclerotinia minor and Sclerotinia sclerotiorum (Lettuce Drop),
Pseudomonas spp. (Bacterial Soft Rot), Botrytis cinerea (Grey
Mold), Verticillium dahlia (Verticillium Wilt), Xanthomonas spp.
(Bacterial Leaf Spot), Microdochium panattonianum (Anthracnose),
Fusarium oxysporum f sp. lactucae, Rhizoctonia solani (Bottom Rot),
Cabbage Loopers, Lettuce Root Aphid, Myzus persicae (Green Peach
Aphid), Liriomyza langei (Pea Leafminer), Liriomyza trifolii
(Serpentine Leafminer), Liriomyza sativae (Vegetable Leafminer),
Foxglove Aphid, Potato Aphid, Beet Armyworm, Bemisia argentifolii
(Silver Whitefly), and/or Aster Yellows. Other resistance genes,
against pathogenic viruses (e.g., Mirafiori Lettuce Big Vein Virus
(LMBVV), Lettuce Infectious Yellows Virus (LIYV), Lettuce Mosaic
Virus (LMV), Lettuce Necrotic Stunt Virus (LNSV), Cucumber Mosaic
Virus (CMV), Tomato Bushy Stunt Virus (Dieback), Tomato Spotted
Wilt Virus (TSWV), Turnip Mosaic Virus, Beet Western Yellows Virus
(BWYV), Alfalfa mosaic virus (AMV)), fungi, bacteria, nematodes,
insects or other pests may also be introduced. In one aspect,
resistance against Nasonovia ribisnigri biotype Nr:0 and/or Nr:1
maybe introduced into the plant disclosed herein. Also, any
resistances to physiological stresses may be introduced into the
plant described herein, or progeny thereof or into a plant
comprising all but 1, 2, or 3 or more of the morphological and
physiological characteristics of said plant (e.g., as listed in
Table 1-3). Resistance against one or more of the following may be
introduced into the plant described herein: Tip burn, Heat,
Drought, Cold, Salt and/or Brown rob (Rib Discoloration/Rib
Blight).
[0172] The disclosure also provides a plant having one, two, or
three physiological and/or morphological characteristics which are
different from those of lettuce variety NUN 09165 LTL, and which
otherwise has all of the physiological and morphological
characteristics of said variety, wherein a representative sample of
seed of said variety has been deposited under Accession Number
NCIMB ______. In particular, variants which differ from lettuce
variety NUN 09165 LTL in none, one, two, or three of the
characteristics mentioned in Table 1-3 are encompassed.
[0173] The disclosure also provides a lettuce plant comprising at
least a set of first set of the chromosomes of lettuce variety NUN
09165 LTL, a sample of seed of said variety has been deposited
under Accession Number NCIMB ______; optionally further comprising
a single locus conversion or mutation, 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, the single locus
conversion or mutation confers a trait, wherein the trait is yield,
color, size, taste, crunchiness, enhance nutritional quality,
post-harvest quality, male sterility, herbicide tolerance, insect
resistance, pest resistance, disease resistance, environmental
stress tolerance, modified carbohydrate metabolism, or modified
protein metabolism.
[0174] In one aspect, the disclosure provides for a haploid plant
and/or a doubled haploid plant of variety NUN 09165 LTL, or a plant
having all but one, two or three physiological and/or morphological
characteristics of lettuce variety NUN 09165 LTL, 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. DH 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.
[0175] In another aspect, the disclosure comprises a method for
producing doubled haploid cells of lettuce variety NUN 09165 LTL,
comprising making doubles haploid cells from haploids cells from
the plant or plant part of lettuce variety NUN 09165 LTL with a
chromosome doubling agent, such as colchicine treatment (see, e.g.,
Nikolova and Niemirowicz-Szczytt (1996) Acta Soc Bot Pol
65:311-317).
