U.S. patent application number 16/579262 was filed with the patent office on 2020-01-09 for novel peppers with unique aroma and taste.
This patent application is currently assigned to Seminis Vegetable Seeds, Inc.. The applicant listed for this patent is Seminis Vegetable Seeds, Inc.. Invention is credited to Swapan K. Chaudhuri, Howard L. Constant, Graeme S. Garvey, Brian J. Just.
Application Number | 20200008383 16/579262 |
Document ID | / |
Family ID | 58797987 |
Filed Date | 2020-01-09 |
United States Patent
Application |
20200008383 |
Kind Code |
A1 |
Chaudhuri; Swapan K. ; et
al. |
January 9, 2020 |
Novel Peppers with Unique Aroma and Taste
Abstract
This disclosure concerns the improvement of flavor in peppers by
addition of aromas from other or the same pepper species. The
disclosure describes novel Capsicum germplasm with aromatic
components from Capsicum chinense and other horticultural traits
from Capsicum annuum. The disclosure also describes a method for
improving the flavor of a pepper through metabolomic-assisted
selection to produce a pepper fruit with desirable aromas. The
disclosure further provides multiple quantitative trait loci
(QTLs), associated markers, and their use for genetic breeding of
pepper plants with desired sweetness and flavor.
Inventors: |
Chaudhuri; Swapan K.;
(Sacramento, CA) ; Constant; Howard L.;
(Sacramento, CA) ; Garvey; Graeme S.; (Woodland,
CA) ; Just; Brian J.; (Fort Myers, FL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Seminis Vegetable Seeds, Inc. |
St. Louis |
MO |
US |
|
|
Assignee: |
Seminis Vegetable Seeds,
Inc.
St. Louis
MO
|
Family ID: |
58797987 |
Appl. No.: |
16/579262 |
Filed: |
September 23, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15368392 |
Dec 2, 2016 |
10463014 |
|
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16579262 |
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62261951 |
Dec 2, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C12Q 1/6895 20130101;
A01H 5/08 20130101; C12Q 2600/13 20130101; A01H 6/822 20180501;
C12Q 2600/156 20130101; A01H 1/04 20130101 |
International
Class: |
A01H 5/08 20060101
A01H005/08; A01H 6/82 20060101 A01H006/82; C12Q 1/6895 20060101
C12Q001/6895 |
Claims
1-20. (canceled)
21. A Capsicum seed, plant grown therefrom, or part of either,
wherein pepper fruit at or near maturity obtained from said seed or
plant or part have a pericarp comprising a total sugar content of
at least 5.5%, and one or more of alpha-Cubebene at a level having
a Relative GC/MS Response Score of at least 0.05, delta-Cadinene at
a level having a Relative GC/MS Response Score of at least 0.1, or
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.05.
22. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said pericarp comprises a titratable
acidity at a higher level than that of the pericarp of hybrid
pepper PS09954859 grown under similar conditions.
23. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said pericarp comprises a titratable
acidity of at least 4.2 mmol H.sup.+/100 grams fresh tissue.
24. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said Capsicum seed is a sweet bell
pepper seed.
25. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said pepper fruit comprises a
pungency of below 5 Scoville Heat Units (SHUs).
26. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said Capsicum seed is a hybrid.
27. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein at least 70% of the nuclear genetic
material of said Capsicum seed is from a C. annuum background.
28. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein less than 30% of the nuclear genetic
material of said Capsicum seed is from a C. chinense
background.
29. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said Capsicum seed is Capsicum
annuum line ZSP8T14-6274, or SVPS2625, wherein a representative
sample of seed of said Capsicum annuum line ZSP8T14-6274 has been
deposited at ATCC under Accession No. PTA-122300, and wherein a
representative sample of seed of said Capsicum annuum line SVPS2625
has been deposited at ATCC under Accession No. PTA-122296.
30. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21; wherein said seed, plant, or part is a
Capsicum annuum seed, plant, or part; wherein said seed or plant or
part is heterozygous for a SMO2 QTL; wherein said SMO2 QTL is
identifiable by flanking markers SEQ ID NO: 1 and 2 and is
obtainable from the Capsicum annum line SVPS2625, a representative
sample seed of the Capsicum annuum line SVPS2625 having been
deposited at ATCC under Accession No. PTA-122296.
31. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21; wherein said seed, plant, or part is a
Capsicum annuum seed, plant, or part; wherein said seed or plant or
part is heterozygous for a SMO3 QTL; wherein said SMO3 QTL is
identifiable by flanking markers SEQ ID NO: 3 and 4 and is
obtainable from the Capsicum annuum line SVPS2625, a representative
sample seed of the Capsicum annuum line SVPS2625 having been
deposited at ATCC under Accession No. PTA-122296.
32. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein said Capsicum seed is an F.sub.1
progeny of a first parent selected from the group consisting of
Capsicum line ZSP8T14-6274 and Capsicum line SVPS2625, wherein a
representative sample of seed of said Capsicum line ZSP8T14-6274
has been deposited at ATCC under Accession No. PTA-122300, and
wherein a representative sample of seed of said Capsicum line
SVPS2625 has been deposited at ATCC under Accession No.
PTA-122296.
33. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 32, wherein said seed, plant grown therefrom, or
part of either is heterozygous for a SMO2 QTL; wherein said SMO2
QTL is identifiable by marker SEQ ID NO: 1 or 2 and is obtainable
from the Capsicum line SVPS2625, a representative sample seed of
the Capsicum line SVPS2625 having been deposited under Accession
No. PTA-122296.
34. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 32, wherein said seed, plant grown therefrom, or
part of either is heterozygous for a SMO3 QTL; wherein said SMO3
QTL is identifiable by marker SEQ ID NO: 3 or 4 and is obtainable
from the Capsicum line SVPS2625, a representative sample seed of
the Capsicum line SVPS2625 having been deposited under Accession
No. PTA-122296.
35. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 32, wherein said part is an ovule or pollen.
36. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 32, wherein the second parent of said F.sub.1
progeny is: a. a plant of an elite variety selected from the group
consisting of Aleppo, Anaheim, ancho, bell, cascabel, cayenne,
chilaca, chiltepin, cubanelle, de arbol, dandicut, Fresno,
guajillo, Hungarian wax, Italian sweet, jalapeno, Japanese,
mirasol, macho, mulato, New Mexico, pasilla, pepperoncini, piquin,
pimento, poblano, puya, Serrano, and Tientsin; or b. a plant of a
Capsicum annuum variety, wherein the pericarp obtained from a fruit
of said Capsicum annuum variety comprises a total sugar content of
at least 5.5%.
37. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein pepper fruit at or near maturity
obtained from said Capsicum seed or plant or part have a pericarp
which comprises two or more of alpha-Cubebene at a level having a
Relative GC/MS Response Score of at least 0.05, delta-Cadinene at a
level having a Relative GC/MS Response Score of at least 0.1, or
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.05.
38. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein pepper fruit at or near maturity
obtained from said Capsicum seed or plant or part have a pericarp
which comprises alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.05, delta-Cadinene at a level having a
Relative GC/MS Response Score of at least 0.1, and 1,4-Cadinadiene
at a level having a Relative GC/MS Response Score of at least
0.05.
39. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein pepper fruit at or near maturity
obtained from said Capsicum seed or plant or part have a pericarp
which comprises one or more of alpha-Cubebene at a level having a
Relative GC/MS Response Score of at least 0.25, delta-Cadinene at a
level having a Relative GC/MS Response Score of at least 0.25, or
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.15.
40. A Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein pepper fruit at or near maturity
obtained from said Capsicum seed or plant or part have a pericarp
which comprises two or more of alpha-Cubebene at a level having a
Relative GC/MS Response Score of at least 0.25, delta-Cadinene at a
level having a Relative GC/MS Response Score of at least 0.25, or
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.15.
41. The Capsicum seed, plant grown therefrom, or part of either,
according to claim 21, wherein pepper fruit at or near maturity
obtained from said Capsicum seed or plant or part have a pericarp
which comprises alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.25, delta-Cadinene at a level having a
Relative GC/MS Response Score of at least 0.25, and 1,4-Cadinadiene
at a level having a Relative GC/MS Response Score of at least 0.15.
Description
REFERENCE TO RELATED APPLICATION
[0001] This application is a continuation of Ser. No. 15/368,392,
filed Dec. 2, 2016, which claims the benefit of U.S. provisional
application No. 62/261,951, filed on Dec. 2, 2015, which are herein
incorporated by reference in their entireties.
INCORPORATION OF SEQUENCE LISTING
[0002] A computer readable form of the Sequence Listing is filed
with this application by electronic submission and is incorporated
into this application by reference in its entirety. The Sequence
Listing is contained in the file created on Sep. 19, 2019, having
the file name P34325US02_.txt, and which is 24,698 bytes in size
(as measured in the MS-Windows.RTM.).
FIELD
[0003] The present disclosure relates to and includes novel pepper
plants and methods of their production. In particular the
disclosure relates to the improvement of flavor in sweet peppers by
addition of desirable aromas from other pepper species. The
disclosure also describes metabolomics-assisted selection of pepper
plants with desirable aromas or flavors.
BACKGROUND
[0004] Peppers are members of the Solanaceae family and the genus
Capsicum, which includes species such as C. annuum, C. baccatum, C.
cardenasii, C. chacoense, C. chinense, C. ciliatum, C. eximium, C.
flexuosum, C. frutescens, C. galapagoense, C. praetermissum, C.
pubescens, and C. tovarii. Peppers are cultivated and used around
the world as sweet peppers, such as bell peppers (C. annuum); or as
pungent chili peppers, jalapeflo peppers, Habanero peppers (C.
chinense) and TABASCO.RTM. peppers (C. frutescens); or as a source
of dried powders of various colors, such as paprika. Cultivated
peppers can be distinguished by their pungency, fruit shape, color
and size (see for example U.S. Pat. No. 6,498,287).
[0005] Some consumers regard certain types of peppers, for example,
sweet bell peppers, as lacking in aroma, or as having a bland
flavor, or as having an undesirable flavor. One approach is to
breed for sweet peppers with more desirable aromas. C. chinense
comprises some extremely pungent peppers and has traditionally been
used as donors for certain disease resistance traits. However, C.
chinense has not been explored as a source for desirable pepper
flavor. Little is known about aromatic components (e.g., aroma
compounds) underlying the C. chinense fruit flavor. No objective
analytical method has been reported to assess C. chinense aroma or
flavor. It is also unclear whether C. chinense fruit aromas are
separable from its high pungency. Numex Suave Orange (C. chinense)
has been reported as a mild habanero-type pepper, but not a sweet
pepper. See Votava and Bosland, HortScience, 39(3):627-28 (2004).
Accordingly, there is a need in pepper breeding to incorporate
desirable aromas or flavor (e.g., aromas characteristic of C.
chinense) into sweet peppers.
SUMMARY
[0006] In an aspect, this disclosure provides a Capsicum seed or
plant grown therefrom, where the Capsicum plant exhibits a pepper
fruit at, or immediately prior to, maturity having pericarp
comprising a total sugar content of at least 5.5% and one or more
aroma molecules characteristic of Capsicum chinense. In an aspect,
a pepper fruit disclosed herein comprises one or more aroma
molecules characteristic of Capsicum chinense selected from the
group consisting of alpha-Cubebene, delta-Cadinene,
1,4-Cadinadiene, and combinations thereof. In an aspect, a pepper
fruit comprises pericarp having a titratable acidity at a level
higher than that of pericarp of hybrid pepper PS09954859 grown
under similar conditions. In another aspect, a pepper fruit
comprises pericarp having a titratable acidity of at least 4.2
millimole (mmol)H.sup.+/100 g fresh tissue.
[0007] In an aspect, a pepper fruit disclosed herein comprises
pericarp comprising alpha-Cubebene at a level having a Relative
GC/MS Response Score of at least 0.25. In another aspect, a pepper
fruit disclosed herein comprises pericarp comprising delta-Cadinene
at a level having a Relative GC/MS Response Score of at least 0.25.
In a further aspect, a pepper fruit disclosed herein comprises
pericarp comprising 1,4-Cadinadiene at a level having a Relative
GC/MS Response Score of at least 0.15.
[0008] In another aspect, this disclosure provides a Capsicum seed
or plant grown therefrom, where the Capsicum plant exhibits a
pepper fruit at, or immediately prior to, maturity having pericarp
comprising a total sugar content of at least 5.5% and one or more
aroma molecules characteristic of Capsicum chinense, where the one
or more aroma molecules are absent from a Capsicum with no Capsicum
chinense in the pedigree, absent from the pericarp of hybrid pepper
PS09954859 or at a higher level than that of pericarp of hybrid
pepper PS09954859 grown under similar conditions.
[0009] In an aspect, a Capsicum seed, plant, or fruit disclosed
herein is a progeny of Capsicum line ZSP8T14-6274 or SVPS2625,
where a representative sample of seed of the Capsicum line
ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, and where a representative sample of seed of the
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
[0010] In another aspect, a Capsicum seed, plant, or fruit
disclosed herein is an F.sub.1 progeny of Capsicum line
ZSP8T14-6274 or SVPS2625, where a representative sample of seed of
the Capsicum line ZSP8T14-6274 has been deposited at ATCC under
Accession No. PTA-122300, and where a representative sample of seed
of the Capsicum line SVPS2625 has been deposited at ATCC under
Accession No. PTA-122296.
[0011] In an aspect, a Capsicum seed, plant, or fruit disclosed
herein is an F.sub.2 progeny of Capsicum line ZSP8T14-6274 or
SVPS2625, where a representative sample of seed of the Capsicum
line ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, and where a representative sample of seed of the
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
[0012] In another aspect, a Capsicum seed, plant, or fruit
disclosed herein is essentially derived from Capsicum line
ZSP8T14-6274 or SVPS2625, where a representative sample of seed of
the Capsicum line ZSP8T14-6274 has been deposited at ATCC under
Accession No. PTA-122300, and where a representative sample of seed
of the Capsicum line SVPS2625 has been deposited at ATCC under
Accession No. PTA-122296.
[0013] In a further aspect, a Capsicum seed, plant, or fruit
disclosed herein comprises one or more introgressed loci from
Capsicum line ZSP8T14-6274 or SVPS2625, where a representative
sample of seed of the Capsicum line ZSP8T14-6274 has been deposited
at ATCC under Accession No. PTA-122300, where a representative
sample of seed of the Capsicum line SVPS2625 has been deposited at
ATCC under Accession No. PTA-122296, and where the one or more
introgressed loci provide the genetic determinant for producing the
one or more aroma molecules characteristic of Capsicum
chinense.
[0014] In an aspect, this disclosure provides a Capsicum seed or
plant grown therefrom, where the Capsicum plant exhibits a fruit
comprising at maturity one or more aroma molecules characteristic
of Capsicum chinense at a level equal to or higher than that of a
Capsicum line when grown under similar conditions, where said
Capsicum line is selected from the group consisting of lines
ZSP8T14-6274 and SVPS2625, where a representative sample of seed of
Capsicum line ZSP8T14-6274 has been deposited at ATCC under
Accession No. PTA-122300, where a representative sample of seed of
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
[0015] In an aspect, a Capsicum seed or plant disclosed herein
exhibits a pepper fruit at, or immediately prior to, maturity
having pericarp comprising a total sugar content of at least 5.5%
and one or more terpene molecules selected from the group
consisting of alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.25, delta-Cadinene at a level having a
Relative GC/MS Response Score of at least 0.25, 1,4-Cadinadiene at
a level having a Relative GC/MS Response Score of at least 0.15,
and any combination thereof.
[0016] In another aspect, this disclosure provides a sweet bell
pepper fruit comprising at maturity one or more aroma molecules
characteristic of Capsicum chinense, where the one or more aroma
molecules are absent from pericarp of hybrid pepper PS09954859 or
at a higher level than that of pericarp of hybrid pepper PS09954859
grown under similar conditions.
[0017] In another aspect, this disclosure provides a method of
producing a pepper fruit, where the method comprises: cultivating a
pepper plant provided herein; and collecting a pepper fruit from
the plant.
[0018] In an aspect, this disclosure provides a method for
producing a Capsicum plant capable of producing a sweet pepper
fruit comprising one or more aroma molecules characteristic of
Capsicum chinense, where the method comprises: (a) crossing a C.
annuum plant or hybrid with a C. chinense plant or hybrid to
produce a population of progeny Capsicum plants; and (b) selecting
a progeny Capsicum plant comprising a C. annuum fruit type and
producing a fruit at, or immediately prior to, maturity comprising
one or more aroma molecules characteristic of C. chinense, where
the one or more aroma molecules are selected from the group
consisting of alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.25; delta-Cadinene at a level having a
Relative GC/MS Response Score of at least 0.25; 1,4-Cadinadiene at
a level having a Relative GC/MS Response Score of at least 0.15;
and any combinations thereof.
[0019] In another aspect, this disclosure provides a method for
producing a Capsicum plant capable of producing a sweet pepper
fruit comprising one or more aroma molecules characteristic of
Capsicum chinense, where the method comprises: (a) crossing a C.
annuum plant or hybrid with a C. chinense plant or hybrid to
produce a population of progeny Capsicum plants; and (b) selecting
a progeny Capsicum plant comprising a C. annuum fruit type and
producing a fruit at, or immediately prior to, maturity comprising
one or more aroma molecules characteristic of C. chinense.
[0020] In another aspect, this disclosure provides a method for
selecting a sweet pepper fruit comprising one or more aroma
molecules characteristic of Capsicum chinense, where the method
comprises: (a) obtaining a sweet pepper fruit; and (b) detecting in
the sweet pepper fruit one or more aroma molecules selected from
the group consisting of alpha-Cubebene, delta-Cadinene,
1,4-Cadinadiene, and combinations thereof.
[0021] In another aspect, this disclosure provides a method for
introducing a desired Capsicum chinense specific aroma molecule
into a sweet pepper plant, where the desired Capsicum chinense
specific aroma molecule is selected from the group consisting of
alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene, and any
combinations thereof, where the method comprises: [0022] a.
crossing a sweet pepper plant with a plant selected from the group
consisting of Capsicum line ZSP8T14-6274, a representative sample
seed of which line having been deposited at ATCC under Accession
No. PTA-122300, a progeny line of Capsicum line ZSP8T14-6274,
Capsicum line SVPS2625, a representative sample of seed of which
line having been deposited at ATCC under Accession No. PTA-122296,
and a progeny line of Capsicum line SVPS2625, [0023] b. selecting
an F.sub.1 progeny pepper plant comprising the desired Capsicum
chinense specific aroma molecule in a fruit at, or immediately
prior to, maturity; [0024] c. backcrossing the F.sub.1 progeny to
the sweet pepper plant; [0025] d. selecting a backcrossed progeny
pepper plant comprising the desired Capsicum chinense specific
aroma molecule; [0026] e. repeating steps (c) and (d) three or more
times in succession to produce selected fourth or higher backcross
progeny that comprise the desired Capsicum chinense specific aroma
molecule.
[0027] In another aspect, this disclosure provides a method for
producing a Capsicum plant exhibiting a sweet pepper fruit
comprising one or more desirable aromas, where the method
comprises: (a) crossing a C. annuum plant or hybrid with a Capsicum
plant or hybrid capable of producing a fruit exhibiting the one or
more desirable aromas to produce a population of progeny Capsicum
plants; and (b) selecting a progeny Capsicum plant comprising a C.
annuum fruit type and exhibiting a fruit comprising the one or more
desirable aromas.
BRIEF DESCRIPTION OF THE FIGURES
[0028] FIG. 1: Sample specifications for the consumer liking test
detailing hybrid pepper SVPS2625, PERO.RTM. brand mini-pointy
yellow, and PERO.RTM. brand mini-pointy orange peppers (PERO.RTM.
Family Farms Food Company, LLC).
[0029] FIG. 2: Flow chart of the preparation steps for diced
peppers used in the consumer liking test.
[0030] FIG. 3: Flow chart of the pepper non-volatile and volatile
sampling process used in the consumer liking test.
[0031] FIG. 4: Hedonic attributes as determined by consumers for
the hybrid pepper SVPS2625, PERO.RTM. brand mini-pointy yellow, and
PERO.RTM. brand mini-pointy orange peppers (PERO.RTM. Family Farms
Food Company, LLC) during a sequential monadic liking test of 142
consumers. Consumers grade each of the attributes on a scale of 1
to 9. A score of 1 meaning "dislike extremely", a score of 2
meaning "dislike very much", a score of 3 meaning "dislike
moderately", a score of 4 meaning "dislike slightly", a score of 5
meaning "neither like nor dislike", a score of 6 meaning "like
slightly", a score of 7 meaning "like moderately", a score of 8
meaning "like very much", and a score of 9 meaning "like
extremely".
DETAILED DESCRIPTION
[0032] Unless defined otherwise herein, terms are to be understood
according to conventional usage by those of ordinary skill in the
relevant art.
[0033] As used herein, terms in the singular and the singular forms
"a," "an," and "the," for example, include plural referents unless
the content clearly dictates otherwise. Thus, for example,
reference to "plant," "the plant," or "a plant" also includes a
plurality of plants; also, depending on the context, use of the
term "plant" can also include genetically similar or identical
progeny of that plant.
[0034] As used herein, "metabolomics assisted selection" (MAS) is a
process by which phenotypes are selected based on a metabolic
profile. "Metabolomics assisted selection breeding" refers to the
process of selecting a desired trait or traits in a plant or plants
by detecting one or more metabolites from the plant, where the
metabolites are associated with the desired trait, and then
selecting the plant or germplasm possessing those one or more
metabolites.
[0035] As used herein, a "pepper" refers to a plant of the genus
Capsicum or a fruit thereof.
[0036] As used herein, a "sweet pepper" refers to a fruit or a
plant of a non-pungent sweet pepper variety. Example sweet peppers
include, but are not limited to, bell peppers (C. annuum), the
"Thai sweet"--also a cultivar of C. annuum, and the "dulce"--a
popular cultivar of C. baccatum.
[0037] As used herein, a "bell pepper" refers to a C. annuum pepper
plant or fruit with a bell-shaped or blocky fruit. For bell
peppers, the fruit length divided by the fruit width is in a range
from 0.8 to 1.2.
[0038] As used herein, a "C. annuum fruit type" refers to a fruit
type having a C. annuum fruit appearance, e.g., a bell-shaped,
blocky, pointed, or round fruit.
[0039] Sweet bell pepper, a cultivar of C. annuum, has a zero
rating on the Scoville scale. Without being bound by any scientific
theory, the lack of capsaicin in sweet bell peppers is reported to
be due to a recessive mutation that eliminates capsaicin. The Pun1
locus, formerly known as the C locus, encodes a putative
acyltransferase AT3, of which the allelic state functions as an
on-off switch for pungency in pepper. Hot (pungent) peppers have a
functional CS (Pun1) allele, but in sweet (non-pungent) peppers the
predominant pun1-1 allele has a deletion in an AT3 gene and, as a
consequence, capsaicinoids cannot be formed. See Wahyuni, Ph.D.
thesis entitled "Breeding for pepper fruit quality: A genetical
metabolomics approach," Wageningen University, Chapter 1, page 6
(2014).
[0040] As used herein, "maturity" or "harvest maturity" refers to a
pepper fruit developmental stage when the pepper fruit has fully
developed (e.g., reached its final size) and is ready for harvest.
The development of pepper fruits shows a growth (between 4-7 weeks
after fruit set) and ripening (7-10 weeks after fruit set) phase.
During growth, fresh weight increases rapidly, while during
ripening the color turns from green to a mature color, for example
red, yellow or orange, which is reflected in a decreased level of
chlorophyll a (i.e., green pigment) and an increased level of
carotenoids (i.e., orange/red pigments). As used herein, "at or
near maturity" or "at or immediately prior to maturity" refers to a
pepper fruit ripening stage where between about 50% and 100% (more
preferably between 75% and 100%) of total fruit surface area
exhibits mature color.
[0041] As used herein, "flavor" refers to the sensory impression of
a pepper fruit or fruit part (fruit flesh) perceived during
consumption. Flavor is determined mainly by the chemical senses of
taste and smell. Flavor can be influenced by volatile and/or
non-volatile chemical components (organic acids, lipids,
carbohydrates, salts, etc.). The composition of non-volatile
compounds influences mainly the sensory perceived taste.
Non-volatile compounds include, but are not limited to, sugars
(e.g., sucrose, glucose, and fructose) and organic acids (e.g.,
ascorbic acid, citric acid, malic acid, oxalic acid, fumaric acid,
shikimic acid, and pyroglutamic acid).
[0042] As used herein, fruit "aroma" refers to the smell of a fruit
and is mainly affected by volatile compounds. Exemplary aromatic
compounds in sweet peppers have been reported, including
2,3-butanedione (caramel odor), 1-penten-3-one (chemical/pungent,
spicy), hexanal (grassy), 3-carene (red bell pepper, rubbery),
(Z)-.beta.-ocimene (rancid, sweaty), octanal (fruity),
2-isobutyl-3-methoxypyrazine (green bell pepper),
(Z)-linalooloxide, (Z)-2-penten-1-ol, (E)-geranylacetone,
(E,Z)-2,6-nonadienal, and (E,E)-decadienal. See, e.g., Selahel et
al., "Postharvest responses of red and yellow sweet peppers grown
under photo-selective nets," Food Chemistry, 173:951-56 (2015);
Junior et al., "Analysis of the volatile compounds of Brazilian
chilli," Food Research International 48:98-107 (2012).
[0043] As used herein, "pepper aroma compounds" refer to compounds
associated with the aroma of a pepper fruit. Concentrations of
pepper aroma compounds can be measured by any known metabolite
profiling methods in the art including, without limitation, gas
chromatography mass spectrometry (GC-MS), nuclear magnetic
resonance spectroscopy, liquid chromatography-linked mass
spectrometry. See The Handbook of Plant Metabolomics, edited by
Weckwerth and Kahl, (Wiley-Blackwell) (May 28, 2013); see also,
Barrett et al., "Color, Flavor, Texture, and Nutritional Quality of
Fresh-Cut Fruits and Vegetables: Desirable Levels, Instrumental and
Sensory Measurement, and the Effects of Processing," Critical
Reviews in Food Science and Nutrition, 50(5):369-89 (2010).
