U.S. patent application number 16/570588 was filed with the patent office on 2020-07-23 for carrot variety purple royale.
The applicant listed for this patent is Sensient Colors, LLC. Invention is credited to Daniel Jeffrey BROTSLAW, Bradley Katsu KAJI, Joerg MEYER.
Application Number | 20200229371 16/570588 |
Document ID | / |
Family ID | 71610241 |
Filed Date | 2020-07-23 |
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United States Patent
Application |
20200229371 |
Kind Code |
A1 |
MEYER; Joerg ; et
al. |
July 23, 2020 |
CARROT VARIETY PURPLE ROYALE
Abstract
The invention relates to the carrot variety designated Purple
Royale. Provided by the invention are the seeds, plants, and
derivatives of the carrot variety Purple Royale. Also provided by
the invention are tissue cultures of the carrot variety Purple
Royale and the plants regenerated therefrom. Still further provided
by the invention are methods for producing carrot plants by
crossing the carrot variety Purple Royale with itself or another
carrot variety and plants produced by such methods.
Inventors: |
MEYER; Joerg; (St. Charles,
MO) ; KAJI; Bradley Katsu; (Livingston, CA) ;
BROTSLAW; Daniel Jeffrey; (Modesto, CA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Sensient Colors, LLC |
Saint Louis |
MO |
US |
|
|
Family ID: |
71610241 |
Appl. No.: |
16/570588 |
Filed: |
September 13, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62794230 |
Jan 18, 2019 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A01H 6/068 20180501;
A01H 5/06 20130101 |
International
Class: |
A01H 6/06 20060101
A01H006/06; A01H 5/06 20060101 A01H005/06 |
Claims
1. A plant of carrot variety Purple Royale, wherein representative
seed of said carrot variety has been deposited under ATCC Accession
No. ______.
2. A plant part of the plant of claim 1, wherein the plant part
comprises at least one cell of said plant.
3. A seed of carrot variety Purple Royale, wherein representative
seed of said carrot variety Purple Royale has been deposited under
ATCC Accession No. ______.
4. A method of producing carrot seed, the method comprising
crossing the plant of claim 1 with itself or a second carrot plant
to produce said carrot seed.
5. The method of claim 4, the method further comprising crossing
the plant of carrot variety Purple Royale with a second,
non-isogenic carrot plant to produce said carrot seed.
6. An F.sub.1 carrot seed produced by the method of claim 5.
7. A carrot plant produced by growing the F.sub.1 carrot seed of
claim 6.
8. A composition comprising the seed of claim 3 comprised in plant
seed growth media.
9. The composition of claim 8, wherein the plant seed growth media
is soil or a synthetic cultivation medium.
10. A plant of carrot variety Purple Royale further comprising a
single locus conversion, wherein said plant otherwise comprises all
of the morphological and physiological characteristics of said
carrot variety when grown under the same environmental conditions,
and wherein representative seed of said carrot variety have been
deposited under ATCC Accession No.
11. A seed that produces the plant of claim 10.
12. The seed of claim 11, wherein the single locus confers a trait
selected from the group consisting of increased anthocyanin
content, increased flower size, multiple petals, broad
environmental adaptation, and insect and pest resistance, and
resistance to bacterial, fungal, or viral disease.
13. The method of claim 5, the method further comprising: a.
crossing a plant grown from said carrot seed with itself or a
different carrot plant to produce seed of a progeny plant of a
subsequent generation; b. 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 to produce seed of a
progeny plant of a further subsequent generation; and c. repeating
step (b) with sufficient inbreeding to produce seed of an inbred
carrot plant that is derived from carrot variety Purple Royale.
14. A method of producing a commodity plant product, the method
comprising producing the commodity plant product from the plant of
claim 1.
15. The method of claim 15, wherein the commodity plant product is
anthocyanin.
Description
[0001] This application claims the benefit of priority of United
States Provisional Application No. 62/794,230, filed Jan. 18, 2019,
the disclosure of which is hereby incorporated by reference as if
written herein in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to the field of plant
breeding. In particular, the invention provides carrot plants with
a high anthocyanin level in the roots. The invention further
provides for a new and distinct carrot variety designated Purple
Royale and for breeding methods with these plants.
BACKGROUND OF THE INVENTION
[0003] Carrot (Daucus carota subsp. sativus), is a root vegetable,
usually orange in color, though purple, red, white, and yellow
varieties exist. It has a crisp texture when fresh. The most
commonly eaten part of a carrot is a taproot, although the greens
are edible as well. It is a domesticated form of the wild carrot
Daucus carota, wildly believed to be from central Asia. Most of the
modern breeding of the domestic carrot originated in Europe and
which has focused on the selection of a greatly enlarged and more
palatable, less woody-textured edible taproot. The annual purple
varieties essentially missed out on those centuries of modern
breeding until approximately the last 20 years. The world
production of carrots and turnips for calendar year 2011 was almost
35.658 million tons (Food and Agriculture Organization of the
United Nations (FAO)).
[0004] The aim of a vegetable breeder is to combine desirable
traits in a single variety. Such desirable traits may include any
trait deemed beneficial by a grower and/or consumer, including
greater yield, resistance to insects or disease, tolerance to
environmental stress, and nutritional value.
[0005] Total anthocyanin level is an example of such a desirable
trait. Anthocyanins are polyphenols with known antioxidant activity
which may be responsible for some biological activities including
the prevention or lowering the risk of cardiovascular disease,
diabetes, arthritis and cancer. Nevertheless, such properties,
their stability and bioavailability depend on their chemical
structure (Miguel MG. Journal of Applied Pharmaceutical Science. 01
(06); 2011: 7-15.
[0006] A uniform population of a breeding line can be obtained by
self-pollination and selection for type. Plants thus obtained
become homozygous at almost all gene loci, i.e. a homozygous plant.
Crossing two such plants of different genotypes produces a uniform
population of plants that are heterozygous for many loci. On the
other hand, a cross of two plants each heterozygous at a number of
loci produces a population of plants that differ genetically and
are not uniform. Due to this non-uniformity, performance of such
plants is unpredictable.
[0007] Thus, a vegetable breeder prefers development of a
homozygous inbred plant that can be crossed to produce uniform
varieties. Pedigree breeding and recurrent selection are examples
of breeding methods that have been used to develop inbred plants
from breeding populations. Those breeding methods combine the
genetic backgrounds from two or more plants or various other
broad-based sources into breeding pools from which new lines
derived therefrom are developed by selfing and selection of desired
phenotypes. The new lines are evaluated to determine which of those
have commercial potential.
[0008] So far, breeding efforts have provided a number of useful
carrot lines with beneficial traits, however, there remains a great
need in the art for new lines with further improved traits. There
is thus a need for new carrot varieties having specific combination
of trait or color.
SUMMARY OF THE INVENTION
[0009] The objective of the invention was to develop an Imperator
type variety with purple coloration throughout and a high
anthocyanin content of the mature root.
[0010] In one aspect, the present invention provides seed of a new
carrot (Daucus carota) variety, designated Purple Royale, having
been deposited under Accession Number ______, a plant, or a part
thereof, produced by growing said seed. The invention also provides
methods and compositions relating to plants, plant parts, seeds and
progenies of carrot variety Purple Royale.
[0011] Variety Purple Royale is most similar to comparison variety
Anthonina or Deep Purple, which is a variety produced and sold by
Bejo. However, Purple Royale differs from Anthonina, Purple Elite,
Purple Haze, and Deep Purple in one or more, e.g., at least two, at
least three, optionally all morphological and/or physiological
characteristics listed in the following (see also Table 1), when
grown under the same environmental conditions:
[0012] an (average) root core thickness (at midpoint of
cross-section) that is at least about 12%, or preferably at least
about 13%, 14%, 15%, 16%, 17%, 18%, or even about 19% bigger than
the core thickness of one or more of black carrot varieties
Anthonina, Purple Elite, Purple Haze, and Deep Purple;
[0013] a (average) root diameter at midpoint that is at least about
7.5%, or preferably at least about 10%, 11%, 12%, 13%, 14%, or even
about 14.9% smaller than the diameter at midpoint of one or more of
black carrot varieties Anthonina, Purple Elite, Purple Haze, and
Deep Purple;
[0014] a (average) carrot root length (minus taproot) that is at
least about 5%, or preferably at least about 5.5%, 6%, 6.5%, 7%,
7.5%, 8%, or even about 8.3% smaller than the carrot length of one
or more of black carrot varieties Anthonina, Purple Elite, Purple
Haze, and Deep Purple;
[0015] a purple cross-section interior color of both core and
cortex, which is darker than the purple cross-section interior
color of Anthonina, Purple Elite, Purple Haze, and Deep Purple,
e.g. N79A for Purple Royale versus the designation listed for the
other carrot varieties in Table 1;
[0016] Other differences between variety Purple Royale and
Anthonina, Purple Elite, Purple Haze, and Deep Purple reveals that
Purple Royale also differs significantly from Anthonina, Purple
Elite, Purple Haze, and Deep Purple in one or more, e.g., at least
two, at least three, optionally all morphological and/or
physiological characteristics listed in the following (see also
Table 1), when grown under the same environmental conditions.
