U.S. patent application number 15/293951 was filed with the patent office on 2018-04-19 for lettuce variety julian.
This patent application is currently assigned to Syngenta Participations AG. The applicant listed for this patent is Syngenta Participations. Invention is credited to Els GROOT.
Application Number | 20180103603 15/293951 |
Document ID | / |
Family ID | 61872967 |
Filed Date | 2018-04-19 |
United States Patent
Application |
20180103603 |
Kind Code |
A1 |
GROOT; Els |
April 19, 2018 |
LETTUCE VARIETY JULIAN
Abstract
The present invention provides novel lettuce cultivar Julian and
plant parts, seed, and tissue culture therefrom. The invention also
provides methods for producing a lettuce plant by crossing the
lettuce plants of the invention with themselves or another lettuce
plant. The invention also provides lettuce plants produced from
such a crossing as well as plant parts, seed, and tissue culture
therefrom.
Inventors: |
GROOT; Els; (Enkhuizen,
NL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Syngenta Participations |
Basel |
|
CH |
|
|
Assignee: |
Syngenta Participations AG
Basel
CH
|
Family ID: |
61872967 |
Appl. No.: |
15/293951 |
Filed: |
October 14, 2016 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12Q 1/6895 20130101;
C12Q 2600/13 20130101; A01H 6/1472 20180501; C12Q 2600/156
20130101; A01H 5/12 20130101 |
International
Class: |
A01H 5/12 20060101
A01H005/12; A01H 1/02 20060101 A01H001/02; A01H 4/00 20060101
A01H004/00; C12N 15/82 20060101 C12N015/82; C12Q 1/68 20060101
C12Q001/68 |
Claims
1. A seed of lettuce cultivar Julian, a representative sample of
seed having been deposited under ATCC Accession No. ______.
2. A plant of lettuce cultivar Julian, a representative sample of
seed having been deposited under ATCC Accession No. ______.
3. A lettuce plant, or a part thereof, having all the physiological
and morphological characteristics of the lettuce plant of claim
2.
4. (canceled)
5. (canceled)
6. A plant part of the lettuce plant of claim 2.
7. The plant part of claim 6, wherein the plant part is a leaf,
pollen, an ovule, an anther, a root, or a cell.
8. A tissue culture of regenerable cells of the plant of claim
2.
9. A lettuce plant regenerated from the tissue culture of claim 8
or a selfed progeny thereof, wherein said lettuce plant expresses
all of the physiological and morphological characteristics of
lettuce cultivar Julian.
10. A processed product from the plant of claim 2, wherein the
processed product comprises cut, sliced, ground, pureed, dried,
canned, jarred, washed, packaged, frozen and/or heated leaves.
11. A method of producing lettuce seed, the method comprising
crossing the plant of claim 2 with itself or a second lettuce plant
and harvesting the resulting seed.
12. A lettuce seed produced by the method of claim 11.
13. A lettuce plant, or part thereof, produced by growing the seed
of claim 12.
14. A doubled haploid plant produced from the lettuce plant of
claim 13.
15. A method for producing a seed of a lettuce plant derived from
the plant of claim 2, the method comprising: (a) crossing a plant
of lettuce cultivar Julian with a second lettuce plant; and (b)
allowing seed to form; (c) growing a plant from the seed of step
(b) to produce a plant derived from lettuce cultivar Julian; (d)
selfing the plant of step (c) or crossing it to a second lettuce
plant to form additional lettuce seed derived from lettuce cultivar
Julian; and (e) optionally repeating steps (c) and (d) one or more
times to generate further derived lettuce seed from lettuce
cultivar Julian, wherein in step (c) a plant is grown from the
additional lettuce seed of step (d) in place of growing a plant
from the seed of step (b).
16. (canceled)
17. (canceled)
18. A method of vegetatively propagating the plant of claim 2, the
method comprising: (a) collecting tissue capable of being
propagated from a plant of lettuce cultivar Julian; (b) cultivating
the tissue to obtain proliferated shoots; (c) rooting the
proliferated shoots to obtain rooted plantlets; and (d) optionally,
growing plants from the rooted plantlets.
19. Lettuce plantlet or plant obtained by the method of claim 18,
wherein the lettuce plantlet or plant expresses all of the
physiological and morphological characteristics of lettuce cultivar
Julian.
20. A method of introducing a desired added trait into lettuce
cultivar Julian, the method comprising: (a) crossing the plant of
claim 2 with a lettuce plant that comprises a desired added trait
to produce F1 progeny; (b) selecting an F1 progeny that comprises
the desired added trait; (c) crossing the selected F1 progeny with
lettuce cultivar Julian to produce backcross progeny; (d) selecting
a backcross progeny comprising the desired added trait and
essentially all of the physiological and morphological
characteristics of the lettuce cultivar Julian; and (e) optionally
repeating steps (c) and (d) one or more times to produce a plant
derived from lettuce cultivar Julian comprising a desired added
trait and essentially all of the physiological and morphological
characteristics of lettuce cultivar Julian, wherein in step (c) the
selected backcross progeny produced in step (d) is used in place of
the selected F1 progeny of step (b).
21. The method of claim 20, wherein the desired added trait is male
sterility, pest resistance, insect resistance, disease resistance,
herbicide resistance, or any combination thereof.
22. A lettuce plant produced by the method of claim 20 or a selfed
progeny thereof, wherein the lettuce plant has the desired added
trait.
23. Seed of the plant of claim 22, wherein the seed produces a
plant that has the desired added trait.
24. Seed that produces the plant of claim 22.
25. A method of producing a plant of lettuce cultivar Julian
comprising a desired added trait, the method comprising introducing
a transgene conferring the desired trait into the plant of claim
2.
26. A lettuce plant produced by the method of claim 25 or a selfed
progeny thereof, wherein the lettuce plant has the desired added
trait.
27. Seed of the plant of claim 26, wherein the seed produces a
plant that has the desired added trait.
28. A method of determining a genotype of lettuce cultivar Julian,
the method comprising: (a) obtaining a sample of nucleic acids from
the plant of claim 2; and (b) detecting a polymorphism in the
nucleic acid sample.
29. A method of producing a lettuce leaf, the method comprising:
(a) growing the lettuce plant according to claim 2 to produce a
lettuce leaf; and (b) harvesting the lettuce leaf.
30. A method of producing a lettuce leaf, the method comprising:
(a) growing the lettuce plant according to claim 22 to produce a
lettuce leaf; and (b) harvesting the lettuce leaf.
Description
FIELD OF THE INVENTION
[0001] This invention is in the field of lettuce plants.
BACKGROUND OF THE INVENTION
[0002] The present invention relates to a butterhead lettuce
(Lactuca sativa L.) variety designated Julian.
[0003] Practically speaking, 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 Cichoreae
tribe of the Asteraceae (Compositae) family. Lettuce is related to
chicory, sunflower, aster, dandelion, artichoke, and chrysanthemum.
Sativa is one of about 300 species in the genus Lactuca. There are
seven different morphological types of lettuce. The crisphead group
includes the iceberg and batavian types. Iceberg lettuce has a
large, firm head with a crisp texture and a white or creamy yellow
interior. The batavian lettuce predates the iceberg type and has a
smaller and less firm head. The butterhead group has a small, soft
head with an almost oily texture. The romaine, also known as cos
lettuce, has elongated upright leaves forming a loose, loaf-shaped
head and the outer leaves are usually dark green. Leaf lettuce
comes in many varieties, none of which form a head, and include the
green oak leaf variety. Latin lettuce looks like a cross between
romaine and butterhead. Stem lettuce has long, narrow leaves and
thick, edible stems. Oilseed lettuce is a type grown for its large
seeds that are pressed to obtain oil. Latin lettuce, stem lettuce,
and oilseed lettuce are seldom seen in the United States.
[0004] Presently, there are over one thousand known lettuce
cultivars. As a crop, lettuce is grown commercially wherever
environmental conditions permit the production of an economically
viable yield.
[0005] Lettuce, in general, and leaf lettuce in particular, is an
important and valuable vegetable crop. Thus, there is an ongoing
need for improved lettuce varieties.
SUMMARY OF THE INVENTION
[0006] According to the invention, there is provided a novel
lettuce cultivar designated Julian, also known as LS14616. Thus,
the invention also encompasses the seeds of lettuce cultivar
Julian, the plants of lettuce cultivar Julian, plant parts of the
lettuce cultivar Julian (including leaves, seed, gametes), methods
of producing seed from lettuce cultivar Julian, and method for
producing a lettuce plant by crossing the lettuce cultivar Julian
with itself or another lettuce plant, methods for producing a
lettuce plant containing in its genetic material one or more
transgenes, and the transgenic lettuce plants produced by that
method. The invention also relates to methods for producing other
lettuce plants derived from lettuce cultivar Julian and to lettuce
plants, parts thereof and seed derived by the use of those methods.
The present invention further relates to hybrid lettuce seeds and
plants (and parts thereof including leaves) produced by crossing
lettuce cultivar Julian with another lettuce plant.
[0007] In another aspect, the present invention provides
regenerable cells for use in tissue culture of lettuce cultivar
Julian. In embodiments, the tissue culture is capable of
regenerating plants having all or essentially all of the
physiological and morphological characteristics of the foregoing
lettuce plant and/or of regenerating plants having the same or
substantially the same genotype as the foregoing lettuce plant. In
exemplary embodiments, the regenerable cells in such tissue
cultures are meristematic cells, cotyledons, hypocotyl, leaves,
pollen, embryos, roots, root tips, anthers, pistils, ovules,
shoots, stems, petiole, pith, flowers, capsules and/or seeds as
well as callus and/or protoplasts derived from any of the
foregoing. Still further, the present invention provides lettuce
plants regenerated from the tissue cultures of the invention.
[0008] As a further aspect, the invention provides a method of
producing lettuce seed, the method comprising crossing a plant of
lettuce cultivar Julian with itself or a second lettuce plant.
Optionally, the method further comprises collecting the seed.
[0009] Another aspect of the invention provides methods for
producing hybrids and other lettuce plants derived from lettuce
cultivar Julian. Lettuce plants derived by the use of those methods
are also part of the invention as well as plant parts, seed,
gametes and tissue culture from such hybrid or derived lettuce
plants.