[0044] As used herein, "C. chinense specific aroma compounds" or
"aroma compounds characteristic of C. chinense" refer to aroma
compounds which are found in C. chinense, but not in C. annuum.
Example C. chinense specific aroma compounds include, but are not
limited to, .alpha.-Cubebene, .delta.-Cadinene, and
1,4-Cadinadiene.
[0045] As used herein, "titratable acidity" refers to the number of
protons recovered during a titration with a strong base to a
specified endpoint. It can also be expressed as a molar quantity
(e.g., millimoles H.sup.+/100 g) and measured by titration of a
sample using a Mettler T90 titrator.
[0046] As used herein, "total sugar content" refers to the total
percentage of fructose and glucose in a pepper sample.
[0047] As used herein, a "Relative GC/MS Response Score" refers to
a relative quantification of a volatile compound using solid-phase
microextraction (SPME) coupled with gas chromatography mass
spectrometry (GC-MS) and an internal deuterated standard (5 ppm,
d3-Ethyl Acetate in methanol) as described in Example 2. In short,
the retention time and unique ions for a compound of interest are
first used to extract the area under its curve. Curve area for a
deuterated internal standard is then calculated using a retention
time of 61.2 seconds and selected ions (46+91 m/z). The relative
GC-MS response score of a compound is then determined using the
following formula: Relative GC-MS Response Score of compound X=Area
of Compound X/Area of Deuterated Internal Standard.
[0048] As used herein, "capsaicinoid" refers to a collection of
compounds including capsaicin [N-vanillyl-8-methyl-6-nonenamide],
dihydrocapsaicin, and other analogs.
[0049] As used herein, "pericarp" refers to the wall of a pepper
fruit, which is the colored, edible part of the pepper fruit.
[0050] A "C. annuum plant" is a plant of the genus Capsicum and of
the species annuum. A C. annuum plant can be a pure C. annuum plant
(e.g., 100% of its genetic material is from C. annuum) or a C.
annuum hybrid. A "C. annuum hybrid" is a C. annuum plant having one
or more segments of nuclear DNA introgressed from another member of
the Capsicum genus, where greater than about 50%, 60%, 75% of the
nuclear DNA is DNA derived from a C. annuum plant.
[0051] A "C. chinense plant" is a plant of the genus Capsicum and
of the species chinense. A C. chinense plant can be a pure C.
chinense plant (e.g., 100% of its genetic material is from C.
chinense) or a C. chinense hybrid. A "C. chinense hybrid" is a C.
chinense plant having one or more segments of nuclear DNA
introgressed from another member of the Capsicum genus, where
greater than about 75% of the nuclear DNA is DNA derived from a C.
chinense plant. In an aspect, a C. chinense hybrid has greater than
80%, 85%, 90%, 95%, 98% or 99% of its nuclear DNA derived from a C.
chinense plant.
[0052] As used herein, "germplasm" refers to living sources of
genetic material. The germplasm can be part of an organism or cell,
or can be separate from the organism or cell. In general, germplasm
provides genetic material with a specific molecular makeup that
provides a physical foundation for some or all of the hereditary
qualities of an organism or cell culture. As used herein, germplasm
includes cells, seed or tissues from which new plants may be grown,
or plant parts, such as leaves, stems, pollen, ovules, or cells
that can be cultured into a whole plant.
[0053] As used herein, the phrase "associated with" refers to a
recognizable and/or assayable relationship between two entities.
For example, the phrase "associated with a C. chinense aroma"
refers to a trait, locus, gene, allele, marker, phenotype,
metabolic profile, etc., or the expression thereof, the presence or
absence of which can influence or indicate an extent, degree,
and/or rate at which a plant or a part of interest thereof has a C.
chinense aroma. As such, a metabolite marker is "associated with" a
trait when it is linked to it and when the presence of the
metabolite marker is an indicator of whether and/or to what extent
the desired trait or trait form will occur in a plant/germplasm
comprising the metabolite marker.
[0054] As used herein, "male sterile" or "male sterility" refers to
plants that are not usually capable of breeding from
self-pollination, but are capable of breeding from
cross-pollination. One type of male sterility is cytoplasmic male
sterility (CMS). Several CMS systems have been reported in pepper
breeding, such as the Peterson CMS system and Baccatum CMS system
(see US 2013/0145489 A1, published Jun. 6, 2013). In the Peterson
CMS system, the male sterility factors are coded in the
mitochondrial DNA. The cytoplasm, including the mitochondria, is
passed from the female parent to its progeny. Progeny plants coming
from a cross between two parents carry the cytoplasm of the female
parental plant. Therefore, if the female parental plant displays a
CMS trait, then all progenies will also carry the CMS trait in the
absence of "Restorer" alleles. In the presence of Restorer alleles,
a plant will be fertile even if its cytoplasm is derived from a CMS
maternal parent.
[0055] As used herein, "genetic element" or "gene" refers to a
heritable sequence of DNA, e.g., a genomic sequence, with
functional significance. The term "gene" can also be used to refer
to, e.g., a cDNA and/or an mRNA encoded by a genomic sequence, as
well as to that genomic sequence. A "genetic determinant" refers to
one or more genes, gene elements, or combinations thereof that are
capable of providing a trait of interest.
[0056] As used herein, "crossed" or "cross" means to produce
progeny via fertilization (e.g. cells, seeds or plants) and
includes crosses between plants (sexual) and self-fertilization
(selfing).
[0057] As used herein, the term "inbred" means a line that has been
bred for genetic homogeneity.
[0058] As used herein, the term "hybrid" means a progeny of mating
between at least two genetically dissimilar parents. Without
limitation, examples of mating schemes include single crosses,
modified single cross, double modified single cross, three-way
cross, modified three-way cross, and double cross wherein at least
one parent in a modified cross is the progeny of a cross between
sister lines.
[0059] As used herein, "introgression" refers to the transmission
of a desired allele of a genetic locus from one genetic background
to another.
[0060] As used herein, the term "chromosome interval" or
"chromosomal interval" designates a contiguous linear span of
genomic DNA that resides on a single chromosome.
[0061] As used herein, "flanked by," when used to describe a
chromosomal interval, refers to two loci physically surrounding the
chromosomal interval, with one locus on each side of the
chromosomal interval. As referenced herein, a chromosomal interval
flanked by two marker loci includes the two marker loci.
[0062] As used herein, "locus" is a chromosome region or
chromosomal region where a polymorphic nucleic acid, trait
determinant, gene, or marker is located. A locus can represent a
single nucleotide, a few nucleotides or a large number of
nucleotides in a genomic region. The loci of this disclosure
comprise one or more polymorphisms in a population (e.g.,
alternative alleles are present in some individuals).
[0063] As used herein, "allele" refers to an alternative nucleic
acid sequence at a particular locus. The length of an allele can be
as small as one nucleotide base. For example, a first allele can
occur on one chromosome, while a second allele occurs on a second
homologous chromosome, e.g., as occurs for different chromosomes of
a heterozygous individual, or between different homozygous or
heterozygous individuals in a population.
[0064] As used herein, "marker assay" means a method for detecting
a polymorphism at a particular locus using a particular method,
e.g. measurement of at least one phenotype (such as seed color,
flower color, or other visually detectable traits), restriction
fragment length polymorphism (RFLP), single base extension,
electrophoresis, sequence alignment, allelic specific
oligonucleotide hybridization (ASO), random amplified polymorphic
DNA (RAPD), microarray-based technologies, and nucleic acid
sequencing technologies, etc.
[0065] As used herein, a "population of plants" or a "population of
seeds" means a set comprising any number, at least two, of
individuals, objects, or data from which samples are taken for
evaluation. Most commonly, the terms relate to a breeding
population of plants from which members are selected and crossed to
produce progeny in a breeding program. A population of plants can
include the progeny of a single breeding cross or a plurality of
breeding crosses, and can be either actual plants or plant derived
material, or in silico representations of the plants or seeds. The
population members need not be identical to the population members
selected for use in subsequent cycles of analyses or those
ultimately selected to obtain final progeny plants or seeds. Often,
a population of plants or seeds is derived from a single biparental
cross, but can also derive from two or more crosses between the
same or different parents. Although a population of plants or seeds
can comprise any number of individuals, those of skill in the art
will recognize that plant breeders commonly use population sizes
ranging from one or two hundred individuals to several thousand,
and that the highest performing 5% to 20% of a population is what
is commonly selected to be used in subsequent crosses in order to
improve the performance of subsequent generations of the
population.
[0066] As used herein, "backcross" and "backcrossing" refer to the
process whereby a progeny plant is repeatedly crossed back to one
of its parents. In a backcrossing scheme, a "donor" parent refers
to the parental plant with the desired gene, locus, or trait to be
introgressed. A "recipient" parent (used one or more times) or
"recurrent" parent (used two or more times) refers to a parental
plant into which the gene, locus, or trait is being introgressed.
For example, see Ragot, M. et al. Marker-assisted Backcrossing: A
Practical Example, in Techniques Et Utilisations Des Marqueurs
Moleculaires Les Colloques, Vol. 72, pp. 45-56 (1995); and Openshaw
et al., Marker-assisted Selection in Backcross Breeding, in
Proceedings Of The Symposium "Analysis of Molecular Marker Data,"
pp. 41-43 (1994). The initial cross gives rise to the F1
generation. The term "BC1" (backcross 1) refers to the second use
of the recurrent parent, "BC2" refers to the third use of the
recurrent parent, and so on. In an aspect, a backcross is performed
repeatedly, with a progeny individual of each successive backcross
generation being itself backcrossed to the same parental
genotype.
[0067] As used herein, "elite line" or "elite variety" means any
line that has resulted from breeding and selection for superior
agronomic performance. Similarly, an "elite germplasm" or elite
strain of germplasm is an agronomically superior germplasm.
Exemplary named pepper types or varieties include Aleppo, Anaheim,
ancho, bell, cascabel, cayenne, chilaca, chiltepin, cubanelle, de
arbol, dandicut, Fresno, guajillo, Hungarian wax, Italian sweet,
jalapeflo, Japanese, mirasol, macho, mulato, New Mexico, pasilla,
pepperoncini (Tuscan), piquin, pimento, poblano, puya, Serrano and
Tientsin (Tien Tsin).
[0068] As used herein, "genotype" or "genetic composition" or
"genetic complement" is the genetic constitution of an individual
(or group of individuals) at one or more genetic loci, as
contrasted with the observable trait (phenotype). Genotype is
defined by the allele(s) of one or more known loci that the
individual has inherited from its parents. The term genotype can be
used to refer to an individual's genetic constitution at a single
locus, at multiple loci, or, more generally, the term genotype can
be used to refer to an individual's genetic make-up for all the
genes in its genome.
[0069] As used herein, "selecting" or "selection" in the context of
metabolomics-assisted selection, marker-assisted selection, or
breeding refer to the act of picking or choosing desired
individuals, normally from a population, based on certain
pre-determined criteria.
[0070] As used herein, the term "trait," "phenotypic trait," or
"phenotype" refer to one or more detectable characteristics of a
cell or organism which can be influenced by genotype. The phenotype
can be observable to the naked eye, or by any other means of
evaluation known in the art, e.g., microscopy, biochemical
analysis, genomic analysis, transcriptional profiling, metabolic
profiling, etc. In some cases, a phenotype is directly controlled
by a single gene or genetic locus, e.g., a "single gene trait." In
other cases, a phenotype is the result of several genes or genetic
loci.
[0071] As used herein, a "centimorgan" (cM) is a unit of measure of
recombination frequency and genetic distance between two loci. One
cM is equal to a 1% chance that a marker at one genetic locus will
be separated from a marker at a second locus due to crossing over
in a single generation.
[0072] As used herein, "linkage" refers to relative frequency at
which types of gametes are produced in a cross. For example, if
locus A has genes "A" or "a" and locus B has genes "B" or "b," then
a cross between parent 1 with AABB and parent 2 with aabb can
produce four possible gametes segregating into AB, Ab, aB and ab
genotypes. The null expectation is that there will be independent
equal segregation into each of the four possible genotypes, i.e. no
linkage between locus A and locus B results in 1/4 of the gametes
from each genotype (AB, Ab, aB, and ab). Segregation of gametes
into genotype ratios differing from 1/4 indicates linkage between
locus A and locus B. As used herein, linkage can be between two
markers, or alternatively between a marker and a phenotype. A
marker locus can be associated with (linked to) a trait, e.g., a
marker locus can be associated with tolerance or improved tolerance
to a plant pathogen when the marker locus is in linkage
disequilibrium (LD) with the tolerance trait. The degree of linkage
of a molecular marker to a phenotypic trait (e.g., a QTL) is
measured, e.g., as a statistical probability of co-segregation of
that molecular marker with the phenotype.
[0073] As used herein, "linkage disequilibrium" (LD) refers to a
non-random segregation of genetic loci or traits (or both). In
either case, linkage disequilibrium implies that the relevant loci
are within sufficient physical proximity along a length of a
chromosome so that they segregate together with greater than random
(i.e., non-random) frequency (in the case of co-segregating traits,
the loci that underlie the traits are in sufficient proximity to
each other). Linked loci cosegregate more than 50% of the time,
e.g., from about 51% to about 100% of the time. Linkage
disequilibrium can be measured using any one of the methods
provided in Hedrick, Gametic disequilibrium measures: proceed with
caution. Genetics, 117:331-41(1987). The term "physically linked"
is sometimes used to indicate that two loci, e.g., two marker loci,
are physically present on the same chromosome. Advantageously, the
two linked loci are located in close proximity such that
recombination between homologous chromosome pairs does not occur
between the two loci during meiosis with high frequency, e.g., such
that linked loci cosegregate at least about 90% of the time, e.g.,
91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.75%, or more
of the time.
[0074] As used herein, the linkage relationship between a molecular
marker and a phenotype is given as the statistical likelihood that
the particular combination of a phenotype and the presence or
absence of a particular marker allele is random. Thus, the lower
the probability score, the greater the likelihood that a phenotype
and a particular marker will cosegregate. In some embodiments, a
probability score of 0.05 (p=0.05, or a 5% probability) of random
assortment is considered a significant indication of
co-segregation. However, the present invention is not limited to
this particular standard, and an acceptable probability can be any
probability of less than 50% (p<0.5). For example, a significant
probability can be less than 0.25, less than 0.20, less than 0.15,
or less than 0.1.
[0075] As used herein, the term "linked" or "genetically linked,"
when used in the context of markers and/or genomic regions, means
that recombination between two linked loci occurs with a frequency
of equal to or less than about 10% (i.e., are separated on a
genetic map by not more than 10 cM). In one aspect, any marker of
the invention is linked (genetically and physically) to any other
marker that is at or less than 50 cM distant. In another aspect,
any marker of the invention is closely linked (genetically and
physically) to any other marker that is in close proximity, e.g.,
at or less than 10 cM distant. Two closely linked markers on the
same chromosome can be positioned 20, 19, 18, 17, 16, 15, 14, 13,
12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.75, 0.5 or 0.25 cM or less
from each other.
[0076] As used herein, "marker," "genetic marker," "molecular
marker," and "marker locus" refer to a nucleotide sequence or
encoded product thereof (e.g., a protein) used as a point of
reference when identifying a linked locus. A marker can be derived
from genomic nucleotide sequence or from expressed nucleotide
sequences (e.g., from a spliced RNA, a cDNA, etc.), or from an
encoded polypeptide, and can be represented by one or more
particular variant sequences, or by a consensus sequence. In
another sense, a marker is an isolated variant or consensus of such
a sequence. The term also refers to nucleic acid sequences
complementary to or flanking the marker sequences, such as nucleic
acids used as probes or primer pairs capable of amplifying the
marker sequence. A "marker probe" is a nucleic acid sequence or
molecule that can be used to identify the presence of a marker
locus, e.g., a nucleic acid probe that is complementary to a marker
locus sequence. Alternatively, in some aspects, a marker probe
refers to a probe of any type that is able to distinguish (i.e.,
genotype) the particular allele that is present at a marker locus.
A "marker locus" is a locus that can be used to track the presence
of a second linked locus, e.g., a linked locus that encodes or
contributes to expression of a phenotypic trait. For example, a
marker locus can be used to monitor segregation of alleles at a
locus, such as a QTL, that are genetically or physically linked to
the marker locus. Thus, a "marker allele," alternatively an "allele
of a marker locus" is one of a plurality of polymorphic nucleotide
sequences found at a marker locus in a population that is
polymorphic for the marker locus.
[0077] As used herein, "polymorphism" means the presence of one or
more variations in a population. A polymorphism can manifest as a
variation in the nucleotide sequence of a nucleic acid or as a
variation in the amino acid sequence of a protein. Polymorphisms
include the presence of one or more variations of a nucleic acid
sequence or nucleic acid feature at one or more loci in a
population of one or more individuals. The variation can comprise,
but is not limited to, one or more nucleotide base changes, the
insertion of one or more nucleotides or the deletion of one or more
nucleotides. A polymorphism can arise from random processes in
nucleic acid replication, through mutagenesis, as a result of
mobile genomic elements, from copy number variation and during the
process of meiosis, such as unequal crossing over, genome
duplication, and chromosome breaks and fusions. The variation can
be commonly found or can exist at low frequency within a
population, the former having greater utility in general plant
breeding and the latter can be associated with rare but important
phenotypic variation. Useful polymorphisms can include a single
nucleotide polymorphisms (SNP), an insertion or deletion in DNA
sequence (indel), a simple sequence repeats of DNA sequence (SSR),
a restriction fragment length polymorphism (RFLP), and a tag SNP. A
genetic marker, a gene, a DNA-derived sequence, a RNA-derived
sequence, a promoter, a 5' untranslated region of a gene, a 3'
untranslated region of a gene, microRNA, small interfering RNA, a
tolerance locus, a satellite marker, a transgene, mRNA,
double-stranded RNA, a transcriptional profile, and a methylation
pattern can also comprise a polymorphism. In addition, the
presence, absence, or variation in copy number of the preceding can
comprise a polymorphism.
[0078] As used herein, "SNP" or "single nucleotide polymorphism"
means a sequence variation that occurs when a single nucleotide (A,
T, C, or G) in the genome sequence is altered or variable. "SNP
markers" exist when SNPs are mapped to sites on the genome.
[0079] As used herein, "quantitative trait locus (QTL)" means a
genetic domain that effects a phenotype that can be described in
quantitative terms and can be assigned a "phenotypic value" which
corresponds to a quantitative value for the phenotypic trait. A QTL
can act through a single gene mechanism or by a polygenic
mechanism. In one embodiment of this invention, the boundaries of
genomic regions are drawn to encompass markers that will be linked
to one or more QTL. In other words, the chromosome interval is
drawn such that any marker that lies within that region (including
the terminal markers that define the boundaries of the region) is
genetically linked to the QTL. Each region comprises at least one
QTL, and furthermore, may indeed comprise more than one QTL. Close
proximity of multiple QTL in the same region may obfuscate the
correlation of a particular marker with a particular QTL, as one
marker may demonstrate linkage to more than one QTL. Conversely,
e.g., if two markers in close proximity show co-segregation with
the desired phenotypic trait, it is sometimes unclear if each of
those markers identifying the same QTL or two different QTL.
Regardless, knowledge of how many QTL are in a particular interval
is not necessary to make or practice the invention.
[0080] As used herein "LOD" means a log.sub.10 of an odds ratio and
is used to estimate a QTLs location. QTL location estimates are
determined empirically. A maximum likelihood estimate (MLE) for the
presence of a marker is calculated, together with an MLE assuming
no QTL effect, to avoid false positives. A log.sub.10 of an odds
ratio (LOD) is then calculated as: LOD=log.sub.10 (MLE for the
presence of a QTL/MLE given no linked QTL). The LOD score
essentially indicates how much more likely the data are to have
arisen assuming the presence of a QTL versus in its absence. The
LOD threshold value for avoiding a false positive with a given
confidence, say 95%, depends on the number of markers and the
length of the genome. Graphs indicating LOD thresholds are set
forth in Lander and Botstein, (Lander and Botstein, Mapping
Mendelian Factors Underlying Quantitative Traits Using RFLP Linkage
Maps. Genetics, 121:185-199 (1989), and further described by Arils
and Moreno-Gonzalez, Plant Breeding, Hayward, Bosemark, Romagosa
(eds.) Chapman & Hall, London, pp. 314-331 (1993).
[0081] As used herein, a "plant" refers to a whole plant or a cell
or tissue culture derived from a plant, comprising any of: whole
plants, plant components or organs (e.g., leaves, stems, roots,
etc.), plant tissues, seeds, plant cells, and/or progeny of the
same. A progeny plant can be from any filial generation, e.g.,
F.sub.1, F.sub.2, F.sub.3, F.sub.4, F.sub.5, F.sub.6, F.sub.7, etc.
A plant cell is a biological cell of a plant, taken from a plant or
derived through culture from a cell taken from a plant.
[0082] As used herein, a "population of plants" or "plant
population" means a set comprising any number, including one, of
individuals, objects, or data from which samples are taken for
evaluation. Most commonly, the terms relate to a breeding
population of plants from which members are selected and crossed to
produce progeny in a breeding program. A population of plants can
include the progeny of a single breeding cross or a plurality of
breeding crosses, and can be either actual plants or plant derived
material, or in silico representations of the plants. The
population members need not be identical to the population members
selected for use in subsequent cycles of analyses or those
ultimately selected to obtain final progeny plants. Often, a plant
population is derived from a single biparental cross, but may also
derive from two or more crosses between the same or different
parents. Although a population of plants may comprise any number of
individuals, those of skill in the art will recognize that plant
breeders commonly use population sizes ranging from one or two
hundred individuals to several thousand, and that the highest
performing 5-20% of a population is what is commonly selected to be
used in subsequent crosses in order to improve the performance of
subsequent generations of the population.
[0083] As used herein, "cultivar" and "variety" are used
synonymously and mean a group of plants within a species (e.g., C.
annuum) that share certain genetic traits that separate them from
other possible varieties within that species. Pepper cultivars can
be inbreds or hybrids.
[0084] As used herein, "introgression" or "introgressing" refers to
the transmission of a desired trait or a desired allele of a
genetic locus from one genetic background to another.
[0085] As used herein, "single gene converted" or "single gene
conversion" refers to plants that are developed using a plant
breeding technique known as backcrossing, or via genetic
engineering, where essentially all of the desired morphological and
physiological characteristics of a variety are recovered in
addition to the single gene transferred into the variety via the
backcrossing technique or via genetic engineering.
[0086] As used herein, "similar conditions" or "similar growth
conditions" refer to similar environmental conditions and agronomic
practices for growing plants and making meaningful comparisons
between two or more plant genotypes so that neither environmental
conditions nor agronomic practices would contribute to or explain
any difference observed between the two or more plant genotypes.
Environmental conditions include, for example, light, temperature,
water (humidity), and nutrition (e.g., nitrogen and phosphorus).
Exemplary ways of cultivating peppers include open field,
greenhouse and shade house production.
[0087] As used herein, "part(s) of a plant" or a "plant part(s)"
includes, without limitation, leaves, calyx, pollen, embryos,
pedicle, peduncle, cotyledon, hypocotyl, meristematic cells, roots,
root tips, anthers, flowers, seeds, stem, and pepper fruit.
[0088] As used herein, "Brix" or "Brix rating" is a measure of the
percent of total soluble solids in a fruit. Brix is measured in
Degree Brix(B) using a refractometer or density meter (e.g.
Refracto 30PX, Mettler-Toledo, Columbus, Ohio). The amount of
sucrose, fructose, vitamins, minerals, proteins, hormones, and
other soluble solids can affect Brix ratings. Typically for
peppers, a Brix rating of 4 is considered poor, a Brix rating of 6
is considered average, a Brix rating of 8 is considered good, and a
Brix rating of 10 is considered excellent. As used here, a Brix
rating (.degree. B) can be higher than a total sugar content (%),
due to the former reflecting more than the reducing sugars
(fructose and glucose) measured by the latter.
[0089] As used herein, "hedonic attribute test" is a form of
sequential monadic liking test used by consumers for the comparison
of sample pepper fruits. Consumers are asked to rate the hedonic
attributes of overall rating, flavor, and sweetness. An exemplary
evaluation process is described in Example 6. On a scale of 1 to 9,
consumers grade each of the attributes in a sample comparison. A
score of 1 meaning "dislike extremely", a score of 2 meaning
"dislike very much", a score of 3 meaning "dislike moderately", a
score of 4 meaning "dislike slightly", a score of 5 meaning
"neither like nor dislike", a score of 6 meaning "like slightly", a
score of 7 meaning "like moderately", a score of 8 meaning "like
very much", and a score of 9 meaning "like extremely".
[0090] Peppers are commonly broken down into three groupings: bell
peppers, sweet peppers, and hot peppers. Most popular pepper
varieties fall into one of these categories, or as a cross between
them. These groupings are not absolute, as both "hot pepper" and
"sweet pepper" encompass members belonging to several different
species. Additionally, members of each of the groups may be
different cultivars of the same species. For example, the bell
pepper, the jalapeno pepper, and the "Thai sweet" all belong to the
species C. annuum L. Hot peppers are grown for edible as well as
ornamental and medicinal uses. While there are pungent (i.e.,
"hot") varieties of C. annuum, many well-known hot peppers are
members of different species. For example, both the cayenne pepper
and the TABASCO.RTM. pepper are varieties of C. frutescens, while
some members of C. chinense, including the habanero and naga
varieties, are well-known hot peppers.
[0091] Depending on its pungency, a pepper fruit may be referred to
as sweet, mild, medium, hot or very hot pepper varieties as defined
in U.S. Pat. No. 8,802,939. For example, a "sweet" pepper has about
0 Scoville Heat Units (SHU) or less than about 0.5 ppm (parts per
million) which is less than about 7 SHU of total capsaicinoids
(capsaicin, norhydrocapsaicin, and dihydrocapsaicin). A "mild"
pepper has greater than about 0.5 ppm (greater than about 7.5 SHU)
to about 4.0 ppm (60 SHU) of total capsaicinoids, or more
preferably about 2 ppm (30 SHU) to 3 ppm (45 SHU) of total
capsaicinoids based upon the weight and capsaicin content of whole
pepper fruit.