[0017] The present invention provides a carrot plant variety
designated, representative seed of said variety having been
deposited under ATCC Accession Number ______, and plant parts of
the new variety such as for example seed, leaf, pollen, an ovule,
taproot, root and a cell of the plant.
[0018] The invention also concerns the seed of the carrot Purple
Royale, representative seed of said variety having been deposited
under ATCC Accession Number ______, a plant, or a part thereof,
produced by growing said seed. Or carrot plants having all or
essentially all the physiological and morphological characteristics
of Purple Royale.
[0019] The invention further relates to breeding methods using
plants or seed of carrot variety Purple Royale.
[0020] Additional objects, features and advantages of the present
invention will become apparent from the following detailed
description. It should be understood, however, that the detailed
description and any specific examples provided, while indicating
specific embodiments of the invention, are given by way of
illustration only, since various changes and modifications within
the spirit and scope of the invention will become apparent to those
skilled in the art from this detailed description.
DETAILED DESCRIPTION OF THE DRAWINGS
[0021] The patent or application file contains at least one drawing
executed in color. Copies of this patent or patent application
publication with color drawing(s) will be provided by the Office
upon request and payment of the necessary fee.
[0022] FIG. 1 shows the presence of anthocyanin in Purple
Royale.
[0023] FIG. 2 shows the presence/absence of anthocyanin in Purple
Royale, Anthonina, Purple Elite, Purple Haze, and Deep Purple
DETAILED DESCRIPTION OF THE INVENTION
[0024] The present invention provides a carrot plant, or part
thereof, the roots of said plant. The invention further relates to
plant parts of these plants such as for example seed, leaf, pollen,
an ovule, taproot, root and a cell of the plant. In another aspect,
the plant parts include leaf, pollen, an ovule, taproot, root and a
cell of the plant. In yet another aspect, the plant part is a
root.
[0025] The invention also includes seed of the plant of the plants
of the invention, i.e seeds from which a carrot plant, the roots of
which comprise, can be grown.
[0026] In another embodiment the invention concerns a plant
according to the invention wherein the genetic elements conferring
said total anthocyanin level is obtainable from carrot variety
Purple Royale, a representative sample of seed of Purple Royale
having been deposited under ATCC Accession Number ______.
[0027] In still another embodiment the invention relates to a plant
according to the invention wherein the genetic elements conferring
said are obtainable from carrot variety Purple Royale, a
representative sample of seed of Purple Royale having been
deposited under ATCC Accession Number ______.
[0028] Anthocyanin content is a function of growth stage and
production environment.
[0029] In another aspect the root of the carrot plant of the
invention has a total average anthocyanin level of at least about
20 ppm or preferably at least about 21 ppm, 22 ppm, 23 ppm, 24 ppm,
25 ppm, or even about 26 ppm. The anthocyanin content of the carrot
variety Purple Royale will exceed any known carrot variety at all
growth stages.
[0030] All parts of the specification which refer herein to variety
Purple Royale, such as breeding, progeny, plant parts, cells,
seeds, development of identification of EDVs, etc. can also be
applied in a more general way, i.e. to methods for transfer the
genetic elements conferring high anthocyanin levels. Thus, the
invention not only relates to the above carrot plant, but also to
cells, tissues, plant parts, seeds, EDVs, progeny, transformed
carrot plants, etc. which comprise the genetic determinants
obtainable from (obtained from; as present in) Purple Royale which
confer the high anthocyanin content and optionally the genetic
elements which confer the faint haloing and zoning.
[0031] The present invention provides a carrot plant variety
designated Purple Royale, representative seed of said variety
having been deposited under ATCC Accession Number and plant parts
of the new variety such as for example seed, leaf, pollen, an
ovule, taproot, root and a cell of the plant. In another aspect,
the plant parts include leaf, pollen, an ovule, taproot, root and a
cell of the plant. In yet another aspect, the plant part is a root.
Also provided are carrot plants having all or essentially all the
physiological and morphological characteristics of such a plant,
i.e. a carrot plant that does not differ (statistically)
significantly from Purple Royale in the morphological and/or
physiological characteristics of Table 1, when grown under the same
conditions.
[0032] The invention also concerns the seed of the carrot Purple
Royale, representative seed of said variety having been deposited
under Accession Number ATCC ______, a plant, or a part thereof
(e.g. a root), produced by growing said seed. Or carrot plants
having all or essentially all the physiological and/or
morphological characteristics of Purple Royale when grown under the
same conditions. The carrot seed of the invention (i.e. seed from
which a plant of variety Purple Royale can be grown) may be
provided as an essentially homogeneous population of carrot seed
according to the invention. Essentially homogeneous populations of
seed are generally free from substantial numbers of other seed.
Therefore, seed of the carrot Purple Royale or carrot plants having
all or essentially all the physiological and/or morphological
characteristics of Purple Royale may be defined as forming at least
about 97% of the total seed, including at least about 98%, 99% or
more of the seed. The seed population may be separately grown to
provide an essentially homogeneous population of carrot plants
according to the invention. Also provided are plants or plant parts
such as seed (produced on the plant Purple Royale, e.g. after
self-pollination or cross-pollination by another carrot plant),
leaf, pollen, an ovule, taproot, roots or a cell produced by
growing the seeds of the invention. In yet another aspect, the
plant part is a root.
[0033] In another aspect the invention relates to a variety of
Purple Royale having at least one, two or three physiological
and/or morphological characteristics which are (statistically
significantly) different from those of Purple Royale and which
otherwise has essentially all physiological and morphological
characteristics of a carrot plant designated Purple Royale, a
representative sample of seeds of which having been deposited under
ATCC Accession Number ______.
[0034] In another aspect, the invention relates to a variety of
Purple Royale having at least one or two physiological and/or
morphological characteristics which are significantly different
from those of Purple Royale and which otherwise comprises at least
3, 4 or 5 or more (or all) of the distinguishing characteristics
1)-5) as defined below, or preferably at least 3, 4, 5, 6, 7, 8, 9,
10, 11, or 12 or all 12 of the distinguishing characteristics
1)-12) of Purple Royale and/or has essentially all physiological
and morphological characteristics of a carrot plant designated
Purple Royale obtainable by selecting a natural or induced mutant,
or a somaclonal variant, or a natural variant from a population of
plants designated Purple Royale.
[0035] In still another aspect, a carrot plant, or a part thereof,
is provided which does not significantly differ in distinguishing
characteristics 1)-5) of carrot plant Purple Royale when grown
under the same conditions. In yet another aspect, the invention
relates to a carrot plant, or a part thereof, which does not
significantly differ in distinguishing characteristics 1)-5) and
additionally comprises (i.e. does not differ statistically
significantly in) at least one, two, three or all of distinguishing
characteristics 6)-12) of carrot plant Purple Royale when grown
under the same conditions.
[0036] In yet another aspect, the invention relates to a carrot
plant, or a part thereof, which does not differ significantly from
carrot plant Purple Royale in any of the physiological and/or
morphological characteristics of Table 1 when grown under the same
conditions.
[0037] In another aspect the invention relates to a carrot plant,
or a part thereof, which does not significantly differ from carrot
plant Purple Royale in any of the distinguishing characteristics
(see USDA descriptors) consisting of 1) average root core thickness
at midpoint of cross-section; 2) average root diameter at midpoint;
3) average carrot length (minus taproot); 4) cross-section interior
color of both core and cortex; 5) haloing and zoning of the root at
market maturity, when grown under the same conditions.
[0038] In still another aspect, a carrot plant is provided, which
statistically significantly differs from Purple Royale in at least
one morphological and/or physiological characteristic, but which
does not differ significantly from Purple Royale in the following
characteristics (see USDA descriptor) when grown under the same
conditions: 1) average root core thickness at midpoint of
cross-section; 2) average root diameter at midpoint; 3) average
carrot length (minus taproot); 4) cross-section interior color of
both core and cortex; 5) haloing and zoning of the root at market
maturity.)
[0039] In a further aspect, a carrot plant is provided, which
statistically significantly differs from Purple Royale in at least
one morphological and/or physiological characteristics, but which
does not differ significantly from Purple Royale in the following
characteristics (see USDA descriptors) when grown under the same
conditions: 1) average root core thickness at midpoint of
cross-section; 2) average root diameter at midpoint; 3) average
carrot length (minus taproot); 4) cross-section interior color of
both core and cortex; 5) haloing and zoning of the root at market
maturity; and which further does not significantly differ from the
plant designated Purple Royale in one, two, three or more of the
following characteristics when grown under the same conditions: 6)
Petiole length from crown to first pinna; 7) average length of
taproot; 8) leaf blade color (at harvest stage); 9) leaf blade
divisions (at harvest stage); 10) leaf petiole anthocyanin level at
harvest stage and petiole pubescence at harvest stage; 11) average
root diameter at shoulder; 12) total anthocyanin level.