[0176] In another aspect, the disclosure provides for haploid
plants and/or doubled haploid plants derived from lettuce variety
NUN 09165 LTL that, when combined, make a set of parents of lettuce
variety NUN 09165 LTL. The haploid plant and/or the doubled haploid
plant of variety NUN 09165 LTL can be used in a method for
generating parental lines of lettuce variety NUN 09165 LTL.
[0177] The disclosure also provides methods for determining the
identity of parental lines of the plant described herein, in
particular the identity of the female line. US 2015/0126380, which
is hereby incorporated by reference, 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
the steps of 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 a plant of lettuce variety NUN 09165 LTL is a progeny of
said variety, because the seed coat of the seed is a maternal
tissue genetically identical to lettuce variety NUN 09165 LTL.
Since lettuce variety NUN 09165 LTL is an inbred variety, with a
very high degree of homozygosity, any F 1 progeny will inherit the
same, predictable, set of chromosomes from its parent. Thus, the
skilled person will also be able to identify maternal tissues of a
seed grown on an F1 progeny of lettuce variety NUN 09165 LTL, using
the methods described in US 2015/0126380. In another particular
aspect, the skilled person can determine the identity of the female
parental line of lettuce variety NUN 09165 LTL 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 lettuce variety NUN
09165 LTL.
[0178] 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 lettuce variety NUN 09165
LTL. A skilled person can take any individual heterozygous plant
(called a "phenotypically superior plant" in Example 2 of US
2015/0245570 hereby incorporated by reference in its entirety;
lettuce variety NUN 09165 LTL is such plant) and generate a
combination of parental lines (reverse breeding parental lines)
that, when crossed, produce the variety NUN 09165 LTL. 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., lettuce variety NUN 09165 LTL), 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.
[0179] In another aspect, the method for producing parental lines
for hybrid organisms, e.g., of lettuce variety NUN 09165 LTL, which
when crossed reconstitute the genome of lettuce variety NUN 09165
LTL, comprising: [0180] a. defining a set genetic a) markers that
are present a heterozygous form (H) in a partially heterozygous
starting organism; [0181] b. producing at least one further
generation from the starting organism by self-pollination (e.g., F2
or F3 generation); [0182] 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 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.
[0183] The disclosure also relates to a method of producing a
combination of parental lines of a plant of variety NUN 09165 LTL,
comprising making double haploid cells from haploid cells form said
plant or a seed of that plant; and optionally crossing these
parental lines to produce and collect 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 09165
LTL, when these parental lines are crossed. In still another
aspect, the disclosure relates to a combination of a parental lines
from which a seed or plant having all physiological and/or
morphological characteristics of lettuce variety NUN 09165 LTL
(when the characteristics are determined at the 5% significance
level for plants grown under the same environmental
conditions).
[0184] The disclosure also provides a combination of parental
lines, which when crossed, produce a seed or plant having all of
the physiological and/or morphological characteristics of lettuce
variety NUN 09165 LTL, but one, two, or three which are different
(when grown under the same environmental conditions).
[0185] In another aspect, a combination of a male and a female
parental line of lettuce variety NUN 09165 LTL can be generated by
methods described herein, for example, through reverse synthesis of
breeding lines.
[0186] In another aspect, the disclosure provides a method of
determining the genotype of a plant of the invention comprising the
step of 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
(AFLPD), polymerase chain reaction (PCR), DNA sequencing, allele
specific oligonucleotide (ASO) probes, and 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.
[0187] Also provided is a plant part obtainable from variety NUN
09165 LTL (or from progeny of said variety or from a plant having
all or all but one, two or three physiological and/or morphological
characteristics which are different from those of lettuce variety
NUN 09165 LTL) or from a vegetatively propagated plant of variety
NUN 09165 LTL (or from its progeny or from a plant having all but
one, two or three physiological and/or morphological
characteristics which are different from those of lettuce variety
NUN 09165 LTL), wherein the plant part is a leaf, a harvested leaf,
a part of a leaf, a head, a harvested head, a part of a head,
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 lettuce variety NUN 09165
LTL, or a hypocotyl, a cotyledon, a pistil, an anther, or a flower
or a part thereof.