[0092] Capsaicin content or SHU units can be determined by methods
known in the art including HPLC methods as described for example in
Garces-Claver et al., "Determination of Capsaicin and
Dihydrocapsaicin in Capsicum Fruit by Liquid
Chromatography-Electrospray/Time-of-Flight Mass Spectrometry," J.
Agric. Food Chem. 54:9303-9311 (2006), hereby incorporated by
reference in its entirety. A skilled artisan will understand that
capsaicin levels can vary, and that the capsaicin content of some
pepper fruit tissues (e.g., placenta) can be higher than that of
other tissues. See, Sung et al., "Capsaicin biosynthesis in
water-stressed hot pepper fruits," Bot. Bull. Acad. Sin. 46:35-42
(2005). The conversion between total capsaicinoid levels and
Scoville Heat Units (SHU) was developed by Wilbur Scoville. The
number of Scoville units equals the approximate number of times a
pepper extract would need to be diluted for the spiciness to be
imperceptible. By definition, one part per million (ppm) of
capsaicin has a pungency of 15 SHU.
[0093] Traditionally, pepper breeding has focused on developing
cultivars with high yield (i.e. larger fruit size and more fruits
per plant), improving quality attributes (such as bright fruit
color and balanced spiciness) and to enhance field performance
(improved pathogen resistance and stress tolerance). As discussed
above, there is a need to identify aromatic components (e.g., aroma
compounds) underlying pepper fruit flavor and aromas. There is also
a need to develop an objective analytical method to assess pepper
(e.g., C. chinense) aroma or flavor. There is also a need in pepper
breeding to incorporate desirable aromas or flavor (e.g., aromas
characteristic of C. chinense) into sweet peppers.
[0094] In an aspect, this disclosure provides a Capsicum seed or
plant grown therefrom, where the Capsicum plant is capable of
producing a pepper fruit having pericarp comprising a total sugar
content of at least 5.5% and one or more aroma molecules
characteristic of C. chinense. In an aspect, this disclosure
provides a Capsicum seed or plant grown therefrom, where the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and one or more aroma molecules characteristic of C.
chinense, where the one or more aroma molecules are absent from
pericarp of hybrid pepper PS09954859 or at a higher level than that
of pericarp of hybrid pepper PS09954859 grown under similar
conditions. In one aspect, total sugar content is quantified using
an Agilent HPLC. In an aspect, a Capsicum seed is a C. annuum seed.
In another aspect, one or more aroma molecules are terpene
molecules. In a further aspect, terpene molecules are selected from
the group consisting of .alpha.-Cubebene, .delta.-Cadinene,
1,4-Cadinadiene, and combinations thereof. In an aspect, a pericarp
of a pepper fruit disclosed herein comprises alpha-Cubebene,
delta-Cadinene, and 1,4-Cadinadiene. In an aspect, a pepper fruit
comprises pericarp having a titratable acidity at a higher level
than that of pericarp of hybrid pepper PS09954859 grown under
similar conditions. In another aspect, a pepper fruit comprises
pericarp having a titratable acidity of at least 4.2, 4.3, 4.4,
4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, or 5.7
millimole (mmol) H.sup.+/100 g fresh tissue. In another aspect, a
pepper fruit comprises pericarp having a titratable acidity of
between 4.2 and 5.5, between 4.2 and 5.4, between 4.2 and 5.3,
between 4.2 and 5.2, between 4.2 and 5.1, between 4.2 and 5.1,
between 4.2 and 5.0, between 4.2 and 4.9, between 4.2 and 4.8,
between 4.2 and 4.7, between 4.2 and 4.6, between 4.2 and 4.5, or
between 4.2 and 4.4 millimole (mmol) H.sup.+/100 g fresh tissue. In
one aspect, titratable acidity is determined using 0.1N NaOH to
obtain an end pH point of 8.1 endpoint (AOAC, 1990). In one aspect,
titratable acidity is determined using a Mettler Toledo T90
automatic titrator.
[0095] Aroma molecules, e.g., terpenes, can be sampled from pepper
fruits harvested at various maturation stages. These stages are
characterized by the percentage of total fruit surface area
exhibiting mature color (e.g., color change from green to red,
orange or yellow). In one aspect, an aroma molecule described here
is detected and measured in a pepper fruit harvested at a stage
having at least about 5%, 10%, 25%, 35%, 50%, 60%, 70%, 75%, 80%,
85%, 90%, 95%, or 100% color change. In another aspect, an aroma
molecule described here is detected and measured in a pepper fruit
harvested at a stage having between 5% and 50%, between 10% and
50%, between 25% and 50%, between 35% and 50%, between 50% and
100%, between 60% and 100%, between 70% and 100%, between 75% and
100%, between 80% and 100%, between 85% and 100%, between 90% and
100%, or between 95% and 100% color change. In a further aspect, an
aroma molecule described here is detected and measured in a pepper
fruit harvested at a stage having between 60% and 95%, between 70%
and 90%, or between 75% and 85% color change. In another aspect, an
aroma molecule described here is detected and measured in a pepper
fruit harvested at a stage having between 5% and 10%, between 10%
and 15%, between 15% and 25%, between 25% and 30%, between 50% and
60%, between 60% and 70%, between 70% and 75%, between 75% and 80%,
between 80% and 85%, between 85% and 90%, or between 90% and 95%
color change. Aroma molecules are typically measured within 1 week
of harvest, more preferably within 5 days, within 3 days, within 1
day, between 1 day and 1 week, between 1 day and 5 days, between 1
day and 3 days, or between 3 days and 5 days of harvesting.
[0096] In an aspect, this disclosure provides a Capsicum seed or
plant grown therefrom, where the Capsicum plant exhibits a pepper
fruit at maturity having pericarp comprising a total sugar content
of at least 5.5% and having a titratable acidity of at least 4.2,
4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5,
5.6, or 5.7 millimole (mmol) H.sup.+/100 g fresh tissue. In one
aspect, a Capsicum seed or plant is a sweet bell pepper. In one
aspect, a Capsicum seed or plant comprises substantially no
pungency. In another aspect, a Capsicum seed or plant comprises a
pungency of substantially zero Scoville Heat Units (SHUs) or a
pungency of below 4000, 3000, 2000, 1000, 500, 250, 200, 150, 100,
50, 40, 30, 20, 10, or 5 SHUs.
[0097] In an aspect, this disclosure provides a Capsicum seed or
plant grown therefrom, where the Capsicum plant exhibits a pepper
fruit at maturity having pericarp having a titratable acidity of at
least 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3,
5.4, 5.5, 5.6, or 5.7 millimole (mmol) H.sup.+/100 g fresh tissue,
and comprising one or more aroma molecules characteristic of C.
chinense, where the one or more aroma molecules are absent from
pericarp of hybrid pepper PS09954859 or at a higher level than that
of pericarp of hybrid pepper PS09954859 grown under similar
conditions. In another aspect, one or more aroma molecules are
selected from the group consisting of .alpha.-Cubebene,
.delta.-Cadinene, 1,4-Cadinadiene, and combinations thereof. In an
aspect, a pericarp of a pepper fruit disclosed herein comprises
alpha-Cubebene, delta-Cadinene, and 1,4-Cadinadiene. In another
aspect, one or more aroma molecules are selected from the group
consisting of alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.25, delta-Cadinene at a level having a
Relative GC/MS Response Score of at least 0.25, 1,4-Cadinadiene at
a level having a Relative GC/MS Response Score of at least 0.15, or
any combination thereof.
[0098] In an aspect, this disclosure provides a Capsicum seed or
plant grown therefrom, where the Capsicum plant exhibits a pepper
fruit at maturity having pericarp comprising a total sugar content
of at least 5.5%, having a titratable acidity of at least 4.2, 4.3,
4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, or
5.7 millimole (mmol) H.sup.+/100 g fresh tissue, comprises a
pungency of substantially zero Scoville Heat Units (SHUs) or below
5 SHUs, and further comprising one or more aroma molecules
characteristic of C. chinense selected from the group consisting of
alpha-Cubebene at a level having a Relative GC/MS Response Score of
at least 0.25, delta-Cadinene at a level having a Relative GC/MS
Response Score of at least 0.25, 1,4-Cadinadiene at a level having
a Relative GC/MS Response Score of at least 0.15, or any
combination thereof.
[0099] In one aspect, a Capsicum seed or plant grown therefrom is
provided, where the Capsicum plant exhibits a pepper fruit at
maturity having pericarp comprising a total sugar content of at
least about 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.4%,
6.5%, 6.6%, 6.8%, 7.0%, 7.2%, 7.4%, 7.6%, 7.8%, 8.0%, 8.2%, 8.4%,
8.6%, 8.8%, or 9.0%. In one aspect, a Capsicum seed or plant grown
therefrom is provided, where the Capsicum plant exhibits a pepper
fruit at, or immediately prior to, maturity having pericarp
comprising a total sugar content of between 5.5% and 9.0%, between
5.6% and 9.0%, between 5.7% and 9.0%, between 5.8% and 9.0%,
between 5.9% and 9.0%, between 6.0% and 9.0%, between 6.1% and
9.0%, between 6.2% and 9.0%, between 6.4% and 9.0%, between 6.5%
and 9.0%, between 6.6% and 9.0%, between 6.8% and 9.0%, between
7.0% and 9.0%, between 7.2% and 9.0%, between 7.4% and 9.0%,
between 7.6% and 9.0%, between 7.8% v, between 8.0% and 9.0%,
between 8.2% and 9.0%, between 8.4% and 9.0%, between 8.6% and
9.0%, or between 8.8% and 9.0%. In another aspect, a Capsicum seed
or plant grown therefrom is provided, where the Capsicum plant
exhibits a pepper fruit at, or immediately prior to, maturity
having pericarp comprising a total sugar content of between 5.5%
and 8.8%, between 5.6% and 8.6%, between 5.7% and 8.4%, between
5.8% and 8.2%, between 5.9% and 8.0%, between 6.0% and 7.8%,
between 6.1% and 7.6%, between 6.2% and 7.4%, between 6.4% and
7.2%, between 6.5% and 7.1%, between 6.6% and 7.0%, or between 6.8%
and 7.0%.
[0100] In another aspect, a pepper disclosed herein comprises
alpha-Cubebene in pericarp at maturity at a level having a Relative
GC/MS Response Score of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18,
0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5,
3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5. In another aspect, a
pepper disclosed herein comprises alpha-Cubebene in pericarp at
maturity at a level having a Relative GC/MS Response Score of
between 0.25 and 17.5, between 0.3 and 17.5, between 0.4 and 17.5,
between 0.5 and 17.5, between 0.6 and 17.5, between 0.7 and 17.5,
between 0.8 and 17.5, between 0.9 and 17.5, between 1.0 and 17.5,
between 1.5 and 17.5 between 2.0 and 17.5, between 2.5 and 17.5,
between 3.0 and 17.5, between 4.0 and 17.5, between 5.0 and 17.5,
between 7.5 and 17.5, between 10.0 and 17.5, between 12.5 and 17.5,
between 15.0 and 17.5, between 0.3 and 15, between 0.4 and 12.5,
between 0.5 and 10, between 0.6 and 7.5, between 0.7 and 5.0,
between 0.8 and 4.0, between 0.9 and 3.0, between 1.0 and 2.0,
between 1.5 and 2.0, between 2.0 and 2.5, between 2.5 and 3.0,
between 3.0 and 4.0, between 4.0 and 5.0, between 5.0 and 7.5,
between 7.5 and 10.0, between 10.0 and 12.5, or between 12.5 and
15. In a further aspect, a pepper disclosed herein comprises
alpha-Cubebene in pericarp at maturity at a level above 1, 2, 3, 4,
5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 60, 100, 150, 200,
300 .mu.g/g fresh fruit. In a further aspect, a pepper disclosed
herein comprises alpha-Cubebene in pericarp at maturity at a level
between 1 and 2, between 2 and 3, between 3 and 4, between 4 and 5,
between 5 and 6, between 6 and 7, between 7 and 8, between 8 and 9,
between 9 and 10, between 10 and 15, between 15 and 20, between 20
and 25, between 25 and 30, between 30 and 35, between 35 and 40,
between 40 and 50, between 50 and 60, between 60 and 100, between
100 and 150, between 150 and 200, or between 200 and 300 .mu.g/g
fresh fruit.
[0101] In an aspect, a pepper disclosed herein comprises
delta-Cadinene in pericarp at maturity at a level having a Relative
GC/MS Response Score of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18,
0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5,
3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5. In another aspect, a
pepper disclosed herein comprises delta-Cadinene in pericarp at
maturity at a level having a Relative GC/MS Response Score of
between 0.25 and 17.5, between 0.3 and 17.5, between 0.4 and 17.5,
between 0.5 and 17.5, between 0.6 and 17.5, between 0.7 and 17.5,
between 0.8 and 17.5, between 0.9 and 17.5, between 1.0 and 17.5,
between 1.5 and 17.5 between 2.0 and 17.5, between 2.5 and 17.5,
between 3.0 and 17.5, between 4.0 and 17.5, between 5.0 and 17.5,
between 7.5 and 17.5, between 10.0 and 17.5, between 12.5 and 17.5,
between 15.0 and 17.5, between 0.3 and 15, between 0.4 and 12.5,
between 0.5 and 10, between 0.6 and 7.5, between 0.7 and 5.0,
between 0.8 and 4.0, between 0.9 and 3.0, between 1.0 and 2.0,
between 1.5 and 2.0, between 2.0 and 2.5, between 2.5 and 3.0,
between 3.0 and 4.0, between 4.0 and 5.0, between 5.0 and 7.5,
between 7.5 and 10.0, between 10.0 and 12.5, or between 12.5 and
15. In another aspect, a pepper disclosed herein comprises
delta-Cadinene in pericarp at maturity at a level above 1, 2, 3, 4,
5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 50, 60, 100, 150, 200,
300 .mu.g/g fresh fruit. In a further aspect, a pepper disclosed
herein comprises delta-Cadinene in pericarp at maturity at a level
between 1 and 2, between 2 and 3, between 3 and 4, between 4 and 5,
between 5 and 6, between 6 and 7, between 7 and 8, between 8 and 9,
between 9 and 10, between 10 and 15, between 15 and 20, between 20
and 25, between 25 and 30, between 30 and 35, between 35 and 40,
between 40 and 50, between 50 and 60, between 60 and 100, between
100 and 150, between 150 and 200, or between 200 and 300 .mu.g/g
fresh fruit.
[0102] In an aspect, a pepper disclosed herein comprises
1,4-Cadinadiene in pericarp at maturity at a level having a
Relative GC/MS Response Score of at least 0.05, 0.08, 0.1, 0.12,
0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5,
2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5. In another
aspect, a pepper disclosed herein comprises 1,4-Cadinadiene in
pericarp at maturity at a level having a Relative GC/MS Response
Score of between 0.15 and 17.5, between 0.18 and 17.5, between 0.2
and 17.5, between 0.25 and 17.5, between 0.3 and 17.5, between 0.4
and 17.5, between 0.5 and 17.5, between 0.6 and 17.5, between 0.7
and 17.5, between 0.8 and 17.5, between 0.9 and 17.5, between 1.0
and 17.5, between 1.5 and 17.5 between 2.0 and 17.5, between 2.5
and 17.5, between 3.0 and 17.5, between 4.0 and 17.5, between 5.0
and 17.5, between 7.5 and 17.5, between 10.0 and 17.5, between 12.5
and 17.5, between 15.0 and 17.5, between 0.15 and 0.18, between
0.18 and 0.2, between 0.2 and 0.25, between 0.25 and 0.3, between
0.3 and 15, between 0.4 and 12.5, between 0.5 and 10, between 0.6
and 7.5, between 0.7 and 5.0, between 0.8 and 4.0, between 0.9 and
3.0, between 0.3 and 0.4, between 0.4 and 0.5, between 0.5 and 0.6,
between 0.6 and 0.7, between 0.7 and 0.8, between 0.8 and 0.9,
between 0.9 and 1.0, between 1.0 and 2.0, between 1.5 and 2.0,
between 2.0 and 2.5, between 2.5 and 3.0, between 3.0 and 4.0,
between 4.0 and 5.0, between 5.0 and 7.5, between 7.5 and 10.0,
between 10.0 and 12.5, or between 12.5 and 15. In another aspect, a
pepper disclosed herein comprises 1,4-Cadinadiene in pericarp at
maturity at a level above 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20,
25, 30, 35, 40, 50, 60, 100, 150, 200, 300 .mu.g/g fresh fruit. In
a further aspect, a pericarp disclosed herein comprises
1,4-Cadinadiene at a level between 1 and 2, between 2 and 3,
between 3 and 4, between 4 and 5, between 5 and 6, between 6 and 7,
between 7 and 8, between 8 and 9, between 9 and 10, between 10 and
15, between 15 and 20, between 20 and 25, between 25 and 30,
between 30 and 35, between 35 and 40, between 40 and 50, between 50
and 60, between 60 and 100, between 100 and 150, between 150 and
200, or between 200 and 300 .mu.g/g fresh fruit. In an aspect,
total sugar content and aroma molecules are measured at a maturity
stage, e.g., substantially near maximum sweetness and flavor
intensity.
[0103] In an aspect, a pepper disclosed herein comprises pericarp
at maturity having alpha-Cubebene at a concentration equal to or
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274 and hybrid pepper SVPS2625, when grown
under similar conditions. In an aspect, a pepper disclosed herein
comprises pericarp at maturity having alpha-Cubebene at a
concentration of at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%,
50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or 300% higher than that
of a pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions. In another aspect, a pepper
disclosed herein comprises pericarp at maturity having
alpha-Cubebene at a concentration of between 10% and 300%, between
20% and 300%, between 30% and 300%, between 40% and 300%, between
50% and 300%, between 75% and 300%, between 100% and 300%, between
150% and 300%, between 200% and 300%, or between 250% and 300%
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions. In a
further aspect, a pepper disclosed herein comprises pericarp at
maturity having alpha-Cubebene at a concentration of about 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 30, or 50 folds higher than that
of a pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions.
[0104] In an aspect, a pepper disclosed herein comprises pericarp
at maturity having delta-Cadinene at a concentration equal to or
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274 and hybrid pepper SVPS2625, when grown
under similar conditions. In an aspect, a pepper disclosed herein
comprises pericarp at maturity having delta-Cadinene at a
concentration of at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%,
50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or 300% higher than that
of a pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions. In another aspect, a pepper
disclosed herein comprises pericarp at maturity having
delta-Cadinene at a concentration of between 10% and 300%, between
20% and 300%, between 30% and 300%, between 40% and 300%, between
50% and 300%, between 75% and 300%, between 100% and 300%, between
150% and 300%, between 200% and 300%, or between 250% and 300%
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions. In a
further aspect, a pepper disclosed herein comprises pericarp at
maturity having delta-Cadinene at a concentration of about 1, 2, 3,
4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 30, or 50 folds higher than that
of a pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions.
[0105] In an aspect, a pepper disclosed herein comprises pericarp
at maturity having 1,4-Cadinadiene at a concentration equal to or
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274 and hybrid pepper SVPS2625, when grown
under similar conditions. In an aspect, a pepper disclosed herein
comprises pericarp at maturity having 1,4-Cadinadiene at a
concentration of at least about 5%, 10%, 15%, 20%, 25%, 30%, 40%,
50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or 300% higher than that
of a pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions. In another aspect, a pepper
disclosed herein comprises pericarp at maturity having
1,4-Cadinadiene at a concentration of between 10% and 300%, between
20% and 300%, between 30% and 300%, between 40% and 300%, between
50% and 300%, between 75% and 300%, between 100% and 300%, between
150% and 300%, between 200% and 300%, or between 250% and 300%
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions. In a
further aspect, a pepper disclosed herein comprises pericarp at
maturity having 1,4-Cadinadiene at a concentration of about 1, 2,
3, 4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 30, or 50 folds higher than
that of a pepper selected from the group consisting of inbred
pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper
PS09954859, when grown under similar conditions.
[0106] In an aspect, a pepper disclosed herein comprises pericarp
at maturity having alpha-Cubebene at a level having a Relative
GC/MS Response Score equal to or at least about 5%, 10%, 15%, 20%,
25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or 300%
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions. In another
aspect, a pepper disclosed herein comprises pericarp at maturity
having alpha-Cubebene at a level having a Relative GC/MS Response
Score of between 10% and 300%, between 20% and 300%, between 30%
and 300%, between 40% and 300%, between 50% and 300%, between 75%
and 300%, between 100% and 300%, between 150% and 300%, between
200% and 300%, or between 250% and 300% higher than that of a
pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions. In a further aspect, a pepper
disclosed herein comprises pericarp at maturity having
alpha-Cubebene at a level having a Relative GC/MS Response Score of
about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 30, or 50 folds
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions.
[0107] In an aspect, a pepper disclosed herein comprises pericarp
at maturity having delta-Cadinene at a level having a Relative
GC/MS Response Score equal to or at least about 5%, 10%, 15%, 20%,
25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or 300%
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions. In another
aspect, a pepper disclosed herein comprises pericarp at maturity
having delta-Cadinene at a level having a Relative GC/MS Response
Score of between 10% and 300%, between 20% and 300%, between 30%
and 300%, between 40% and 300%, between 50% and 300%, between 75%
and 300%, between 100% and 300%, between 150% and 300%, between
200% and 300%, or between 250% and 300% higher than that of a
pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions. In a further aspect, a pepper
disclosed herein comprises pericarp at maturity having
delta-Cadinene at a level having a Relative GC/MS Response Score of
about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 30, or 50 folds
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions.
[0108] In an aspect, a pepper disclosed herein comprises pericarp
at maturity having 1,4-Cadinadiene at a level having a Relative
GC/MS Response Score equal to or at least about 5%, 10%, 15%, 20%,
25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, 200%, or 300%
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions. In another
aspect, a pepper disclosed herein comprises pericarp at maturity
having 1,4-Cadinadiene at a level having a Relative GC/MS Response
Score of between 10% and 300%, between 20% and 300%, between 30%
and 300%, between 40% and 300%, between 50% and 300%, between 75%
and 300%, between 100% and 300%, between 150% and 300%, between
200% and 300%, or between 250% and 300% higher than that of a
pepper selected from the group consisting of inbred pepper
ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid pepper PS09954859,
when grown under similar conditions. In a further aspect, a pepper
disclosed herein comprises pericarp at maturity having
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 30, or 50 folds
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions.
[0109] In an aspect, a pepper disclosed herein further comprises
one or more aroma compounds selected from the group consisting of
2,3-butanedione (caramel odor), 1-penten-3-one (chemical/pungent,
spicy), hexanal (grassy), 3-carene (red bell pepper, rubbery),
(Z)-.beta.-ocimene (rancid, sweaty), octanal (fruity),
2-isobutyl-3-methoxypyrazine (green bell pepper),
(Z)-linalooloxide, (Z)-2-penten-1-ol, (E)-geranylacetone,
(E,Z)-2,6-nonadienal, and (E,E)-decadienal. In another aspect, a
pepper disclosed herein comprises pericarp at maturity having one
or more foregoing aroma compounds at a concentration equal to or
higher than that of a pepper selected from the group consisting of
inbred pepper ZSP8T14-6274, hybrid pepper SVPS2625, and hybrid
pepper PS09954859, when grown under similar conditions.
[0110] In an aspect, a Capsicum seed or plant provided herein
exhibits or produces a pepper fruit comprising no capsaicinoid. In
an aspect, a pepper fruit provided herein comprises no capsaicinoid
at maturity. In an aspect, a pepper fruit from a pepper seed or
plant provided herein comprises an equivalent level of capsaicinoid
compared to C. annuum grown under similar growth conditions. In
another aspect, a pepper fruit provided herein comprises a total
sugar content of at least 5.5% and a lower level of capsaicinoid
compared to C. chinense grown under similar growth conditions. In a
further aspect, a pepper fruit from a pepper seed or plant provided
herein comprises a pungency of substantially zero Scoville Heat
Units (SHUs). In another aspect, a pepper fruit from a pepper seed
or plant provided herein comprises a pungency of below 4000, 3000,
2000, 1000, 500, 250, 200, 150, 100, 50, 40, 30, 20, 10, or 5 SHUs.
In an aspect, the pericarp of a pepper fruit from a pepper seed or
plant provided herein comprises a comparable or higher level of one
or more aroma molecules selected from the group consisting of
alpha-Cubebene, delta-Cadinene, and 1,4-Cadinadiene than C.
chinense pericarp grown under similar conditions. In an aspect, the
pericarp of a pepper fruit from a pepper seed or plant provided
herein comprises a higher level of one or more aroma molecules
selected from the group consisting of alpha-Cubebene,
delta-Cadinene, and 1,4-Cadinadiene than C. annuum pericarp grown
under similar conditions.
[0111] In an aspect, a Capsicum seed or plant provided herein is a
hybrid. In another aspect, a Capsicum seed or plant provided herein
is an inbred. In a further aspect, a Capsicum seed or plant
provided herein is capable of producing a seedless pepper fruit. In
an aspect, a Capsicum seed or plant provided herein is transgenic.
In another aspect, a Capsicum seed or plant provided herein is an
elite line. In a further aspect, a Capsicum seed or plant provided
herein is a single gene or locus conversion. In another aspect, a
Capsicum seed or plant provided herein has multiple genes or loci
(e.g., QTLs) introgressed from a C. chinense background. In a
further aspect, a Capsicum seed or plant provided herein is a C.
annuum line that has been altered with gene editing methods to
contain C. chinense alleles associated with the aroma molecules
described herein.