[0040] In a further embodiment a carrot plant is provided, which
(statistically significantly) differs from the carrot plant
designated Purple Royale, representative seeds of said carrot plant
having been deposited under ATCC accession number ______, in at
least one, two, three, four, or five morphological and/or
physiological characteristics when grown under the same
environmental conditions, whereby the morphological and/or
physiological characteristics are those of Table 1. The carrot
plant does, thus, not differ in a statistically significant way
from Purple Royale in any of the morphological and/or physiological
characteristics of Table 1 when grown under the same conditions, or
only differs is one, two, three, four or five of the morphological
and/or physiological characteristics of Table 1, while there is no
significant difference in the other characteristics.
[0041] In one embodiment a carrot plant is provided, designated
Purple Royale, which does not (statistically significantly) differ
in any of the morphological and/or physiological characteristics of
Table 1 from plants grown from seeds deposited under ATCC accession
number ______ when grown under the same environmental
conditions.
[0042] In yet another aspect of the invention, a tissue culture or
cell-culture of regenerable cells of a carrot plant according to
the invention is provided. The tissue culture or cell-culture will
preferably be capable of regenerating carrot plants capable of
expressing all of the physiological and/or morphological
characteristics of the starting plant, and of regenerating plants
having substantially the same genotype as the starting plant.
Examples of some of the physiological and/or morphological
characteristics of the carrot Purple Royale include those traits
set forth in Table 1 herein when grown under the environmental
conditions outlined herein with reference to the data of Table 1.
The regenerable cells in such tissue or cell cultures may be
derived, for example, from carrot explants, such as embryos,
meristems, petioles, cuttings, protoplasts, cotyledons, pollen,
leaves, nodes, anthers, roots, taproots, root tips, pistils,
flowers, seed and stems. Still further, the present invention
provides carrot plants regenerated from a tissue culture or cell
culture of the invention. These plants have all the physiological
and/or morphological characteristics of a plant according to the
invention.
[0043] In another aspect, the above described carrot plants are
obtainable from in vitro cell or tissue cultures. As already
elsewhere in this application, in vitro cell or tissue cultures are
known in the art and can be used to either vegetatively reproduce
the plant from which the cells or tissues were obtained or to
identify and/or select a phenotypic variant (such as an EDV), and
to regenerate such a variant. The phenotypic variant may, for
example, be a somaclonal variant, mutant or off-type, but is
preferably genetically stable. Thus, the variant phenotype is
preferably genetically stable, also in the mature plants
regenerated from the cell or tissue culture. That means, the
phenotypic variant does not show variation in phenotype which are
transient and are not genetically stable. Once selected, such
selected variants can then in turn also be reproduced true to type
using in vitro cell or tissue culture or by propagation via
seed.
[0044] Thus, in one aspect, a carrot plant is provided which is
clonally propagated (it is a vegetative reproduction) from Purple
Royale cells or tissue and which comprises all the distinguishing
characteristics of Purple Royale when grown under the same
environmental conditions. In another aspect it further comprises
one or more of the further distinguishing characteristics. In yet
another aspect it comprises all morphological and/or physiological
characteristics of Purple Royale as given in Table 1. And in yet a
further aspect it comprises all morphological and/or physiological
characteristics of Purple Royale as given in Table 1, except that
it significantly differs from Purple Royale in one, two, three,
four, or five of the morphological and/or physiological
characteristics of Table 1.
[0045] The invention also concerns methods of vegetatively
propagating a plant of the invention. In certain embodiments, the
method comprises the steps of: (a) collecting tissue or cells
capable of being propagated from a plant of the invention; (b)
cultivating said tissue or cells to obtain proliferated shoots; and
(c) rooting said proliferated shoots, to obtain rooted plantlets.
Steps (b) and (c) 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 plants. In one embodiment, the
method further comprises step (d) growing plants from said rooted
plantlets.
[0046] In still yet another aspect of the invention, processes are
provided for producing carrot seeds, plants and roots, which
processes generally comprise crossing a first parent carrot plant
with a second parent carrot plant, wherein at least one of the
first or second parent carrot plants is a plant according to the
invention.
[0047] One embodiment of the invention refers to a method of
producing a carrot plant comprising crossing a carrot plant of
variety Purple Royale with a second carrot plant one or more times
such as one, two, three, four, five, six or more times. This method
comprises in one embodiment selecting progeny from said
crossing.
[0048] These processes may be further exemplified as processes for
preparing the carrot seed or plants, wherein a first carrot plant
is crossed with a second carrot plant of a different, distinct
genotype to provide a plant that has, as one of its parents, a
plant of Purple Royale.
[0049] The present invention also provides the carrot seeds and
plants produced by a process that comprises crossing a first parent
carrot plant with a second parent carrot plant, wherein at least
one of the first or second parent carrot plants is a plant provided
herein, such as from the variety Purple Royale. In another
embodiment of the invention, carrot seed and plants produced by the
process are first filial generation (F.sub.1) carrot seed and
plants produced by crossing a plant in accordance with the
invention with another, distinct plant. The present invention
further contemplates plant parts of such an F.sub.1 carrot plant,
and methods of use thereof. Therefore, certain exemplary
embodiments of the invention provide an F.sub.1 carrot plant and
seed thereof.
[0050] In another embodiment of the invention, carrot variety
Purple Royale is crossed to produce seed of the variety designated
Purple Royale. In any cross herein, either parent may be the male
or female parent. In these processes, crossing will result in the
production of seed. The seed production occurs regardless of
whether the seed is collected or not.
[0051] In certain embodiments, the invention provides methods of
introducing a desired trait into a carrot plant comprising the
steps of: (a) crossing a plant of variety Purple Royale with a
second carrot plant that comprises a desired trait to produce
F.sub.1 progeny, (b) selecting an F.sub.1 progeny that comprises
the desired trait(s), e.g., one, two, three or more desired
trait(s), (c) optionally selfing the F.sub.1 progeny one or more
times to produce F.sub.2, F.sub.3, or further generation selfing
progeny, (d) crossing the selected F.sub.1 progeny or the selfing
progeny with a plant of variety Purple Royale to produce backcross
progeny, and (e) selecting backcross progeny comprising the desired
trait(s) and which otherwise has all or essentially all the
physiological and morphological characteristics of carrot variety
Purple Royale, (f) optionally, steps (d) and (e) can be repeated
one or more times, e.g., three or more times such as three, four,
five, six or seven times, in succession to produce higher backcross
progeny (e.g., selected fourth, fifth, sixth, seventh or eighth or
higher backcross progeny) that comprises the desired trait. The
invention also provides carrot plants produced by these methods; a
representative sample of seed of Purple Royale having been
deposited under ATCC Accession Number ______.
[0052] In another aspect of the invention, a carrot plant of
variety Purple Royale comprising an added heritable trait (in
addition to all the morphological and/or physiological
characteristics of Purple Royale) or a modified heritable trait is
provided, e.g., an Essentially Derived Variety of Purple Royale
(such as a mutant, or off-type, natural variant or somaclonal
variant) having one, two or three physiological and/or
morphological characteristics which are different from those of
Purple Royale and which otherwise has all the physiological and
morphological characteristics of Purple Royale, wherein a
representative sample of seed of variety Purple Royale has been
deposited under ATCC Accession Number ______. The heritable trait
may comprise a genetic locus that is, for example, a dominant or
recessive allele. In one embodiment of the invention, a plant of
the invention is defined as comprising a single locus conversion.
For example, one, two, three or more heritable traits may be
introgressed at any particular locus using a different allele that
confers the new trait or traits of interest. In specific
embodiments of the invention, the single locus conversion confers
one or more traits such as, for example, herbicide tolerance,
insect resistance, disease resistance and modulation of plant
metabolism and metabolite profiles. In further embodiments, the
trait may be conferred by a naturally occurring gene introduced
into the genome of the variety by backcrossing, a natural or
induced mutation, a somaclonal variant, an off-type, or a transgene
introduced through genetic transformation techniques into the plant
or a progenitor of any previous generation thereof. When introduced
through transformation, a genetic locus may comprise one or more
genes integrated at a single chromosomal location. Thus, the
invention comprises a method of producing a plant comprising an
added desired trait, the method comprising introducing a transgene
conferring a desired trait into a plant of carrot variety Purple
Royale.
[0053] In still yet another aspect, the invention provides a method
of determining the genotype of a plant of the invention comprising
detecting in the genome of the plant at least a first polymorphism.
The method may, in certain embodiments, comprise detecting a
plurality of polymorphisms in the genome of the plant. For example,
a sample of nucleic acid is obtained from a plant and a
polymorphism or a plurality of polymorphisms is detected in said
nucleic acids. The method may further comprise storing the results
of the step of detecting the plurality of polymorphisms on a
computer readable medium.
[0054] In one embodiment of the invention, the invention provides a
method for producing a seed of a variety derived from Purple Royale
comprising the steps of (a) crossing a carrot plant of variety
Purple Royale with a second carrot plant; and (b) allowing seed of
a variety Purple Royale-derived carrot plant to form. This method
can further comprise steps of (c) crossing a plant grown from said
variety Purple Royale-derived carrot seed with itself or a second
carrot plant to yield additional variety Purple Royale-derived
carrot seed; (d) growing said additional variety Purple
Royale-derived carrot seed of step (c) to yield additional variety
Purple Royale-derived carrot plants; and optionally (e) repeating
the crossing and growing steps of (c) and (d) to generate further
variety Purple Royale-derived carrot plants, e.g. one or more times
such as two times, three times, three or more times such as four
times, five times, six times, seven times or even more times. For
example, the second carrot plant is of an inbred carrot variety, or
alternatively, the second carrot plant in step c) is Purple Royale
or the male or female parent plant of Purple Royale.