[0188] A part of lettuce variety NUN 09165 LTL (or of progeny of
said variety or of a plant having all physiological and/or
morphological characteristics but one, two or three which are
different from those of said variety) encompasses any cells,
tissues, organs obtainable from the seedlings or plants, such as
but not limited to: a lettuce head or a part thereof, a leaf or a
part thereof, a cutting, hypocotyl, cotyledon, seed coat, pollen
and the like. Such parts can be stored and/or processed
further.
[0189] The disclosure also provides for a food product, a feed
product, or a processed product comprising or consisting of a plant
part described herein. Preferably, the plant part is a lettuce head
or leaf or a part thereof and/or an extract from a leaf or another
plant part described herein comprising at least one cell of lettuce
variety NUN 09165 LTL. 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, pureed or
concentrated, juiced, pickled, canned, steamed, boiled, fried,
blanched and/or frozen, etc.
[0190] Such plant part of lettuce variety NUN 09165 LTL can be
stored and/or processed further. The disclosure thus also provides
for a food or feed products comprising one or more of such parts,
such as canned, chopped, cooked, roasted, in a sauce, in a
sandwich, pasted, pureed or concentrated, juiced, frozen, dried,
pickled, or powdered lettuce heads or leaves from lettuce variety
NUN 09165 LTL or from progeny of said variety, or from a derived
variety, such as a plant having all or all but one, two, or three
of the physiological and/or morphological characteristics of
lettuce variety NUN 09165 LTL.
[0191] The disclosure further provides for food or feed products
comprising part, or part of progeny of lettuce variety NUN 09165
LTL, or part of a plant having all but one, two, or three of the
physiological and morphological characteristics of lettuce variety
NUN 09165 LTL, comprising one or more of such parts, optionally
processed (e.g., canned, chopped, cooked, roasted, in a sauce, in a
sandwich, pasted, pureed or concentrated, juiced, frozen, dried,
pickled, or powdered).
[0192] In another aspect, the plant, plant part, or seed of lettuce
variety NUN 09165 LTL is inside 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 part of a plant
(fresh and/or processed) or seed of lettuce variety NUN 09165 LTL.
In a particular aspect, the container comprises a plurality of
seeds of lettuce variety NUN 09165 LTL, or a plurality of plant
parts of lettuce variety NUN 09165 LTL. In one aspect, a seed or a
plurality of seeds of said variety are packaged into a container of
any size or type (e.g., bags, cartons, cans, etc.). The seeds may
be pelleted prior to packing (to form pills or pellets) and/or may
be disinfected, primed and/or treated with various compounds, such
as seed coating or crop protection compounds. The seed produces a
plant of lettuce variety NUN 09165 LTL.
[0193] In another aspect, the disclosure provides for a lettuce
head or leaf of variety NUN 09165 LTL, or a part of a head or leaf
of said variety. The head or leaf 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 lettuce heads or leaves or parts of lettuce heads or
leaves of said variety, or lettuce heads or leaves of progeny
thereof, or lettuce heads or leaves of a derived variety.
[0194] Marketable lettuce heads or leaves are generally sorted by
size and quality after harvest. Alternatively, the lettuce heads or
leaves can be sorted by leaf size, shape, texture, glossiness, or
color.