[0112] In an aspect, a Capsicum seed or plant provided herein
comprises at least about 70%, 75%, 77.5%, 80%, 82.5%, 85%, 86%,
87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99%, 99.5% of its nuclear
genetic material from C. annuum. In another aspect, a Capsicum seed
or plant provided herein comprises equal to or less than about 30%,
25%, 20%, 15%, 14%, 13%, 12.5%, 10%, 7.5%, 5%, 2.5%, 2%, 1%, 0.5%
of its nuclear genetic material from a C. chinense background. In a
further aspect, a Capsicum seed or plant provided herein further
comprises one or more traits selected from the group consisting of
resistance to geminivirus, resistance to Xanthomonas, resistance to
aphids, resistance to powdery mildew, ease of fruit shedding, cold
tolerance, having leaf hairs, thick fruit walls, flavor
differences, and the clustering of seed-bearing placental tissue
close to the stem of a fruit.
[0113] In an aspect, a pepper fruit from a pepper seed or plant
disclosed herein comprises a shape selected from the group
consisting of block, pointed, and round. In another aspect, a
pepper fruit from a pepper seed or plant disclosed herein comprises
a color selected from the group consisting of green, yellow, red,
purple, black, brown, white, and orange.
[0114] In an aspect, a Capsicum seed or plant provided herein is a
progeny of Capsicum line ZSP8T14-6274 or SVPS2625, where a
representative sample of seed of the Capsicum line ZSP8T14-6274 has
been deposited at ATCC under Accession No. PTA-122300, and where a
representative sample of seed of the Capsicum line SVPS2625 has
been deposited at ATCC under Accession No. PTA-122296.
[0115] In another aspect, a Capsicum seed or plant provided herein
is an F.sub.1 progeny of Capsicum line ZSP8T14-6274 or SVPS2625,
where a representative sample of seed of the Capsicum line
ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, and where a representative sample of seed of the
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
[0116] In a further aspect, a Capsicum seed or plant provided
herein is an F.sub.2 progeny of Capsicum line ZSP8T14-6274 or
SVPS2625, where a representative sample of seed of the Capsicum
line ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, and where a representative sample of seed of the
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
[0117] In an aspect, a Capsicum seed or plant provided herein is
essentially derived from Capsicum line ZSP8T14-6274 or SVPS2625,
where a representative sample of seed of the Capsicum line
ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, and where a representative sample of seed of the
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
[0118] In another aspect, a Capsicum seed or plant provided herein
comprises one or more introgressed loci from Capsicum line
ZSP8T14-6274 or SVPS2625, where a representative sample of seed of
the Capsicum line ZSP8T14-6274 has been deposited at ATCC under
Accession No. PTA-122300, where a representative sample of seed of
the Capsicum line SVPS2625 has been deposited at ATCC under
Accession No. PTA-122296, and where the one or more introgressed
loci provide genetic determinant for producing the one or more
aroma molecules characteristic of Capsicum chinense.
[0119] In a further aspect, the instant disclosure provides a
Capsicum seed or plant grown therefrom, where the Capsicum plant
exhibits a pepper fruit having pericarp comprising a total sugar
content of at least 5.5%, alpha-Cubebene at a level having a
Relative GC/MS Response Score of at least 0.25, delta-Cadinene at a
level having a Relative GC/MS Response Score of at least 0.25, and
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.15.
[0120] In another aspect, the instant disclosure provides a sweet
bell pepper fruit comprising one or more aroma molecules
characteristic of C. chinense, where the one or more aroma
molecules are absent from pericarp of hybrid pepper PS09954859 or
at a higher level than that of pericarp of hybrid pepper PS09954859
grown under similar conditions. In an aspect, a sweet bell pepper
fruit disclosed herein comprises a total sugar content of at least
5.5%. In another aspect, a sweet bell pepper fruit disclosed herein
comprises one or more terpene molecules at a higher level than
hybrid pepper PS09954859 grown under similar conditions. In an
aspect, a sweet bell pepper fruit disclosed herein comprises one or
more molecules selected from the group consisting of
alpha-Cubebene, delta-Cadinene, and 1,4-Cadinadiene. In another
aspect, a sweet bell pepper fruit disclosed herein comprises a
higher level of alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene, or
any combination thereof than hybrid pepper PS09954859 grown under
similar conditions.
[0121] In an aspect, a Capsicum plant or seed disclosed herein is
capable of producing a pepper fruit comprising a total sugar
content of at least 5.5%, 6.0%, 6.5%, or 7.0% and a Brix rating of
at least 8.0, 8.5, 9.0, or 9.5. In an aspect, a pepper fruit is
preferred by 30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65% more
consumers when compared to a commercially available or
morphologically comparable sweet pepper fruit in a test of consumer
preference. In an aspect, the produced pepper fruit is described by
at least 70%, 75%, or 80% of consumers in a test of consumer
preference as more flavorful, more sweet, and less bitter when
compared to a commercially available or morphologically comparable
sweet pepper fruit. In an aspect, a produced pepper fruit scores at
least 20% higher than a commercially available or morphologically
comparable sweet pepper fruit as based on a consumer graded hedonic
attribute test for one or more categories selected from the group
consisting of overall rating, flavor, and sweetness. In an aspect,
a produced pepper fruit scores at least 5.5, 6.0, 6.5, or greater
for one or more categories selected from the group consisting of
overall rating, flavor, and sweetness as based on a consumer graded
hedonic attribute test. In an aspect, a plant or seed capable of
producing a pepper fruit is heterozygous for a QTL SMO2
identifiable by markers SEQ ID NO: 1 or 2 and heterozygous for a
QTL SMO3 identifiable by markers SEQ ID NO: 3 or 4.
[0122] In an aspect, a pepper fruit is provided comprising a total
sugar content of at least 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%,
or 9.0% and a Brix rating of at least 8.0, 8.5, 9.0, 9.5, 10.0, or
10.5. In an aspect, a pepper fruit is preferred by 30%, 35%, 40%,
45%, 50%, 55%, 60%, or 65% more consumers when compared to a
commercially available or morphologically comparable sweet pepper
fruit in a test of consumer preference. In an aspect, a pepper
fruit is described by at least 70%, 75%, or 80% of consumers in a
test of consumer preference as more flavorful, more sweet, and less
bitter when compared to a commercially available or morphologically
comparable sweet pepper fruit. In an aspect, a pepper fruit scores
at least 20% higher than a commercially available or
morphologically comparable sweet pepper fruit as based on a
consumer graded hedonic attribute test for one or more categories
selected from the group consisting of overall rating, flavor, and
sweetness. In an aspect, a pepper fruit scores at least 5.5, 6.0,
6.5, or greater for one or more categories selected from the group
consisting of overall rating, flavor, and sweetness as based on a
consumer graded hedonic attribute test. In an aspect, a plant or
seed capable of producing a pepper fruit is heterozygous for a QTL
SMO2 identifiable by markers SEQ ID NO: 1 or 2 and heterozygous for
a QTL SMO3 identifiable by markers SEQ ID NO: 3 or 4.
[0123] In one aspect, a Capsicum seed or plant grown therefrom is
provided capable of producing a pepper fruit comprising at, or
immediately prior to, maturity a pericarp comprising a total sugar
content of at least 5.5%, 6.0%, 6.5%, or 7.0% and one or more aroma
molecules characteristic of Capsicum chinense, wherein a Capsicum
plant shares a genetic determinant with Capsicum line SVPS2625;
wherein a genetic determinant provides a total sugar level of at
least 5.5%, 6.0%, 6.5%, or 7.0%, a desired Capsicum chinense
specific aroma molecule, or both. In one aspect, the Capsicum seed
or plant grown therefrom, contains a genetic determinant also
present in a sample seed of SVPS2625, that sample seed having been
deposited at ATCC under Accession No. PTA-122296. In one aspect, a
Capsicum seed or plant grown therefrom, has a desired Capsicum
chinense specific aroma molecule selected from the group consisting
of alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene.
[0124] In one aspect, a Capsicum seed or plant grown therefrom is
provided capable of producing a pepper fruit exhibiting at, or
immediately prior to, maturity a pericarp comprising one or more
aroma molecules characteristic of Capsicum chinense, wherein a
Capsicum plant is a progeny of Capsicum line ZSP8T14-6274. A
representative sample seed Capsicum line ZSP8T14-6274 has been
deposited at ATCC under Accession No. PTA-122300. In one aspect, a
Capsicum seed or plant grown therefrom further comprises a total
sugar content of at least 5.5%, 6.0%, 6.5%, or 7.0%. In one aspect,
a Capsicum seed or plant grown therefrom shares a genetic
determinant with ZSP8T14-6274 for a desired Capsicum chinense
specific aroma molecule. In one aspect, a desired Capsicum chinense
specific aroma molecule is selected from the group consisting of
alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene.
[0125] In one aspect, a Capsicum plant, or part thereof, disclosed
herein is provided capable of producing a pepper fruit exhibiting
at, or immediately prior to, maturity a pericarp comprising a total
sugar content of at least 5.5%, 6.0%, 6.5%, or 7.0%, wherein a
Capsicum plant is a progeny of Capsicum line SMO8T14-6275. In one
aspect, Capsicum line SMO8T14-6275 provides a genetic determinant
for a total sugar level of at least 5.5%, 6.0%, 6.5%, or 7.0%. In
one aspect, Capsicum line SMO8T14-6275 is homozygous for SMO2 and
SMO3 QTLs, wherein a SMO2 QTL is identifiable by markers SEQ ID NO:
1 or 2, and wherein a SMO3 QTL is identifiable by markers SEQ ID
NO: 3 or 4. In an aspect, a progeny plant of Capsicum line
SMO8T14-6275 is heterozygous for SMO2 and SMO3 QTLs, wherein a SMO2
QTL is identifiable by markers SEQ ID NO: 1 or 2, and wherein a
SMO3 QTL is identifiable by markers SEQ ID NO: 3 or 4.
[0126] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SMO2 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 1 and SEQ ID NO: 2, and selecting a
Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 1 or 2, thereby selecting a SMO2
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
1 and SEQ ID NO: 2. In an aspect, a marker molecule can be within a
genomic region flanked by SEQ ID NOs: 1 and 2.
[0127] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SMO3 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 3 and SEQ ID NO: 4, and selecting a
Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 3 or 4, thereby selecting a SMO3
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
3 and SEQ ID NO: 4. In an aspect, a marker molecule can be within a
genomic region flanked by SEQ ID NOs: 3 and 4.
[0128] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SMO8 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 5 and SEQ ID NO: 6, and selecting a
Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 5 or 6, thereby selecting a SMO8
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
5 and SEQ ID NO: 6. In an aspect, a marker molecule can be within a
genomic region flanked by SEQ ID NOs: 5 and 6.
[0129] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SZZ3 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 7 and SEQ ID NO: 8, and selecting a
Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 7 or 8, thereby selecting a SZZ3
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
7 and SEQ ID NO: 8. In an aspect, a marker molecule can be within a
genomic region flanked by SEQ ID NOs: 7 and 8.
[0130] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SZZ4 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 9 and SEQ ID NO: 10, and selecting a
Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 9 or 10, thereby selecting a SZZ4
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
9 and SEQ ID NO: 10. In an aspect, a marker molecule can be within
a genomic region flanked by SEQ ID NOs: 9 and 10.
[0131] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SZZ11 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 11 and SEQ ID NO: 12, and selecting
a Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 11 or 12, thereby selecting a SZZ11
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
11 and SEQ ID NO: 12. In an aspect, a marker molecule can be within
a genomic region flanked by SEQ ID NOs: 11 and 12.
[0132] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SHY1 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 13 and SEQ ID NO: 14, and selecting
a Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 13 or 14, thereby selecting a SHY1
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
13 and SEQ ID NO: 14. In an aspect, a marker molecule can be within
a genomic region flanked by SEQ ID NOs: 13 and 14.
[0133] In one aspect, a method is disclosed herein to obtain a
Capsicum plant by isolating a nucleic acid molecule from a Capsicum
plant, assaying the isolated nucleic acid molecule for a marker
molecule associated with said SHY12 QTL, where a marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 15 and SEQ ID NO: 16, and selecting
a Capsicum plant based on the genotyping results that comprises a
marker molecule SEQ ID NO: 15 or 16, thereby selecting a SHY12
containing Capsicum plant. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
15 and SEQ ID NO: 16. In an aspect, a marker molecule can be within
a genomic region flanked by SEQ ID NOs: 15 and 16.
[0134] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SMO2 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 1 and SEQ ID NO: 2,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2. In an aspect, a marker
molecule can be within a genomic region flanked by SEQ ID NOs: 1
and 2.
[0135] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SMO3 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 3 and SEQ ID NO: 4,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4. In an aspect, a marker
molecule can be within a genomic region flanked by SEQ ID NOs: 3
and 4.
[0136] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SMO8 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 5 and SEQ ID NO: 6,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6. In an aspect, a marker
molecule can be within a genomic region flanked by SEQ ID NOs: 5
and 6.
[0137] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SZZ3 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 7 and SEQ ID NO: 8,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8. In an aspect, a marker
molecule can be within a genomic region flanked by SEQ ID NOs: 7
and 8.
[0138] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SZZ4 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 9 and SEQ ID NO: 10,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10. In an aspect, a
marker molecule can be within a genomic region flanked by SEQ ID
NOs: 9 and 10.
[0139] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SZZ11 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 11 and SEQ ID NO: 12,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12. In an aspect, a
marker molecule can be within a genomic region flanked by SEQ ID
NOs: 11 and 12.
[0140] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SHY1 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 13 and SEQ ID NO: 14,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14. In an aspect, a
marker molecule can be within a genomic region flanked by SEQ ID
NOs: 13 and 14.
[0141] In one aspect, a method is disclosed herein to obtain a
population of Capsicum plants comprising genotyping a population
using a marker molecule associated with a SHY12 QTL, where a marker
molecule is within 10 centimorgans or less from a marker selected
from the group consisting of SEQ ID NO: 15 and SEQ ID NO: 16,
selecting a Capsicum plant based on the genotyping results, and
producing a progeny from the selected Capsicum plant. In an aspect,
a marker molecule can be within 8 centimorgans, within 5
centimorgans, within 3 centimorgans, within 2 centimorgans, or
within 1 centimorgan or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16. In an aspect, a
marker molecule can be within a genomic region flanked by SEQ ID
NOs: 15 and 16.
[0142] In an aspect, a method is disclosed herein to identify a
plant or progeny plant of SVPS2625 by obtaining a Capsicum plant
and genotyping a Capsicum plant using a marker molecule associated
with a SMO2 QTL, where a marker molecule is within 10 centimorgans
or less from a marker selected from the group consisting of SEQ ID
NO: 1 and SEQ ID NO: 2. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
1 and SEQ ID NO: 2. In an aspect, a marker molecule can be within a
genomic region flanked by SEQ ID NOs: 1 and 2.
[0143] In an aspect, a method is disclosed herein to identify a
plant or progeny plant of SVPS2625 by obtaining a Capsicum plant
and genotyping a Capsicum plant using a marker molecule associated
with a SMO3 QTL, where a marker molecule is within 10 centimorgans
or less from a marker selected from the group consisting of SEQ ID
NO: 3 and SEQ ID NO: 4. In an aspect, a marker molecule can be
within 8 centimorgans, within 5 centimorgans, within 3
centimorgans, within 2 centimorgans, or within 1 centimorgan or
less from a marker selected from the group consisting of SEQ ID NO:
3 and SEQ ID NO: 4. In an aspect, a marker molecule can be within a
genomic region flanked by SEQ ID NOs: 3 and 4.
[0144] In an aspect, a pepper seed or pepper plant disclosed herein
is capable of producing a pepper having one or more desirable aroma
compounds and further exhibits one or more traits selected from the
group consisting of: [0145] a. plants with prostrate, compact,
erect growth habits; [0146] b. plants that have glabrous stems or
have sparse, intermediate or abundant stem pubescens; [0147] c.
plants that have glabrous leaves or have sparse, intermediate or
abundant leaf pubescens; [0148] d. plants with green or purple
stems; [0149] e. plants that have pendant, intermediate, or erect
pedicle position at anthesis; [0150] f. plants that have white,
green-white, lavender, blue or violet corolla color; [0151] g.
plants with yellow, pale blue, blue, or purple anthers; [0152] h.
plants with white or blue filament colors; [0153] i. plants having
a stigma included within the anthers, at the same level as the
anthers, or exerted beyond the anthers at full anthesis; [0154] j.
plants that are male sterile or male fertile; [0155] k. plants that
have low, intermediate or high fruit set; [0156] l. plants with
white, straw or cream, yellow, brown, dark brown, or black seeds;
[0157] m. plants that have smooth, intermediate, or dentate calyx
margins; [0158] n. plants that have or lack an annular constriction
at the junction of the calyx and peduncle; [0159] o. plants that
have declining, intermediate, or erect fruit position; [0160] p.
plants that have green, yellow, orange, red, purple, brown, white,
or black immature fruit; [0161] q. plants that have green, yellow,
orange, red, purple, brown, white or black mature fruit; [0162] r.
plants with pepper fruit that is sweet, or has low (i.e. mild),
intermediate (i.e. medium) or high (i.e. hot or very hot) pungency;
[0163] s. plants that have an average fruit length at ripeness that
is very short (less than about one cm), short (about 5 cm or about
2 to about 7 cm), medium (about 10 cm or about 7 to about 12 cm),
long (about 15 cm or about 13 to about 25 cm) or very long (greater
than 25 cm or about 25 cm to about 40 cm); [0164] t. plants with a
fruit wall thickness (measured halfway between the point of
attachment of the stem and the blossom end) from about 0.5 to 1.5
mm or from about 1 to about 2.5 mm or from about 1.5 to about 4 mm
or from about 2 to about 5 mm, or from about 3 to about 6 mm, or
from about 3.5 mm to about 7.5 mm; [0165] u. plants that have an
average fruit width at ripeness that is about 0.3 to 1 cm, about 1
to 2 cm, about 2 to 4 cm, about 3 to 7, about 6 to 10, about 7 to
11 or greater than about 11 cm; [0166] v. plants without persistent
fruit or plants with persistent fruit (fruit that persists and
maintains an attachment to the plant after ripening); [0167] w.
plants with pepper fruit having an average weight at ripeness from
about 1 to 5 g, 5 to 25 g, 25 to 50 g, 50 to 100 g, 100 to 250 g,
150 to 450 g, 200 to 500 g or 300 to 550 g. [0168] x. plants with
pepper fruit that is elongate, oblate, round, conical or pointed,
campanulate, or bell/blocky; [0169] y. plants where the pepper
fruit shape at the point of attachment is acute, obtuse, truncate,
cordate, or lobate; [0170] z. plants where the pepper fruit has or
lacks a neck at the base of the fruit; [0171] aa. plants where the
blossom end is pointed, blunt, or sunken; [0172] bb. plants where
the pepper fruit has a smooth, slightly corrugated, intermediate,
or very corrugated cross section; [0173] cc. plants with resistance
to one or more pests (e.g., nematodes and aphids); dd. plants with
resistance to diseases caused by one or more bacteria or fungi
(e.g., Xanthamonas sp. and Leveillula taurica); [0174] ee. plants
with resistance to diseases caused by one or more viruses (e.g.,
geminivirus, tobamovirus); [0175] ff. plants having or lacking
anthocyanins in unripe pepper fruit; [0176] gg. plants having or
lacking anthocyanins in ripe pepper fruit; [0177] hh. plants that
are resistant or susceptible to low temperature; [0178] ii. plants
that are resistant or susceptible to high temperature; [0179] jj.
plants that are resistant or susceptible to drought; [0180] kk.
plants that are resistant or susceptible to high salt; [0181] ll.
plants that are resistant or susceptible to flooding; [0182] mm.
plants that are resistant or susceptible to heavy metal; [0183] nn.
plants that are resistant or susceptible to high soil moisture;
[0184] oo. plants that are resistant or susceptible to high
humidity; and [0185] pp. plants that shed fruit easily or do not
shed fruit easily.
[0186] In another aspect, this disclosure provides a container of
pepper seeds described herein and a population of pepper plants
described herein. A container of pepper seeds of the instant
disclosure may contain any number, weight or volume of seeds. For
example, a container can contain at least, or greater than, about
10, 25, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000,
1500, 2000, 2500, 3000, 3500, 4000 or more seeds. Alternatively,
the container can contain at least, or greater than, about 5 grams,
10 grams, 15 grams, 20 grams, 25 grams, 50 grams, 100 grams, 250
grams, 500 grams, or 1000 grams of seeds. Alternatively, the
container can contain at least, or greater than, about 1 ounce, 5
ounces, 10 ounces, 1 pound, 2 pounds, 3 pounds, 4 pounds, 5 pounds
or more seeds. Containers of pepper seeds may be any container
available in the art. By way of non-limiting example, a container
may be a box, a bag, a can, a packet, a pouch, a tape roll, a pail,
a tube, or a bottle.
[0187] In an aspect, methods, pepper plants or seeds disclosed
herein are used in combination with one or more pesticides
including, but not limited to, herbicides, fungicides,
insecticides, microbiocides, nematicides, insect repellents,
bactericides, and other substances used to control pests. In
another aspect, methods or pepper plants disclosed herein are used
in combination with one or more triazoles, strobilurins, acylamino
acids, pyrimidines, pyridines, arylphenyl ketones, amides,
benzanilides, imidazoles, dinitrophenols, morpholines,
phenylsulfamides and organophosphorus cpds, derivatives thereof and
combinations thereof which may be applied as seed, foliar, drench
or drip treatments.
[0188] In one aspect, pepper seeds disclosed herein are untreated.
In another aspect, pepper seeds disclosed herein can be subjected
to various treatments. For example, the seeds can be treated to
improve germination by priming the seeds or by disinfection to
protect against seedborne pathogens. In another aspect, seeds can
be coated with any available coating to improve, for example,
plantability, seed emergence, and protection against soilborne
pathogens. Seed coating can be any form of seed coating including,
but not limited to, pelleting, film coating, and encrustments.
[0189] In one aspect, this disclosure provides pepper plant cells,
tissues, and organs that are not reproductive material and do not
mediate the natural reproduction of the plant. In another aspect,
this disclosure also provides pepper plant cells, tissues, and
organs that are reproductive material and mediate the natural
reproduction of the plant. In another aspect, this disclosure
provides pepper plant cells, tissues, and organs that cannot
maintain themselves via photosynthesis. In another aspect, this
disclosure provides somatic pepper plant cells. Somatic cells,
contrary to germline cells, do not mediate plant reproduction.
[0190] The provided cells, tissues and organs can be from seed,
fruit, leaf, cotyledon, hypocotyl, meristem, embryos, endosperm,
root, shoot, stem, pod, flower, inflorescence, stalk, pedicel,
style, stigma, receptacle, petal, sepal, pollen, anther, filament,
ovary, ovule, pericarp, phloem, bud, or vascular tissue. In another
aspect, this disclosure provides a pepper plant chloroplast. In a
further aspect, this disclosure provides epidermal cells, stomata
cell, leaf or root hairs, a storage root, or a tuber. In another
aspect, this disclosure provides a pepper protoplast, mitochondria,
or callus.
[0191] Skilled artisans understand that pepper plants naturally
reproduce via seeds, not via asexual reproduction or vegetative
propagation. In one aspect, this disclosure provides pepper
endosperm. In another aspect, this disclosure provides pepper
endosperm cells. In a further aspect, this disclosure provides a
male or female sterile pepper plant, which cannot reproduce without
human intervention.
[0192] In a further aspect, this disclosure provides processed
products made from the disclosed pepper plants. Such products
include, but are not limited to, prepared or cut pericarp, meal,
oil, plant extract, starch, or fermentation or digestion
products.
[0193] In another aspect, the instant disclosure also provides a
method of producing a pepper fruit, the method comprising: (a)
cultivating a pepper plant disclosure herein; and (b) collecting a
pepper fruit from the pepper plant.
[0194] In a further aspect, the instant disclosure also provides a
method for producing a Capsicum plant capable of producing a sweet
pepper fruit comprising one or more aroma molecules characteristic
of C. chinense, where the method comprises: (a) crossing a C.
annuum plant or hybrid with a C. chinense plant or hybrid to
produce a population of progeny Capsicum plants; and (b) selecting
a progeny Capsicum plant comprising a C. annuum fruit type and
producing a fruit comprising one or more aroma molecules
characteristic of C. chinense. In an aspect, a method further
comprises: (c) backcrossing a selected progeny Capsicum plant with
a C. annuum plant to produce a further progeny generation of
Capsicum plant. In another aspect, a method further comprises: (d)
a further progeny Capsicum plant comprising a C. annuum fruit type
and producing a fruit comprising one or more aroma molecules
characteristic of C. chinense.
[0195] In an aspect, a C. annuum plant or hybrid produced from or
used in a method disclosed herein is male sterile. In another
aspect, a C. annuum plant or hybrid produced from or used in a
method disclosed herein is cytoplasmic male sterile. In an aspect,
a C. chinense plant or hybrid produced from or used in a method
disclosed herein is male sterile. In another aspect, a C. chinense
plant or hybrid produced from or used in a method disclosed herein
is cytoplasmic male sterile.
[0196] In an aspect, a C. annuum hybrid produced from or used in a
method disclosed herein has equal to or greater than about 75%,
77.5%, 80%, 85%, 86%, 87.5%, 90%, 95%, 98% or 99% of its nuclear
DNA derived from a C. annuum plant. In an aspect, a C. annuum
hybrid provided herein comprises an introgression from another
Capsicum species. In an aspect, an introgression found in a C.
annuum hybrid is an introgression from one or more Capsicum species
selected from the group consisting of C. chinense, C. baccatum, C.
praetermissum, C. frutescens, C. galapagoense, and C. eximium. In
another aspect, an introgression found in a C. annuum hybrid is an
introgression from Capsicum chinense. The fraction of the genome
that is derived from C. chinense or C. annuum can be established by
any method known in the art including, but not limited to, the
detection of markers or sequencing to determine where
introgressions have occurred.