[0055] In still yet another aspect, the present invention provides
a method of producing a plant or a seed derived from variety Purple
Royale, the method comprising the steps of: (a) preparing a progeny
plant derived from said variety by crossing a plant of variety
Purple Royale with a second plant; and (b) allowing seed of a
variety Purple Royale-derived carrot plant to form. In one
embodiment, the second plant is a plant of an inbred line or of a
wild accession of Daucus carota.
[0056] The method may additionally comprise: (c) crossing a plant
grown from said variety Purple Royale-derived carrot seed with
itself or a second carrot plant to yield additional variety Purple
Royale-derived carrot seed; (d) growing said additional variety
Purple Royale-derived carrot seed of step (c) to yield additional
variety Purple Royale-derived carrot plants; and optionally (e)
repeating the crossing and growing steps of (c) and (d) to generate
further variety Purple Royale-derived carrot plants. For example,
steps (c) and (d) may be repeated one or more times such as one,
two, three, four five or six, seven, eight, nine, ten, or more
times to produce a further plant derived from the aforementioned
starting variety. The further plant derived from variety Purple
Royale may be an inbred variety, and the aforementioned repeated
crossing steps may be defined as comprising sufficient inbreeding
to produce the inbred variety. In the method, it may be desirable
to select particular plants resulting from step (c) for continued
crossing according to steps (b) and (c). By selecting plants having
one or more desirable traits, a plant is obtained which possesses
some of the desirable traits of the starting plant as well as
potentially other selected traits.
[0057] In certain embodiments, the present invention provides a
method of producing a carrot root comprising: (a) obtaining a plant
of the invention, wherein the plant has been cultivated to
maturity, and (b) collecting the carrot root from said plant.
[0058] The invention also provides for a food or feed product
comprising or consisting of a plant part described herein
preferably a carrot root or part thereof and/or an extract from a
plant part described herein. The food or feed product may be fresh
or processed, e.g., canned, steamed, boiled, fried, blanched and/or
frozen, etc.
[0059] In still another embodiment the invention relates to a seed
or plant produced by selfing a plant of the invention.
[0060] In another aspect the invention refers to packages, e.g., a
container, a bag and the like, comprising at least one of the
following: seeds or seed pellets of carrot variety designated
Purple Royale, carrot plant(s) designated Purple Royale, parts
thereof (e.g. roots), progeny of a carrot plant designated Purple
Royale, parts thereof, EDV of a plant designated Purple Royale or
parts thereof.
[0061] In one embodiment any of the plant of the invention
comprises at least 3, 4, 5 or more of the (average) morphological
and/or physiological characteristics as described in Table 1 for
the carrot variety Purple Royale.
[0062] Another aspect refers to a carrot plant, or a part thereof,
having all or essentially all the physiological and/or
morphological characteristics of a carrot plant of carrot variety
Purple Royale when grown under the same conditions.
[0063] Also provided are one or more progeny plants (offspring or
descendants) of a carrot plant designated Purple Royale obtained by
further breeding with said variety designated Purple Royale. Said
progeny plant(s) has/have essentially all physiological and/or
morphological characteristics of variety Purple Royale when grown
under the same environmental conditions. In one embodiment, said
progeny plant(s) has/have 3, 4, 5, 6, 7, 8, or more, or all of, the
following (average) characteristics (see USDA descriptors) as
described in Table 1.
[0064] Any embodiment discussed herein with respect to one aspect
of the invention applies to other aspects of the invention as well,
unless specifically noted.
[0065] In the description and tables herein, a number of terms are
used. In order to provide a clear and consistent understanding of
the specification and claims, the following definitions are
provided:
[0066] The term "about" is used to indicate that a value includes
the standard deviation of error for the device or method being
employed to determine the value.
[0067] The use of the term "or" in the claims is used to mean
"and/or" unless explicitly indicated to refer to alternatives only
or the alternatives are mutually exclusive, although the disclosure
supports a definition that refers to only alternatives and to
"and/or."
[0068] When used in conjunction with the word "comprising" or other
open language in the claims, the words "a" and "an" denote "one or
more" unless specifically noted.
[0069] The terms "comprise," "have" and "include" are open-ended
linking verbs. Any forms or tenses of one or more of these verbs,
such as "comprises," "comprising," "has," "having," "includes" and
"including," are also open-ended. For example, any method that
"comprises," "has" or "includes" one or more steps is not limited
to possessing only those one or more steps and also covers other
unlisted steps. Similarly, any plant that "comprises," "has" or
"includes" one or more traits is not limited to possessing only
those one or more traits and covers other unlisted traits. The
terms mentioned above also comprise the term "contain" which is
limited to specific embodiments. Thus, one embodiment of the
invention, when the terms "comprise," "have" and "include" are used
to describe a plant, part thereof or a process, refers to an
embodiment wherein the limiting term "contain" is used.
[0070] "Carrot" refers herein to a plant of the species Daucus
carota and parts thereof, e.g., the (edible) root. The most
commonly eaten part of a carrot is a root, although the greens are
edible as well. A carrot is a root vegetable plant, the root
(carrot root) is usually orange in color, though purple, red,
white, and yellow varieties exist, as well. At the tip of a carrot
root is a thin taproot while at the other end (base) of a carrot
root the green is attached.
[0071] "Cultivated carrot" refers to plants of Daucus carota, i.e.
varieties, breeding lines or cultivars of the species Daucus
carota, cultivated by humans and having good agronomic
characteristics; preferably such plants are not "wild plants", i.e.
plants which generally have much poorer yields and poorer agronomic
characteristics than cultivated plants and e.g. grow naturally in
wild populations. "Wild plants" include for example ecotypes, PI
(Plant Introduction) lines, landraces or wild accessions or wild
relatives of a species.
[0072] "USDA descriptors" are the plant variety descriptors
described for carrot in the "Objective description of Variety
Carrot Daucus carota)", ST-470-78 (as published by U.S. Department
of Agriculture, Agricultural Marketing Service, Science and
Technology, Plant Variety Protection Office, Beltsville, Md.
20705.
[0073] "UPOV descriptors" are the plant variety descriptors
described for carrot in the "Guidelines for the Conduct of Tests
for Distinctness, Uniformity and Stability, TG/49/8 (Geneva 2007),
as published by UPOV (International Union for the Protection of New
Varieties and Plants) and is herein incorporated by reference in
its entirety.
[0074] "Core" refers to the phloem and xylem of the root, i.e. the
central vascular tissue. "Cortex" refers to the non-vascular tissue
surrounding the core tissue.
[0075] "Genotype" refers to the genetic composition of a cell or
organism.
[0076] "Phenotype" refers to the detectable characteristics of a
cell or organism, which characteristics are the manifestation of
gene expression.
[0077] As used herein, the term "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, such as
plant organs (e.g. harvested or non-harvested carrot root), plant
cells, plant protoplasts, plant cell and/or tissue cultures from
which whole plants can be regenerated, plant calli, plant cell
clumps, plant transplants, seedlings, hypocotyl, cotyledon, plant
cells that are intact in plants, plant clones or micropropagations,
or parts of plants (e.g. harvested tissues or organs), such as
plant cuttings, vegetative propagations, embryos, pollen, ovules,
flowers, leaves, seeds, clonally propagated plants, roots,
taproots, stems, root tips, grafts, parts of any of these and the
like. Also, any developmental stage is included, such as seedlings,
cuttings prior or after rooting, mature plants, roots or leaves.
Alternatively, plant part may also include a plant seed which
comprises one or two sets of chromosomes derived from the parent
plant.
[0078] "Harvested plant material" refers herein to plant parts
(e.g. a root detached from the whole plant) which have been
collected for further storage and/or further use.
[0079] "Harvested seeds" refers to seeds harvested from a line or
variety, e.g. produced after self-fertilization or
cross-fertilization and collected.
[0080] A plant having "(essentially) all the physiological and/or
morphological characteristics" means a plant having essentially all
or all the physiological and/or morphological characteristics when
grown under the same environmental conditions of the plant of
Purple Royale from which it was derived, e.g. the progenitor plant,
the parent, the recurrent parent, the plant used for tissue- or
cell culture, etc. The skilled person will understand that a
comparison between carrot varieties should occur when said
varieties are grown under the same environmental conditions. For
example, the plant may have all characteristics mentioned in Table
1 when grown under the environmental conditions outlined herein
with reference to the data of Table 1. In certain embodiments, the
plant having "essentially all the physiological and/or
morphological characteristics" are plants having all the
physiological and/or morphological characteristics of Table 1,
except for certain characteristics, such as one, two or three,
mentioned, e.g. the characteristic(s) derived from a converted or
introduced gene or trait and/or except for the characteristics
which differ in an EDV. So, the plant may have all characteristics
mentioned in Table 1, except for one, two or three characteristics
of Table 1, in which the plant may thus differ.