[0010] In representative embodiments, a lettuce plant derived from
lettuce cultivar Julian comprises cells comprising at least one set
of chromosomes derived from lettuce cultivar Julian. In
embodiments, a lettuce plant or population of lettuce plants
derived from lettuce cultivar Julian comprises, on average, at
least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,
70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles (i.e.,
theoretical allelic content; TAC) from lettuce cultivar Julian,
e.g., at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%,
55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the
genetic complement of lettuce cultivar Julian. In embodiments, the
lettuce plant derived from lettuce cultivar Julian is one, two,
three, four, five or more breeding crosses removed from lettuce
cultivar Julian.
[0011] In embodiments, a hybrid or derived plant from lettuce
cultivar Julian comprises a desired added trait(s). In
representative embodiments, a lettuce plant derived from lettuce
cultivar Julian comprises all of the morphological and
physiological characteristics of lettuce cultivar Julian (e.g., as
described in Table 1). In embodiments, the lettuce plant derived
from lettuce cultivar Julian comprises essentially all of the
morphological and physiological characteristics of lettuce cultivar
Julian (e.g., as described in Table 1), with the addition of a
desired added trait(s).
[0012] The invention also relates to methods for producing a
lettuce plant comprising in its genetic material one or more
transgenes and to the transgenic lettuce plant produced by those
methods (and progeny lettuce plants comprising the transgene). Also
provided are plant parts, seed and tissue culture from such
transgenic lettuce plants, optionally wherein one or more cells in
the plant part, seed, or tissue culture comprises the transgene.
The transgene can be introduced via plant transformation and/or
breeding techniques.
[0013] In another aspect, the present invention provides for single
gene converted plants of lettuce cultivar Julian. Plant parts,
seed, and tissue culture from such single gene converted plants are
also contemplated by the present invention. The single transferred
gene may be a dominant or recessive allele. In representative
embodiments, the single transferred gene confers such traits as
male sterility, herbicide resistance, pest resistance (e.g., insect
and/or nematode resistance), modified fatty acid metabolism,
modified carbohydrate metabolism, disease resistance (e.g., for
bacterial, fungal and/or viral disease), male fertility, enhanced
nutritional quality, improved appearance (e.g., color), improved
salt tolerance, industrial usage, or any combination thereof. The
single gene may be a naturally occurring lettuce gene or a
transgene introduced into lettuce through genetic engineering
techniques.
[0014] The invention further provides methods for developing
lettuce plants in a lettuce plant breeding program using plant
breeding techniques including, for example, recurrent selection,
backcrossing, pedigree breeding, double haploid techniques,
restriction fragment length polymorphism enhanced selection,
genetic marker enhanced selection and/or transformation. Seeds,
lettuce plants, and parts thereof, produced by such breeding
methods are also part of the invention.
[0015] The invention also provides methods of multiplication or
propagation of lettuce plants of the invention, which can be
accomplished using any method known in the art, for example, via
vegetative propagation and/or seed.
[0016] The invention further provides a method of producing food or
feed comprising (a) obtaining a lettuce plant of the invention,
optionally wherein the plant has been cultivated to maturity, and
(b) collecting at least one lettuce plant or part thereof (e.g.,
leaves) from the plant.
[0017] Additional aspects of the invention include harvested
products and processed products from the lettuce plants of the
invention. A harvested product can be a whole plant or any plant
part, as described herein. Thus, in some embodiments, a
non-limiting example of a harvested product includes a seed, a leaf
and/or a stem.
[0018] In representative embodiments, a processed product includes,
but is not limited to: cut, sliced, ground, pureed, dried, canned,
jarred, washed, packaged, frozen and/or heated leaves and/or seeds
of the lettuce plants of the invention, or any other part thereof.
In embodiments, a processed product includes a sugar or other
carbohydrate, fiber, protein and/or aromatic compound that is
extracted, purified or isolated from a lettuce plant of the
invention. In embodiments, the processed product includes washed
and packaged leaves (or parts thereof) of the invention.
[0019] The seed of the invention can optionally be provided as an
essentially homogenous population of seed of a single plant or
cultivar. Essentially homogenous populations of seed are generally
free from substantial numbers of other seed, e.g., at least about
90%, 95%, 96%, 97%, 98% or 99% pure.
[0020] In representative embodiments, the invention provides a seed
of lettuce cultivar Julian.
[0021] As a further aspect, the invention provides a plant of
lettuce cultivar Julian.
[0022] As an additional aspect, the invention provides a lettuce
plant, or a part thereof, having all or essentially all of the
physiological and morphological characteristics of a plant of
lettuce cultivar Julian.
[0023] As another aspect, the invention provides leaves and/or seed
of the lettuce plants of the invention and a processed product from
the leaves and/or seed of the inventive lettuce plants.
[0024] As still another aspect, the invention provides a method of
producing lettuce seed, the method comprising crossing a lettuce
plant of the invention with itself or a second lettuce plant. The
invention also provides seed produced by this method and plants
produced by growing the seed.
[0025] As yet a further aspect, the invention provides a method for
producing a seed of a lettuce plant derived from lettuce cultivar
Julian, the method comprising: (a) crossing a lettuce plant of
lettuce cultivar Julian with a second lettuce plant; and (b)
allowing seed of a lettuce plant derived from lettuce cultivar
Julian to form. In embodiments, the method further comprises: (c)
growing a plant from the seed derived from lettuce cultivar Julian
of step (b); (d) selfing the plant grown from the lettuce seed
derived from lettuce cultivar Julian or crossing it to a second
lettuce plant to form additional lettuce seed derived from lettuce
cultivar Julian, and (e) repeating steps (c) and (d) 0 or more
times to generate further derived lettuce seed. Optionally, the
method comprises: (e) repeating steps (c) and (d) one or more times
(e.g., one to three, one to five, one to six, one to seven, one to
ten, three to five, three to six, three to seven, three to eight or
three to ten times) to generate further derived lettuce plants. As
another option, the method can comprise collecting the seed. The
invention also provides seed produced by these methods and plants
produced by growing the seed.
[0026] As another aspect, the invention provides a method of
producing lettuce leaves, the method comprising: (a) obtaining a
plant of lettuce cultivar Julian, optionally wherein the plant has
been cultivated to maturity; and (b) collecting leaves from the
plant. The invention also provides the leaves produced by this
method.
[0027] Still further, as another aspect, the invention provides a
method of vegetatively propagating a plant of lettuce cultivar
Julian. In a non-limiting example, the method comprises: (a)
collecting tissue capable of being propagated from a plant of
lettuce cultivar Julian; (b) cultivating the tissue to obtain
proliferated shoots; and (c) rooting the proliferated shoots to
obtain rooted plantlets. Optionally, the invention further
comprises growing plants from the rooted plantlets. The invention
also encompasses the plantlets and plants produced by these
methods.
[0028] As an additional aspect, the invention provides a method of
introducing a desired added trait into lettuce cultivar Julian, the
method comprising: (a) crossing a first plant of lettuce cultivar
Julian with a second lettuce plant that comprises a desired trait
to produce F.sub.1 progeny; (b) selecting an F.sub.1 progeny that
comprises the desired trait; (c) crossing the selected F.sub.1
progeny with lettuce cultivar Julian to produce backcross progeny;
and (d) selecting backcross progeny comprising the desired trait to
produce a plant derived from lettuce cultivar Julian comprising a
desired trait. In embodiments, the selected progeny comprises all
or essentially all the morphological and physiological
characteristics of the first plant of lettuce cultivar Julian.
Optionally, the method further comprises: (e) repeating steps (c)
and (d) one or more times in succession (e.g., one to three, one to
five, one to six, one to seven, one to ten, three to five, three to
six, three to seven, three to eight or three to ten times) to
produce a plant derived from lettuce cultivar Julian comprising the
desired trait.
[0029] In representative embodiments, the invention also provides a
method of producing a plant of lettuce cultivar Julian comprising a
desired added trait, the method comprising introducing a transgene
conferring the desired trait into a plant of lettuce cultivar
Julian. The transgene can be introduced by transformation methods
(e.g., genetic engineering) or breeding techniques. In embodiments,
the plant comprising the transgene comprises all or essentially all
of the morphological and physiological characteristics of lettuce
cultivar Julian.
[0030] The invention also provides lettuce plants produced by the
methods of the invention, wherein the lettuce plant has the desired
added trait as well as seed from such lettuce plants.
[0031] According to the foregoing methods, the desired added trait
can be any suitable trait known in the art including, for example,
male sterility, male fertility, herbicide resistance, insect or
pest (e.g., insect and/or nematode) resistance, modified fatty acid
metabolism, modified carbohydrate metabolism, disease resistance
(e.g., for bacterial, fungal and/or viral disease), enhanced
nutritional quality, increased sweetness, increased flavor,
improved ripening control, improved salt tolerance, industrial
usage, or any combination thereof.
[0032] In representative embodiments, a transgene conferring
herbicide resistance confers resistance to glyphosate,
sulfonylurea, imidazolinone, dicamba, glufosinate, phenoxy
proprionic acid, L-phosphinothricin, cyclohexone, cyclohexanedione,
triazine, benzonitrile, or any combination thereof.
[0033] In representative embodiments, a transgene conferring pest
resistance (e.g., insect and/or nematode resistance) encodes a
Bacillus thuringiensis endotoxin.
[0034] In representative embodiments, transgenic plants,
transformed plants, hybrid plants and lettuce plants derived from
lettuce cultivar Julian have at least 3, 4, 5, 6, 7, 8, 9, 10 or
more of the morphological and physiological characteristics of
lettuce cultivar Julian (for example, as described in Table 1), or
even all of the morphological and physiological characteristics of
lettuce cultivar Julian, so that said plants are not significantly
different for said traits than lettuce cultivar Julian, as
determined at the 5% significance level when grown in the same
environmental conditions; optionally, with the presence of one or
more desired additional traits (e.g., male sterility, disease
resistance, pest or insect resistance, herbicide resistance, and
the like).
[0035] The invention also encompasses plant parts, plant material,
pollen, ovules, leaves, fruit and seed from the lettuce plants of
the invention. Also provided is a tissue culture of regenerable
cells from the lettuce plants of the invention, where optionally,
the regenerable cells are: (a) embryos, meristem, leaves, pollen,
cotyledons, hypocotyls, roots, root tips, anthers, flowers,
pistils, ovules, seed, shoots, stems, stalks, petioles, pith and/or
capsules; or (b) callus or protoplasts derived from the cells of
(a). Further provided are lettuce plants regenerated from a tissue
culture of the invention.