[0197] In an aspect, a method disclosed herein further comprises
detecting one or more aroma molecules in fruits of the population
of progeny Capsicum plants via GC/MS. In another aspect, one or
more aroma molecules detected in a method disclosed herein are
terpene molecules. In another aspect, one or more aroma molecules
detected in a method disclosed herein are selected from the group
consisting of alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene, and
combinations thereof. In an aspect, a fruit of a selected progeny
Capsicum plant comprises alpha-Cubebene at a level having a
Relative GC/MS Response Score of at least 0.05, 0.08, 0.1, 0.12,
0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5,
2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5. In another
aspect, a fruit of a selected progeny Capsicum plant comprises
delta-Cadinene at a level having a Relative GC/MS Response Score of
at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4,
0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5,
10.0, 12.5, 15.0, or 17.5. In a further aspect, a fruit of a
selected progeny Capsicum plant comprises 1,4-Cadinadiene at a
level having a Relative GC/MS Response Score of at least 0.05,
0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0,
or 17.5. In another aspect, a selected progeny Capsicum plant is
capable of producing a fruit having a total sugar content of at
least 5.5%.
[0198] In an aspect, the instant disclosure further provides a
method for selecting a sweet pepper fruit comprising one or more
aroma molecules characteristic of C. chinense, where the method
comprises: (a) obtaining a sweet pepper fruit; (b) detecting in the
sweet pepper fruit one or more aroma molecules selected from the
group consisting of alpha-Cubebene, delta-Cadinene,
1,4-Cadinadiene, and combinations thereof.
[0199] In another aspect, the instant disclosure further provides a
method for introducing a desired C. chinense specific aroma
molecule into a sweet pepper plant, the method comprising: (a)
crossing a sweet pepper plant with a plant selected from the group
consisting of Capsicum line ZSP8T14-6274, a representative sample
seed of which line having been deposited at ATCC under Accession
No. PTA-122300, a progeny line of Capsicum line ZSP8T14-6274,
Capsicum line SVPS2625, a representative sample of seed of which
line having been deposited at ATCC under Accession No. PTA-122296,
and a progeny line of Capsicum line SVPS2625; (b) selecting an
F.sub.1 progeny pepper plant comprising the desired C. chinense
specific aroma molecule; (c) backcrossing the F.sub.1 progeny to
the sweet pepper plant; (d) selecting a backcrossed progeny pepper
plant comprising the desired C. chinense specific aroma molecule;
and (e) repeating steps (c) and (d) three or more times in
succession to produce selected fourth or higher backcross progeny
that comprise the desired C. chinense specific aroma molecule. In
an aspect, the desired C. chinense specific aroma molecule is
selected from the group consisting of alpha-Cubebene,
delta-Cadinene, 1,4-Cadinadiene, and combinations thereof.
[0200] In an aspect, the instant disclosure provides a method for
producing a Capsicum plant exhibiting a sweet pepper fruit
comprising one or more desirable aromas, where said method
comprises: (a) crossing a C. annuum plant or hybrid with a Capsicum
plant or hybrid capable of producing a fruit exhibiting said one or
more desirable aromas to produce a population of progeny Capsicum
plants; and (b) selecting a progeny Capsicum plant comprising a C.
annuum fruit type and exhibiting a fruit comprising said one or
more desirable aromas. In an aspect, a method disclosed herein
comprises detecting said one or more desirable aromas via GC/MS. In
another aspect, one or more desirable aromas comprise terpene
molecules. In a further aspect, one or more desirable aromas
comprising a terpene are selected from the group consisting of
alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene, and combinations
thereof. In another aspect, one or more desirable aromas comprise
alpha-Cubebene, delta-Cadinene, and 1,4-Cadinadiene. In another
aspect, a method disclosed herein further comprises genotyping the
Pun1 locus in a selected progeny Capsicum plant. In a further
aspect, genotyping further comprises detecting a punt-1 allele. In
a further aspect, progeny in a breeding cross are selected based on
the outcome of detection of the pun1-1 allele.
[0201] Pepper plants, seeds, or fruits disclosed herein can be
identified by assessing their genetic complements. Plant genetic
complements may be assessed by genetic marker profiles, and by the
expression of phenotypic traits that are characteristic of the
expression of the genetic complement, e.g., isozyme typing
profiles. Genetic marker assays include many well-known techniques,
such as Simple Sequence Length Polymorphisms (SSLPs), 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), Simple Sequence
Repeat (SSR), INDEL (INsertion/DELetion) and Single Nucleotide
Polymorphisms (SNPs).
[0202] Pepper plants or seeds disclosed herein (e.g., line
ZSP8T14-6274 and hybrids SVPS2625, 2626, and 2646) can be used as
starting varieties to develop or derive new pepper varieties. The
development of new varieties using one or more starting varieties
is well known in the art. For example, novel varieties may be
created by crossing a pepper disclosed herein with a different
pepper plant followed by multiple generations of breeding according
to well-known methods.
[0203] In selecting such a second plant to cross for the purpose of
developing novel lines or varieties, it may be desired to choose
those plants which either themselves exhibit one or more selected
desirable characteristics or which exhibit the desired
characteristic(s) when in hybrid combination. Examples of desirable
traits include: high seed yield, high seed germination rate,
seedling vigor, early fruit maturity, high fruit yield, ease of
fruit setting, disease tolerance or resistance, and adaptability
for soil and climate conditions. Consumer-driven traits, such as a
preference for a given fruit size, shape, color, and texture are
other traits that may be incorporated into new lines of sweet
pepper plants developed by this disclosure.
[0204] Particularly desirable traits that may be incorporated by
this disclosure are improved resistance to different viral, fungal,
and bacterial pathogens. Anthracnose and Phytophthora blight are
fungal diseases affecting various species of pepper. Fruit lesions
and fruit rot are the commercially important aspects of these
diseases. Bacterial leaf spot and bacterial wilt are other diseases
affecting pepper plants, especially during the wet season. Viral
pathogens affecting pepper plants include Tomato spotted wilt
virus, Cucumber mosaic virus, Chili veinal mottle virus,
Geminiviruses, Potyviruses, Pepper mosaic virus, and Tobacco mosaic
virus.
[0205] Improved resistance to insect pests is another desirable
trait that may be incorporated into new lines of pepper plants
developed by this disclosure. Insect pests affecting the various
species of pepper include the European corn borer, corn earworm,
aphids, flea beetles, whiteflies, mites, broad mites, the pepper
weevil and thrips.
[0206] Once initial crosses have been made, inbreeding and
selection take place to produce new varieties. Molecular markers or
metabolic profiling can be used to assist selection of desired
progenies and further breeding. For development of a uniform line,
often five or more generations of selfing and selection are
involved.
[0207] Uniform lines of new pepper varieties may also be developed
by way of double-haploids. Pepper is naturally a diploid and the
basic chromosome number of the genus is x=12, most are 2n=2x=24,
including the cultivated ones. A few wild species have 2n=26.
Ploidy changes (both tetraploidy and haploidy) are relatively easy
to induce in Capsicum species. In fact, the inducement of an
octaploid C. annuum was reported (Panda et al., Theor. Appl. Genet.
68: 567-570 (1984)). Doubled haploids have proved particularly
valuable in breeding. This technique allows the creation of true
breeding lines without the need for multiple generations of selfing
and selection. In this manner true breeding lines can be produced
in as little as one generation.
[0208] Haploid embryos may be produced from microspores, pollen,
anther cultures, or ovary cultures. The haploid embryos may then be
doubled autonomously, or by chemical treatments (e.g. colchicine
treatment). Alternatively, haploid embryos may be grown into
haploid plants and treated to induce chromosome doubling. In either
case, fertile homozygous plants are obtained. In accordance with
the disclosure, any of such techniques may be used to achieve a
homozygous line.
[0209] Besides doubled haploid, backcrossing can also be used to
improve an inbred plant. Backcrossing transfers a specific
desirable trait from one inbred or non-inbred source to an inbred
that lacks that trait. This can be accomplished, for example, by
first crossing a superior inbred (A) (recurrent parent) to a donor
inbred (non-recurrent parent), which carries the appropriate locus
or loci for the trait in question. The progeny of this cross are
then mated back to the superior recurrent parent (A) followed by
selection in the resultant progeny 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 are heterozygous for loci controlling the characteristic
being transferred, but are like the superior parent for most or
almost all other loci in the pepper genome. The last backcross
generation would be selfed to give pure breeding progeny for the
trait being transferred.
[0210] In a further aspect, pepper plants disclosed herein can also
be subject to additional breeding using one or more known methods
in the art, e.g., pedigree breeding, recurrent selection, mass
selection, and mutation breeding. Pedigree breeding starts with the
crossing of two genotypes, such as a pepper variety comprising a
desirable aroma disclosed herein and another pepper variety lacking
such an aroma. If the two original parents do not provide all the
desired characteristics, other sources can be included in the
breeding population. In the pedigree method, superior plants are
selfed and selected in successive filial generations. In the
succeeding filial generations the heterozygous condition gives way
to homogeneous varieties as a result of self-pollination and
selection. Typically in the pedigree method of breeding, five or
more successive filial generations of selfing and selection is
practiced: F.sub.1 to F.sub.2; F.sub.2 to F.sub.3; F.sub.3 to
F.sub.4; F.sub.4 to F.sub.5, etc. After a sufficient amount of
inbreeding to fix loci in the variety, successive filial
generations will serve to increase seed of the developed variety.
The developed variety may comprise homozygous alleles at about 95%
or more of its loci.
[0211] In addition to being used to create a backcross conversion,
backcrossing can also be used in combination with pedigree
breeding. As discussed previously, backcrossing can be used to
transfer one or more specifically desirable traits from one
variety, the donor parent, to a developed variety called the
recurrent parent, which has overall good agronomic characteristics
yet lacks that desirable trait or traits. However, the same
procedure can be used to move the progeny toward the genotype of
the recurrent parent but at the same time retain many components of
the non-recurrent parent by stopping the backcrossing at an early
stage and proceeding with selfing and selection. For example, a
pepper variety may be crossed with another variety to produce a
first generation progeny plant. The first generation progeny plant
may then be backcrossed to one of its parent varieties to create a
BC1 or BC2. Progeny are selfed and selected so that the newly
developed variety has many of the attributes of the recurrent
parent and yet one or several of the desired attributes of the
non-recurrent parent. This approach leverages the value and
strengths of the recurrent parent for use in new pepper
varieties.
[0212] Recurrent selection is a method used in a plant breeding
program to improve a population of plants. The method entails
individual plants cross pollinating with each other to form
progeny. The progeny are grown and the superior progeny selected by
any number of selection methods, which include individual plant,
half-sib progeny, full-sib progeny and selfed progeny. The selected
progeny are cross pollinated with each other to form progeny for
another population. This population is planted and again superior
plants are selected to cross pollinate with each other. Recurrent
selection is a cyclical process and therefore can be repeated as
many times as desired. The objective of recurrent selection is to
improve the traits of a population. The improved population can
then be used as a source of breeding material to obtain new
varieties for commercial or breeding use, including the production
of a synthetic line. A synthetic line is the resultant progeny
formed by the intercrossing of several selected varieties.
[0213] Mass selection is another useful technique when used in
conjunction with metabolic marker enhanced selection. In mass
selection seeds from individuals are selected based on phenotype or
genotype. These selected seeds are then bulked and used to grow the
next generation. Bulk selection requires growing a population of
plants in a bulk plot, allowing the plants to self-pollinate,
harvesting the seed in bulk and then using a sample of the seed
harvested in bulk to plant the next generation. Also, instead of
self-pollination, directed pollination could be used as part of the
breeding program.
[0214] Mutation breeding can also be used to introduce new traits
into pepper plants disclosed herein. Mutations that occur
spontaneously or are artificially induced can be useful sources of
variability for a plant breeder. The goal of artificial mutagenesis
is to increase the rate of mutation for a desired characteristic.
Mutation rates can be increased by many different means including
temperature, long-term seed storage, tissue culture conditions,
radiation; such as X-rays, Gamma rays (e.g. cobalt 60 or cesium
137), neutrons, (product of nuclear fission by uranium 235 in an
atomic reactor), Beta radiation (emitted from radioisotopes such as
phosphorus 32 or carbon 14), or ultraviolet radiation (from 2500 to
2900 nm), or chemical mutagens (such as base analogues
(5-bromo-uracil), related compounds (8-ethoxy caffeine),
antibiotics (streptonigrin), alkylating agents (sulfur mustards,
nitrogen mustards, epoxides, ethylenamines, sulfates, sulfonates,
sulfones, lactones), azide, hydroxylamine, nitrous acid, or
acridines). Transposon- or T-DNA-based mutagenesis is also
encompassed by the present disclosure. Once a desired trait is
observed through mutagenesis the trait may then be incorporated
into existing germplasm by traditional breeding techniques. A
pepper plant or seed provided herein can also be produced by one or
more genome engineering techniques or subject to further genomic
editing. For example, one or more genes or loci can be introduced
into a C. annuum background. Exemplary genome engineering
techniques include meganucleases, zinc-finger nucleases, TALENs,
and CRISPR/Cas9 systems. See, e.g., Gaj et al., ZFN, TALEN, and
CRISPR/Cas-based methods for genome engineering. Trends in
Biotechnology, 31:397-405 (2013).
[0215] Pepper plants or lines disclosed herein can also be
genetically engineered to express various phenotypes of agronomic
interest. Exemplary genes implicated in this regard include, but
are not limited to, genes that confer resistance to pests or
disease, genes that confer resistance or tolerance to an herbicide,
genes that control male sterility, genes that affect abiotic stress
resistance (e.g., against salt, heavy metal, flooding), and other
genes and transcription factors that affect plant growth and
agronomic traits such as yield, flowering, plant growth or plant
architecture.
[0216] The following are exemplary embodiments of the present
application:
Embodiment 1
[0217] A Capsicum seed or plant grown therefrom, wherein the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and one or more aroma molecules characteristic of
Capsicum chinense.
Embodiment 2
[0218] A Capsicum seed or plant grown therefrom, wherein the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and one or more aroma molecules characteristic of
Capsicum chinense, wherein the one or more aroma molecules are
absent from pericarp of hybrid pepper PS09954859 or at a higher
level than that of pericarp of hybrid pepper PS09954859 grown under
similar conditions.
Embodiment 3
[0219] The Capsicum seed or plant grown therefrom, according to
Embodiment 1, wherein the pericarp comprises a titratable acidity
at a higher level than that of pericarp of hybrid pepper PS09954859
grown under similar conditions.
Embodiment 4
[0220] The Capsicum seed or plant grown therefrom, according to
Embodiment 1, wherein the pericarp comprises a titratable acidity
of at least 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2,
5.3, 5.4, 5.5, 5.6, or 5.7 mmol H.sup.+/100 g fresh tissue.
Embodiment 5
[0221] The Capsicum seed or plant grown therefrom, according to
Embodiment 1, wherein the Capsicum seed is a Capsicum annuum
seed.
Embodiment 6
[0222] The Capsicum seed or plant grown therefrom, according to
Embodiment 1, wherein the one or more aroma molecules are terpene
molecules.
Embodiment 7
[0223] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the terpene molecules are selected from the
group consisting of alpha-Cubebene, delta-Cadinene,
1,4-Cadinadiene, and combinations thereof.
Embodiment 8
[0224] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises alpha-Cubebene,
delta-Cadinene, and 1,4-Cadinadiene.
Embodiment 9
[0225] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises alpha-Cubebene at a
level having a Relative GC/MS Response Score of at least 0.05,
0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0,
or 17.5.
Embodiment 10
[0226] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises alpha-Cubebene at a
level above 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40,
50, 60, 100, 150, 200, 300 .mu.g/g fresh fruit.
Embodiment 11
[0227] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises delta-Cadinene at a
level having a Relative GC/MS Response Score of at least 0.05,
0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0,
or 17.5.
Embodiment 12
[0228] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises delta-Cadinene at a
level above 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40,
50, 60, 100, 150, 200, 300 .mu.g/g fresh fruit.
Embodiment 13
[0229] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises 1,4-Cadinadiene at a
level having a Relative GC/MS Response Score of at least 0.05,
0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0,
or 17.5.
Embodiment 14
[0230] The Capsicum seed or plant grown therefrom, according to
Embodiment 6, wherein the pericarp comprises 1,4-Cadinadiene at a
level above 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40,
50, 60, 100, 150, 200, 300 .mu.g/g fresh fruit.
Embodiment 15
[0231] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit comprises no
capsaicinoid.
Embodiment 16
[0232] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit comprises a
lower level of capsaicinoid compared to Capsicum annuum.
Embodiment 17
[0233] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit comprises a
lower level of capsaicinoid compared to Capsicum chinense.
Embodiment 18
[0234] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit comprises a
pungency of zero Scoville Heat Unit (SHU).
Embodiment 19
[0235] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit comprises a
pungency of below 4000, 3000, 2000, 1000, 500, 250, 200, 150, 100,
50, 40, 30, 20, 10, or 5 Scoville Heat Units (SHUs).
Embodiment 20
[0236] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pericarp has a comparable
level of the one or more aroma molecules than Capsicum chinense
pericarp grown under similar conditions.
Embodiment 21
[0237] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pericarp has a higher level
of the one or more aroma molecules than Capsicum chinense pericarp
grown under similar conditions.
Embodiment 22
[0238] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is a
hybrid.
Embodiment 23
[0239] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is an
inbred.
Embodiment 24
[0240] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed or plant
yields a seedless fruit.
Embodiment 25
[0241] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is
transgenic.
Embodiment 26
[0242] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein at least 70%, 75%, 77.5%, 80%,
82.5%, 85%, 86%, 87.5%, 90%, 92.5%, 95%, 97.5%, 98%, 99%, 99.5% of
the Capsicum seed's nuclear genetic material is from a C. annuum
background.
Embodiment 27
[0243] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein less than 30%, 25%, 20%, 15%,
14%, 13%, 12.5%, 10%, 7.5%, 5%, 2.5%, 2%, 1%, 0.5% of the Capsicum
seed's nuclear genetic material is from a C. chinense
background.
Embodiment 28
[0244] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum plant comprises
one or more traits selected from the group consisting of resistance
to geminivirus, resistance to Xanthomonas, resistance to aphids,
resistance to powdery mildew, ease of fruit shedding, cold
tolerance, having leaf hairs, thick fruit walls, flavor
differences, and the clustering of seed-bearing placental tissue
close to the stem of a fruit.
Embodiment 29
[0245] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit has a shape
selected from the group consisting of block, pointed, and
round.
Embodiment 30
[0246] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the pepper fruit has a color
selected from the group consisting of green yellow, red, purple,
black, brown, white, and orange
Embodiment 31
[0247] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is a progeny
of Capsicum line ZSP8T14-6274 or SVPS2625, wherein a representative
sample of seed of the Capsicum line ZSP8T14-6274 has been deposited
at ATCC under Accession No. PTA-122300, and wherein a
representative sample of seed of the Capsicum line SVPS2625 has
been deposited at ATCC under Accession No. PTA-122296.
Embodiment 32
[0248] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is a F.sub.1
progeny of Capsicum line ZSP8T14-6274 or SVPS2625, wherein a
representative sample of seed of the Capsicum line ZSP8T14-6274 has
been deposited at ATCC under Accession No. PTA-122300, and wherein
a representative sample of seed of the Capsicum line SVPS2625 has
been deposited at ATCC under Accession No. PTA-122296.
Embodiment 33
[0249] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is a F.sub.2
progeny of Capsicum line ZSP8T14-6274 or SVPS2625, wherein a
representative sample of seed of the Capsicum line ZSP8T14-6274 has
been deposited at ATCC under Accession No. PTA-122300, and wherein
a representative sample of seed of the Capsicum line SVPS2625 has
been deposited at ATCC under Accession No. PTA-122296.
Embodiment 34
[0250] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed is
essentially derived from Capsicum line ZSP8T14-6274 or SVPS2625,
wherein a representative sample of seed of the Capsicum line
ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, and wherein a representative sample of seed of the
Capsicum line SVPS2625 has been deposited at ATCC under Accession
No. PTA-122296.
Embodiment 35
[0251] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum seed comprises one
or more introgressed loci from Capsicum line ZSP8T14-6274 or
SVPS2625, wherein a representative sample of seed of the Capsicum
line ZSP8T14-6274 has been deposited at ATCC under Accession No.
PTA-122300, wherein a representative sample of seed of the Capsicum
line SVPS2625 has been deposited at ATCC under Accession No.
PTA-122296, and wherein the one or more introgressed loci provide
genetic determinant for producing the one or more aroma molecules
characteristic of Capsicum chinense.
Embodiment 36
[0252] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 1 to 14, wherein the Capsicum plant exhibits a
fruit comprising at maturity one or more aroma molecules
characteristic of Capsicum chinense at a level equal to or higher
than that of a Capsicum line when grown under similar conditions,
wherein the Capsicum line is selected from the group consisting of
lines ZSP8T14-6274 and SVPS2625, wherein a representative sample of
seed of the Capsicum line ZSP8T14-6274 has been deposited at ATCC
under Accession No. PTA-122300, wherein a representative sample of
seed of the Capsicum line SVPS2625 has been deposited at ATCC under
Accession No. PTA-122296.
Embodiment 37
[0253] A Capsicum seed or plant grown therefrom, wherein the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2,
0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0,
4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5.
Embodiment 38
[0254] A Capsicum seed or plant grown therefrom, wherein the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and delta-Cadinene at a level having a Relative GC/MS
Response Score of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2,
0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0,
4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5.
Embodiment 39
[0255] A Capsicum seed or plant grown therefrom, wherein the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and 1,4-Cadinadiene at a level having a Relative GC/MS
Response Score of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2,
0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0,
4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5.
Embodiment 40
[0256] A Capsicum seed or plant grown therefrom, wherein the
Capsicum plant exhibits a pepper fruit at, or immediately prior to,
maturity having pericarp comprising a total sugar content of at
least 5.5% and one or more terpene molecules selected from the
group consisting of alpha-Cubebene at a level having a Relative
GC/MS Response Score of at least 0.25, delta-Cadinene at a level
having a Relative GC/MS Response Score of at least 0.25,
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.15, and any combination thereof.
Embodiment 41
[0257] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 37 to 40, wherein the Capsicum is Capsicum
annuum.
Embodiment 42
[0258] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 37 to 40, wherein the pericarp comprises a
titratable acidity at a higher level than that of pericarp of
hybrid pepper PS09954859 grown under similar conditions.
Embodiment 43
[0259] The Capsicum seed or plant grown therefrom, according to any
one of Embodiments 37 to 40, wherein the pericarp comprises a
titratable acidity of at least 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,
4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, or 5.7 mmol H.sup.+/100 g
fresh tissue.
Embodiment 44
[0260] A container of Capsicum seeds according to any one of
Embodiments 1 to 14.
Embodiment 45
[0261] A population of Capsicum plants according to any one of
Embodiments 1 to 14.
Embodiment 46
[0262] A part of the Capsicum plant according to any one of
Embodiments 1 to 14, wherein the part is selected from a cell, a
protoplast, a leaf, pollen, an embryo, a root, a root tip, anther,
a flower, a fruit, a pistil, a petiole, a meristem, a cotyledon, a
hypocotyl, and a seed.
Embodiment 47
[0263] A sweet bell pepper fruit comprising at maturity one or more
aroma molecules characteristic of Capsicum chinense, wherein the
one or more aroma molecules are absent from pericarp of hybrid
pepper PS09954859 or at a higher level than that of pericarp of
hybrid pepper PS09954859 grown under similar conditions.
Embodiment 48
[0264] The sweet bell pepper fruit according to Embodiment 47,
wherein the fruit comprises a total sugar content of at least
5.5%.
Embodiment 49
[0265] The sweet bell pepper fruit according to Embodiment 47,
wherein the one or more aroma molecules are terpene molecules.
Embodiment 50
[0266] The sweet bell pepper fruit according to Embodiment 47,
wherein the terpene molecules are selected from the group
consisting of alpha-Cubebene, delta-Cadinene, and
1,4-Cadinadiene.
Embodiment 51
[0267] The sweet bell pepper fruit according to Embodiment 47,
wherein the fruit comprises a titratable acidity at a higher level
than that of a fruit of hybrid pepper PS09954859 grown under
similar conditions.
Embodiment 52
[0268] The sweet bell pepper fruit according to Embodiment 47,
wherein the fruit comprises a titratable acidity of at least 4.2,
4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5,
5.6, or 5.7 mmol H.sup.+/100 g fresh tissue.
Embodiment 53
[0269] A method of producing a pepper fruit, the method comprising:
[0270] a. cultivating a plant according to any one of Embodiments 1
to 14; and [0271] b. collecting a pepper fruit from the plant.
Embodiment 54
[0272] A method for producing a Capsicum plant capable of producing
a sweet pepper fruit comprising one or more aroma molecules
characteristic of Capsicum chinense, wherein the method comprises:
[0273] a. crossing a C. annuum plant or hybrid with a C. chinense
plant or hybrid to produce a population of progeny Capsicum plants;
and [0274] b. selecting a progeny Capsicum plant comprising a C.
annuum fruit type and producing a fruit at, or immediately prior
to, maturity comprising one or more aroma molecules characteristic
of C. chinense, wherein the one or more aroma molecules are
selected from the group consisting of alpha-Cubebene at a level
having a Relative GC/MS Response Score of at least 0.05, 0.08, 0.1,
0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,
1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5;
delta-Cadinene at a level having a Relative GC/MS Response Score of
at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4,
0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5,
10.0, 12.5, 15.0, or 17.5; 1,4-Cadinadiene at a level having a
Relative GC/MS Response Score of at least 0.05, 0.08, 0.1, 0.12,
0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5,
2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5; and any
combinations thereof.
Embodiment 55
[0275] A method for producing a Capsicum plant capable of producing
a sweet pepper fruit comprising one or more aroma molecules
characteristic of Capsicum chinense, wherein the method comprises:
[0276] c. crossing a C. annuum plant or hybrid with a C. chinense
plant or hybrid to produce a population of progeny Capsicum plants;
and [0277] d. selecting a progeny Capsicum plant comprising a C.
annuum fruit type and producing a fruit at, or immediately prior
to, maturity comprising one or more aroma molecules characteristic
of C. chinense.
Embodiment 56
[0278] The method of Embodiment 55, wherein the method further
comprises detecting one or more aroma molecules in fruits of the
population of progeny Capsicum plants via GC/MS.
Embodiment 57
[0279] The method of Embodiment 55, wherein the one or more aroma
molecules are terpene molecules.