[0081] A plant having one or more or all "essential physiological
and/or morphological characteristics" or one or more
"distinguishing characteristics" (such as one, two, three, four or
five) refers to a plant having (or retaining) one or more, or all,
or retaining all except one, two or three of the distinguishing
characteristics mentioned in Table 1 when grown under the same
environmental conditions that distinguish Purple Royale from most
similar varieties Anthonina, Purple Elite, Purple Haze, and Deep
Purple. For example, such distinguishing characteristics being
selected from (but not limited to): 1) average root core thickness
at midpoint cross-section, 2) average root diameter at midpoint, 3)
carrot length (minus taproot), 4) cross-section interior color of
core and cortex, 5) haloing and zoning of the root at market
maturity, 6) Petiole length from crown to first pinna, 7) average
length of taproot, 8) blade color of leaf at harvest stage, 9)
blade divisions of leaf at harvest stage, 10) leaf petiole
anthocyanin level and petiole pubescence at harvest stage, 11)
average rood diameter at shoulder (see USDA descriptors, and 12)
total anthocyanin levels.
[0082] The physiological and/or morphological characteristics
mentioned above are commonly evaluated at significance levels of
1%, 5%, 8% or 10% significance level, when measured under the same
environmental conditions. For example, a progeny plant of Purple
Royale may have one or more (or all, or all except one, two or
three) of the essential physiological and/or morphological
characteristics of Purple Royale listed in Table 1, or one or more
or all (or all except one, two or three) of the distinguishing
characteristics of Purple Royale listed in Table 1 and above, as
determined at the 1% or 5% significance level when grown under the
same environmental conditions.
[0083] Physiological and/or morphological characteristics which are
"substantially equivalent" or "not significantly different" or "not
significantly differ" refers to a characteristic that, when
compared, does not show a statistically significant difference
(e.g., p>0.05 using ANOVA) from the mean. Vice versa,
"significantly different" or "statistically significantly
different" refers to a characteristic that, when compared, does
show a statistically significant difference (e.g., p<0.05 using
ANOVA) from the mean.
[0084] As used herein, the term "variety" or "cultivar" means a
plant grouping within a single botanical taxon of the lowest known
rank, which grouping, irrespective of whether the conditions for
the grant of a breeder's right are fully met, can be defined by the
expression of the characteristics resulting from a given genotype
or combination of genotypes, distinguished from any other plant
grouping by the expression of at least one of the said
characteristics and considered as a unit with regard to its
suitability for being propagated unchanged.
[0085] The terms "gene converted" or "conversion plant" in this
context refer to carrot plants which are often developed by
backcrossing wherein essentially all of the desired morphological
and/or physiological characteristics of parent are recovered in
addition to the one or more genes transferred into the parent via
the backcrossing technique or via genetic engineering. Likewise, a
"Single Locus Converted (Conversion) Plant" refers to plants which
are often developed by plant breeding techniques comprising or
consisting of backcrossing, wherein essentially all of the desired
morphological and physiological characteristics of a carrot variety
are recovered in addition to the characteristics of the single
locus having been transferred into the variety via, e.g., the
backcrossing technique and/or by genetic transformation. Likewise,
a double-loci converted plant/a triple loci converted plant refers
to plants having essentially all of the desired morphological
and/or physiological characteristics of given variety, expect that
at two or three loci, respectively, it contains the genetic
material (e.g., an allele) from a different variety.
[0086] A variety is referred to as an "Essentially Derived Variety"
(EDV) i.e., shall be deemed to be essentially derived from another
variety, "the initial variety" when (i) it is predominantly derived
from the initial variety, or from a variety that is itself
predominantly derived from the initial variety, while retaining the
expression of the essential characteristics that result from the
genotype or combination of genotypes of the initial variety; (ii)
it is clearly distinguishable from the initial variety; and (iii)
except for the differences which result from the act of derivation,
it conforms to the initial variety in the expression of the
essential characteristics that result from the genotype or
combination of genotypes of the initial variety. Thus, an EDV may
be obtained for example by the selection of a natural or induced
mutant, or of a somaclonal variant, or an off-type, or the
selection of a (natural) variant individual from plants of the
initial variety, backcrossing, or transformation by genetic
engineering. Such a variant may be selected at any time, e.g. in
the field or greenhouse, during breeding, during or after in vitro
culture of cells or tissues, during regeneration of plants, etc.
The term EDV, thus, also encompassed a "phenotypic variant" derived
from Purple Royale seed, plant tissue or cells. In one embodiment,
an EDV is a gene converted plant.
[0087] "Plant line" is for example a breeding line which can be
used to develop one or more varieties.
[0088] "Progeny" (or "descendants") as used herein refers to plants
derived from a plant designated Purple Royale. Progeny may be
derived by regeneration of cell culture or tissue culture or parts
of a plant designated Purple Royale or selfing of a plant
designated Purple Royale or by producing seeds of a plant
designated Purple Royale. In further embodiments, progeny may also
encompass plants derived from crossing of at least one plant
designated Purple Royale with another carrot plant of the same or
another variety or (breeding) line, or with a wild carrot plant,
backcrossing, inserting of a locus into a plant or selecting a
plant comprising a mutation or selecting a variant. A progeny is,
e.g., a first generation progeny, i.e. 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
crossing) or regeneration. However, the term "progeny" generally
encompasses further generations such as second, third, fourth,
fifth, sixth, seventh or more generations, i.e., generations of
plants which are derived from, obtained from, obtainable from or
derivable from the former generation by, e.g., traditional breeding
methods, regeneration or genetic transformation techniques. For
example, a second generation progeny can be produced from a first
generation progeny by any of the methods mentioned above.
Especially progeny of Purple Royale which are EDVs or which retain
all (or all except 1, 2 or 3) physiological and/or morphological
characteristics of Purple Royale listed in Table 1, or which retain
all (or all except 1, 2, or 3) of the distinguishing
characteristics of Purple Royale described elsewhere herein and in
Table 1, are encompassed herein.
[0089] The term "traditional breeding techniques" encompasses
herein crossing, selfing, selection, double haploid production,
embryo rescue, protoplast fusion, marker assisted selection,
mutation breeding etc. as known to the breeder (i.e. methods other
than genetic modification/transformation/transgenic methods), by
which, for example, a genetically heritable trait can be
transferred from one carrot line or variety to another.
[0090] "Crossing" refers to the mating of two parent plants.
[0091] "Cross-pollination" refers to the fertilization by the union
of two gametes from different plants.
[0092] "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 progeny plants. 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".
[0093] "Selfing" refers to self-pollination of a plant, i.e., the
transfer of pollen from the anther to the stigma of the same
plant.
[0094] "Regeneration" refers to the development of a plant from
cell culture or tissue culture or vegetative propagation.
[0095] "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 where plant part is, e.g., defined as or
derived from (e.g. by cutting of) leaf, pollen, embryo, cotyledon,
hypocotyl, cells, nodes, protoplasts, meristematic cell, root, root
tip, pistil, anther, flower, shoot tip, shoot, stem, petiole, etc.
When a whole plant is regenerated by vegetative propagation, it is
also referred to as a vegetative propagation.
[0096] The term "locus" (loci plural) means a specific place or
places or a site on a chromosome where for example a gene or
genetic marker is found. "Linkage" or "genetic linkage" is the
tendency of genes genes or molecular markers that are located
proximal to each other on a chromosome chromosome to be inherited
together during meiosis. Genes or molecular markers whose loci are
nearer to each other are less likely to be separated onto different
chromatids chromatids during chromosomal crossover chromosomal
crossover and are therefore said to be genetically linked.
[0097] "Marker" or "molecular marker" refers to a readily
detectable DNA sequence or nucleotide, which may be genetically
closely linked to a gene or locus. Such closely linked markers can
be used in MAS (marker assisted selection) of the gene or
locus.
[0098] "Marker assisted selection" or "MAS" is a process of using
the presence of molecular markers, which are genetically linked to
a particular locus or to a particular chromosome region, to select
plants for the presence of the specific locus or region.
[0099] The term "allele(s)" means any of one or more alternative
forms of a gene at a particular locus, all of which alleles relate
to one trait or characteristic at a specific locus. In a diploid
cell of an organism, alleles of a given gene are located at a
specific location, or locus (loci plural) on a chromosome. One
allele is present on each chromosome of the pair of homologous
chromosomes. A diploid plant species may comprise a large number of
different alleles at a particular locus. These may be identical
alleles of the gene (homozygous) or two different alleles
(heterozygous).
[0100] 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. These terms
are also used to describe plants showing some symptoms but that are
still able to produce marketable product with an acceptable
yield.
[0101] "Tissue Culture" refers to a composition comprising isolated
cells or tissues of the same or a different type or a collection of
such cells organized into parts of a plant.
[0102] "Average" refers herein to the arithmetic mean.
[0103] The term "mean" refers to the arithmetic mean of several
measurements. The skilled person understands that the appearance of
a plant depends to some extent on the growing conditions of said
plant. Thus, the skilled person will know typical growing
conditions for carrot described herein. The mean, if not indicated
otherwise within this application, refers to the arithmetic mean of
measurements on at least 10 different, randomly selected plants or
plant parts of a variety or line.