[0036] In still yet another aspect, the invention provides a method
of determining a genetic characteristic of lettuce cultivar Julian
or a progeny thereof, e.g., a method of determining a genotype of
lettuce cultivar Julian or a progeny thereof. In embodiments, the
method comprises detecting in the genome of a Julian plant, or a
progeny plant thereof, at least a first polymorphism. To
illustrate, in embodiments, the method comprises obtaining a sample
of nucleic acids from the plant and detecting at least a first
polymorphism in the nucleic acid sample (e.g., using one or more
molecular markers). Optionally, the method may comprise detecting a
plurality of polymorphisms (e.g., two or more, three or more, four
or more, five or more, six or more, eight or more or ten or more
polymorphisms, etc.) in the genome of the plant. In representative
embodiments, the method further comprises storing the results of
the step of detecting the polymorphism(s) on a computer readable
medium. The invention further provides a computer readable medium
produced by such a method.
[0037] In addition to the exemplary aspects and embodiments
described above, the invention is described in more detail in the
description of the invention set forth below.
DETAILED DESCRIPTION OF THE INVENTION
[0038] It should be appreciated that the invention can be embodied
in different forms and should not be construed as limited to the
embodiments set forth herein. Rather, these embodiments are
provided so that this disclosure will be thorough and complete, and
will fully convey the scope of the invention to those skilled in
the art.
[0039] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. The
terminology used in the description of the invention herein is for
the purpose of describing particular embodiments only and is not
intended to be limiting of the invention.
[0040] Unless the context indicates otherwise, it is specifically
intended that the various features and embodiments of the invention
described herein can be used in any combination.
[0041] Moreover, the present invention also contemplates that in
some embodiments of the invention, any feature or combination of
features set forth herein can be excluded or omitted. To
illustrate, if the specification states that a composition
comprises components A, B and C, it is specifically intended that
any of A, B or C, or a combination thereof, can be omitted and
disclaimed singularly or in any combination.
[0042] Unless otherwise defined, all technical and scientific terms
used herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. The
terminology used in the description of the invention herein is for
the purpose of describing particular embodiments only and is not
intended to be limiting of the invention.
[0043] All publications, patent applications, patents, and other
references mentioned herein are incorporated by reference in their
entirety.
Definitions
[0044] In the description and tables that follow, a number of terms
are used. In order to provide a clear and consistent understanding
of the specification and claims, including the scope to be given
such terms, the following definitions are provided:
[0045] As used in the description of the invention and the appended
claims, the singular forms "a," "an" and "the" are intended to
include the plural forms as well, unless the context clearly
indicates otherwise.
[0046] As used herein, "and/or" refers to and encompasses any and
all possible combinations of one or more of the associated listed
items, as well as the lack of combinations when interpreted in the
alternative ("or").
[0047] The term "about," as used herein when referring to a
measurable value such as a dosage or time period and the like, is
meant to encompass variations of .+-.20%, +10%, .+-.5%, +1%, +0.5%,
or even .+-.0.1% of the specified amount.
[0048] The term "comprise," "comprises" and "comprising" as used
herein, specify the presence of the stated features, integers,
steps, operations, elements, and/or components, but do not preclude
the presence or addition of one or more other features, integers,
steps, operations, elements, components, and/or groups thereof.
[0049] As used herein, the transitional phrase "consisting
essentially of" means that the scope of a claim is to be
interpreted to encompass the specified materials or steps recited
in the claim "and those that do not materially affect the basic and
novel characteristic(s)" of the claimed invention. See, In re Herz,
537 F.2d 549, 551-52, 190 U.S.P.Q. 461, 463 (CCPA 1976) (emphasis
in the original); see also MPEP .sctn. 2111.03. Thus, the term
"consisting essentially of" when used in a claim or the description
of this invention is not intended to be interpreted to be
equivalent to "comprising."
[0050] "Allele". An allele is any of one or more alternative forms
of a gene, all of which relate to a trait or characteristic. In a
diploid cell or organism, the two alleles of a given gene occupy
corresponding loci on a pair of homologous chromosomes.
[0051] "Backcrossing". Backcrossing is a process in which a breeder
repeatedly crosses hybrid progeny back to one of the parents, for
example, a first generation hybrid F.sub.1 with one of the parental
genotype of the F.sub.1 hybrid.
[0052] "Big Vein virus". Big vein is a disease of lettuce caused by
Lettuce Mirafiori Big Vein Virus which is transmitted by the fungus
Olpidium virulentus, with vein clearing and leaf shrinkage
resulting in plants of poor quality and reduced marketable
value.
[0053] "Bolting". The premature development of a flowering stalk,
and subsequent seed, before a plant produces a food crop. Bolting
is typically caused by late planting when temperatures are low
enough to cause vernalization of the plants.
[0054] "Bremia lactucae". An Oomycete that causes downy mildew in
lettuce in cooler growing regions.
[0055] "Core length". Length of the internal lettuce stem measured
from the base of the cut and trimmed head to the tip of the
stem.
[0056] "Corky root". A disease caused by the bacterium Sphingomonas
suberifaciens, which causes the entire taproot to become brown,
severely cracked, and non-functional.
[0057] "Cotyledon". One of the first leaves of the embryo of a seed
plant; typically one or more in monocotyledons, two in
dicotyledons, and two or more in gymnosperms.
[0058] "Double haploid line". A stable inbred line achieved by
doubling the chromosomes of a haploid line, e.g., from anther
culture. For example, some pollen grains (haploid) cultivated under
specific conditions develop plantlets containing In chromosomes.
The chromosomes in these plantlets are then induced to "double"
(e.g., using chemical means) resulting in cells containing 2n
chromosomes. The progeny of these plantlets are termed "double
haploid" and are essentially not segregating any more (e.g., are
stable). The term "double haploid" is used interchangeably herein
with "dihaploid."
[0059] "Essentially all the physiological and morphological
characteristics". A plant having "essentially all the physiological
and morphological characteristics" means a plant having the
physiological and morphological characteristics of the recurrent
parent, except for the characteristics derived from the converted
gene(s).
[0060] "First water date". The date the seed first receives
adequate moisture to germinate. This can and often does equal the
planting date.
[0061] "Gene". As used herein, "gene" refers to a segment of
nucleic acid comprising an open reading frame. A gene can be
introduced into a genome of a species, whether from a different
species or from the same species, using transformation or various
breeding methods.
[0062] "Head diameter". Diameter of the cut and trimmed head,
sliced vertically, and measured at the widest point perpendicular
to the stem.
[0063] "Head height". Height of the cut and trimmed head, sliced
vertically, and measured from the base of the cut stem to the cap
leaf.
[0064] "Head weight". Weight of saleable lettuce head, cut and
trimmed to market specifications.
[0065] "Inbred line": As used herein, the phrase "inbred line"
refers to a genetically homozygous or nearly homozygous population.
An inbred line, for example, can be derived through several cycles
of sib crossing and/or selfing and/or via double haploid
production. In some embodiments, inbred lines breed true for one or
more traits of interest. An "inbred plant" or "inbred progeny" is
an individual sampled from an inbred line.
[0066] "Lettuce Mosaic virus". A disease that can cause a stunted,
deformed, or mottled pattern in young lettuce and yellow, twisted,
and deformed leaves in older lettuce.
[0067] "Maturity date". Maturity refers to the stage when the
plants are of full size and/or optimum weight and/or in marketable
form to be of commercial or economic value.
[0068] "Nasonovia ribisnigri". A lettuce aphid that colonizes the
innermost leaves of the lettuce plant, contaminating areas that
cannot be treated easily with insecticides.
[0069] "Plant." As used herein, the term "plant" includes plant
cells, plant protoplasts, plant cell tissue cultures from which
plants can be regenerated, plant calli, plant clumps, and plant
cells that are intact in plants or parts of plants, such as leaves,
pollen, embryos, cotyledons, hypocotyl, roots, root tips, anthers,
pistils, flowers, ovules, seeds, fruit, stems, and the like.
[0070] "Plant material". The terms "plant material" and "material
obtainable from a plant" are used interchangeably herein and refer
to any plant material obtainable from a plant including without
limitation, leaves, stems, roots, flowers or flower parts, fruits,
pollen, ovules, zygotes, seeds, cuttings, cell or tissue cultures,
or any other part or product of the plant.
[0071] "Plant part". As used herein, a "plant part" includes any
part, organ, tissue or cell of a plant including without limitation
an embryo, meristem, leaf, pollen, cotyledon, hypocotyl, root, root
tip, anther, flower, flower bud, pistil, ovule, seed, shoot, stem,
stalk, petiole, pith, capsule, a scion, a rootstock and/or a fruit
including callus and protoplasts derived from any of the
foregoing.
[0072] "Quantitative Trait Loci". Quantitative Trait Loci (QTL)
refers to genetic loci that control to some degree, numerically
representable traits that are usually continuously distributed.
[0073] "Ratio of head height/diameter". Head height divided by the
head diameter is an indication of the head shape; <1 is
flattened, 1=round, and >1 is pointed.
[0074] "Regeneration". Regeneration refers to the development of a
plant from tissue culture.
[0075] "Resistance". As used herein the terms "resistance" and
"tolerance" (and grammatical variations thereof) are used
interchangeably to describe plants that show reduced or essentially
no symptoms to a specific biotic (e.g., a pest, pathogen or
disease) or abiotic (e.g., exogenous or environmental, including
herbicides) factor or stressor. In some embodiments, "resistant" or
"tolerant" plants show some symptoms but are still able to produce
marketable product with an acceptable yield, e.g., the yield may
still be reduced and/or the plants may be stunted as compared with
the yield or growth in the absence of the biotic and/or abiotic
factor or stressor. Those skilled in the art will appreciate that
the degree of resistance or tolerance may be assessed with respect
to a plurality or even an entire field of plants. A lettuce plant
may be considered "resistant" or "tolerant" if resistance/tolerance
is observed over a plurality of plants (e.g., an average), even if
particular individual plants may be susceptible to the biotic or
abiotic factor or stressor.