Embodiment 58
[0280] The method of Embodiment 55, wherein the one or more aroma
molecules are selected from the group consisting of alpha-Cubebene,
delta-Cadinene, 1,4-Cadinadiene, and combinations thereof.
Embodiment 59
[0281] The method of Embodiment 55, wherein the one or more aroma
molecules are alpha-Cubebene, delta-Cadinene, and
1,4-Cadinadiene.
Embodiment 60
[0282] The method of Embodiment 55, wherein the fruit of the
selected progeny Capsicum plant comprises alpha-Cubebene at a level
having a Relative GC/MS Response Score of at least 0.05, 0.08, 0.1,
0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,
1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or
17.5.
Embodiment 61
[0283] The method of Embodiment 55, wherein the fruit of the
selected progeny Capsicum plant comprises delta-Cadinene at a level
having a Relative GC/MS Response Score of at least 0.05, 0.08, 0.1,
0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,
1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or
17.5.
Embodiment 62
[0284] The method of Embodiment 55, wherein the fruit of the
selected progeny Capsicum plant comprises 1,4-Cadinadiene at a
level having a Relative GC/MS Response Score of at least 0.05,
0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7,
0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0,
or 17.5.
Embodiment 63
[0285] The method of Embodiment 55, wherein the selected progeny
Capsicum plant is capable of producing a fruit having a total sugar
content of at least 5.5%.
Embodiment 64
[0286] The method of Embodiment 55, wherein the method further
comprises: [0287] e. backcrossing the selected progeny Capsicum
plant with the C. annuum plant to produce a population of further
progeny Capsicum plants.
Embodiment 65
[0288] The method of Embodiment 55, wherein the method further
comprises: [0289] f. selecting a further progeny Capsicum plant
comprising a C. annuum fruit type and producing a fruit comprising
one or more aroma molecules characteristic of C. chinense.
Embodiment 66
[0290] The method of Embodiment 55, wherein the C. annuum plant or
hybrid is male sterile.
Embodiment 67
[0291] The method of Embodiment 55, wherein the C. annuum plant or
hybrid is cytoplasmic male sterile.
Embodiment 68
[0292] The method of Embodiment 55, wherein the C. chinense plant
or hybrid is male sterile.
Embodiment 69
[0293] The method of Embodiment 55, wherein the C. chinense plant
or hybrid is cytoplasmic male sterile.
Embodiment 70
[0294] A method for selecting a sweet pepper fruit comprising one
or more aroma molecules characteristic of Capsicum chinense,
wherein the method comprises: [0295] a. obtaining a sweet pepper
fruit; [0296] b. detecting in the sweet pepper fruit one or more
aroma molecules selected from the group consisting of
alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene, and combinations
thereof.
Embodiment 71
[0297] A method for introducing a desired Capsicum chinense
specific aroma molecule into a sweet pepper plant, wherein the
desired Capsicum chinense specific aroma molecule is selected from
the group consisting of alpha-Cubebene, delta-Cadinene,
1,4-Cadinadiene, and any combinations thereof, the method
comprising: [0298] a. Crossing a sweet pepper plant with a plant
selected from the group consisting of Capsicum line ZSP8T14-6274, a
representative sample seed of which line having been deposited at
ATCC under Accession No. PTA-122300, a progeny line of Capsicum
line ZSP8T14-6274, Capsicum line SVPS2625, a representative sample
of seed of which line having been deposited at ATCC under Accession
No. PTA-122296, and a progeny line of Capsicum line SVPS2625,
[0299] b. selecting an F.sub.1 progeny pepper plant comprising the
desired Capsicum chinense specific aroma molecule in a fruit at, or
immediately prior to, maturity; [0300] c. backcrossing the F.sub.1
progeny to the sweet pepper plant; [0301] d. selecting a
backcrossed progeny pepper plant comprising the desired Capsicum
chinense specific aroma molecule; [0302] e. repeating steps (c) and
(d) three or more times in succession to produce selected fourth or
higher backcross progeny that comprise the desired Capsicum
chinense specific aroma molecule.
Embodiment 72
[0303] The method of Embodiment 71, wherein the desired Capsicum
chinense specific aroma molecule is selected from the group
consisting of alpha-Cubebene at a level having a Relative GC/MS
Response Score of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2,
0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0,
4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5; delta-Cadinene at a level
having a Relative GC/MS Response Score of at least 0.05, 0.08, 0.1,
0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9,
1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5, 10.0, 12.5, 15.0, or 17.5;
1,4-Cadinadiene at a level having a Relative GC/MS Response Score
of at least 0.05, 0.08, 0.1, 0.12, 0.15, 0.18, 0.2, 0.25, 0.3, 0.4,
0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 7.5,
10.0, 12.5, 15.0, or 17.5; and any combinations thereof.
Embodiment 73
[0304] A method for producing a Capsicum plant exhibiting a sweet
pepper fruit comprising one or more desirable aromas, wherein the
method comprises: [0305] c. crossing a C. annuum plant or hybrid
with a Capsicum plant or hybrid capable of producing a fruit
exhibiting the one or more desirable aromas to produce a population
of progeny Capsicum plants; and [0306] d. selecting a progeny
Capsicum plant comprising a C. annuum fruit type and exhibiting a
fruit comprising the one or more desirable aromas.
Embodiment 74
[0307] The method of Embodiment 73, wherein the method comprises
detecting the one or more desirable aromas via GC/MS.
Embodiment 75
[0308] The method of Embodiment 73, wherein the one or more
desirable aromas comprise terpene molecules.
Embodiment 76
[0309] The method of Embodiment 73, wherein the one or more
desirable aromas comprise a terpene are selected from the group
consisting of alpha-Cubebene, delta-Cadinene, 1,4-Cadinadiene, and
combinations thereof.
Embodiment 77
[0310] The method of Embodiment 73, wherein the one or more
desirable aromas comprise alpha-Cubebene, delta-Cadinene, and
1,4-Cadinadiene.
Embodiment 78
[0311] The method of Embodiment 73, wherein the method further
comprises genotyping the Pun1 locus in the selected progeny
Capsicum plant.
Embodiment 79
[0312] The method of Embodiment 73, wherein the genotyping further
comprises detecting a pun1-1 allele.
Embodiment 80
[0313] A Capsicum plant or seed capable of producing a pepper fruit
comprising at or near maturity a total sugar content of at least
5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, or 9% and a Brix rating
of at least 8.0, 8.5, 9.0, or 9.5.
Embodiment 81
[0314] The Capsicum plant or seed capable of producing a pepper
fruit of Embodiment 80, wherein the fruit is preferred by at least
30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65% more consumers when
compared to a commercially available or morphologically comparable
sweet pepper fruit in a test of consumer preference.
Embodiment 82
[0315] The Capsicum plant or seed capable of producing a pepper
fruit of Embodiment 81, wherein the fruit is described by at least
30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 75%, or 80% more consumers
in a test of consumer preference as more flavorful, more sweet, and
less bitter when compared to a commercially available or
morphologically comparable sweet pepper fruit.
Embodiment 83
[0316] The Capsicum plant or seed capable of producing a pepper
fruit of Embodiment 80, wherein the fruit scores at least 20%
higher than a commercially available or morphologically comparable
sweet pepper fruit based on a consumer graded hedonic attribute
test for one or more categories selected from the group consisting
of overall rating, flavor, and sweetness.
Embodiment 84
[0317] The Capsicum plant or seed capable of producing a pepper
fruit of Embodiment 80, wherein the fruit scores at least 5.5, 6.0,
or 6.5 for one or more categories selected from the group
consisting of overall rating, flavor, and sweetness based on a
consumer graded hedonic attribute test.
Embodiment 85
[0318] The Capsicum fruit comprising a total sugar content of at
least 5.5%, 6.0%, 6.5%, or 7.0% and a Brix rating of at least 8.0,
8.5, 9.0, or 9.5 at or near maturity.
Embodiment 86
[0319] The Capsicum fruit of Embodiment 85, wherein the fruit is
preferred by at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, or 65%
more consumers when compared to a commercially available or
morphologically comparable sweet pepper fruit in a test of consumer
preference.
Embodiment 87
[0320] The Capsicum fruit of Embodiment 86, wherein the fruit is
described by at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 70%, 75%,
or 80% more consumers in a test of consumer preference as more
flavorful, more sweet, and less bitter when compared to a
commercially available or morphologically comparable sweet pepper
fruit.
Embodiment 88
[0321] The Capsicum fruit of Embodiment 85, wherein the fruit
scores at least 20% higher than a commercially available sweet
pepper fruit based on a consumer graded hedonic attribute test for
one or more categories selected from the group consisting of
overall rating, flavor, and sweetness.
Embodiment 89
[0322] The Capsicum fruit of Embodiment 85, wherein the fruit
scores at least 5.5, 6.0, or 6.5 for one or more categories
selected from the group consisting of overall rating, flavor, and
sweetness based on a consumer graded hedonic attribute test.
Embodiment 90
[0323] A Capsicum seed or plant grown therefrom, capable of
producing a pepper fruit comprising at, or immediately prior to,
maturity a pericarp comprising a total sugar content of at least
5.5%, 6.0%, 6.5%, 7.5%, 8.5%, 9%, or 9.5% and one or more aroma
molecules characteristic of Capsicum chinense, wherein the Capsicum
plant shares a genetic determinant with Capsicum line SVPS2625, a
sample seed of which having been deposited at ATCC under Accession
No. PTA-122296; wherein the genetic determinant provides a total
sugar level of at least 5.5%, 6.0%, 6.5%, 7.5%, 8.5%, 9%, or 9.5%,
a desired Capsicum chinense specific aroma molecule, or both.
Embodiment 91
[0324] A Capsicum seed or plant grown therefrom, capable of
producing a pepper fruit comprising at, or immediately prior to,
maturity a pericarp comprising a total sugar content of at least
5.5%, 6.0%, 6.5%, 7.5%, 8.5%, 9%, or 9.5% and one or more aroma
molecules characteristic of Capsicum chinense, wherein a genetic
determinant providing said total sugar level of at least 5.5% or
said desired Capsicum chinense specific aroma molecule is also
present in SVPS2625, a sample seed of which having been deposited
at ATCC under Accession No. PTA-122296.
Embodiment 92
[0325] The Capsicum seed or plant grown therefrom, of Embodiment 90
or 91, wherein the desired Capsicum chinense specific aroma
molecule is selected from the group consisting of alpha-Cubebene,
delta-Cadinene, 1,4-Cadinadiene.
Embodiment 93
[0326] A Capsicum seed or plant grown therefrom, capable of
producing a pepper fruit exhibiting at, or immediately prior to,
maturity a pericarp comprising one or more aroma molecules
characteristic of Capsicum chinense, wherein the Capsicum plant is
a progeny of Capsicum line ZSP8T14-6274, wherein a representative
sample seed Capsicum line ZSP8T14-6274 has been deposited at ATCC
under Accession No. PTA-122300.
Embodiment 94
[0327] The Capsicum seed or plant grown therefrom, of Embodiment
93, further comprising a total sugar content of at least 5.5%,
6.0%, 6.5%, 7.5%, 8.5%, 9%, or 9.5%.
Embodiment 95
[0328] The Capsicum seed or plant grown therefrom of Embodiment 93,
wherein the Capsicum seed or plant shares with ZSP8T14-6274 a
genetic determinant providing for the desired Capsicum chinense
specific aroma molecule.
Embodiment 96
[0329] The Capsicum seed or plant grown therefrom of Embodiment 95,
wherein the desired Capsicum chinense specific aroma molecule is
selected from the group consisting of alpha-Cubebene,
delta-Cadinene, 1,4-Cadinadiene.
Embodiment 97
[0330] A Capsicum plant, or part thereof, capable of producing a
pepper fruit at, or immediately prior to, maturity comprising a
pericarp comprising a total sugar content of at least 5.5%, 6.0%,
6.5%, 7.5%, 8.5%, 9%, or 9.5%, wherein the Capsicum plant is a
progeny of Capsicum line SMO8T14-6275.
Embodiment 98
[0331] The Capsicum plant, or part thereof, of Embodiment 97,
wherein the Capsicum plant comprises a genetic determinant from
SMO8T14-6275 providing for a total sugar level of at least 5.5%,
6.0%, 6.5%, 7.5%, 8.5%, 9%, or 9.5%.
Embodiment 99
[0332] The Capsicum plant, or part thereof, of Embodiment 97,
wherein the Capsicum plant is homozygous for the SMO2 and SMO3
QTLs, wherein the SMO2 QTL is identifiable by markers SEQ ID NO: 1
or 2, and wherein the SMO3 QTL is identifiable by markers SEQ ID
NO: 3 or 4.
Embodiment 100
[0333] The Capsicum plant, or part thereof, of Embodiment 97,
wherein the Capsicum plant is heterozygous for the SMO2 and SMO3
QTLs, wherein the SMO2 QTL is identifiable by markers SEQ ID NO: 1
or 2, and wherein the SMO3 QTL is identifiable by markers SEQ ID
NO: 3 or 4.
Embodiment 101
[0334] The Capsicum plant, or part thereof, according to Embodiment
80 or 85, wherein the pepper plant is heterozygous for the QTL SMO2
identifiable by markers SEQ ID NO: 1 or 2.
Embodiment 102
[0335] The Capsicum plant, or part thereof, according to Embodiment
80 or 85, wherein the pepper plant is heterozygous for the QTL SMO3
identifiable by markers SEQ ID NO: 3 or 4.
Embodiment 103
[0336] A method to obtain a Capsicum plant, comprising: [0337] a.
isolating a nucleic acid molecule from a Capsicum plant, [0338] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SMO2 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2, and [0339] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SMO2 containing Capsicum
plant.
Embodiment 104
[0340] The method of Embodiment 103, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 105
[0341] The method of Embodiment 103, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 106
[0342] The method of Embodiment 103, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 107
[0343] The method of Embodiment 103, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 108
[0344] The method of Embodiment 103, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 109
[0345] A method to obtain a Capsicum plant, comprising: [0346] a.
isolating a nucleic acid molecule from a Capsicum plant, [0347] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SMO3 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4, and [0348] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SMO3 containing Capsicum
plant.
Embodiment 110
[0349] The method of Embodiment 109, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 111
[0350] The method of Embodiment 109, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 112
[0351] The method of Embodiment 109, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 113
[0352] The method of Embodiment 109, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 114
[0353] The method of Embodiment 109, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 115
[0354] A method comprising: [0355] a. obtaining a population of
Capsicum plants, [0356] b. genotyping the population using a marker
molecule associated with a SMO2 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 1 and SEQ ID NO: 2, [0357] c.
selecting a Capsicum plant based on the genotyping, and [0358] d.
producing a progeny from the selected Capsicum plant.
Embodiment 116
[0359] The method of Embodiment 115, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 117
[0360] The method of Embodiment 115, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 118
[0361] The method of Embodiment 115, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 119
[0362] The method of Embodiment 115, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 120
[0363] The method of Embodiment 115, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 121
[0364] A method comprising: [0365] a. obtaining a population of
Capsicum plants, [0366] b. genotyping the population using a marker
molecule associated with a SMO3 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 3 and SEQ ID NO: 4, [0367] c.
selecting a Capsicum plant based on the genotyping, and [0368] d.
producing a progeny from the selected Capsicum plant.
Embodiment 122
[0369] The method of Embodiment 121 wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 123
[0370] The method of Embodiment 121, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 124
[0371] The method of Embodiment 121, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 125
[0372] The method of Embodiment 121, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 126
[0373] The method of Embodiment 121, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 127
[0374] A method comprising: [0375] a. obtaining a Capsicum plant of
Embodiment 80 or 85, and [0376] b. genotyping the Capsicum plant
using a marker molecule associated with a SMO2 QTL, where the
marker molecule is within 10 centimorgans or less from a marker
selected from the group consisting of SEQ ID NO: 1 and SEQ ID NO:
2.
Embodiment 128
[0377] The method of Embodiment 127, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 129
[0378] The method of Embodiment 127, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 130
[0379] The method of Embodiment 127, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 131
[0380] The method of Embodiment 127, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 132
[0381] The method of Embodiment 127, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 1 and SEQ ID NO: 2.
Embodiment 133
[0382] A method comprising: [0383] a. obtaining a Capsicum plant of
Embodiment 80 or 85, and [0384] b. genotyping the Capsicum plant
using a marker molecule associated with a SMO3 QTL, where the
marker molecule is within 10 centimorgans or less from a marker
selected from the group consisting of SEQ ID NO: 3 and SEQ ID NO:
4.
Embodiment 134
[0385] The method of Embodiment 133, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 135
[0386] The method of Embodiment 133, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 136
[0387] The method of Embodiment 133, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 137
[0388] The method of Embodiment 133, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 138
[0389] The method of Embodiment 133, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 3 and SEQ ID NO: 4.
Embodiment 139
[0390] The method of Embodiment 103, 115, or 127, wherein the
marker molecule is in a genomic region flanked by SEQ ID NOs: 1 and
2.
Embodiment 140
[0391] The method of Embodiment 109, 121, or 133, wherein the
marker molecule is in a genomic region flanked by SEQ ID NOs: 3 and
4.
Embodiment 141
[0392] A method to obtain a Capsicum plant, comprising: [0393] a.
isolating a nucleic acid molecule from a Capsicum plant, [0394] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SMO8 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6, and [0395] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SMO8 containing Capsicum
plant.
Embodiment 142
[0396] The method of Embodiment 141, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 143
[0397] The method of Embodiment 141, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 144
[0398] The method of Embodiment 141, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 145
[0399] The method of Embodiment 141, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 146
[0400] The method of Embodiment 141, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 147
[0401] The method of Embodiment 141, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 5 and 6.
Embodiment 148
[0402] A method to obtain a Capsicum plant, comprising: [0403] a.
isolating a nucleic acid molecule from a Capsicum plant, [0404] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SZZ3 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8, and [0405] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SZZ3 containing Capsicum
plant.
Embodiment 149
[0406] The method of Embodiment 148, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 150
[0407] The method of Embodiment 148, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 151
[0408] The method of Embodiment 148, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 152
[0409] The method of Embodiment 148, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 153
[0410] The method of Embodiment 148, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 154
[0411] The method of Embodiment 148, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 7 and 8.
Embodiment 155
[0412] A method to obtain a Capsicum plant, comprising: [0413] a.
isolating a nucleic acid molecule from a Capsicum plant, [0414] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SZZ4 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10, and [0415] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SZZ4 containing Capsicum
plant.
Embodiment 156
[0416] The method of Embodiment 155, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 157
[0417] The method of Embodiment 155, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 158
[0418] The method of Embodiment 155, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 159
[0419] The method of Embodiment 155, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 160
[0420] The method of Embodiment 155, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 161
[0421] The method of Embodiment 155, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 9 and 10.
Embodiment 162
[0422] A method to obtain a Capsicum plant, comprising: [0423] a.
isolating a nucleic acid molecule from a Capsicum plant, [0424] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SZZ11 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12, and [0425] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SZZ11 containing Capsicum
plant.
Embodiment 163
[0426] The method of Embodiment 162, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 164
[0427] The method of Embodiment 162, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 165
[0428] The method of Embodiment 162, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 166
[0429] The method of Embodiment 162, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 167
[0430] The method of Embodiment 162, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 168
[0431] The method of Embodiment 162, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 11 and 12.
Embodiment 169
[0432] A method to obtain a Capsicum plant comprising: [0433] a.
isolating a nucleic acid molecule from a Capsicum plant, [0434] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SHY1 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14, and [0435] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SHY1 containing Capsicum
plant.
Embodiment 170
[0436] The method of Embodiment 169, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 171
[0437] The method of Embodiment 169, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 172
[0438] The method of Embodiment 169, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 173
[0439] The method of Embodiment 169, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 174
[0440] The method of Embodiment 169, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 175
[0441] The method of Embodiment 169, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 13 and 14.
Embodiment 176
[0442] A method to obtain a Capsicum plant, comprising: [0443] a.
isolating a nucleic acid molecule from a Capsicum plant, [0444] b.
assaying the isolated nucleic acid molecule for a marker molecule
associated with a SHY12 QTL, where the marker molecule is within 10
centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16, and [0445] c.
selecting a Capsicum plant based on the genotyping that comprises
the marker molecule, thereby selecting a SHY12 containing Capsicum
plant.
Embodiment 177
[0446] The method of Embodiment 176, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 178
[0447] The method of Embodiment 176, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 179
[0448] The method of Embodiment 176, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 180
[0449] The method of Embodiment 176, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 181
[0450] The method of Embodiment 176, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 182
[0451] The method of Embodiment 176, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 15 and 16.
Embodiment 183
[0452] A method comprising: [0453] a. obtaining a population of
Capsicum plants, [0454] b. genotyping the population using a marker
molecule associated with a SMO8 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 5 and SEQ ID NO: 6, [0455] c.
selecting a Capsicum plant based on the genotyping, and [0456] d.
producing a progeny from the selected Capsicum plant.
Embodiment 184
[0457] The method of Embodiment 183, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 185
[0458] The method of Embodiment 183, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 186
[0459] The method of Embodiment 183, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 187
[0460] The method of Embodiment 183, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 188
[0461] The method of Embodiment 183, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 5 and SEQ ID NO: 6.
Embodiment 189
[0462] The method of Embodiment 183, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 5 and 6.
Embodiment 190
[0463] A method comprising: [0464] a. obtaining a population of
Capsicum plants, [0465] b. genotyping the population using a marker
molecule associated with a SZZ3 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 7 and SEQ ID NO: 8, [0466] c.
selecting a Capsicum plant based on the genotyping, and [0467] d.
producing a progeny from the selected Capsicum plant.
Embodiment 191
[0468] The method of Embodiment 190, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 192
[0469] The method of Embodiment 190, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 193
[0470] The method of Embodiment 190, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 194
[0471] The method of Embodiment 190, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 195
[0472] The method of Embodiment 190, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 7 and SEQ ID NO: 8.
Embodiment 196
[0473] The method of Embodiment 190, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 7 and 8.
Embodiment 197
[0474] A method comprising: [0475] a. obtaining a population of
Capsicum plants, [0476] b. genotyping the population using a marker
molecule associated with a SZZ4 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 9 and SEQ ID NO: 10, [0477] c.
selecting a Capsicum plant based on the genotyping, and [0478] d.
producing a progeny from the selected Capsicum plant.
Embodiment 198
[0479] The method of Embodiment 197, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 199
[0480] The method of Embodiment 197, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 200
[0481] The method of Embodiment 197, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 201
[0482] The method of Embodiment 197, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 202
[0483] The method of Embodiment 197, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 9 and SEQ ID NO: 10.
Embodiment 203
[0484] The method of Embodiment 197, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 9 and 10.
Embodiment 204
[0485] A method comprising: [0486] a. obtaining a population of
Capsicum plants, [0487] b. genotyping the population using a marker
molecule associated with a SZZ11 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 11 and SEQ ID NO: 12, [0488] c.
selecting a Capsicum plant based on the genotyping, and [0489] d.
producing a progeny from the selected Capsicum plant.
Embodiment 205
[0490] The method of Embodiment 204, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 206
[0491] The method of Embodiment 204, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 207
[0492] The method of Embodiment 204, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 208
[0493] The method of Embodiment 204, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 209
[0494] The method of Embodiment 204, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 11 and SEQ ID NO: 12.
Embodiment 210
[0495] The method of Embodiment 204, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 11 and 12.
Embodiment 211
[0496] A method comprising: [0497] a. obtaining a population of
Capsicum plants, [0498] b. genotyping the population using a marker
molecule associated with a SHY1 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 13 and SEQ ID NO: 14, [0499] c.
selecting a Capsicum plant based on the genotyping, and [0500] d.
producing a progeny from the selected Capsicum plant.
Embodiment 212
[0501] The method of Embodiment 211, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 213
[0502] The method of Embodiment 211, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 214
[0503] The method of Embodiment 211, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 215
[0504] The method of Embodiment 211, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 216
[0505] The method of Embodiment 211, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 13 and SEQ ID NO: 14.
Embodiment 217
[0506] The method of Embodiment 211, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 13 and 14.
Embodiment 218
[0507] A method comprising: [0508] a. obtaining a population of
Capsicum plants, [0509] b. genotyping the population using a marker
molecule associated with a SHY12 QTL, where the marker molecule is
within 10 centimorgans or less from a marker selected from the
group consisting of SEQ ID NO: 15 and SEQ ID NO: 16, [0510] c.
selecting a Capsicum plant based on the genotyping, and [0511] d.
producing a progeny from the selected Capsicum plant.
Embodiment 219
[0512] The method of Embodiment 218, wherein the marker molecule is
within 8 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 220
[0513] The method of Embodiment 218, wherein the marker molecule is
within 5 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 221
[0514] The method of Embodiment 218, wherein the marker molecule is
within 3 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 222
[0515] The method of Embodiment 218, wherein the marker molecule is
within 2 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 223
[0516] The method of Embodiment 218, wherein the marker molecule is
within 1 centimorgans or less from a marker selected from the group
consisting of SEQ ID NO: 15 and SEQ ID NO: 16.
Embodiment 224
[0517] The method of Embodiment 218, wherein the marker molecule is
in a genomic region flanked by SEQ ID NOs: 15 and 16.
EXAMPLES
Example 1. Preparation of C. annuum Plants Producing Fruits with a
C. chinense Specific Aroma
[0518] Sweet bell peppers belong to Capsicum annuum (C. annuum).
Some consumers assess bell peppers as bland and desire more aromas
or flavors in sweet bell peppers. Capsicum chinense (C. chinense),
on the other hand, comprises some extremely pungent peppers. C.
chinense also comprises some desirable flavors, although
traditionally C. chinense has only been used as donors for certain
disease resistance traits. Not much has been explored to
incorporate desirable flavors from C. chinense into other peppers,
e.g., C. annuum. Here, a genetic introgression approach is used to
breed C. annuum plants with C. chinense specific aromas or
flavors.