[0195] All documents (e.g., patent publications) are herein
incorporated by reference in their entirety, including the
following cited references: [0196] Naktuinbow and NARO,
"Calibration Manual: DUS Test for Lettuce," world-wide web at
naktuinbow.nl [0197] US Department of Agriculture, Agricultural
Marketing Service, "Objective Description of Variety--Lettuce
(Lactuca sativa L.)", world wide web at
ams.usda.gov/services/plant-variety-protection/pvpo-c-forms, under
lettuce. [0198] UPOV, "Guidelines for the Conduct of Tests for
Distinctness, Uniformity and Stability", TG/13/11 (Geneva 2006,
last updated Apr. 5, 2017), world-wide web at upov.int under
edocs/tgdocs/en/tg013 .pdf. [0199] World Seed, Guidelines for the
Handling of a Dispute on Essential Derivation in Lettuce [0200]
Bertier, L. D., et. al., "High-Resolution Analysis of the
Efficiency, Heritability, and Editing Outcomes of
CRISPR/Cas9-Induced Modifications of NCED4 in Lettuce (Lactuca
sativa)," G3: Genes, Genomes, Genetics, 2018, vol. 8, pp.
1513-1521. [0201] Brotman, Y., et. al., "Resistance Gene Homologues
in Melon are Linked to Genetic Loci Conferring Disease and Pest
Resistance", Theor Appl Genet, 2002, vol. 104, pp. 1055-1063, DOI
10.1007/s00122-001-0808-x [0202] Gonai, T., et al., "Abscisic Acid
in the Thermoinhibition of Lettuce Seed Germination and Enhancement
of its Catabolism by Gibberellin", Journal of Experimental Botany,
2004, vol. 55(394), pp. 111-118. [0203] Hunter, P. J., et. al.,
"Oxidative Discolouration in Whole-head and Cut Lettuce:
Biochemical and Environmental Influences on a Complex Phenotype and
Potential Breeding Strategies to Improve Shelf-life," Euphytica,
2017, vol. 213(180), DOI 10.1007/s10681-017-1964-7. [0204]
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. [0205] Nikolova, V., et. al., "Diploidization of Cucumber
(Cucumis sativus L.) Haploids by Colchini Treatment", Acta Societas
Botanicorum Poloniae, 1996, vol. 65, pp. 311-317. [0206] Smith, et.
al., "Leaf Lettuce Production in California," 2011, University of
California Agricultural and Natural Resources, Publication 7216,
pp. 1-6. [0207] Songstad, D. D., et. al., "Genome Editing of
Plants," Critical Reviews in Plant Sciences, vol. 36, no. 1, pp
1-23. [0208] Teng, W., et al., "Rapid Regeneration of Lettuce from
Suspension Culture", HortScience, 1992, vol. 27(9), pp. 1030-1032.
[0209] Teng, W., et al., "Regenerating Lettuce from Suspension
Culture in a 2-Liter Bioreactor", HortScience, 1993, vol. 28(6),
pp. 669-671. [0210] Turini, et. al., "Iceberg Lettuce Production in
California," 2011, University of California Agricultural and
Natural Resources, Publication 7215, pp. 1-6. [0211] Vos, P., et
al., "AFLP: A New Technique for DNA Fingerprinting", Nucleic Acids
Research, 1995, vol. 23(21), pp. 4407-4414. [0212] Wijnker, E., et
al., "Hybrid Recreation by Reverse breeding in Arabidopsis
thaliana", Nature Protocols, 2014, vol. 9, pp. 761-772. DOI: doi:
10.1038/nprot.2014.049 [0213] Zhang, X., et al., "Genotypic Effects
on Tissue Culture Response of Lettuce Cotyledons", Journal of
Genetics and Breeding, 1992, vol. 46, pp. 287-290. [0214] US
2008/0222949 [0215] EP 1 197 137 A1 [0216] US 2015/0126380 [0217]
WO 2017/144669 [0218] WO 2008/092505 [0219] U.S. Pat. No.
8,237,019
Development of Lettuce Variety NUN 09165 LTL
[0220] The inbred variety NUN 09165 LTL was developed from an
initial cross between lettuce lines. The female and male parents
were crossed to produce seeds. After the cross, progeny was
self-pollinated or backcrossed, followed by pedigree selection and
line selection. Lettuce variety NUN 09165 LTL can be propagated by
seeds or vegetatively, or by regeneration of a tissue culture. The
seeds of lettuce variety NUN 09165 LTL can be grown to produce
inbred plants and parts thereof (e.g., lettuce heads and
leaves).