[0076] "RHS". RHS refers to the Royal Horticultural Society of
England which publishes an official botanical color chart
quantitatively identifying colors according to a defined numbering
system. The chart may be purchased from Royal Horticulture Society
Enterprise Ltd., RHS Garden; Wisley, Woking; Surrey GU236QB,
UK.
[0077] "Single gene converted". A single gene converted or
conversion plant refers to a plant that is developed by plant
breeding techniques (e.g., backcrossing) or via genetic engineering
wherein essentially all of the desired morphological and
physiological characteristics of a line are recovered in addition
to the single gene transferred into the line via the plant breeding
technique or via genetic engineering.
[0078] "Substantially equivalent characteristic". A characteristic
that, when compared, does not show a statistically significant
difference (e.g., p=0.05) from the mean.
[0079] "Tip burn". Means a browning of the edges or tips of lettuce
leaves that is a physiological response to a lack of calcium.
[0080] "Tomato Bushy Stunt". Also called "lettuce necrotic stunt".
A disease that causes stunting of growth and leaf mottling.
[0081] "Transgene". A nucleic acid of interest that can be
introduced into the genome of a plant by genetic engineering
techniques (e.g., transformation) or breeding. The transgene can be
from the same or a different species. If from the same species, the
transgene can be an additional copy of a native coding sequence or
can present the native sequence in a form or context (e.g.,
different genomic location and/or in operable association with
exogenous regulatory elements such as a promoter) than is found in
the native state. The transgene can comprise an open reading frame
encoding a polypeptide or can encode a functional non-translated
RNA (e.g., RNAi).
Botanical Description of the Lettuce Cultivar Julian.
[0082] Lettuce variety Julian is a butterhead lettuce. Julian is a
brilliant green butterhead variety, with an attractive look and
good volume and bottom quality. This variety has medium bolting
resistance, and is suitable for spring and autumn sowings.
[0083] Lettuce cultivar Julian has the following morphologic and
other characteristics, described in Table 1.
TABLE-US-00001 TABLE 1 Variety Description Information. Comparison
with similar variety. Characteristic in State of Denomination which
the similar State of expression of similar variety is expression of
candidate variety different of similar variety variety (Julian)
Servis Plant: diameter Medium to Large Medium Servis Leaf:
intensity of Medium to Dark Medium to Dark color of outer (darker)
leaves
[0084] Additional Information.
TABLE-US-00002 Trait Name Value Variety code LS14616 Multiplication
indicator Open-pollinated Type of culture Open field Period of
Growing Spring and autumn Seed: color White Seedling: anthocyanin
coloration Absent Leaf: attitude (10 to 12 leaf stage) Semi-erect
Leaf blade: division (10 to 12 leaf Entire stage) Plant: fasciation
(at flowering stage) Present Plant: intensity of fascination Weak
(flowering plant) Plant: diameter Medium Plant: head formation
Closed head (overlapping) Head: degree of overlapping of upper
Medium part of leaves Head: density Dense Head: size Medium Head:
shape in longitudinal section Broad elliptic Plant: type Butterhead
Time of harvest maturity Early to Medium Leaf: thickness Medium
Leaf: attitude at 10-12 leaf stage Semi-erect Leaf: shape Circular
to Transverse broad elliptic Leaf: shape of tip Rounded Leaf: hue
of green color of outer Absent to Yellowish leaves Leaf: color of
outer leaves (at harvest Green maturity) Leaf: intensity of color
of outer leaves Medium to Dark Leaf: anthocyanin coloration Absent
Leaf: glossiness of upper side Weak to Medium Leaf: blistering Weak
to Medium Leaf: size of blisters Medium Leaf blade: degree of
undulation of Very weak to Weak margin Leaf blade: incisions of
margin on Absent apical part Leaf blade: venation Not flabellate
Axillary sprouting Weak to Medium Time of harvest maturity Early to
Medium Time of beginning of bolting (under Medium long day
conditions)
[0085] Resistances to Pests and Diseases.
[0086] Bremia lactucae (downy mildew): Highly resistant to races
1-31
[0087] Lettuce mosaic virus (LMV) strain Ls-1: Highly resistant
[0088] Nasovonia ribisnigri biotype 0 (Nr: 0): Highly resistant
[0089] The cultivar has shown uniformity and stability for the
expressed traits, within the limits of environmental influence for
the traits. No variant traits have been observed or are expected in
cultivar Julian.
Further Embodiments of the Invention
[0090] With the advent of molecular biological techniques that have
allowed the isolation and characterization of genes that encode
specific protein products, scientists in the field of plant biology
developed a strong interest in engineering the genome of plants to
contain and express foreign nucleic acids including additional or
modified versions of native (endogenous) nucleic acids (optionally
driven by a non-native promoter) in order to alter the traits of a
plant in a specific manner. Any nucleic acid sequences, whether
from a different species or from the same species, which are
introduced into the genome using transformation or various breeding
methods, are referred to herein collectively as "transgenes." Over
the last fifteen to twenty years, several methods for producing
transgenic plants have been developed, and in particular
embodiments the present invention also relates to transformed
versions of lettuce plants disclosed herein.
[0091] Genetic engineering techniques can be used (alone or in
combination with breeding methods) to introduce one or more desired
added traits into plant, for example, lettuce cultivar Julian or
progeny or lettuce plants derived thereof.
[0092] Plant transformation generally involves the construction of
an expression vector that will function in plant cells. Optionally,
such a vector comprises one or more nucleic acids comprising a
coding sequence for a polypeptide or an untranslated functional RNA
under control of, or operatively linked to, a regulatory element
(for example, a promoter). In representative embodiments, the
vector(s) 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 as described herein to
incorporate transgenes into the genetic material of the lettuce
plant.
[0093] Additional methods include, but are not limited to,
expression vectors introduced into plant tissues using a direct
nucleic acid transfer method, such as microprojectile-mediated
delivery (e.g., with a biolistic device), DNA injection,
Agrobacterium-mediated transformation, electroporation, and the
like. Transformed plants obtained from the plants (and parts and
tissue culture thereof) of the invention are intended to be within
the scope of this invention.
Expression Vectors for Plant Transformation--Selectable
Markers.
[0094] Expression vectors typically include at least one nucleic
acid comprising or encoding a selectable marker, operably linked to
a regulatory element (for example, a promoter) that allows
transformed cells containing the marker to be either recovered by
negative selection, e.g., inhibiting growth of cells that do not
contain the selectable marker, or by positive selection, e.g.,
screening for the product encoded by the selectable marker. Many
commonly used selectable markers for plant transformation are well
known in the transformation art, and include, for example, nucleic
acids that code for enzymes that metabolically detoxify a selective
chemical agent which may be an antibiotic or an herbicide, or
nucleic acids that encode an altered target which is insensitive to
the inhibitor. Positive selection methods are also known in the
art.
[0095] One commonly used selectable marker for plant transformation
is a neomycin phosphotransferase II (nptII) coding sequence, for
example, isolated from transposon Tn5, which when placed under the
control of plant regulatory signals confers resistance to
kanamycin. Fraley, et al., PNAS, 80:4803 (1983). Another commonly
used selectable marker is hygromycin phosphotransferase, which
confers resistance to the antibiotic hygromycin. Vanden Elzen, et
al., Plant Mol. Biol., 5:299 (1985).
[0096] Additional selectable markers of bacterial origin that
confer resistance to antibiotics include gentamycin acetyl
transferase, streptomycin phosphotransferase,
aminoglycoside-3'-adenyl transferase, the bleomycin resistance
determinant. Hayford, et al., Plant Physiol., 86:1216 (1988);
Jones, et al., Mol. Gen. Genet., 210:86 (1987); Svab, et al., Plant
Mol. Biol., 14:197 (1990); Hille, et al., Plant Mol. Biol., 7:171
(1986). Other selectable markers confer resistance to herbicides
such as glyphosate, glufosinate, or bromoxynil. Comai, et al.,
Nature, 317:741-744 (1985); Gordon-Kamm, et al., Plant Cell,
2:603-618 (1990); and Stalker, et al., Science, 242:419-423
(1988).
[0097] Selectable markers for plant transformation that are not of
bacterial origin include, for example, mouse dihydrofolate
reductase, plant 5-enolpyruvylshikimate-3-phosphate synthase, and
plant acetolactate synthase. Eichholtz, et al., Somatic Cell Mol.
Genet., 13:67 (1987); Shah, et al., Science, 233:478 (1986); and
Charest, et al., Plant Cell Rep., 8:643 (1990).
[0098] Another class of selectable marker for plant transformation
involves screening of presumptively transformed plant cells rather
than direct genetic selection of transformed cells for resistance
to a toxic substance such as an antibiotic. These selectable
markers are particularly useful to quantify or visualize the
spatial pattern of expression of a transgene in specific tissues
and are frequently referred to as a reporter gene because they can
be fused to transgene or regulatory sequence for the investigation
of nucleic acid expression. Commonly used reporters for screening
presumptively transformed cells include alpha-glucuronidase (GUS),
alpha-galactosidase, luciferase and chloramphenicol,
acetyltransferase. Jefferson, R. A., Plant Mol. Biol., 5:387
(1987); Teeri, et al., EMBO J., 8:343 (1989); Koncz, et al., PNAS,
84:131 (1987); and DeBlock, et al., EMBO J., 3:1681 (1984).
[0099] In vivo methods for visualizing GUS activity that do not
require destruction of plant tissues are available. Molecular
Probes, Publication 2908, IMAGENE GREEN, pp. 1-4 (1993) and
Naleway, et al., J. Cell Biol., 115:151a (1991).
[0100] Green Fluorescent Protein (GFP) is also utilized as a marker
for nucleic acid expression in prokaryotic and eukaryotic cells.
Chalfie, et al., Science, 263:802 (1994). GFP and mutants of GFP
may be used as screenable markers.
Expression Vectors for Plant Transformation-Promoters.
[0101] Transgenes included in expression vectors are generally
driven by a nucleotide sequence comprising a regulatory element
(for example, a promoter). Numerous types of promoters are well
known in the transformation arts, as are other regulatory elements
that can be used alone or in combination with promoters.
[0102] As used herein, "promoter" includes reference to a region of
DNA upstream from the start of transcription and involved in
recognition and binding of RNA polymerase and other proteins to
initiate transcription. A "plant promoter" is a promoter capable of
initiating transcription in plant cells.