[0519] In general, a Capsicum line is developed as follows: C.
annuum with aroma profiles typical of C. annuum (recipient parent)
are crossed to a C. chinense with unusual aroma (donor parent). The
hybrid plants are either self-pollinated (to generate a segregating
F.sub.2) or are crossed again to a C. annuum with a typical aroma
profile (to generate a segregating BC.sub.1F.sub.1 population or a
segregating modified BC.sub.1F.sub.1 population). The segregating
populations are grown and selected for fertility, lack of pungency,
and presence of aroma from the donor parent. The individual plants
in these segregating populations are then self-pollinated and the
resulting families (F.sub.2 or BC.sub.1F.sub.2 or modified
BC.sub.1F.sub.2) are planted and again, individual plants are
selected for self-pollination. This process is repeated several
times until fixed lines are created.
[0520] For example, a C. annuum plant with an aroma profile typical
of C. annuum (recipient parent or C. annuum parent, comprising an
approximately 100% C. annuum genome) is crossed with a donor parent
having a C. chinense specific aroma profile. The donor parent
comprises approximately 75% C. annuum genome and approximately 25%
C. chinense genome (e.g., line ZSP8T14-6274, deposited at ATCC
under accession no. PTA-122300) which is derived from a mild orange
C. chinense landrace. A landrace is a dynamic population of a
cultivated plant that has historical origin, distinct identity and
lacks formal crop improvement, as well as often being genetically
diverse, locally adapted and associated with traditional farming
systems. See Camacho Villa et al., Plant Genetic Resources:
Characterization and Utilization 3(3):373-84 (2006). An exemplary
C. annuum line used in the cross is SMO8T14-6275 which deposited at
the ATCC under accession no. PTA-122298). From the cross, three
separate hybrid plants are produced. They are hybrid SVPS2625,
hybrid SVPS2646, and hybrid SVPS2626. They all comprise a C. annuum
fruit type and aromas unique to C. chinense. Hybrid SVPS2625 is
deposited at ATCC under accession no. PTA-122296.
Example 2. Origin and Breeding History of Parent Lines ZSP8T14-6274
and SMO8T14-6275
[0521] Hybrid SVPS2625 is an F.sub.1 progeny of a cross between two
inbreds SMO8T14-6275 and ZSP8T14-6274. Inbred ZSP8T14-6274 is the
male parent and is developed using pedigree breeding from a three
way cross. First, an F.sub.1 hybrid is made between AP 2384 (a
Capsicum chinense variety producing medium sized 2 cm.times.2 cm
fruit that ripens from green to orange) and SMO-28-1284 (a green to
orange mini blocky bell pepper). The F.sub.1 hybrid is then crossed
to 05LB LBGH 0550-M (a green to yellow mini blocky bell pepper that
has a resistance gene conferring Tobamovirus (P0)) and single plant
pedigree selection is initiated for a number of generations
bringing the generation to F.sub.6. The F6 seeds are bulked and
subsequently named ZSP8T14-6274.
[0522] ZSP8T14-6274 develops a medium sized plant that produces
small sized, yellow-orange mini fruit with deep interloculary
grooves. The fruit matures from a medium green to yellow-orange.
ZSP8T14-6274 contains a resistance gene conferring resistance to
Tobamo virus (P0) and is easily crossed to Capsicum annuum.
ZSP8T14-6274 has been observed as uniform and stable during the
years 2010 through 2015, and any variation is within commercially
acceptable limits.
[0523] Inbred SMO8T14-6275 is the female parent and is developed
using the doubled haploid technique. Specifically, an F.sub.1
hybrid is made between SMO-28-1234 and SBY-29-469 and is used as a
donor for anthers for anther culture to grow haploid plants. The
haploid plants are doubled to create homozygous diploid plants
(doubled haploids, or DH), from which seeds are collected. Doubled
haploids are planted in a trial and observed for horticultural
characteristics. The DH variety is bulked, used in test crosses,
and subsequently named SMO8T14-6275.
[0524] SMO8T14-6275 develops a medium sized anthocyaninless plant
that produces medium sized, midi-blocky, fruit. The fruit matures
from a medium green to orange. The line contains the L2 resistance
gene conferring resistance to Tobamo virus (P0,1). SMO8T14-6275 has
been observed as uniform and stable during the years 2010 through
2015, and any variation is within commercially acceptable limits.
SMO-28-1234 is a green to orange mini blocky bell pepper and
contains the L2 resistance gene conferring resistance to Tobamo
virus (P0, 1) and has purple anthers. SBY-29-469 is a green to
yellow large deep blocky pepper and has anthocyaninless
anthers.
Example 3. Determination of Aromatic Molecules Contributing to C.
chinense Aromas Via Metabolic Profiling
[0525] A comparative metabolic profiling approach is used to
identify aromatic molecules underlying a C. chinense flavor
profile. These aromatic molecules can also serve as metabolic
fingerprints for identifying and tracking C. chinense aromas during
pepper selection and breeding. Specifically, solid-phase
microextraction (SPME) and gas chromatography mass spectrometry
(GC-MS) are performed to analyze volatile metabolites from pepper
fruits. The peppers analyzed include C. annuum peppers with an
aroma profile typical of C. annuum, a C. chinense aroma donor
plant, and a set of fixed progeny lines from Example 1 that
comprise C. annuum horticultural traits and a C. chinense aroma
profile. Volatile metabolites shared between pepper plants with a
C. chinense flavor but absent from C. annuum are good candidate C.
chinense aroma compounds.
[0526] To prepare samples for SPME and GC-MS analyses, leaf
materials are first removed from pepper fruits which are then cut
in half to remove seeds and rib materials. The remaining pepper
materials (pericarp) are cut into small pieces and further shredded
using a food chopper. The shredded peppers are mixed with an equal
volume of water and homogenized in a blender to form a pepper
puree. Twenty grams of the homogenized pepper puree are transferred
into a 50 mL centrifuge tube, followed by adding 5 mL of a 200 mM
EDTA-NaOH solution (pH 7.5) and 5 mL of a CaCl.sub.2) solution (0.8
g/mL) to the tube. After mixing the solutions by inverting the tube
for multiple times, 100 .mu.L of a working deuterated internal
standard solution (5 ppm, d3-Ethyl Acetate in methanol) is added
into the mixture, followed by adding 100 .mu.L of Methanol and
further mixing by inverting the tube to form a processed pepper
puree. Six grams of the processed pepper puree are transferred to a
SPME headspace vial and subject to SPME volatile extraction on a
Gerstel MPS Rail or similar autosampler with parameters shown in
Table 1. After the SPME volatile extraction, a Thermo GC-MS is then
used for the separation and detection of the volatile metabolites.
The GC-MS uses a SPME injection liner coupled with a Merlin
Microseal. The parameters used for GC-MS analysis are shown in
Table 2.
[0527] Compound detection and relative response is performed using
unique ions and retention times related to the compounds of
interest. Signal extraction and compound identification is
performed in XCalibur with the NIST database. Signal areas are
integrated to determine system response. Data processing for
calculation of relative response is performed in Microsoft
Excel.
[0528] A relative response to a known concentration of
deuterated-internal standard is used to avoid sampling variability.
First, the retention time and unique ions for each compound of
interest are used to extract the area under its curve. Curve area
for the deuterated internal standard is also calculated using a
retention time of 61.2 seconds and selected ions (46+91 m/z). The
relative GC-MS response score of a compound is then determined
using the following formula: Relative GC-MS Response Score of
compound X=Area of Compound X/Area of Deuterated Internal
Standard.
TABLE-US-00001 TABLE 1 Representative SPME parameters used for
volatile extraction. Parameter Value SPME Fiber 65 .mu.m PDMS-DVB
23 gauge needle Incubation Temp 50.degree. C. Incubation Time 4 min
Extraction Time 10 min Desorption Time 60 sec.
TABLE-US-00002 TABLE 2 Representative GC/MS Parameters for
metabolic profiling of pepper aromatic molecules. Parameter Value
Column VF-5MS (20M .times. 0.15 mm ID .times. 0.15 .mu.m df)
Injection Temperature 220.degree. C. Injection Type Splitless
Column Flow 2.3 mL/min Split Flow (ml/min) 46 mL/min Flow Control
Constant Flow Oven Program 60.degree. C. (2 min) -> 10.degree.
C./min to 160.degree. C. -> 20.degree. C./min to 260.degree. C.
-> 260.degree. C. (1 min) Transfer Line 250.degree. C. Mass
Spectrometer Parameters Acquisition Range 40-300 amu Scan Rate 500
amu/sec Ion Source Temp 200.degree. C.
Example 4. Identification of Three Terpenes as C. chinense Aroma
Fingerprints
[0529] Through the comparative metabolic profiling approach
described in Example 2, predominant volatiles are identified in
line ZSP8T14-6274 but not found in C. annuum parent. These
volatiles include three terpenes (.alpha.-Cubebene,
.delta.-Cadinene, and 1,4-Cadinadiene). The same terpenes are also
present in F.sub.1 hybrid progenies of ZSP8T14-6274 and C.
annuum.
[0530] To evaluate whether any of these volatiles correlate to
unique aromas from C. chinense, the volatile profiles of a
collection of 18 C. annuum genotypes are determined by SPME coupled
with GC-MS. This more extensive survey reveals that the three
terpenes (.alpha.-Cubebene, .delta.-Cadinene, and 1,4-Cadinadiene)
appear unique to line ZSP8T14-6274 and its hybrid progenies.
Therefore, these terpenes represent volatile fingerprints for C.
chinense aromas and apparently are inheritable from line
ZSP8T14-6274 into its hybrid progenies (Table 3). The same three
terpenes are found in very low abundance in only one C. annuum
(hybrid pepper PS09954859, published in U.S. Pat. No. 8,471,113,
deposited at the American Type Culture Collection (ATCC) as
accession No. PTA-11514), out of the 18 genotypes surveyed.
PS09954859 is a C. annuum line with C. chinense in the pedigree
from a disease resistance introgression cross. PS09954859 has two
inbred parent lines SMR 99-1275 and SMY 99-1322, having ATCC
Accession Nos. PTA-11520 and PTA-11517, respectively.
[0531] A trial from the year 2015 also confirms the identified
three terpenes (.alpha.-Cubebene, 6-Cadinene, and 1,4-Cadinadiene)
as volatile fingerprints of C. chinense aromas and their
inheritability from line ZSP8T14-6274 into a hybrid progeny (Table
4).
[0532] Although the field conditions of the 2015 trial appears not
optimal, the pepper plants are still substantially uniform and
stable with some variations that are within commercially acceptable
limits. As is true with most pepper breeding, a small percentage of
variants can occur within commercially acceptable limits for almost
any characteristic during the course of repeated multiplication of
an inbred or its hybrid progeny.
[0533] To assess the best timing for sampling C. chinense signature
aroma molecules, hybrid SVPS2625 pepper fruits are harvested at
various maturation stages together with a control pepper line
(Table 3a). These stages are characterized by the percentage of
total fruit surface area exhibiting mature color (e.g., color
change from green to red). For example, "75% color" stands for a
pepper fruit having about 75% of total fruit surface area
exhibiting mature color. In this test, peppers are harvested and
immediately shipped on day 1 and received at the lab on day 2.
Samples are prepared on day 2 and subject to GC-MS analysis on day
3. The observed amounts of signature volatiles (.alpha.-Cubebene,
.delta.-Cadinene, and 1,4-Cadinadiene) are highest in fruits with
between about 25% and 50% mature color, depending on plant
condition. See Samples 1 and 2 in Table 3a. These volatiles are
also detected in fruits with around 75% mature color and up to 100%
mature color, albeit of lower levels. Therefore, to achieve the
most reliable measurement of the identified three C. chinense
signature aroma molecules (.alpha.-Cubebene, .delta.-Cadinene, and
1,4-Cadinadiene), fruits are preferably picked or harvested around
25%, around 50%, around 75%, between 25% and 50%, or between 50%
and 75% color change and subject to flesh sample testing
immediately or shortly thereafter (preferably within 3 days or 1
day of harvest). Fruit color continues to change to a higher
percent mature color after harvest. Samples assayed outside of the
desired time period may result in signature volatile levels lower
than the detectable limit. See e.g., fruits in Example 6 below.
TABLE-US-00003 TABLE 3 Relative abundance of three terpene
volatiles (.alpha.-Cubebene, .delta.-Cadinene, and 1,4-Cadinadiene)
that are associated with C. chinense aromas (data from a 2014
trial). Relative GC-MS Response Score alpha- delta- 1,4- Plot
Capsicum line Cubebene Cadinene Cadinadiene location Line
ZSP8T14-6274 2.19 4.60 2.11 Woodland (male parent of SVPS2625)
SMO8T14-6275 0.001 0.001 0.001 Woodland (female parent of SVPS2625)
SVPS2625 0.32 0.34 0.19 San Juan Bautista Hybrid 2646 0.68 1.04
0.55 San Juan Bautista Hybrid 2626 3.4 4.62 3.06 San Juan Bautista
Hybrid 2663 5.95 10.31 7.35 San Juan Bautista Hybrid 2661 13.48
22.05 16.39 San Juan Bautista PS09954859 0.19 0.20 0.09 Woodland
PS09954859(resubmit) 0.001 0.09 0.001 Woodland PS09954859 0.001
0.08 0.001 San Juan Bautista
TABLE-US-00004 TABLE 3a Evaluation of C. chinense signature aroma
molecules across fruit maturation stages. .alpha.- .delta.- 1,4-
Cubebene Cadinene Cadinadiene Variety N (ratio to IS) (ratio to IS)
(ratio to IS) Control Green 6 ND ND ND Control 75% Color 8 ND ND ND
Control 100% Color 3 ND ND ND Sample-1 Green 12 0.439 0.125 0.077
Sample-1 25% Color 11 0.446 0.160 0.087 Sample-1 50% Color 11 0.332
0.092 0.061 Sample-1 75% Color 13 0.207 0.062 0.037 Sample-1 100%
Color 12 0.224 0.086 0.042 Sample-2 Green 15 0.043 0.024 ND
Sample-2 25% Color 13 0.056 0.018 ND Sample-2 50% Color 15 0.217
0.070 ND Sample-2 75% Color 13 0.124 0.039 ND Sample-2 100% Color
20 0.056 0.030 ND Samples 1 and 2 are hybrid SVPS2625 fruits
harvested from two different plots where Sample 2 plants appear
more stressed than Sample 1 plants. Control is a hybrid pepper line
related to hybrid PS09954859 described in Example 4. ND stands for
"not detected". Average abundance of each volatile is shown for
each harvest. N refers to the number of pepper fruits sampled for
each stage.
TABLE-US-00005 TABLE 4 Relative abundance of .alpha.-Cubebene,
.delta.-Cadinene, and 1,4-Cadinadiene in a 2015 field trial (s.d.
representing standard deviation). Relative GC-MS Response Score
Total Sugars Titratible Acidity alpha- delta- 1,4- (%) (millimoles
H.sup.+/100 g) Number Capsicum line Cubebene Cadinene Cadinadiene
Average s.d. Average s.d. of plots PS09954859 0.001 0.001 0.001 6.3
n/a 3.4 n/a 1 SVPS2625 0.056 0.103 0.010 6.5 0.38 4.7 0.25 8 Line
ZSP8T14-6274 6.832 17.537 9.951 6.8 0.21 4.6 0.24 7 (male parent of
SVPS2625) SMO8T14-6275 0.001 0.001 0.001 6.4 0.24 5.0 0.17 8
(female parent of SVPS2625)
Example 5. Development of Additional C. annuum Plants Having a C.
chinense Aroma Via Genetic Introgression
[0534] Additional C. annuum plants having a C. chinense aroma are
produced by crossing a C. annuum parent plant with a donor parent
having a C. chinense specific aroma profile. For example, the donor
parent can be a C. chinense plant; line ZSP8T14-6274, deposited at
ATCC under accession no. PTA-122300; or SVPS2625, deposited at ATCC
under accession no. PTA-122296. The donor plant can also be any
other C. chinense with high levels of one or more aroma molecules
identified herein (e.g., .alpha.-Cubebene, .delta.-Cadinene, and
1,4-Cadinadiene). An F.sub.1 hybrid plant from this cross is either
self-pollinated (to generate a segregating F.sub.2) or is crossed
again to the C. annuum parent plant (to generate a segregating
BC.sub.1F.sub.1 population or a segregating modified BC iFi
population). The segregating populations are grown and selected for
fertility, lack of pungency, and presence of aroma from the donor
parent. Volatile profiling can be performed as described in Example
2 to select progenies comprising signature volatile molecules that
are associated with a C. chinense specific flavor (e.g.,
.alpha.-Cubebene, .delta.-Cadinene, and 1,4-Cadinadiene). Plants
are also tested for their fruit sweetness (e.g., total sugar
content) and sweet individuals are selected. Selected individual
plants are then self-pollinated and the resulting families (F.sub.2
or BC.sub.1F.sub.2 or modified BC.sub.1F.sub.2) are planted and
individual plants are again selected for self-pollination. This
process is repeated several times until fixed progeny lines are
created which comprise essentially all the traits of C. annuum
(e.g., sweet) and an aroma profile similar to the donor parent
(e.g., a C. chinense specific flavor profile).
Example 6. Consumer Liking Test of Hybrid SVPS2625
[0535] Consumer preference is tested for hybrid SVPS2625 compared
to PERO.RTM. brand mini-pointy yellow and PERO.RTM. brand
mini-pointy orange peppers (PERO.RTM. Family Farms Food Company,
LLC). For this example, none of the peppers tested have detectable
C. chinense volatiles likely due to the harvesting time as
described in Example 4 (FIG. 1). Commercially available peppers are
purchased locally and hybrid SVPS2625 peppers are harvested at
maturity and shipped overnight to the testing facility. The average
total sugar content of the hybrid SVPS2625 pepper is 7.1% compared
to 5.1% and 5.0% for the orange and yellow retail peppers,
respectively (FIG. 1). The Brix rating for hybrid SVPS2625 is 10.3
which is significantly higher than the commercially available
orange (9.0) and the commercially available yellow (8.8) (FIG.
1).
[0536] All peppers are prepared on the day of testing. Before
testing, all abnormal peppers are removed. Test peppers are washed
in 8.degree. C. tap water for one minute. The peppers are then
submerged in a wash solution containing 200 ppm CHLOROX.RTM.
regular bleach for five minutes (Chlorox Co., Oakland, Calif.). The
peppers are then washed again in fresh 8.degree. C. tap water for
one minute before air drying.
[0537] Before sample preparation, peppers are sorted by color. The
top and bottom 10% of each fruit is cut off. A pepper greater than
3.8 cm is cut in half from stem to peduncle and the seeds and
placenta are removed using the pointed end of a knife. A pepper
less than 3.8 cm in length is not cut and the seeds and placenta
are removed using the pointed end of a knife. The peppers are diced
using a NEMCO.RTM. easy chopper into 0.95 cm cubes (NEMCO.RTM.
model 55500-2, Hicksville, Ohio). The diced peppers are mixed using
a stainless steel spoon and separated for consumer testing and
analytical sampling (FIG. 2). Pepper non-volatile and volatile
sampling is performed for each group of peppers. 250 grams of diced
peppers are blended with 250 grams of water and separated into
sample tubes for testing; two replicates for both non-volatiles and
volatiles for all three pepper preparations are collected (FIG.
3).
[0538] Consumer testing of 142 people is conducted in a supervised
sensory lab setting. A paired preference format is used to make two
comparisons: 1) hybrid SVPS2625 versus the commercially available
mini-pointy yellow pepper and 2) hybrid SVPS2625 versus the
commercially available mini-pointy orange peppers. Consumer testing
is conducted under red light to reduce the impact of color
differences on outcomes. For the test, consumers are asked to:
identify which pepper is preferred (if there is no preference then
pick a forced preference) and to identify the reason for a
preference with a fill-in-the-blank questionnaire. Demographic,
behavioral, and general taste preference questions are asked in
follow up.
[0539] Consumers prefer the taste of SVPS2625 (84%) over
commercially available mini-pointy orange peppers (16%), citing
reasons such as more sweet, more flavorful, and less bitter.
Consumers prefer the taste of SVPS2625 (71%) over commercially
available mini-pointy yellow peppers (29%), citing reasons such as
more sweet, more flavorful, and less bitter. In a sequential
monadic liking test, consumers are given all three samples and
asked to rate each sample on a 9-point hedonic scale. The hedonic
attribute scoring demonstrates that the hybrid SVPS2625 pepper is
moderately liked overall, for flavor, and for sweetness. The
commercially available orange and yellow peppers have significantly
lower scores in all three categories (FIG. 4).
Example 7. Identification of Quantitative Trait Loci from the
SMO-28-1234 Parental Line
[0540] The SMO-28-1234 line is described in Example 2 as a
progenitor to the hybrid pepper line SVPS2625. Use of this line is
described below in the identification of genomic regions that
control variation in total sugar because of its higher sugar,
slightly higher acid, and better flavor rating compared to modern
commercial bell peppers. Bi-parental F.sub.2:F.sub.3 and double
haploid mapping populations are used for QTL identification. The
SMO-28-1234 parental line is crossed to SBY-29-469 for linkage
mapping. F.sub.2:F.sub.3 families are phenotyped in a randomized
complete block design with four replicate plots of up to ten plants
per F.sub.3 family. Double haploid lines are generated from the
F.sub.1 plants of these crosses (DH.sub.1 generation). DH.sub.1
lines are sown in Felda, Fla. in a randomized complete block design
with three replicate plots per DH.sub.1 line for phenotypic data
collection. A subset of each DH.sub.1 line is sampled for
genotyping and linkage mapping. Genotyping for F.sub.2:F.sub.3 and
DH.sub.1 lines is performed using over 130 markers from a pepper
Taqman library to minimize gaps greater than 20 cM.
[0541] Phenotypic data is collected by pooling fruits from multiple
plants per plot per sample. Fruits are assessed for total sugars
(fructose+sucrose), fructose, glucose, titratable acidity (TA), and
a second measure of titratable acidity (TA7) in a vegetable quality
lab in Woodland, Calif. Plants at the Felda, Fla. location are also
measured for perceived sweetness (sugartaste), acidity (acidtaste),
and skin thickness (skin) by an individual tester on a 1-9 scale
where 1 corresponds to the sweetest, highest acid, thinnest skin,
and most favorable texture, respectively and 9 corresponds to the
least sweet, lowest acid, thickest skin, and least pleasant
texture. To describe the estimated properties of QTLs, tables are
presented with data on the estimated QTL effect size (i.e. how much
the QTL is expected to influence the trait), the proportion of
total phenotypic variation in the population explained by a QTL,
and its dominance estimate. The effect size is one half of the
difference between the two contrasting homozygous QTL genotypes.
The proportion of phenotypic variation describes how much of the
variation present in the experiment is explained by a single QTL.
Dominance is the phenotypic difference between the heterozygote QTL
genotype and the average of the two homozygous QTL genotypes.
Traits that are strongly influenced by the SMO2, SMO3, and SMO8
QTLs are shown in Tables 5, 9, and 12.
[0542] QTL identification is performed with the rqtl package
implemented in the R platform and three QTL harboring intervals,
SMO2, SMO3, and SMOG, are identified in the SMO-28-1234 parental
line. To describe the genetic location of QTLs, the LOD profile is
used to determine the statistical significance of a QTL result. The
significance of a LOD score is determined from 1,000 permutations
of the data. Because QTL peak estimates are not perfect, tables are
provided with the confidence interval for each QTL. The confidence
intervals are estimated using a 1.5 LOD interval calculated by
moving down the LOD profile from the peak 1.5 `steps` (-1.5) and up
from the peak 1.5 steps (+1.5) to identify the chromosome positions
(Table 6, 7, 8, 10, 11, 13, and 14).
[0543] The SMO2 interval containing one or more QTLs influencing
sugar content is discovered in a Woodland, Calif. field trial and
subsequently identified in Culiacan, MX and Felda, Fla. field
trials. The SMO2 interval is broad, ranging from 65 cM to 113 cM.
In addition, a QTL for perceived sweetness flavor was also
identified in Felda, Fla. within this interval. Consequently, there
is strong support for a QTL influencing both sugar and the
perceived flavor of sugar in the interval harboring SMO2. QTLs for
sugars and acids are detected in Woodland, C A and Culiacan, MX
(Table 5). QTLs for sugars and acid are found on chromosome 2 in
the Felda, Fla. location (Table 5). The QTL confidence intervals
for Woodland, Calif. (Table 6), Culiacan, MX (Table 7), and Felda,
Fla. are shown (Table 8). The Capsicum line SMO8T14-6275 is tested
and confirmed to comprise a homozygous SMO2 QTL. The hybrid
Capsicum line SVPS2625 is the progeny of SMO8T14-6275 and is
heterozygous at the SMO2 QTL.