[0221] The Applicant concluded that lettuce variety NUN 09165 LTL
is uniform and stable. This has been established through evaluation
of horticultural characteristics. Several seed production events
resulted in no observable deviation in genetic stability. Lettuce
variety NUN 09165 LTL has been assigned an inventory ID of
219406394.
Deposit Information
[0222] A total of 2500 seeds of variety NUN 09165 LTL have been
deposited according to the Budapest Treaty by Nunhems B.V. on
______ at the NCIMB Ltd., Ferguson Building, Craibstone Estate,
Bucksburn, Aberdeen AB21 9YA, United Kingdom (NCIMB). The deposit
has been assigned NCIMB ______. A statement indicating the
viability of the sample has been provided. A deposit of lettuce
variety NUN 09165 LTL is also maintained at Nunhems B.V. The seed
lot number for lettuce variety NUN 09165 LTL is 30331901002.
[0223] The deposit will be maintained in NCIMB 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. .sctn. 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 this application
or under the Plant Variety Protection Act (7 U.S.C. .sctn. 2321 et
seq.). Accordingly, the requirements of 37 CFR .sctn. 1.801-1.809
have been satisfied.
Characteristics of Lettuce Variety NUN 09165 LTL
[0224] The most similar variety to NUN 09165 LTL is referred to as
NUN 09074 LTL, a variety from Nunhems B.V. with the commercial name
Skrunch Green 74.
[0225] In Table 1, the characteristics of lettuce variety NUN 09165
LTL are shown based on trials conducted in Salinas, Calif., USA
between March to September 2019. For numerical characteristics,
averages were calculated. For non-numerical characteristics, the
type/degree were determined.
[0226] In Tables 2 and 3, a comparison between lettuce variety NUN
09165 LTL and Reference Variety is shown based on a trial in the
USA. Trial location: Salinas, Calif., USA; Seeding date: May 12,
2020; Harvesting date: Jul. 9, 2020. In Table 4, the distinguishing
characteristics between lettuce variety NUN 09165 LTL and the
Reference Variety are shown.
[0227] One replication of 30 plants of each variety, from which at
least 15 plants or plant parts were randomly selected and were used
to measure characteristics. For numerical characteristics, averages
were calculated. For non-numerical characteristics, the type/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 when grown under the same
environmental conditions. 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 T-test, a standard method known to the
skilled person. A non-numerical characteristics is considered to be
"the same" when the values have the same "degree" or "type" when
scored using USDA and/or UPOV descriptors, for plants grown under
the same environmental conditions. In one aspect, a statistical
analysis of the quantitative characteristics showing the degree of
significance at 5% is provided (see, Tables 5-11).
[0228] In another aspect, the disclosure provides a plant having
all of the physiological and morphological characteristics of
lettuce variety NUN 09165 LTL as presented in Table 1-3, when grown
under the same environmental conditions.