[0103] Examples of promoters under developmental control include
promoters that preferentially initiate transcription in certain
tissues, such as leaves, roots, seeds, fibers, xylem vessels,
tracheids, or sclerenchyma. Such promoters are referred to as
"tissue-preferred." Promoters that initiate transcription only in
certain tissue are referred to as "tissue-specific." A "cell type"
specific promoter preferentially drives expression in certain cell
types in one or more organs, for example, vascular cells in roots
or leaves. An "inducible" promoter is a promoter that is under
environmental control. Examples of environmental conditions that
may affect transcription by inducible promoters include anaerobic
conditions or the presence of light. Tissue-specific,
tissue-preferred, cell type specific, and inducible promoters
constitute the class of "non-constitutive" promoters. A
"constitutive" promoter is a promoter that is active under most
environmental conditions.
A. Inducible Promoters:
[0104] An inducible promoter is operably linked to a nucleic acid
for expression in a plant. Optionally, the inducible promoter is
operably linked to a nucleotide sequence encoding a signal sequence
which is operably linked to a nucleic acid for expression in the
plant. With an inducible promoter, the rate of transcription
increases in response to an inducing agent.
[0105] Any inducible promoter can be used in the instant invention.
See Ward, et al., Plant Mol. Biol., 22:361-366 (1993). Exemplary
inducible promoters include, but are not limited to, that from the
ACEI system which responds to copper (Melt, et al., PNAS,
90:4567-4571 (1993)); promoter from the In2 gene from maize which
responds to benzenesulfonamide herbicide safeners (Hershey, et al.,
Mol. Gen. Genet., 227:229-237 (1991) and Gatz, et al., Mol. Gen.
Genet., 243:32-38 (1994)) or Tet repressor from Tn10 (Gatz, et al.,
Mol. Gen. Genet., 227:229-237 (1991)). A representative inducible
promoter is a promoter that responds to an inducing agent to which
plants do not normally respond. An exemplary inducible promoter is
the inducible promoter from a steroid hormone gene, the
transcriptional activity of which is induced by a
glucocorticosteroid hormone. Schena, et al., PNAS, 88:0421
(1991).
B. Constitutive Promoters:
[0106] A constitutive promoter is operably linked to a nucleic acid
for expression in a plant or the constitutive promoter is operably
linked to a nucleotide sequence encoding a signal sequence which is
operably linked to a nucleic acid for expression in a plant.
[0107] Many different constitutive promoters can be utilized in the
instant invention. Exemplary constitutive promoters include, but
are not limited to, the promoters from plant viruses such as the
35S promoter from CaMV (Odell, et al., Nature, 313:810-812 (1985))
and the promoters from such genes as rice actin (McElroy, et al.,
Plant Cell, 2:163-171 (1990)); ubiquitin (Christensen, et al.,
Plant Mol. Biol., 12:619-632 (1989) and Christensen, et al., Plant
Mol. Biol., 18:675-689 (1992)); pEMU (Last, et al., Theor. Appl.
Genet., 81:581-588 (1991)); MAS (Velten, et al., EMBO J.,
3:2723-2730 (1984)) and maize H3 histone (Lepetit, et al., Mol.
Gen. Genet., 231:276-285 (1992) and Atanassova, et al., Plant J., 2
(3):291-300 (1992)). The ALS promoter, XbaI/NcoI fragment 5' to the
Brassica napus ALS3 structural gene (or a nucleotide sequence
similarity to said XbaI/NcoI fragment), represents a particularly
useful constitutive promoter. See PCT Application No. WO
96/30530.
C. Tissue-Specific or Tissue-Preferred Promoters:
[0108] A tissue-specific promoter is operably linked to a nucleic
acid for expression in a plant. Optionally, the tissue-specific
promoter is operably linked to a nucleotide sequence encoding a
signal sequence which is operably linked to a nucleic acid for
expression in a plant. Plants transformed with a nucleic acid of
interest operably linked to a tissue-specific promoter transcribe
the nucleic acid of interest exclusively, or preferentially, in a
specific tissue.
[0109] Any tissue-specific or tissue-preferred promoter can be
utilized in the instant invention. Exemplary tissue-specific or
tissue-preferred promoters include, but are not limited to, a
root-preferred promoter, such as that from the phaseolin gene
(Murai, et al., Science, 23:476-482 (1983) and Sengupta-Gopalan, et
al., PNAS, 82:3320-3324 (1985)); a leaf-specific and light-induced
promoter such as that from cab or rubisco (Simpson, et al., EMBO
J., 4(11):2723-2729 (1985) and Timko, et al., Nature, 318:579-582
(1985)); an anther-specific promoter such as that from LAT52
(Twell, et al., Mol. Gen. Genet., 217:240-245 (1989)); a
pollen-specific promoter such as that from Zm13 (Guerrero, et al.,
Mol. Gen. Genet., 244:161-168 (1993)) or a microspore-preferred
promoter such as that from apg (Twell, et al., Sex. Plant Reprod.,
6:217-224 (1993)).
Signal Sequences for Targeting Proteins to Subcellular
Compartments.
[0110] Transport of polypeptides produced by transgenes to a
subcellular compartment such as the chloroplast, vacuole,
peroxisome, glyoxysome, cell wall, or mitochondrion, or for
secretion into the apoplast, is generally accomplished by means of
operably linking a nucleotide sequence encoding a signal sequence
to the 5' and/or 3' region of a nucleic acid encoding the
polypeptide of interest. Signal sequences at the 5' and/or 3' end
of the coding sequence target the polypeptide to particular
subcellular compartments.
[0111] The presence of a signal sequence can direct a polypeptide
to either an intracellular organelle or subcellular compartment or
for secretion to the apoplast. Many signal sequences are known in
the art. See, for example, Becker, et al., Plant Mol. Biol., 20:49
(1992); Close, P. S., Master's Thesis, Iowa State University
(1993); Knox, C., et al., "Structure and Organization of Two
Divergent Alpha-Amylase Genes from Barley," Plant Mol. Biol.,
9:3-17 (1987); Lerner, et al., Plant Physiol., 91:124-129 (1989);
Fontes, et al., Plant Cell, 3:483-496 (1991); Matsuoka, et al.,
PNAS, 88:834 (1991); Gould, et al., J. Cell. Biol., 108:1657
(1989); Creissen, et al., Plant J, 2:129 (1991); Kalderon, et al.,
A short amino acid sequence able to specify nuclear location, Cell,
39:499-509 (1984); and Steifel, et al., Expression of a maize cell
wall hydroxyproline-rich glycoprotein gene in early leaf and root
vascular differentiation, Plant Cell, 2:785-793 (1990).
Foreign Polypeptide Transgenes and Agronomic Transgenes.
[0112] With transgenic plants according to the present invention, a
foreign protein can be produced in commercial quantities. Thus,
techniques for the selection and propagation of transformed plants,
which are well understood in the art, yield a plurality of
transgenic plants which are harvested in a conventional manner, and
a foreign polypeptide then can be extracted from a tissue of
interest or from total biomass. Protein extraction from plant
biomass can be accomplished by known methods which are discussed,
for example, by Heney and Orr, Anal. Biochem., 114:92-6 (1981).
[0113] According to a representative embodiment, the transgenic
plant provided for commercial production of foreign protein is a
lettuce plant of the invention. In another embodiment, the biomass
of interest is seed. For the relatively small number of transgenic
plants that show higher levels of expression, a genetic map can be
generated, for example via conventional RFLP, PCR, and SSR
analysis, which identifies the approximate chromosomal location of
the integrated DNA molecule. For exemplary methodologies in this
regard, see Methods in Plant Molecular Biology and Biotechnology,
Glick and Thompson Eds., 269:284, CRC Press, Boca Raton (1993). Map
information concerning chromosomal location is useful for
proprietary protection of a subject transgenic plant. If
unauthorized propagation is undertaken and crosses made with other
germplasm, the map of the integration region can be compared to
similar maps for suspect plants, to determine if the latter have a
common parentage with the subject plant. Map comparisons can
involve hybridizations, RFLP, PCR, SSR, and sequencing, all of
which are conventional techniques.
[0114] Likewise, by means of the present invention, agronomic
transgenes and other desired added traits can be expressed in
transformed plants (and their progeny, e.g., produced by breeding
methods). More particularly, plants can be genetically engineered
to express various phenotypes of agronomic interest or other
desired added traits. Exemplary nucleic acids of interest in this
regard conferring a desired added trait(s) include, but are not
limited to, those categorized below:
A. Transgenes that Confer Resistance to Pests or Disease:
[0115] 1. Plant disease resistance transgenes. Plant defenses are
often activated by specific interaction between the product of a
disease resistance gene (R) in the plant and the product of a
corresponding avirulence (Avr) gene in the pathogen. A plant line
can be transformed with a cloned resistance transgene to engineer
plants that are resistant to specific pathogen strains. See, for
example, Jones, et al., Science, 266:789 (1994) (cloning of the
tomato Cf-9 gene for resistance to Cladosporium fulvum); Martin, et
al., Science, 262:1432 (1993) (tomato Pto gene for resistance to
Pseudomonas syringae pv. tomato encodes a protein kinase); and
Mindrinos, et al., Cell, 78:1089 (1994) (Arabidopsis RSP2 gene for
resistance to Pseudomonas syringae).
[0116] 2. A Bacillus thuringiensis protein, a derivative thereof,
or a synthetic polypeptide modeled thereon. See, for example,
Geiser, et al., Gene, 48:109 (1986), who disclose the cloning and
nucleotide sequence of a Bt delta-endotoxin gene. Moreover, DNA
molecules encoding delta-endotoxin transgenes can be purchased from
American Type Culture Collection, Manassas, Va., for example, under
ATCC Accession Nos. 40098, 67136, 31995, and 31998.
[0117] 3. A lectin. See, for example, the disclosure by Van Damme,
et al., Plant Mol. Biol., 24:25 (1994), who disclose the nucleotide
sequences of several Clivia miniata mannose-binding lectin
transgenes.
[0118] 4. A vitamin-binding protein such as avidin. See, e.g., PCT
Application No. US 93/06487. The application teaches the use of
avidin and avidin homologues as larvicides against insect
pests.