[0104] As used herein, the term "anthocyanin" refers to
anthocyanidins with sugar group(s), mostly 3-glucosides of the
anthocyanidins, though other 3-glycosides and 5-glycosides are also
known (Stintzing et al. Agric Food Chem. 2002. Vol 50. 6172-6181).
The anthocyanins can be subdivided into the sugar-free
anthocyanidin aglycone and the anthocyanin glycosides. The
difference in chemical structure that occurs in response to changes
in pH is the reason why anthocyanins are often used as pH
indicators, as they change from red in acids to blue in bases (i.e.
basic environment such as pH>7 or >8 or even pH>9 or
>10 or >11).
[0105] The term "total anthocyanin level" or "total anthocyanin
content" refers to the amount of anthocyanins in the root of the
carrot at market maturity. Whenever reference is made to "total
anthocyanin level", the combined level of all glycosides of
anthocyanidin together is being referred to, without identifying
individual anthocyanins. Different parts of the root of a carrot
may contain different levels of anthocyanin e.g. sampling root
shoulder vs root tips will give different results. The middle
section (between top and shoulder) of a carrot gives a good sample
of the anthocyanin level of the major part of a carrot. Total
anthocyanin levels may also depend somewhat depending on the trial
location and sampling moment (e.g. stage of growth), however when
measured under the same conditions, the variation between genotypes
remains relatively stable and the ranking of genotypes does not
change.
[0106] The total anthocyanin level per carrot can be determined
using methods known in the art for example by purifying fresh
carrot (whole carrot root, excluding tap root), optionally add an
internal standard like malvidin-3-galactoside chloride to determine
extraction efficacy, extract with 10% formic acid in methanol five
times, centrifuge the extracts and combine the supernatants.
Anthocyanins can then be determined by HPLC e.g. by using a C18
column and a mobile phase A of 10% formic acid in water and a
mobile phase B of methanol using a gradient of B from 0 to 55%.
Anthocyanins can be identified by comparing retention times and UV
or MS spectra with known anthocyanins. A standard response curve
for quantifying the anthocyanin may be prepared from
cyanidin-3-glucoside.
[0107] The term "genetic elements conferring the total anthocyanin
level" refers to all genes responsible for the total anthocyanin
level in the root of a carrot plant.
[0108] The term "genetic elements conferring the level of haloing
and zoning" refers to all genes responsible for the level of
haloing and zoning in the root of a carrot plant.
[0109] Genetic elements conferring a specific trait may for example
be determined using methods known in the art such as whole exome
sequencing (also known as targeted exome capture) which is an
approach that involves using sequencing technology and sequence
assembly tools to piece together all coding portions of the genome.
The sequence is then compared to a reference genome and any
differences are noted. After filtering out all known polymorphisms,
synonymous changes, and intronic changes (that do not affect splice
sites), only potentially trait affecting variants will be left.
This technique can be combined with other techniques to further
exclude potentially trait-affecting variants should more than one
be identified. Alternatively, genetic approaches, such as gene
mapping or Quantitative Trait Analysis (using e.g. Bulk Segregant
Analysis, as described by Michelmore et al. Proc Natl Acad Sci USA.
1991 Nov. 1; 88(21): 9828-9832 may be used to identify the number
and location of genes which determine the trait and molecular
markers linked to the genes or genome regions which determine the
trait.
[0110] Seeds of the Carrot Variety Purple Royale.
[0111] Also provided are seeds of carrot variety Purple Royale,
i.e. seeds from which the variety can be grown.
[0112] In one embodiment, a plurality of Purple Royale seeds are
packaged into small and/or large containers (e.g., bags, cartons,
cans, etc.). Seeds may be treated with one or more chemical
compounds and/or biological control agents (e.g. to improved
germination, insecticidal-, acaricidal-, nematicidal- or
fungicidal-compounds or compositions, etc.) and/or seeds may be
primed. Biological control agents are one or more microorganisms
which protect the seed or seedling against pathogens. For example,
strains of bacteria and/or fungi, such as bacteria of the species
of Streptomyces, Pseudomonas, Bacillus and Enterobacter or fungi of
the species Phomopsis, Ectomycorrhizae, Trichoderma, Cladosporium
and Gliocladium.
[0113] Priming is a water-based process that is performed on seeds
to increase uniformity of germination and emergence from the soil,
and thus enhance vegetable stand establishment. Methods how to
prime carrot seeds are well known in the art, see WO2008/107097,
describing different priming methods, such as hydro-priming
(including drum-priming), osmo-priming and solid-matrix priming,
which can be used. The priming process may also be combined with
the chemical compounds or compositions and/or biological control
agent treatment, so seeds may e.g. be hydrated in a first step,
dried in a second step and treated in a third step with one or more
seed treatment compounds or compositions. Priming is also sometimes
referred to as seed conditioning.
[0114] Hydropriming includes those techniques in which seeds are
allowed to take up water for a short period or at low temperatures,
mostly at ample water supply. These techniques are sometimes also
referred to as soaking or steeping. With osmo-priming, the seeds
are exposed to an osmotic solution (see e.g. WO2008/107097).
[0115] With solid matrix priming (SMP), seeds are mixed with water
and solid carriers. Examples of solid carriers are vermiculite and
diatomaceous silica products. The water is taken up by the seeds as
well as absorbed on the solid particle surfaces, which in this way
control the water uptake of the seeds. In addition to using
particle-like carriers, SMP can be carried out using, amongst
others, moist towels, gunny bags, moist sand, sterilized compost or
press mud as well.
[0116] So, in one aspect seeds of Purple Royale are provided
wherein said seeds are primed seeds and/or chemically and/or
biologically treated seeds, comprising one or more chemical
compounds or compositions and/or one or more biological control
agents, selected from the group consisting of: a compound that
improves germination, an insecticidal compound, an acaricidal
compound, a nematicidal compound, and a fungicidal compound.
Breeding with Carrot Plants of the Invention
[0117] One aspect of the current invention concerns methods for
crossing a carrot variety provided herein with itself or a second
carrot plant and the seeds and plants produced by such methods.
These methods can be used for propagation of a variety provided
herein or can be used to produce carrot seeds and the plants grown
therefrom. Such seed can be produced by crossing two parent lines
or varieties to produce the variety.
[0118] In one embodiment of the invention, the first step in
"crossing" comprises planting seeds of a first and a second parent
carrot 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.
[0119] Also in accordance with the invention, novel varieties may
be created by crossing a plant of the invention followed by
multiple generations of breeding according to such well known
methods. New varieties may be created by crossing with any second
plant. In selecting such a second plant to cross for the purpose of
developing novel varieties, it may be desired to choose those
plants that either themselves exhibit one or more selected
desirable characteristics or that exhibit the desired
characteristic(s). Once initial crosses have been made, inbreeding
and selection take place to produce new varieties. For development
of a uniform variety, often five or more generations of selfing and
selection are involved.
[0120] Uniform varieties of new varieties may also be developed by
way of double-haploids. This technique allows the creation of true
breeding varieties without the need for multiple generations of
selfing and selection. In this manner, true breeding varieties can
be produced in as little as one generation. Haploid cells, such as
microspores, pollen, anther cultures, or ovary cultures can be
used. The chromosomes of the haploid cells may then be doubled
autonomously, or by chemical treatments (e.g. colchicine
treatment). From the double-haploid cells, fertile homozygous
plants are obtained. In accordance with the invention, any of such
techniques may be used in connection with a plant of the invention
and progeny thereof to achieve a homozygous variety.
[0121] Backcrossing can also be used to improve an inbred plant.
Backcrossing transfers one or more heritable traits from one inbred
or non-inbred source to an inbred that lacks those traits. The
exact backcrossing protocol will depend on the characteristic(s) or
trait(s) being altered to determine an appropriate testing
protocol. When the term variety Purple Royale is used in the
context of the present invention, this also includes plants
modified to include at least a first desired heritable trait such
as one, two or three desired heritable trait(s).
[0122] This can be accomplished, for example, by first crossing a
superior inbred (recurrent parent) to a donor inbred (non-recurrent
parent), which carries the appropriate genetic information (e.g.,
an allele) at the locus or loci relevant to the trait in question.
The progeny of this cross is then mated back to the recurrent
parent followed by selection in the resultant progeny (first
backcross generation, or BC.sub.1) for the desired trait to be
transferred from the non-recurrent parent. After five or more
backcross generations with selection for the desired trait, the
progeny is heterozygous at loci controlling the characteristic
being transferred but are like the superior parent for most or
almost all other loci. The last backcross generation would be
selfed to give pure breeding progeny for the trait being
transferred.
[0123] The parental carrot plant which contributes the desired
characteristic or characteristics is termed the non-recurrent
parent because it can be used one time in the backcross protocol
and therefore need not recur. The parental carrot plant to which
the locus or loci from the non-recurrent parent are transferred is
known as the recurrent parent as it is used for several rounds in
the backcrossing protocol.