TABLE-US-00001 TABLE 1 Characteristics of Lettuce Variety NUN 09165
LTL based on Salinas, USA trials between March to September 2019
Application Variety Characteristics (NUN 09165 LTL) Vigor/Size:
small volume; normal; large volume Large Color: light green, normal
green, darker green Normal green Closed head: open habit even at
mature stage; Open habit normal; very closed Head shape: not market
acceptable; acceptable, Acceptable very nice shape Head health: big
damage or infections; normal; No damage no damage or infections
Frame: lack of frame, normal, good frame Normal Base shape: very
veiny or shoulders, open base; Normal and normal and acceptable
base; closed base, not acceptable base shoulders and flat Base
health: very sensitive for Botrytis cinerea Strong for (Grey Mold),
Sclerotinia minor and Sclerotinia base diseases sclerotiorum
(Lettuce Drop), Rhizoctonia solani (Bottom Rot); normal; strong for
base diseases Compactness: loose open; normal; very compact Loose
open External tip burn: tip burn; normal; no tip burn No tip burn
Internal tip burn: tip burn and rotting inside the No tip burn
head; normal; no tip burn Bolting resistance: bolting fast; normal;
strong Normal for bolting Taste: bitter; normal; sweet Normal
TABLE-US-00002 TABLE 2 Characteristics of Lettuce Variety NUN 09165
LTL and the Reference Variety (Exhibit C-USDA descriptors) based on
Salinas California, USA Trial, May-July 2020 Application Variety
Reference Variety Characteristics (NUN 09165 LTL) (NUN 09074 LTL)
UPOV Morphology: Seed: Seed color: Black White white, yellow,
brown, black Plant: Degree of overlapping of upper part of leaves:
Absent or weak Absent or weak absent or weak, medium, strong Number
of leaves (only for varieties with degree Medium Medium of
overlapping of upper part of leaves absent or weak): very few, very
few to few, few, few to medium, medium, medium to many, many, many
to very many, very many Mature Leaves: Leaf attitude: Semi-erect
Semi-erect erect, erect to semi-erect, semi-erect, semi-erect to
horizontal, horizontal Number of divisions: Medium Many absent or
very few, very few to few, few, few to medium, medium, medium to
many, many, many to very many, very many Anthocyanin coloration:
Absent or very weak Absent or very weak absent or very weak, very
weak to weak, weak, weak to medium, medium, medium to strong,
strong, strong to very strong, very strong Color: Green Green
green, yellowish green, greyish green Intensity of green color:
Dark Medium very light, very light to light, light, light to
medium, medium, medium to dark, dark, dark to very dark, very dark
Glossiness of upper side: Medium Medium absent or very weak, very
weak to weak, weak, weak to medium, medium, medium to strong,
strong, strong to very strong, very strong Thickness: Medium Medium
very thin, thin, medium, thick, very thick Blistering: Absent or
very weak Absent or very weak absent or very weak, very weak to
weak, weak, weak to medium, medium, medium to strong, strong,
strong to very strong, very strong Undulation of margin: Strong
Strong absent or very weak, very weak to weak, weak, weak to
medium, medium, medium to strong, strong, strong to very strong,
very strong Type of incisions of margin: Tridentate Tridentate
crenate, regularly dentate, irregularly dentate, bidentate,
tridentate Depth of incisions of margin: Deep Medium absent or very
shallow, very shallow to shallow, shallow, shallow to medium,
medium, medium to deep, deep, deep to very deep, very deep Depth of
secondary incisions of margin (only Deep Deep for varieties with
type of incisions of margin irregularly dentate, bi-or
tri-dentate): very shallow, very shallow to shallow, shallow,
shallow to medium, medium, medium to deep, deep, deep to very deep,
very deep Density of incisions of margin: Dense Very dense very
sparse, very sparse to sparse, sparse, sparse to medium, medium,
medium to dense, dense, dense to very dense, very dense Venation:
Semi-flabellate Semi-flabellate not flabellate, semi-flabellate,
flabellate Resistance: Resistance to Bremia lactucae Isolate Bl:
16EU Present Present Resistance to Bremia lactucae Isolate Bl: 17EU
Present Present Resistance to Bremia lactucae Isolate Bl: 18EU
Present Present Resistance to Bremia lactucae Isolate Bl: 20EU
Present Present Resistance to Bremia lactucae Isolate Bl: 21EU
Present Present Resistance to Bremia lactucae Isolate Bl: 22EU
Present Present Resistance to Bremia lactucae Isolate Bl: 23EU
Present Present Resistance to Bremia lactucae Isolate Bl: 24EU