[0119] 5. An enzyme inhibitor, for example, a protease or
proteinase inhibitor, or an amylase inhibitor. See, for example,
Abe, et al., J. Biol. Chem., 262:16793 (1987) (nucleotide sequence
of rice cysteine proteinase inhibitor); Huub, et al., Plant Mol.
Biol., 21:985 (1993) (nucleotide sequence of cDNA encoding tobacco
proteinase inhibitor I); and Sumitani, et al., Biosci. Biotech.
Biochem., 57:1243 (1993) (nucleotide sequence of Streptomyces
nitrosporeus alpha-amylase inhibitor).
[0120] 6. An insect-specific hormone or pheromone, such as an
ecdysteroid and juvenile hormone, a variant thereof, a mimetic
based thereon, or an antagonist or agonist thereof. See, for
example, the disclosure by Hammock, et al., Nature, 344:458 (1990),
of baculovirus expression of cloned juvenile hormone esterase, an
inactivator of juvenile hormone.
[0121] 7. An insect-specific peptide or neuropeptide which, upon
expression, disrupts the physiology of the affected pest. For
example, see the disclosures of Regan, J. Biol. Chem., 269:9 (1994)
(expression cloning yields DNA coding for insect diuretic hormone
receptor) and Pratt, et al., Biochem. Biophys. Res. Comm., 163:1243
(1989) (an allostatin is identified in Diploptera puntata). See
also, U.S. Pat. No. 5,266,317 to Tomalski, et al., who disclose
transgenes encoding insect-specific, paralytic neurotoxins.
[0122] 8. An insect-specific venom produced in nature, by a snake,
a wasp, etc. For example, see Pang, et al, Gene, 116:165 (1992),
for disclosure of heterologous expression in plants of a transgene
coding for a scorpion insectotoxic peptide.
[0123] 9. An enzyme responsible for a hyper-accumulation of a
monoterpene, a sesquiterpene, a steroid, hydroxamic acid, a
phenylpropanoid derivative, or another non-protein molecule with
insecticidal activity.
[0124] 10. An enzyme involved in the modification, including the
post-translational modification, of a biologically active molecule;
for example, a glycolytic enzyme, a proteolytic enzyme, a lipolytic
enzyme, a nuclease, a cyclase, a transaminase, an esterase, a
hydrolase, a phosphatase, a kinase, a phosphorylase, a polymerase,
an elastase, a chitinase, and a glucanase, whether natural or
synthetic. See PCT Application No. WO 93/02197 in the name of
Scott, et al., which discloses the nucleotide sequence of a callase
transgene. DNA molecules which contain chitinase-encoding sequences
can be obtained, for example, from the ATCC under Accession Nos.
39637 and 67152. See also, Kramer, et al., Insect Biochem. Mol.
Biol., 23:691 (1993), who teach the nucleotide sequence of a cDNA
encoding tobacco hornworm chitinase, and Kawalleck, et al., Plant
Mol. Biol., 21:673 (1993), who provide the nucleotide sequence of
the parsley ubi4-2 polyubiquitin transgene.
[0125] 11. A molecule that stimulates signal transduction. For
example, see the disclosure by Botella, et al., Plant Mol. Biol.,
24:757 (1994), of nucleotide sequences for mung bean calmodulin
cDNA clones, and Griess, et al., Plant Physiol., 104:1467 (1994),
who provide the nucleotide sequence of a maize calmodulin cDNA
clone.
[0126] 12. A hydrophobic moment peptide. See PCT Application No. WO
95/16776 (disclosure of peptide derivatives of tachyplesin which
inhibit fungal plant pathogens) and PCT Application No. WO 95/18855
(teaches synthetic antimicrobial peptides that confer disease
resistance).
[0127] 13. A membrane permease, a channel former, or a channel
blocker. For example, see the disclosure of Jaynes, et al., Plant
Sci., 89:43 (1993), of heterologous expression of a cecropin-beta,
lytic peptide analog to render transgenic tobacco plants resistant
to Pseudomonas solanacearum.
[0128] 14. A viral-invasive protein or a complex toxin derived
therefrom. For example, the accumulation of viral coat proteins in
transformed plant cells imparts resistance to viral infection
and/or disease development effected by the virus from which the
coat protein transgene is derived, as well as by related viruses.
See Beachy, et al., Ann. Rev. Phytopathol., 28:451 (1990). Coat
protein-mediated resistance has been conferred upon transformed
plants against alfalfa mosaic virus, cucumber mosaic virus, tobacco
streak virus, potato virus X, potato virus Y, tobacco etch virus,
tobacco rattle virus, and tobacco mosaic virus. Id.
[0129] 15. An insect-specific antibody or an immunotoxin derived
therefrom. Thus, an antibody targeted to a critical metabolic
function in the insect gut would inactivate an affected enzyme,
killing the insect. See Taylor, et al., Abstract #497, Seventh
Int'l Symposium on Molecular Plant-Microbe Interactions, Edinburgh,
Scotland (1994) (enzymatic inactivation in transgenic tobacco via
production of single-chain antibody fragments).
[0130] 16. A virus-specific antibody. See, for example,
Tavladoraki, et al., Nature, 366:469 (1993), who show that
transgenic plants expressing recombinant antibody transgenes are
protected from virus attack.
[0131] 17. A developmental-arrestive protein produced in nature by
a pathogen or a parasite. Thus, fungal
endo-alpha-1,4-D-polygalacturonases facilitate fungal colonization
and plant nutrient released by solubilizing plant cell wall
homo-alpha-1,4-D-galacturonase. See Lamb, et al., Bio/technology,
10:1436 (1992). The cloning and characterization of a transgene
which encodes a bean endopolygalacturonase-inhibiting protein is
described by Toubart, et al., Plant J., 2:367 (1992).
[0132] 18. A developmental-arrestive protein produced in nature by
a plant. For example, Logemann, et al., Bio/technology, 10:305
(1992), have shown that transgenic plants expressing the barley
ribosome-inactivating transgene have an increased resistance to
fungal disease.
[0133] 19. A lettuce mosaic potyvirus (LMV) coat protein transgene
introduced into Lactuca sativa in order to increase its resistance
to LMV infection. See Dinant, et al., Mol. Breeding, 3:1, 75-86
(1997).
[0134] Any disease or present resistance transgenes, including
those exemplified above, can be introduced into a lettuce plant of
the invention through a variety of means including but not limited
to transformation and breeding.
B. Transgenes that Confer Resistance to an Herbicide:
[0135] Exemplary polynucleotides encoding polypeptides that confer
traits desirable for herbicide resistance include acetolactate
synthase (ALS) mutants that lead to herbicide resistance such as
the S4 and/or Hra mutations ((resistance to herbicides including
sulfonylureas, imidazolinones, triazolopyrimidines, pyrimidinyl
thiobenzoates); glyphosate resistance (e.g.,
5-enol-pyrovyl-shikimate-3-phosphate-synthase (EPSPS) transgene,
including but not limited to those described in U.S. Pat. Nos.
4,940,935, 5,188,642, 5,633,435, 6,566,587, 7,674,598 as well as
all related application; or the glyphosate N-acetyltransferase
(GAT) transgene, described in Castle et al., Science, 2004,
304:1151-1154; and in U.S. Patent Application Publication Nos.
20070004912, 20050246798, and 20050060767)); glufosinate resistance
(e.g., BAR; see e.g., U.S. Pat. No. 5,561,236); 2,4-D resistance
(e.g., aryloxy alkanoate dioxygenase or AAD-1, AAD-12, or AAD-13),
HPPD resistance (e.g., Pseudomonas HPPD) and PPO resistance (e.g.,
fomesafen, acifluorfen-sodium, oxyfluorfen, lactofen,
fluthiacet-methyl, saflufenacil, flumioxazin, flumiclorac-pentyl,
carfentrazone-ethyl, sulfentrazone,); a cytochrome P450 or variant
thereof that confers herbicide resistance or tolerance to, inter
alia, HPPD-inhibiting herbicides, PPO-inhibiting herbicides and
ALS-inhibiting herbicides (U.S. Patent Application Publication No.
20090011936; U.S. Pat. Nos. 6,380,465; 6,121,512; 5,349,127;
6,649,814; and 6,300,544; and PCT International Publication No. WO
2007/000077); dicamba resistance (e.g., dicamba monoxygenase), and
traits desirable for processing or process products such as high
oil (e.g., U.S. Pat. No. 6,232,529); modified oils (e.g., fatty
acid desaturase transgenes (U.S. Pat. No. 5,952,544; PCT
International Publication No. WO 94/11516)); modified starches
(e.g., ADPG pyrophosphorylases (AGPase), starch synthases (SS),
starch branching enzymes (SBE), and starch debranching enzymes
(SDBE)); and polymers or bioplastics (e.g., U.S. Pat. No.
5,602,321; beta-ketothiolase, polyhydroxybutyrate synthase, and
acetoacetyl-CoA reductase (Schubert et al., J. Bacteriol., 1988,
170:5837-5847) facilitate expression of polyhydroxyalkanoates
(PHAs)).
[0136] In embodiments, the polynucleotide encodes a polypeptide
conferring resistance to an herbicide selected from glyphosate,
sulfonylurea, imidazolinone, dicamba, glufosinate, phenoxy
proprionic acid, L-phosphinothricin, cyclohexone, cyclohexanedione,
triazine, and benzonitrile.
[0137] Any transgene conferring herbicide resistance, including
those exemplified above, can be introduced into the lettuce plants
of the invention through a variety of means including, but not
limited to, transformation (e.g., genetic engineering techniques)
and crossing.
C. Transgenes that Confer or Contribute to a Value-Added Trait:
[0138] 1. Increased iron content of the lettuce, for example, by
introducing into a plant a soybean ferritin transgene as described
in Goto, et al., Acta Horticulturae., 521, 101-109 (2000).
[0139] 2. Decreased nitrate content of leaves, for example, by
introducing into a lettuce a transgene coding for a nitrate
reductase. See, for example, Curtis, et al., Plant Cell Rep.,
18:11, 889-896 (1999).
[0140] 3. Increased sweetness of the lettuce by introducing a
transgene coding for monellin that elicits a flavor 100,000 times
sweeter than sugar on a molar basis. See Penarrubia, et al.,
Bio/technology, 10:561-564 (1992).
[0141] 4. Modified fatty acid metabolism, for example, by
introducing into a plant an antisense sequence directed against
stearyl-ACP desaturase to increase stearic acid content of the
plant. See Knultzon, et al., PNAS, 89:2625 (1992).