[0124] Many single locus traits have been identified that are not
regularly selected for in the development of a new inbred but that
can be improved by backcrossing techniques. Single locus traits may
or may not be transgenic; examples of these traits include, but are
not limited to, male sterility, herbicide resistance, resistance to
bacterial, fungal, or viral disease, insect resistance, restoration
of male fertility, modified fatty acid or carbohydrate metabolism,
and enhanced nutritional quality. These comprise genes generally
inherited through the nucleus.
[0125] Direct selection or screening may be applied where the
single locus (e.g. allele) acts in a dominant fashion. For example,
when selecting for a dominant allele providing resistance to a
bacterial disease, the progeny of the initial cross can be
inoculated with bacteria prior to the backcrossing. The inoculation
then eliminates those plants which do not have the resistance, and
only those plants which have the resistance allele are used in the
subsequent backcross. This process is then repeated for all
additional backcross generations.
[0126] Although backcrossing methods are simplified when the
characteristic being transferred is a dominant allele, recessive,
co-dominant and quantitative alleles may also be transferred. In
this instance, it may be necessary to introduce a test of the
progeny to determine if the desired locus has been successfully
transferred. In the case where the non-recurrent variety was not
homozygous, the F.sub.1 progeny would not be equivalent. F.sub.1
plants having the desired genotype at the locus of interest could
be phenotypically selected if the corresponding trait was
phenotypically detectable in a heterozygous or hemizygous state. In
the case where a recessive allele is to be transferred and the
corresponding trait is not phenotypically detectable in the
heterozygous of hemizygous state, the resultant progeny can be
selfed, or crossed back to the donor to create a segregating
population for selection purposes. Non-phenotypic tests may also be
employed. Selected progeny from the segregating population can then
be crossed to the recurrent parent to make the first backcross
generation (BC.sub.1).
[0127] Molecular markers may also be used to aid in the
identification of the plants containing both a desired trait and
having recovered a high percentage of the recurrent parent's
genetic complement. Selection of carrot plants for breeding is not
necessarily dependent on the phenotype of a plant and instead can
be based on genetic investigations. For example, one can utilize a
suitable genetic marker which is closely genetically linked to a
trait of interest. One of these markers can be used to identify the
presence or absence of a trait in the offspring of a particular
cross and can be used in selection of progeny for continued
breeding. This technique is commonly referred to as marker assisted
selection. Any other type of genetic marker or other assay that is
able to identify the relative presence or absence of a trait of
interest in a plant can also be useful for breeding purposes.
Procedures for marker assisted selection applicable to the breeding
of carrot are well known in the art. Such methods will be of
particular utility in the case of recessive traits and variable
phenotypes, or where conventional assays may be more expensive,
time consuming or otherwise disadvantageous. Types of genetic
markers which could be used in accordance with the invention
include, but are not necessarily limited to, Simple Sequence Length
Polymorphisms (SSLPs), Simple Sequence Repeats (SSR), Randomly
Amplified Polymorphic DNAs (RAPDs), DNA Amplification
Fingerprinting (DAF), Sequence Characterized Amplified Regions
(SCARs), Arbitrary Primed Polymerase Chain Reaction (AP-PCR),
Amplified Fragment Length Polymorphisms (AFLPs), and Single
Nucleotide Polymorphisms (SNPs).
[0128] Carrot varieties can also be developed from more than two
parents. The technique, known as modified backcrossing, uses
different recurrent parents during the backcrossing. Modified
backcrossing may be used to replace the original recurrent parent
with a variety having certain more desirable characteristics or
multiple parents may be used to obtain different desirable
characteristics from each.
[0129] The variety of the present invention are particularly well
suited for the development of new varieties based on the elite
nature of the genetic background of the variety. In selecting a
second plant to cross with Purple Royale for the purpose of
developing novel carrot varieties, it will typically be preferred
to choose those plants that either themselves exhibit one or more
selected desirable characteristics or that exhibit the desired
characteristic(s). Examples of desirable characteristics may
include, but are not limited to herbicide tolerance, pathogen
resistance (e.g., insect resistance, nematode resistance,
resistance to bacterial, fungal, and viral disease), cytoplasmic
male sterility (CMS), improved harvest characteristics, enhanced
nutritional quality, increased antioxidant content, improved
processing characteristics, high yield, improved characteristics
related to the carrot root flavor, texture, size, shape,
durability, shelf life, and yield, increased soluble solids
content, uniform ripening, delayed or early ripening, seedling
vigor, adaptability for soil conditions, and adaptability for
climate conditions.
DEPOSIT INFORMATION
[0130] Although the foregoing invention has been described in some
detail by way of illustration and example for purposes of clarity
and understanding, it will be obvious that certain changes and
modifications may be practiced within the scope of the invention,
as limited only by the scope of the appended claims.
[0131] All references cited herein are hereby expressly
incorporated herein by reference.
EXAMPLES
Example 1
Development of Carrot Variety Purple Royale
[0132] The variety Purple Royale can be developed from two
replications of 50 plants each, from which 15 plants or plant parts
can be randomly selected and used to measure characteristics. Table
1 provides the USDA descriptors for Purple Royale (this patent
application), and of Anthonina, Purple Elite, Purple Haze, and Deep
Purple (commercial reference varieties).
[0133] In accordance with one aspect of the present invention,
provided is a plant having essentially all physiological and/or
morphological characteristics of a carrot variety Purple Royale. A
description of the physiological and/or morphological
characteristics of carrot variety Purple Royale is presented in
Table 1.
[0134] A. Characteristics of Purple Royale
TABLE-US-00001 TABLE 1 The USDA descriptors for Purple Royale,
Anthonina, Purple Elite, Purple Haze, and Deep Purple a field
trial. The values are mean values. Purple Purple Purple Deep USDA
Descriptor Royale Anthonina Elite Haze Purple 1. TYPE: 6 6 6 6 6 1
= Amsterdam; 2 = Flakee; 3 = Berlicum; 4 = Chantenay; 5 = Danvers;
6 = Imperator; 7 = Nantes; 8 = Other (Specify) 2. REGION OF
ADAPTATION 7 7 7 7 7 IN THE UNITED STATES OF AMERICA: 1 =
Northeast; 2 = Northwest; 3 = Southeast; 4 = Southwest; 5 = North
Central; 6 = South Central; 7 = Most Regions 3. MARKET MATURITY: NA
NA NA NA NA No. Days from Seeding to Harvest 4. PLANT TOP: (At
Harvest NA NA NA NA NA Stage) Habit: 2 2 2 2 2 1 = Erect; 2 =
Semi-erect; 3 = Prostrate Plant Top Height (from 33.44 cm 40.69 cm
39.39 cm 39.46 cm 40.15 cm Shoulder to Top of Crown) Plant Top Neck
Diameter 1.23 cm 1.62 cm 1.41 cm 1.55 cm 1.41 cm Top Attachment: 1
1 1 1 1 1 = Single; 2 = Multiple 5. LEAF: (At Harvest Stage) Name
of Color Chart: RHS Blade Color: 1 = Light Green; 1 2 2 2 2 2 =
Medium Green; 3 = Dark Green; 4 = Other (Specify) Color Chart Value
Blue Green Green Green Green Green Color Chart Value Number 120C
N134B N134B 139B 135B Blade Divisions: 2 2 2 2 2 1 = Fine; 2 =
Medium; 3 = Coarse Blade Length (Without 20.54 cm 22.88 cm 23.53 cm
20.73 cm 20.20 cm Petiole) Petiole Length from Crown 14.64 cm 22.75
cm 22.48 cm 21.69 cm 24.25 cm to First Pinna 5.50 cm 7.71 cm 7.74
cm 7.73 cm 6.80 cm Petiole Anthocyanin: 2 2 2 2 2 1 = Absent; 2 =
Present Petiole Pubescence 2 1 (very 2 2 1 (very (heavy) slight)
slight) 1 = Absent; 2 = Present 6. ROOT: (At Market Maturity)
Cortex Thickness (Midpoint 0.50 cm 0.57 cm 0.72 cm 0.72 cm 0.56 cm
X-Section) Core Thickness (Midpoint 1.23 cm 1.13 cm 1.16 cm 0.98 cm
1.12 cm X-Section) Carrot Length (Minus Taproot) 7.78 cm 9.76 cm
15.07 cm 10.64 cm 9.42 cm Length of Taproot 11.51 cm 12.88 cm 6.66
cm 5.01 cm 8.35 cm Diameter at Shoulder 2.19 cm 2.23 cm 2.55 cm
2.43 cm 2.17 cm Diameter at Midpoint 0.59 cm 1.38 cm 1.80 cm 1.93
cm 1.52 cm Amount Exposed (Above 1 1 1 1 1 Ground): 1 = None; 2 =
1-10%; 3 = 11-20%; 4 = 21-30%; 5 = 31-40%; 6 = >40% Shape: 2 2 2
2 2 1 = Round; 2 = Conic; 3 = Cylindrical Collar: 2 2 2 2 2 1 =
Sunken; 2 = Level; 3 = Square Shoulder: 1 1 1 1 1 1 = Rounded; 2 =
Sloping; 3 = Square Base: 1 1 2 2 1 1 = Pointed; 2 = Medium; 3 =
Blunt Surface Smoothness: 2 2 2 2 2 1 = Very Smooth; 2 = Dimpled or
Corrugated Number of Secondary Root 2 2 2 2 2 Scars: 1 = None; 2 =
Few; 3 = Many Appearance of Secondary Root 2 2 2 2 2 Scars: 1 = Not
Prominent; 2 = Prominent Halo: 2 3 3 3 3 1 = None; 2 = Faint; 3 =
Prominent Zoning: 2 3 2 2 3 1 = None; 2 = Faint; 3 = Prominent
Flavor Harshness: NA NA NA NA NA 1 = Very Harsh; Moderately Harsh;
3 = Mildly Harsh Flavor Sweetness: NA NA NA NA NA 1 = Not Sweet; 2
= Moderately Sweet; 3 = Very Sweet 7. COLORS: Color choices: Name
of Color Chart: RHS Below Ground Exterior Color N187A N187A N79A
N79B N79A Shoulder N187A N187A N79A N79B N79A Skin N187A N187A N79A
N79B N79A Cross-Section Interior Color NA NA NA NA NA Core N79A 6D
5C N25C No color to describe ** Cortex 79A 79A 79A N79A 79A 8.