Present Present Resistance to Bremia lactucae Isolate Bl: 25EU
Present Present Resistance to Bremia lactucae Isolate Bl: 26EU
Present Present Resistance to Bremia lactucae Isolate Bl: 27EU
Present Absent Resistance to Bremia lactucae Isolate Bl: 28EU
Present Present Resistance to Bremia lactucae Isolate Bl: 29EU
Present Absent Resistance to Bremia lactucae Isolate Bl: 30EU
Present Absent Resistance to Bremia lactucae Isolate Bl: 31EU
Present Present Resistance to Bremia lactucae Isolate Bl: 32EU
Present Present Resistance to Bremia lactucae Isolate Bl: 33EU
Present Absent Resistance to Bremia lactucae Isolate Bl: 34EU
Present Absent Resistance to Bremia lactucae Isolate Bl: 35EU
Present Absent Resistance to Bremia lactucae Isolate Bl: 36EU
Present Absent Resistance to Nasonovia ribisnigri (Nr) Biotype
Present Present Nr: 0 Resistance to Fusarium oxysporum sp. lactucae
Absent Absent Race 1 Resistance to Lettuce Mosaic Virus (LMV)
Absent Absent Pathotype II Resistance to Lettuce Root Aphids Absent
Absent Resistance to Rhizomonas subefaciens (Corky Absent Absent
Root) Resistance to Lettuce Necrotic Stunt Virus Absent Absent
(LNSV) USDA Morphology: Plant type: Butterhead, Novita Type,
Iceberg Type, Batavia Leaf Leaf Type, Frisee d'Amerique Type, Lollo
Type, Oakloaf Type, Multi-divided Type, Frillice Type, Cos or
Romaine, Bibb/Gem, Stem, Cutting/Whole Leaf, Latin, Other (specify)
Spread of frame leaves, cm: 17.22 cm 16.98 cm Butt: Midrib:
Flattened Flattened flattened, moderately raised, prominently
raised
TABLE-US-00003 TABLE 3 Characteristics of Lettuce Variety NUN 09165
LTL and the Reference Variety (Non- USDA descriptors) based on
Salinas California, USA Trial, May-July 2020 Application Variety
Reference Variety Characteristics (NUN 09165 LTL) (NUN 09074 LTL)
Plant: Height, cm: 7.92 cm 8.30 cm Weight, g: 80.40 g 85.20 g Head
formation: No head No head no head, open head, closed head
Diameter: Medium Medium small, medium, large, very large Firmness:
Loose Loose Core: Diameter at base of head, mm (core width): 11.76
mm 10.05 mm Height from base of head to apex (mm), average 15.07 mm
12.16 mm (core length): Height from base of head to apex (mm),
range 10.91-18.56 mm 9.47-15.03 mm (min-max): Mature leaves
(harvest mature outer leaves): Length, mm: 101.67 mm 100.96 mm
Width, mm: 106.45 mm 128.28 mm Leaf size: Medium Medium small,
medium, large Leaf trichomes: Absent Absent absent, present
Incision of margin on apical part: Present Present Hue of green
color of outer leaves: Absent Absent Leaf attitude at 10-12 leaf
stage: Semi-erect Semi-erect erect, semi-erect, horizontal Leaf
blade division at 10-12 leaf stage: Divided Divided entire, lobed,
divided
TABLE-US-00004 TABLE 4 Distinguishing Characteristics between
Lettuce Variety NUN 09165 LTL and the Reference Variety Application
Variety Reference Variety Characteristics (NUN 09165 LTL) (NUN
09074 LTL) Seed: Seed color: Black White white, yellow, brown,
black Plant: Core: Diameter at base of head, mm (core width): 11.76
mm 10.05 mm Height from base of head to apex (mm), 15.07 mm 12.16
mm average (core length): Mature leaves: Number of divisions:
Medium Many absent or very few, very few to few, few, few to
medium, medium, medium to many, many, many to very many, very many
Intensity of green color: Dark Medium very light, very light to
light, light, light to medium, medium, medium to dark, dark, dark
to very dark, very dark Depth of incisions of margin: Deep Medium
absent or very shallow, very shallow to shallow, shallow, shallow
to medium, medium, medium to deep, deep, deep to very deep, very
deep Density of incisions of margin: Dense Very dense very sparse,
very sparse to sparse, sparse, sparse to medium, medium, medium to
dense, dense, dense to very dense, very dense Width, mm: 106.45 mm
128.28 mm Resistance: Resistance to Bremia lactucae Isolate Bl:
Present Absent 27EU Resistance to Bremia lactucae Isolate Bl:
Present Absent 29EU Resistance to Bremia lactucae Isolate Bl:
Present Absent 30EU Resistance to Bremia lactucae Isolate Bl:
Present Absent 33EU Resistance to Bremia lactucae Isolate Bl:
Present Absent 34EU Resistance to Bremia lactucae Isolate Bl:
Present Absent 35EU Resistance to Bremia lactucae Isolate Bl:
Present Absent 36EU
[0229] The results of the T-Test show significant differences at 5%
significance level between lettuce variety NUN 09165 LTL and the
Reference Variety for height from base of head to apex (core
length), diameter at based of head (core width), and mature leaf
width as shown in Tables 5-7.