[0142] 5. Modified carbohydrate composition effected, for example,
by introducing into plants a transgene coding for an enzyme that
alters the branching pattern of starch. See Shiroza, et al., J.
Bacteria, 170:810 (1988) (nucleotide sequence of Streptococcus
mutants fructosyltransferase transgene); Steinmetz, et al., Mol.
Gen. Genet., 20:220 (1985) (nucleotide sequence of Bacillus
subtilis levansucrase transgene); Pen, et al., Bio/technology,
10:292 (1992) (production of transgenic plants that express
Bacillus lichenifonnis alpha-amylase); Elliot, et al., Plant Mol.
Biol., 21:515 (1993) (nucleotide sequences of tomato invertase
transgenes); Sogaard, et al., J. Biol. Chem., 268:22480 (1993)
(site-directed mutagenesis of barley alpha-amylase transgene); and
Fisher, et al., Plant Physiol., 102:1045 (1993) (maize endosperm
starch branching enzyme II).
[0143] Any transgene that confers or contributes a value-added
trait, including those exemplified above, can be introduced into
the lettuce plants of the invention through a variety of means
including, but not limited to, transformation (e.g., genetic
engineering techniques) and crossing.
D. Transgenes that Control Male-Sterility:
[0144] 1. Introduction of a deacetylase transgene under the control
of a tapetum-specific promoter and with the application of the
chemical N-Ac-PPT. See, e.g., International Publication WO
01/29237.
[0145] 2. Introduction of various stamen-specific promoters. See,
e.g., International Publications WO 92/13956 and WO 92/13957.
[0146] 3. Introduction of the barnase and the barstar transgenes.
See, e.g., Paul, et al., Plant Mol. Biol., 19:611-622 (1992).
[0147] Any transgene that controls male sterility, including those
exemplified above, can be introduced into the lettuce plants of the
invention through a variety of means including, but not limited to,
transformation (e.g., genetic engineering techniques) and
crossing.
Methods for Plant Transformation.
[0148] Numerous methods for plant transformation have been
developed, including biological and physical, plant transformation
protocols. See, for example, Miki, et al., "Procedures for
Introducing Foreign DNA into Plants" in Methods in Plant Molecular
Biology and Biotechnology, Glick and Thompson Eds., CRC Press,
Inc., Boca Raton, pp. 67-88 (1993). In addition, expression vectors
and in vitro culture methods for plant cell or tissue
transformation and regeneration of plants are available. See, for
example, Gruber, et al., "Vectors for Plant Transformation" in
Methods in Plant Molecular Biology and Biotechnology, Glick and
Thompson Eds., CRC Press, Inc., Boca Raton, pp. 89-119 (1993).
A. Agrobacterium-Mediated Transformation.
[0149] One method for introducing an expression vector into plants
is based on the natural transformation system of Agrobacterium.
See, for example, Horsch, et al., Science, 227:1229 (1985); Curtis,
et al., Journal of Experimental Botany, 45:279, 1441-1449 (1994);
Torres, et al., Plant Cell Tissue and Organ Culture, 34:3, 279-285
(1993); and Dinant, et al., Molecular Breeding, 3:1, 75-86 (1997).
A. tumefaciens and A. rhizogenes are plant pathogenic soil bacteria
which genetically transform plant cells. The Ti and Ri plasmids of
A. tumefaciens and A. rhizogenes, respectively, carry genes
responsible for genetic transformation of the plant. See, for
example, Kado, C. I., Crit. Rev. Plant Sci., 10:1 (1991).
Descriptions of Agrobacterium vector systems and methods for
Agrobacterium-mediated transgene transfer are provided by Gruber,
et al., supra, Miki, et al., supra, and Moloney, et al., Plant Cell
Rep., 8:238 (1989). See also, U.S. Pat. No. 5,591,616 issued Jan.
7, 1997.
B. Direct Transgene Transfer.
[0150] Several methods of plant transformation collectively
referred to as direct transgene transfer have been developed as an
alternative to Agrobacterium-mediated transformation. A generally
applicable method of plant transformation is
microprojectile-mediated transformation wherein DNA is carried on
the surface of microprojectiles measuring 1 micron to 4 micron. The
expression vector is introduced into plant tissues with a biolistic
device that accelerates the microprojectiles to speeds of 300 m/s
to 600 m/s which is sufficient to penetrate plant cell walls and
membranes. Russell, D. R., et al., Plant Cell Rep., 12 (3,
January), 165-169 (1993); Aragao, F. J. L., et al., Plant Mol.
Biol., 20 (2, October), 357-359 (1992); Aragao, F. J. L., et al.,
Plant Cell Rep., 12 (9, July), 483-490 (1993); Aragao, Theor. Appl.
Genet., 93:142-150 (1996); Kim, J., Minamikawa, T., Plant Sci.,
117:131-138 (1996); Sanford, et al., Part. Sci. Technol., 5:27
(1987); Sanford, J. C., Trends Biotech., 6:299 (1988); Klein, et
al., Bio/technology, 6:559-563 (1988); Sanford, J. C., Physiol.
Plant, 7:206 (1990); Klein, et al., Bio/technology, 10:268
(1992).
[0151] Another method for physical delivery of DNA to plants is
sonication of target cells. Zhang, et al., Bio/technology, 9:996
(1991). Alternatively, liposome and spheroplast fusion have been
used to introduce expression vectors into plants. Deshayes, et al.,
EMBO J., 4:2731 (1985) and Christou, et al., PNAS, 84:3962 (1987).
Direct uptake of DNA into protoplasts using CaCl.sub.2
precipitation, polyvinyl alcohol, or poly-L-ornithine has also been
reported. Hain, et al., Mol. Gen. Genet., 199:161 (1985) and
Draper, et al., Plant Cell Physiol., 23:451 (1982). Electroporation
of protoplasts and whole cells and tissues have also been
described. Saker, M., Kuhne, T., Biologia Plantarum, 40(4):507-514
(1997/98); Donn, et al., In Abstracts of VIIth International
Congress on Plant Cell and Tissue Culture IAPTC, A2-38, p. 53
(1990); D'Halluin, et al., Plant Cell, 4:1495-1505 (1992); and
Spencer, et al., Plant Mol. Biol., 24:51-61 (1994). See also
Chupean, et al., Bio/technology, 7:5, 503-508 (1989).
[0152] Following transformation of plant target tissues, expression
of the above-described selectable marker transgenes allows for
preferential selection of transformed cells, tissues and/or plants,
using regeneration and selection methods now well known in the
art.
[0153] The foregoing methods for transformation would typically be
used for producing a transgenic lettuce line. The transgenic
lettuce line could then be crossed with another (non-transformed or
transformed) line in order to produce a new transgenic lettuce
line. Alternatively, a genetic trait that has been engineered into
a particular plant cultivar using the foregoing transformation
techniques could be introduced into another line using traditional
breeding (e.g., backcrossing) techniques that are well known in the
plant breeding arts. For example, a backcrossing approach could be
used to move an engineered trait from a public, non-elite inbred
line into an elite inbred line, or from an inbred line containing a
foreign transgene in its genome into an inbred line or lines which
do not contain that transgene. As used herein, "crossing" can refer
to a simple X by Y cross, or the process of backcrossing, depending
on the context.
Gene Conversions.
[0154] When the term "lettuce plant" is used in the context of the
present invention, this term also includes any gene conversions of
that plant or variety. The term "gene converted plant" as used
herein refers to those lettuce plants that are developed, for
example, by backcrossing, genetic engineering and/or mutation,
wherein essentially all of the desired morphological and
physiological characteristics of a variety (e.g., as shown in Table
1) are recovered in addition to the one or more genes transferred
into the variety. To illustrate, backcrossing methods can be used
with the present invention to improve or introduce a characteristic
into the variety. The term "backcrossing" as used herein refers to
the repeated crossing of a hybrid progeny back to the recurrent
parent, e.g., backcrossing 1, 2, 3, 4, 5, 6, 7, 8, 9, or more times
to the recurrent parent. The parental plant that contributes the
gene for the desired characteristic is termed the "nonrecurrent" or
"donor parent." This terminology refers to the fact that the
nonrecurrent parent is generally used one time in the breeding
e.g., backcross) protocol and therefore does not recur. The gene
that is transferred can be a native gene, a mutated native gene or
a transgene introduced by genetic engineering techniques into the
plant (or ancestor thereof). The parental plant into which the
gene(s) from the nonrecurrent parent are transferred is known as
the "recurrent" parent as it is used for multiple rounds in the
backcrossing protocol. Poehlman & Sleper (1994) and Fehr
(1993). In a typical backcross protocol, the original variety of
interest (recurrent parent) is crossed to a second variety
(nonrecurrent parent) that carries the gene(s) of interest to be
transferred. The resulting progeny from this cross are then crossed
again to the recurrent parent and the process is repeated until a
plant is obtained wherein essentially all of the desired
morphological and physiological characteristics of the recurrent
parent are recovered in the converted plant in addition to the
transferred gene(s) and associated trait(s) from the nonrecurrent
parent.
[0155] Many gene traits have been identified that are not regularly
selected in the development of a new line but that can be improved
by backcrossing techniques. Gene traits may or may not be
transgenic. Examples of these traits include, but are not limited
to, male sterility, modified fatty acid metabolism, modified
carbohydrate metabolism, herbicide resistance, pest or disease
resistance (e.g., resistance to bacterial, fungal, or viral
disease), insect resistance, enhanced nutritional quality,
increased sweetness, increased flavor, improved ripening control,
improved salt tolerance, industrial usage, yield stability, and
yield enhancement. These genes are generally inherited through the
nucleus.
[0156] Tissue Culture.