FLOWER: NA NA NA NA NA 9. DISEASE REACTIONS: NA NA NA NA NA 1 =
Susceptible; 2 = Resistant; give races (if known) 10. INSECT
REACTIONS: NA NA NA NA NA 1 = Susceptible; 2 = Resistant; give
races (if known) 11. PHYSIOLOGICAL NA NA NA NA NA REACTIONS: 1 =
Susceptible; 2 = Resistant Brix NA NA NA NA NA 1 = Susceptible and
2 = Resistant 12: TOTAL ANTHOCYANIN 26.26 ppm 4.01 ppm 0.97 ppm
1.53 ppm 2.93 ppm LEVEL:
[0135] B. Brix *These are typical values. Values may vary due to
environment. Other values that are substantially equivalent are
also within the scope of the disclosure. (--- equals not
measured).
[0136] The degree Brix can be determined in Purple Royale and in
the most similar varieties Anthonina, Purple Elite, Purple Haze,
and Deep Purple.
[0137] The method used was as follows:
[0138] C. Total Anthocyanin Level
[0139] The anthocyanin level of Purple Royale, Anthonina, Purple
Elite, Purple Haze, and Deep Purple was determined using the
following protocols:
[0140] Anthocyanin Extraction
[0141] The skin will be peeled off the root slug and the root will
be cut in half in a longitudinal direction. Half of the root will
be diced into small pieces. About 1.+-.0.0.1 g of the diced root
will be weighed in an extraction bag and 5 mL of 0.025 M potassium
chloride buffer (pH 1.0) will be added to the bag. Next, the root
pieces will be grinded using a pulverizer until chunks are not
visible. 1 mL of the juice will be pipetted into a micro centrifuge
tube. It will be centrifuged at 10,000.times.g for 10 minutes at
room temperature. The supernatant will be used for anthocyanin
analysis.
[0142] Total Anthocyanin Measurement Using Spectrophotometer
[0143] Cyanidin-3-glucoside will be used as a standard to determine
the extinction coefficient of total anthocyanin. The supernatant
from purple carrot extraction will be diluted in 0.025M potassium
chloride buffer (pH 1.0) and absorbance will be read at 510 and 700
nm using UV-VIS spectrophotometer with a sipper attachment. The
anthocyanin concentration will be calculated based on the
absorbance level.
[0144] A separate trial will be performed to quantitatively
determine anthocyanin levels in Purple Royale, Anthonina, Purple
Elite, Purple Haze, and Deep Purple using this protocol. Plants
will be harvested before bolting started. The recorded anthocyanin
levels are the average of about 30 individual roots.
[0145] Dry matter (DM) was determined by i) drying an empty
aluminum pan in the oven for 1 hour and cool down in a desiccator
cabinet; ii) weigh the empty aluminum pan (A=pan wt.); iii) weigh
1.+-.0.0.1 g of diced root in the pan (B=pan+fresh root wt.); iv)
dry in the convection oven at 100.degree. C. for 24 hours or in the
vacuum oven at 90.degree. C. for 18 hours; v) cool in the
desiccator cabinet; vi) weigh the pan with the dried root
(C=pan+dry root wt.); vii) calculate % dry matter
({(C-A)/(B-A)}*100%).
Example 2
Breeding History for Carrot Variety Purple Royale
[0146] Years 2014 and 2015--
[0147] Four different samples of Turkish origin dark purple carrots
were provided by Sensient Colors in early 2014 for testing in
Livingston, Calif. All subsequent years of production were also
grown at the same trial site with essentially the same cultural
practices. Based upon previous experience with carrots with an
annual growth habit, plots (length=50', bed width=40'') of these
four lots were planted on 20 Sep. 2014 in the Livingston Research
Nursery. Annual carrots typically are best suited for planting and
growing during periods of shortening daylength to reduce premature
bolting. When daylength begins to rapidly lengthen in the early
spring, bolting frequently occurs rapidly and roots convert fleshy
tissue into a lignified taproot. Initial observations of these
plots noted extremely tall vegetative top growth, highly pubescent
leaves, and multiple growing points at the crown. Plots were lifted
on 25 Feb. 2015 for steckling selections for potential breeding
use. A wide array of root colors and shapes were observed with many
forked and split roots and high levels of lenticels. Root colors
ranged from a light tan/yellow to violet to extremely dark purple.
Intense selection pressure was placed upon selecting a straight,
non-forked, smooth root with an Imperator type shape. Roots than
met these criteria were further selected for color intensity with
the most desirable selections having an almost black coloration
across both phloem & xylem. Candidate roots were cut
approximately 6'' beneath the crown and visually sorted for color
content at the tip. Elite roots (approximately 10%) were then
laboratory screened for anthocyanin content. Exactly 2 grams of
tissue was grated and anthocyanin determined using a proprietary
rapid extraction process. Approximately the highest 20%
(anthocyanin content) were selected for self-pollination net cages
and immediately replanted. The lot designated `A` had the highest
visual levels of pigment and virtually all selections were made
therefrom. A total of 28 cages were planted in a fashion where the
primary king umbel was enclosed in the cage to create a
self-pollination and other umbels were left on the outside to
randomly pollinate with other purple/black carrots to create
half-sib populations. At bloom, pupae of blue-bottle flies were
inserted into these cages for pollination. Four larger cages
composed of at least 50 roots/cage were also planted from roots
with less extensive selection pressure. All seeds were harvested in
mid-July 2015.
[0148] Years 2015 and 2016--
[0149] On Sep. 30, 2015, 24 plots (10'.times.40'') were
planted.
[0150] Twenty-two were from self-pollinations of elite roots and
two were from half-sib populations. Additionally, 0.2 acres were
bulk planted from the visually selected large cage 510 and two
larger plots (50'.times.40'') with remnant seed of the original
seed provided by Sensient Colors. Foliage for plots derived from
self-pollination cages was distinctly more compact and consistent
than the larger tops exhibited by the original population, the
half-sibs, or the less aggressively selected larger cages. Plots
were lifted for evaluation and selection on Mar. 17, 2016. Plot
number C60212 was identified as having the best combination of a
long, straight, smooth root combined with consistent dark pigment,
which was further confirmed by laboratory analysis. Most other
plots derived from self- or mass-pollinations demonstrated
inconsistent root shapes and color segregation ranging from
tan/yellow roots to violet to dark purple/black pigment.
Altogether, eight individual cages for additional self-pollination
and one large cage (10'.times.20') were replanted on 25 Mar. 2016
for regrowth and seed production. Seed from all cages was harvested
in late July 2016.
[0151] Years 2016 and 2017--
[0152] Plots (50'.times.40'') of harvested 2016 seed were planted
on 16 Sep. 16, 2016. Growing conditions that winter were extremely
challenging. Near record rainfall kept soil moisture at or above
saturation and created disease pressure and delayed growth.
Stecklings were not lifted until 2 Apr. 2017, which was the latest
possible date due to the onset of bolting. Only limited selections
were made. Relevant, however, was the anthocyanin test performed
comparing the elite cage 668 vs. the original unselected Turkish
material. This advanced selection contained 51% higher anthocyanin
content than the source population, better overall color
uniformity, and more consistent root shape. Overall, four large
cages (2 @ 20'.times.40' and 2 @ 10.times.20') were planted to
increase seed of this elite selection.
[0153] Years 2017 and 2018--
[0154] 2 acres of bulk increase of seed from the four 2017 cages
was planted on Sep. 26, 2017 to produce stecklings for large scale
open multiplication (0.6 acres) that was harvested in July 2018
with a yield of approximately 600 lbs. that was subsequently
designated `Purple Royale`. Photos below provide reference to the
visual appearance of selected stecklings and the subsequent 2018
seed production. Steckling photos demonstrate the color uniformity
and intensity obtained by selection program as well as improvements
in root shape consistency. The current breeding generation is SM3
indicating one generation of self-pollination followed by three
generations of mass pollination.
* * * * *