[0230] Table 5 shows a significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p<0.001) for
height from base of head to apex (core length, mm) based on the
results of the trial conducted in the US during the trial season
2020.
TABLE-US-00005 TABLE 5 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 10.91 9.47 Max 18.56 15.03 Median 15.26 12.0
Mean 15.07 12.16 Standard deviation 2.23 1.41
[0231] Table 6 shows a significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p<0.001) for
diameter at based of head (core width, mm) based on the results of
the trial conducted in the US during the trial season 2020.
TABLE-US-00006 TABLE 6 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 9.94 7.58 Max 14.87 11.62 Median 11.30 10.22
Mean 11.76 10.05 Standard deviation 1.49 1.07
[0232] Table 7 shows a significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p<0.001) for
mature leaf width (mm) based on the results of the trial conducted
in the US during the trial season 2020.
TABLE-US-00007 TABLE 7 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 90.10 112.01 Max 130.15 155.17 Median 105.96
125.51 Mean 106.45 128.28 Standard deviation 10.64 11.29
[0233] The results of the T-Test show no significant differences at
5% significance level between lettuce variety NUN 09165 LTL and the
Reference Variety for plant spread of frame leaves, plant height,
plant weight, and mature leaf length as shown in Tables 8-11.
[0234] Table 8 shows no significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p=0.511) for plant
spread of frame leaves (cm) based on the results of the trial
conducted in the US during the trial season 2020.
TABLE-US-00008 TABLE 8 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 14.70 15.40 Max 19.40 19.50 Median 17.20 16.95
Mean 17.22 16.98 Standard deviation 1.23 1.05
[0235] Table 9 shows no significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p=0.256) for plant
height (cm) based on the results of the trial conducted in the US
during the trial season 2020.
TABLE-US-00009 TABLE 9 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 6.30 6.80 Max 9.40 10.40 Median 7.80 8.10 Mean
7.92 8.30 Standard deviation 1.06 1.03
[0236] Table 10 shows no significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p=0.374) for plant
weight (g) based on the results of the trial conducted in the US
during the trial season 2020.
TABLE-US-00010 TABLE 10 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 56.0 52.0 Max 116.0 114.0 Median 79.0 87.0 Mean
80.40 85.20 Standard deviation 16.30 16.30
[0237] Table 11 shows no significant difference between lettuce
variety NUN 09165 LTL and the Reference Variety (p=0.801) for
mature leaf length (mm) based on the results of the trial conducted
in the US during the trial season 2020.
TABLE-US-00011 TABLE 11 Application Variety Reference Variety
Statistical Parameter (NUN 09165 LTL) (NUN 09074 LTL) Number of
samples 20 20 Min. 82.25 89.91 Max 117.30 127.07 Median 100.34
99.86 Mean 101.67 100.96 Standard deviation 9.45 8.15
* * * * *