[0157] Further reproduction of lettuce plants variety can occur by
tissue culture and regeneration. Tissue culture of various tissues
of lettuce and regeneration of plants therefrom is well known and
widely published. For example, reference may be had to Teng, et
al., HortScience, 27:9, 1030-1032 (1992); Teng, et al.,
HortScience, 28:6, 669-1671 (1993); Zhang, et al., Journal of
Genetics and Breeding, 46:3, 287-290 (1992); Webb, et al., Plant
Cell Tissue and Organ Culture, 38:1, 77-79 (1994); Curtis, et al.,
Journal of Experimental Botany, 45:279, 1441-1449 (1994); Nagata,
et al., Journal for the American Society for Horticultural Science,
125:6, 669-672 (2000); and Ibrahim, et al., Plant Cell Tissue and
Organ Culture, 28(2), 139-145 (1992). It is clear from the
literature that the state of the art is such that these methods of
obtaining plants are routinely used and have a very high rate of
success. Thus, another aspect of this invention is to provide cells
which upon growth and differentiation produce lettuce plants having
desired characteristics of lettuce cultivar Julian (e.g., one or
more of the characteristics shown in Table 1). Optionally, lettuce
plants can be regenerated from the tissue culture of the invention
comprising all or essentially all of the physiological and
morphological characteristics of lettuce cultivar Julian.
[0158] As used herein, the term "tissue culture" indicates a
composition comprising isolated cells of the same or a different
type or a collection of such cells organized into parts of a plant.
Exemplary types of tissue cultures are protoplasts, calli,
meristematic cells, and plant cells that can generate tissue
culture that are intact in plants or parts of plants, such as
leaves, pollen, embryos, roots, root tips, anthers, pistils,
flowers, seeds, petioles, suckers, and the like. Means for
preparing and maintaining plant tissue culture are well known in
the art. By way of example, a tissue culture comprising organs has
been used to produce regenerated plants. U.S. Pat. Nos. 5,959,185,
5,973,234, and 5,977,445 describe certain techniques.
Additional Breeding Methods.
[0159] This invention is also directed to methods for producing a
lettuce plant by crossing a first parent lettuce plant with a
second parent lettuce plant wherein the first or second parent
lettuce plant is a plant of lettuce cultivar Julian. Further, both
first and second parent lettuce can come from lettuce cultivar
Julian. Thus, any of the following exemplary methods using lettuce
cultivar Julian are part of this invention: selfing, backcrosses,
hybrid production, crosses to populations, double haploid
production, and the like. All plants produced using lettuce
cultivar Julian as at least one parent are within the scope of this
invention, including those developed from lettuce plants derived
from lettuce cultivar Julian. Advantageously, lettuce cultivar
Julian can be used in crosses with other, different, lettuce plants
to produce the first generation (F.sub.1) lettuce hybrid seeds and
plants with desirable characteristics. The lettuce plants of the
invention can also be used for transformation where exogenous
transgenes are introduced and expressed by the plants of the
invention. Genetic variants created either through traditional
breeding methods or through transformation of the cultivars of the
invention by any of a number of protocols known to those of skill
in the art are intended to be within the scope of this
invention.
[0160] The following describes exemplary breeding methods that may
be used with lettuce cultivar Julian in the development of further
lettuce plants. One such embodiment is a method for developing
lettuce cultivar Julian progeny lettuce plants in a lettuce plant
breeding program comprising: obtaining a plant, or a part thereof,
of lettuce cultivar Julian, utilizing said plant or plant part as a
source of breeding material, and selecting a lettuce cultivar
Julian progeny plant with molecular markers in common with lettuce
cultivar Julian and/or with some, all or essentially all
morphological and/or physiological characteristics of lettuce
cultivar Julian (see, e.g., Table 1). In representative
embodiments, the progeny plant has at least 1, 2, 3, 4, 5, 6, 7, 8,
9, 10 or more of the morphological and physiological
characteristics of lettuce cultivar Julian (e.g., as described in
Table 1), or even all of the morphological and physiological
characteristics of lettuce cultivar Julian so that said progeny
lettuce plant is not significantly different for said traits than
lettuce cultivar Julian, as determined at the 5% significance level
when grown in the same environmental conditions; optionally, with
the presence of one or more desired additional traits (e.g., male
sterility, disease resistance, pest or insect resistance, herbicide
resistance, and the like). Breeding steps that may be used in the
breeding program include pedigree breeding, backcrossing, mutation
breeding and/or recurrent selection. In conjunction with these
steps, techniques such as RFLP-enhanced selection, genetic marker
enhanced selection (for example, SSR markers) and/or and the making
of double haploids may be utilized.
[0161] Another representative method involves producing a
population of lettuce cultivar Julian progeny plants, comprising
crossing lettuce cultivar Julian with another lettuce plant,
thereby producing a population of lettuce plants that, on average,
derives at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its
alleles (i.e., TAC) from lettuce cultivar Julian, e.g., at least
about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,
70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the genetic
complement of lettuce cultivar Julian. One embodiment of this
invention is the lettuce plant produced by this method and that has
obtained at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its
alleles from lettuce cultivar Julian. A plant of this population
may be selected and repeatedly selfed or sibbed with a lettuce
plant resulting from these successive filial generations. Another
approach is to make double haploid plants to achieve
homozygosity.
[0162] One of ordinary skill in the art of plant breeding would
know how to evaluate the traits of two plant varieties to determine
if there is no significant difference between the two traits
expressed by those varieties. For example, see Fehr and Walt,
Principles of Cultivar Development, pp. 261-286 (1987). In
embodiments, the invention encompasses progeny plants having a
combination of at least 2, 3, 4, 5, 6, 7, 8, 9, 10 or more of the
characteristics as described herein for lettuce cultivar Julian, so
that said progeny lettuce plant is not significantly different for
said traits than lettuce cultivar Julian, as determined at the 5%
significance level when grown in the same environmental conditions.
Using techniques described herein and those known in the art,
molecular markers may be used to identify said progeny plant as
progeny of lettuce cultivar Julian. Mean trait values may be used
to determine whether trait differences are significant, and
optionally the traits are measured on plants grown under the same
environmental conditions.
[0163] Progeny of lettuce cultivar Julian may also be characterized
through their filial relationship with lettuce cultivar Julian, as
for example, being within a certain number of breeding crosses of
lettuce cultivar Julian. A breeding cross is a cross made to
introduce new genetics into the progeny, and is distinguished from
a cross, such as a self or a sib cross or a backcross to Julian as
a recurrent parent, made to select among existing genetic alleles.
The lower the number of breeding crosses in the pedigree, the
closer the relationship between lettuce cultivar Julian and its
progeny. For example, progeny produced by the methods described
herein may be within 1, 2, 3, 4, 5 or more breeding crosses of
lettuce cultivar Julian.
[0164] In representative embodiments, a lettuce plant derived from
lettuce cultivar Julian comprises cells comprising at least one set
of chromosomes derived from lettuce cultivar Julian. In
embodiments, the lettuce plant or population of lettuce plants
derived from lettuce cultivar Julian comprises, on average, at
least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,
70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of its alleles (i.e.,
TAC) from lettuce cultivar Julian, e.g., at least about 6.25%,
12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%,
90%, 95%, 96%, 97%, 98% or 99% of the genetic complement of lettuce
cultivar Julian. In embodiments, the lettuce plant derived from
lettuce cultivar Julian is one, two, three, four, five or more
breeding crosses removed from lettuce cultivar Julian.
[0165] In representative embodiments, a plant derived from lettuce
cultivar Julian is a double haploid plant, a hybrid plant or an
inbred plant.
[0166] In embodiments, a hybrid or derived plant from lettuce
cultivar Julian comprises a desired added trait. In representative
embodiments, a lettuce plant derived from lettuce cultivar Julian
comprises all of the morphological and physiological
characteristics of lettuce cultivar Julian (e.g., as described in
Table 1). In embodiments, the lettuce plant derived from lettuce
cultivar Julian comprises essentially all of the morphological and
physiological characteristics of lettuce cultivar Julian (e.g., as
described in Table 1), with the addition of a desired added
trait.
[0167] Those skilled in the art will appreciate that any of the
traits described above with respect to plant transformation methods
can be introduced into a plant of the invention (e.g., lettuce
cultivar Julian and hybrid lettuce plants and other lettuce plants
derived therefrom) using breeding techniques.
Genetic Analysis of Lettuce Cultivar Julian.
[0168] The invention further provides a method of determining a
genetic characteristic of lettuce cultivar Julian or a progeny
thereof, e.g., a method of determining a genotype of lettuce
cultivar Julian or a progeny thereof. In embodiments, the method
comprises detecting in the genome of a Julian plant, or a progeny
plant thereof, at least a first polymorphism (e.g., using one or
more molecular markers). To illustrate, in embodiments, the method
comprises obtaining a sample of nucleic acids from the plant and
detecting at least a first polymorphism in the nucleic acid sample.
Optionally, the method may comprise detecting a plurality of
polymorphisms (e.g., two or more, three or more, four or more, five
or more, six or more, eight or more or ten or more polymorphisms,
etc.) in the genome of the plant. In representative embodiments,
the method further comprises storing the results of the step of
detecting the polymorphism(s) on a computer readable medium. The
invention further provides a computer readable medium produced by
such a method.
DEPOSIT INFORMATION
[0169] Applicants have made a deposit of at least 2500 seeds of
lettuce cultivar Julian with the American Type Culture Collection
(ATCC), 10801 University Boulevard, Manassas, Va., 20110-2209
U.S.A. under ATCC Deposit No. ______ on ______. This deposit of
lettuce variety Julian will be maintained in the ATCC depository,
which is a public depository, for a period of 30 years, or 5 years
after the most recent request, or for the effective life of the
patent, whichever is longer, and will be replaced if any of the
deposited seed becomes nonviable during that period. Additionally,
Applicants have satisfied all the requirements of 37 C.F.R.
.sctn..sctn. 1.801-1.809, including providing an indication of the
viability of the samples. Access to this deposit will be made
available during the pendency of this application to the
Commissioner upon request. Upon the issuance of a patent on the
variety, the variety will be irrevocably and without restriction
released to the public by providing access to the deposit of at
least 2500 seeds of the variety with the ATCC. Applicants impose no
restrictions on the availability of the deposited material from the
ATCC; however, Applicants have no authority to waive any
restrictions imposed by law on the transfer of biological material
or its transportation in commerce. Applicants do not waive any
infringement of its rights granted under this patent or under the
Plant Variety Protection Act (7 USC .sctn. 2321 et seq.).
[0170] The foregoing invention has been described in detail by way
of illustration and example for purposes of clarity and
understanding. However, it will be apparent that certain changes
and modifications such as single gene modifications and mutations,
somaclonal variants, variant individuals selected from large
populations of the plants of the instant inbred and the like may be
practiced within the scope of the invention.
* * * * *