TABLE-US-00006 TABLE 5 SMO2 phenotype parameter estimates from SBY-
29-469/SMO-28-1234 F.sub.2:F.sub.3 and DH.sub.1 populations
Position % of Size of Name Trait Chr in cM variance Effect
Woodland, CA; Summer 2010 SMO2-sugar Sugar 2 77.2 26.7 -0.41
SMO2-sugar Fructose 2 74.2 22.6 -0.19 SMO2-sugar Glucose 2 98.2
27.8 -0.19 SMO2-acid TA 2 98.2 26.6 -0.28 SMO2-acid TA7 2 102.2
27.1 -0.26 Culiacan, MX; Winter 2010 SMO2-sugar Sugar 2 100.2 23
-0.49 SMO2-sugar Fructose 2 100.2 13.5 -0.2 SMO2-sugar Glucose 2
100.2 32.2 -0.3 SMO2-acid TA 2 76.2 13.7 -0.27 SMO2-acid TA7 2
100.2 22.6 -0.23 Felda, FL; Winter 2011 SMO2-sugar Sugar 2 109 14.4
-0.28 SMO2-sugar Glucose 2 108 21.6 -0.18 SMO2-acid TA7 2 111.7
17.1 -0.17 SMO2-sugar SugarTaste 2 100 20.6 0.62 SMO2-acid
AcidTaste 2 99 18.2 0.57 SMO2-thickness Skin 2 55 21.2 0.64
TABLE-US-00007 TABLE 6 SMO2 1.5 LOD QTL confidence interval
Woodland, CA, summer 2010 LOD chr pos Sugar chr pos Fructose chr
pos Glucose -1.5 2 67.17 10.61 2 63.17 8.82 2 70.17 11.60 peak 2
77.17 12.31 2 74.17 10.40 2 98.17 13.18 +1.5 2 100.17 10.80 2 98.17
8.76 2 105.17 11.34 LOD chr pos TA chr pos TA7 -1.5 2 76.17 10.81 2
95.17 11.99 peak 2 98.17 12.33 2 102.17 13.92 +1.5 2 103.58 10.67 2
107.17 12.28
TABLE-US-00008 TABLE 7 SMO2 1.5 LOD QTL confidence interval
Culiacan, MX, winter 2010 LOD chr pos Sugar chr pos Fructose chr
pos Glucose -1.5 2 95.17 6.43 2 95.17 3.34 2 96.17 11.31 peak 2
100.17 8.92 2 100.17 4.99 2 100.17 13.17 +1.5 2 110.17 7.41 2
113.17 3.12 2 107.17 11.66 LOD chr pos TA chr pos TA7 -1.5 2 50.17
3.64 2 70.17 6.87 peak 2 76.17 5.22 2 99.17 8.56 +1.5 2 104.17 3.71
2 106.17 6.99
TABLE-US-00009 TABLE 8 SMO2 1.5 LOD QTL confidence interval Felda,
FL, winter 2010 LOD chr pos Sugar chr pos Glucose chr pos
SugarTaste -1.5 2 72.03 1.16 2 72.03 2.37 2 26.03 1.67 peak 2
109.03 2.71 2 108.03 3.94 2 100.03 3.18 +1.5 2 113.62 2.45 2 113.62
3.45 2 113.62 1.80 LOD chr pos AcidTaste chr pos Skin -1.5 2 26.03
0.25 2 43.03 1.71 peak 2 100.03 3.10 2 55.03 3.43 +1.5 2 113.62
1.10 2 86.03 1.92
[0544] The SMO3 interval is identified as a large but variable QTL
on chromosome 3 influencing variation in sugars and acids in plants
grown in Woodland, Calif. and acids in Culiacan, MX (Table 9). The
QTL is detected in Woodland, Calif. and in Culiacan, MX field plots
at a relaxed significance threshold. The SMO3 interval spans 143 to
184 cM on chromosome 3. The QTL confidence intervals are shown for
Woodland, Calif. (Table 10) and Culiacan, MX (Table 11). The
Capsicum line SMO8T14-6275 is tested and confirmed to comprise a
homozygous SMO3 QTL. The hybrid Capsicum line SVPS2625 is the
progeny of SMO8T14-6275 and is heterozygous at the SMO3 QTL.
TABLE-US-00010 TABLE 9 SMO3 phenotype parameter estimates from
SBY-29-469/SMO-28-1234 F.sub.2:F.sub.3 populations Position % of
Size of Name Trait Chr in cM variance Effect Woodland, CA; summer
2010 SMO3-sugar Sugar 3 159.9 10.8 -0.22 SMO3-sugar Fructose 3
155.9 14.2 -0.12 SMO3-sugar Glucose 3 159.9 8.6 -0.098 SMO3-acid TA
3 161.9 20.5 -0.25 SMO3-acid TA7 3 161.9 21.2 -0.23 Culiacan, MX;
Winter 2010 SMO3-acid TA 3 184.4 14.7 -0.22
TABLE-US-00011 TABLE 10 SMO3 1.5 LOD QTL confidence interval
Woodland, CA, summer 2010 LOD chr pos Sugar chr pos Fructose chr
pos Glucose -1.5 3 143.88 2.94 3 147.88 4.43 3 141.88 2.06 peak 3
156.88 4.49 3 155.88 6.09 3 160.88 3.62 +1.5 3 177.88 2.82 3 174.88
4.42 3 180.88 1.93 LOD chr pos TA chr pos TA7 -1.5 3 152.88 7.71 3
151.88 8.02 peak 3 161.88 9.33 3 161.88 9.82 +1.5 3 172.46 7.64 3
170.88 8.09
TABLE-US-00012 TABLE 11 SMO3 1.5 LOD QTL confidence interval
Culiacan, MX, winter 2010 LOD chr pos Sugar chr pos Glucose -1.5 3
176.88 3.40 3 154.88 11.00 peak 3 184.44 5.02 3 159.94 12.84 +1.5 3
184.44 5.02 3 164.88 11.06
[0545] The SMO8 interval contains a QTL on chromosome 8 influencing
variation in sugars and acids and is detected in Woodland, C A and
Felda, Fla. (Table 12). The SMO8 interval spans 37 to 63 cM on
chromosome 8. The QTL confidence intervals are shown for Woodland,
Calif. (Table 13) and Felda, Fla. (Table 14).
TABLE-US-00013 TABLE 12 SMO8 phenotype parameter estimates from
SBY- 29-469/SMO-28-1234 F.sub.2:F.sub.3 and DH.sub.1 populations
Position % of Size of Name Trait Chr in cM variance Effect
Woodland, CA; Summer 2010 SMO8-sugar Sugar 8 59.4 10.5 -0.21
SMO8-sugar Fructose 8 59.4 9 -0.09 SMO8-sugar Glucose 8 58.7 11
-0.11 SMO8-acid TA 8 56.7 17 -0.25 SMO8-acid TA7 8 59.4 10.7 -0.17
Felda, FL; Winter 2010 SMO8-sugar Fructose 8 45.7 26.8 -0.21
SMO8-acid TA 8 54.7 33.8 -0.37
TABLE-US-00014 TABLE 13 SMO8 1.5 LOD QTL confidence interval
Woodland, CA, summer 2010 LOD chr pos Sugar chr pos Fructose chr
pos Glucose -1.5 8 50.73 2.95 8 50.73 2.10 8 49.73 3.36 peak 8
59.37 4.61 8 59.37 3.85 8 58.73 4.91 +1.5 8 64.73 3.00 8 67.23 2.29
8 64.73 2.93 LOD chr pos TA chr pos TA7 -1.5 8 50.73 6.10 8 49.73
2.85 peak 8 57.73 7.63 8 59.37 4.55 +1.5 8 62.73 5.36 8 63.73
2.93
TABLE-US-00015 TABLE 14 SMO8 1.5 LOD QTL confidence interval Felda,
FL, winter 2010 LOD chr pos TA chr pos Fructose -1.5 8 42.73 5.03 8
37.73 3.18 peak 8 54.73 6.76 8 45.73 4.73 +1.5 8 61.73 5.11 8 62.73
3.03
Example 8. Identification of Additional Flavor and Taste
Quantitative Trait Loci
[0546] To identify genomic regions that control variation in total
sugar the SZZ-8T10901 line is chosen for its higher sugar, slightly
higher acid, and better flavor rating compared to modern commercial
bell peppers. Bi-parental F.sub.2:F.sub.3 and double haploid
mapping populations are used for QTL identification. The
SZZ-8T10901 parental line is crossed to SBY-29-469 for linkage
mapping. F.sub.2:F.sub.3 families are phenotyped in a randomized
complete block design with four replicate plots of up to ten plants
per F.sub.3 family. Double haploid lines are generated from the
F.sub.1 plants of these crosses (DH.sub.1 generation). DH.sub.1
lines are sown in Felda, Fla. in a randomized complete block design
with three replicate plots per DH.sub.1 line for phenotypic data
collection. A subset of each DH.sub.1 line is sampled for
genotyping and linkage mapping. Genotyping is performed using over
130 markers from a pepper Taqman library to minimize gaps greater
than 20 cM.
[0547] Phenotypic data is collected by pooling fruits from multiple
plants per plot per sample. Fruits are assessed for total sugars
(fructose+sucrose), fructose, glucose, titratable acidity (TA), and
a second measure of titratable acidity (TA7) in a vegetable quality
lab in Woodland, Calif. Plants at the Felda, Fla. location are also
measured for perceived sweetness (sugartaste) and skin thickness
(skin) by an individual tester on a 1-9 scale where 1 corresponds
to the sweetest, highest acid, thinnest skin, and most favorable
texture, respectively and 9 corresponds to the least sweet, lowest
acid, thickest skin, and least pleasant texture. To describe the
estimated properties of QTLs, tables are presented with data on the
estimated QTL effect size (i.e. how much the QTL is expected to
influence the trait), the proportion of total phenotypic variation
in the population explained by a QTL, and its dominance estimate.
The effect size is one half of the difference between the two
contrasting homozygous QTL genotypes. The proportion of phenotypic
variation describes how much of the variation present in the
experiment is explained by a single QTL. Dominance is the
phenotypic difference between the heterozygote QTL genotype and the
average of the two homozygous QTL genotypes. Traits that are
strongly influenced by the SZZ3, SZZ4, and SZZ11 QTLs are shown in
Tables 15, 18, and 20.
[0548] QTL identification is performed with the rqtl package
implemented in the R platform and three QTL harboring intervals,
SZZ3, SZZ4, and SZZ11, are identified in the SZZ-8T10901 parental
line. To describe the genetic location of QTLs, the LOD profile is
used to determine the statistical significance of a QTL result. The
significance of a LOD score is determined from 1,000 permutations
of the data. Because QTL peak estimates are not perfect, tables are
provided with the confidence interval for each QTL. The confidence
intervals are estimated using a 1.5 LOD interval calculated by
moving down the LOD profile from the peak 1.5 `steps` (-1.5) and up
from the peak 1.5 steps (+1.5) to identify the chromosome positions
(Table 16, 17, 19, 21, 22, and 23).
[0549] The SZZ3 interval contains one to two QTLs influencing
sugars on chromosome 3 and is detected at the Felda, Fla. and
Almeria, ES locations (Table 15). The perceived sweetness
(sugartaste) data from Felda, Fla. also implicates the SZZ3
interval (Table 15). There is a first potential QTL delimited by a
confidence interval spanning 63-115 cM that is detected in Felda,
Fla. (Table 17). A second potential QTL is within the interval of
158-186 cM, which is a more significant QTL at the Almeria, ES
location (Table 16). Both the Felda, Fla. and Almeria, ES sites
show two distinct peaks in the LOD profile on chromosome 3,
although in Felda, Fla. the second peak in the 158-186 cM region is
not significant. The perceived sweetness data from Felda, Fla. also
exhibits a QTL that overlaps with the 63-115 cM QTL (Table 17). For
these reasons, the sugar QTL in the 63-118 cM interval is
designated SZZ3 due to its repeated observations in two locations
and co-location with a perceived sweetness QTL (Table 16 and
17).
TABLE-US-00016 TABLE 15 SZZ3 phenotype parameter estimates from
SBY- 29-469/SZZ-8T10901 F.sub.2:F.sub.3 and DH.sub.1 populations
Position % of Size of Name Trait Chr in cM variance Effect Almeria,
ES SZZ3-sugar Sugar 3 156.4 15 -0.24 SZZ3-sugar Fructose 3 156.4
15.3 -0.15 SZZ3-sugar Glucose 3 107.4 13.3 -0.11 SZZ3-acid TA 3
185.4 14.4 -0.19 SZZ3-acid TA7 3 185.4 12.4 -0.14 Felda, Fl; winter
2010 SZZ3-sugar Sugar 3 82.2 17.5 -0.35 SZZ3-sugar Fructose 3 82.2
18.3 -0.21 SZZ3-sugar SugarTaste 3 27.7 26.8 0.66
TABLE-US-00017 TABLE 16 SZZ3 1.5 LOD QTL confidence interval
Almeria, ES, winter 2010 LOD chr pos Sugar chr pos Fructose chr pos
Glucose -1.5 3 99.43 3.80 3 146.43 4.46 3 85.43 3.20 peak 3 156.43
5.35 3 156.43 5.97 3 107.43 4.81 +1.5 3 167.43 3.84 3 166.43 4.30 3
185.91 3.22 LOD chr pos TA chr pos TA7 -1.5 3 158.43 3.36 3 162.43
2.71 peak 3 185.43 4.97 3 185.43 4.25 1.5 3 185.91 4.89 3 185.91
4.17
TABLE-US-00018 TABLE 17 SZZ3 1.5 LOD QTL confidence interval Felda,
FL, winter 2010 LOD chr pos Sugar chr pos Fructose chr pos
SugarTaste -1.5 3 63.90 1.48 3 64.71 1.59 3 15.71 3.30 peak 3 82.21
2.99 3 82.21 3.13 3 27.71 5.02 +1.5 3 115.71 1.41 3 115.71 1.56 3
79.71 3.49
[0550] The SZZ4 interval contains a QTL on chromosome 4 influencing
sugars and is detected in Woodland, Calif. but there was no
evidence for a QTL at this position in the Almeria, ES trial (Table
18). However, the SZZ4 interval for fructose had a p value of 0.09
in the Felda trial, and because the Woodland trial used a
relatively stringent false discovery rate of 5%, the FDR rate was
relaxed to 10% at which point the fructose LOD peak for SZZ4
becomes significant for the Felda, Fla. population. SZZ4 is
detected within the interval of 84-107 cM on chromosome 4. The QTL
confidence interval is shown for Woodland, Calif. (Table 19).
TABLE-US-00019 TABLE 18 SZZ4 phenotype parameter estimates from
SBY-29-469/ SZZ-8T10901 F.sub.2:F.sub.3 and DH.sub.1 populations
Woodland, CA; Summer 2010 Position % of Size of Name Trait Chr in
cM variance Effect SZZ4-sugar Sugar 4 95 15.7 -0.28 SZZ4-sugar
Fructose 4 95.1 17.1 -0.16
TABLE-US-00020 TABLE 19 SZZ4 1.5 LOD QTL confidence interval
Woodland, CA, summer 2010 LOD chr pos Sugar chr pos Fructose -1.5 4
84.05 3.83 4 84.05 4.66 peak 4 95.05 5.51 4 95.05 6.35 +1.5 4
107.05 3.92 4 108.05 4.64
[0551] The SZZ11 interval on chromosome 11 influences sugars and
acids at Woodland, Calif. and Almeria, ES and acids and skin
thickness at Felda, Fla. (Table 20). The consistent acid QTL
detected in all three locations within the interval of 50-73 cM on
chromosome 11. The QTL confidence intervals are shown for Woodland,
Calif. (Table 21), Almeria, ES (Table 22), and Felda, Fla. (Table
23).
TABLE-US-00021 TABLE 20 SZZ11 phenotype parameter estimates from
SBY- 29-469/SZZ-8T10901 F.sub.2:F.sub.3 and DH.sub.1 populations
Position % of Size of Name Trait Chr in cM variance Effect
Woodland, CA; Summer 2010 SZZ11-sugar Sugar 11 59.3 13.8 -0.23
SZZ11-sugar Fructose 11 57.9 15.3 -0.12 SZZ11-acid TA 11 57.9 43.6
-0.26 SZZ11-acid TA7 11 59.3 28 -0.16 Almeria, ES; Winter 2010
SZZ11-sugar Fructose 11 38.9 10.9 -0.11 SZZ11-acid TA 11 60.9 13.1
-0.2 SZZ11-acid TA7 11 59.9 13.8 -0.16 Felda, FL; Winter 2010
SZZ11-acid TA 11 66.9 38.4 -0.43 SZZ11-acid TA7 11 65.9 39 -0.34
SZZ11-thickness Skin 11 50.6 20.3 -0.68
TABLE-US-00022 TABLE 21 SZZ4 1.5 LOD QTL confidence interval
Woodland, CA, summer 2010 LOD chr pos Sugar chr pos Fructose chr
pos TA -1.5 11 39.91 3.16 11 38.91 3.79 11 51.91 16.79 peak 11
59.91 4.75 11 57.91 5.40 11 57.91 18.37 +1.5 11 80.91 3.20 11 71.91
3.87 11 65.91 16.54 LOD chr pos TA7 -1.5 11 54.24 8.36 peak 11
59.31 10.41 +1.5 11 66.91 8.87
TABLE-US-00023 TABLE 22 SZZ11 1.5 LOD QTL confidence interval
Almeria, ES, winter 2010 LOD chr pos TA chr pos TA7 -1.5 11 35.91
2.62 11 35.91 2.99 peak 11 60.91 4.29 11 59.91 4.59 +1.5 11 70.91
2.65 11 70.91 3.00
TABLE-US-00024 TABLE 23 SZZ11 1.5 LOD QTL confidence interval
Felda, FL, winter 2010 LOD chr pos TA chr pos TA7 chr pos Skin -1.5
11 50.91 6.07 11 50.91 6.12 11 42.91 1.94 peak 11 66.91 7.66 11
65.91 7.69 11 50.61 3.53 +1.5 11 73.41 5.98 11 73.41 5.94 11 70.91
1.89
Example 9. Identification of Further Additional Flavor and Taste
Quantitative Trait Loci
[0552] To identify genomic regions that control variation in total
acids and aroma the SHY2761 line is chosen for its acidity and
aroma compared to modern commercial bell peppers. Bi-parental
F.sub.2:F.sub.3 and double haploid mapping populations are used for
QTL identification. The SHY2761 parental line is crossed to
SIT-27-500 for linkage mapping. Double haploid lines are generated
from the F.sub.1 plants of these crosses (DH.sub.1 generation).
DH.sub.1 lines are sown in Felda, Fla. in a randomized complete
block design with three replicate plots per DH.sub.1 line for
phenotypic data collection. A subset of each DH.sub.1 line is
sampled for genotyping and linkage mapping. Genotyping for DH.sub.1
lines is performed using over 130 markers from a pepper Taqman
library to minimize gaps greater than 20 cM.
[0553] Phenotypic data is collected by pooling fruits from multiple
plants per plot per sample. Fruits are assessed for total sugars
(fructose+sucrose), fructose, glucose, titratable acidity (TA), and
a second measure of titratable acidity (TA7) in a vegetable quality
lab in Woodland, Calif. (Table 24). Plants at the Felda, Fla.
location are also measured for perceived sweetness (sugartaste),
acidity (acidtaste), and skin thickness (skin) by an individual
tester on a 1-9 scale where 1 corresponds to the sweetest, highest
acid, thinnest skin, and most favorable texture, respectively and 9
corresponds to the least sweet, lowest acid, thickest skin, and
least pleasant texture (Table 24). To describe the estimated
properties of QTLs, tables are presented with data on the estimated
QTL effect size (i.e. how much the QTL is expected to influence the
trait) and the proportion of total phenotypic variation in the
population explained by a QTL. The effect size is one half of the
difference between the two contrasting homozygous QTL genotypes.
The proportion of phenotypic variation describes how much of the
variation present in the experiment is explained by a single
QTL.
[0554] The SHY1 interval is identified from the SHY2761 parental
line with the rqtl package implemented in the R platform. To
describe the genetic location of QTLs, the LOD profile is used to
determine the statistical significance of a QTL result. The
significance of a LOD score is determined from 1,000 permutations
of the data. Because QTL peak estimates are not perfect, tables are
provided with the confidence interval for each QTL. The confidence
intervals are estimated using a 1.5 LOD interval calculated by
moving down the LOD profile from the peak 1.5 `steps` (-1.5) and up
from the peak 1.5 steps (+1.5) to identify the chromosome positions
(Table 25).
[0555] One to two QTLs affecting aroma and longipinene were
identified on chromosome 1 in Felda, Fla. field trials (Table 24).
The region separating these QTLs has no genetic markers. Therefore,
it is not clear if these two peaks are a consequence of this
marker-gap or represent two true QTLs. The SHY1 aroma QTL is
detected within the interval of 38-130 cM on chromosome 1. The
chromosome 1 QTL confidence interval for the Felda, Fla. population
is shown in Table 25. Within this same population, composite
interval mapping detected a QTL for aroma and longipinene on
chromosome 12 from 95-106 cM (longipinene) and 97-106 cM (aroma)
that was not detected by interval mapping.
TABLE-US-00025 TABLE 24 SHY1 phenotype parameter estimates from
SIT-27-500/SHY-27-61 DH.sub.1 population Felda, FL; Winter 2010
Position % of Size of Name Trait Chr in cM variance Effect
SHY1-sugar Fructose 1 44.8 19.9 -0.19 SHY1-thickness Skin 1 72.8
39.6 -1.1 SHY1-aroma Aroma 1 113.6 25 0.82 SHY1-aroma Longipinene 1
56.4 21.8 -10496
TABLE-US-00026 TABLE 25 SHY1 1.5 LOD QTL confidence interval Felda,
FL, winter 2010 LOD chr pos Fructose chr pos Skin chr pos Aroma
-1.5 1 28.78 1.85 1 45.78 5.20 1 38.78 1.74 peak 1 44.78 3.43 1
72.78 6.73 1 113.62 3.80 +1.5 1 115.78 1.92 1 91.78 5.22 1 129.78
1.80 LOD chr pos Longipinene -1.5 1 29.78 1.15 peak 1 56.37 3.23
+1.5 1 130.78 1.15
Example 10. Development of Additional C. annuum Plants Having
Increased Sugars and Acids Via Genetic Introgression
[0556] Additional C. annuum plants having increased sugars and
acids are produced by crossing a C. annuum parent plant with a
donor having one or more QTLs for increased sugars, acids, or both
(Table 26). For Example, the donor parent can be SMO-28-1234,
SZZ-8T10901, SHY27-61, a progeny thereof, or any other plant with
high levels of sugars or acids. An F.sub.1 hybrid plant from this
cross is either self-pollinated (to generate a segregating F.sub.2)
or is crossed again to the C. annuum parent plant (to generate a
segregating BC.sub.1F.sub.1 population or a segregating modified
BC.sub.1F.sub.1 population). The segregating populations are grown
and selected for fertility and checked for the desired traits from
the donor parent. Plants are tested for their fruit sweetness
(e.g., total sugar content) and sweet individuals are selected.
Selected individual plants are then self-pollinated and the
resulting families (F.sub.2 or BC.sub.1F.sub.2 or modified
BC.sub.1F.sub.2) are planted and individual plants are again
selected for self-pollination. This process is repeated several
times until fixed progeny lines are created which comprise
essentially all the desired traits of C. annuum parents (e.g.,
sweetness or acidity).
TABLE-US-00027 TABLE 26 Summary of Sugar, Acid, and Aroma QTLs
presented Underlying Flanking Marker QTL Chromosome Interval Trait
SEQ ID NO SMO2 2 65-113 cM total sugar SEQ ID NO: 1-2 content SMO3
3 143-184 cM total sugar SEQ ID NO: 3-4 content SMO8 8 37-63 cM
total sugar SEQ ID NO: 5-6 content SZZ3 3 63-118 cM total sugar SEQ
ID NO: 7-8 content SZZ4 4 84-107 cM total sugar SEQ ID NO: 9-10
content SZZ11 11 50-73 cM total acidity SEQ ID NO: 11-12 content
SHY1 1 38-130 cM total aroma SEQ ID NO: 13-14 SHY12 12 95-106 cM
aroma- SEQ ID NO: 15-16 longipinene
[0557] Additional C. annuum plants having increased sugars, acids,
C. chinense aroma, or a combination of these traits are produced by
crossing a C. annuum parent plant introgressed with the desired
sugar or acid QTLs described above with a donor parent having a C.
chinense specific aroma profile. For example, the donor parent can
be a C. chinense plant; line ZSP8T14-6274, deposited at ATCC under
accession no. PTA-122300; or SVPS2625, deposited at ATCC under
accession no. PTA-122296. The donor plant can also be any other C.
chinense with high levels of one or more aroma molecules identified
herein (e.g., .alpha.-Cubebene, .delta.-Cadinene, and
1,4-Cadinadiene). Breeding with a C. chinense donor parent is
described in example 5.
[0558] As various modifications could be made in the constructions
and methods herein described and illustrated without departing from
the scope of the disclosure, it is intended that the foregoing
description shall be interpreted as illustrative rather than
limiting. The breadth and scope of the present disclosure should
not be limited by any of the above-described exemplary embodiments,
but should be defined only in accordance with the following claims
appended hereto and their equivalents. All patent and non-patent
documents, publications, and references cited in this specification
are incorporated herein by reference in their entireties.
Deposit Information
[0559] Applicant has made a deposit of at least 2500 seeds for each
of pepper lines SVPS2625, SMO8T14-6275, and ZSP8T14-6274 disclosed
herein with the American Type Culture Collection (ATCC), 10801
University Boulevard, Manassas, Va. 20110-2209 USA. The deposit
accession numbers for lines SVPS2625, SMO8T14-6275, and
ZSP8T14-6274 are ATCC Accession Nos. PTA-122296, PTA-122298, and
PTA-122300, respectively. The date of deposit was Jul. 10, 2015. A
deposit of pepper hybrid PS09954859 and inbred parent lines SMR
99-1275 and SMY 99-1322 was also made with the ATCC on Dec. 1,
2010, under ATCC Accession Nos. PTA-11514, PTA-11520, and
PTA-11517, respectively. Access to the deposits will be available
during the pendency of the application to the Commissioner of
Patents and Trademarks and persons determined by the Commissioner
to be entitled thereto upon request. The deposits will be
maintained for a period of 30 years, or 5 years after the most
recent request, or for the enforceable life of the patent,
whichever is longer, and will be replaced if they become nonviable
during that period. Applicant does not waive any infringement of
rights granted under this patent or under the Plant Variety
Protection Act (7 U.S.C. 2321 et seq.).
Sequence CWU 1
1
161692DNACapsicum 1gatagtgaaa tagtctttgt tcagttatta tgcgctaaaa
gaagaaacat agtcccccaa 60tcaatgtttg ttaggctggt ttaaggatcc gtaagacata
gaagagcttg gaacaattca 120caacatgtta aatcagaata agtaaatacg
gatagtggaa ttgaaaatta tgataaagta 180aaatagatat agtgattaca
atgtaaatca aatgcaaagt tatgctgact tatttcccta 240tactggattt
cttgtttcct attcctaagc atggcaaatc attcatagcg cttggaaata
300tttaacatat caattagtca acactatcta ttgtttagtg tcgccctatt
caagaaggag 360cccgtgggtg aagaggcgca caccgtagtg ccttggtttc
tgcactgaag tgccacctaa 420gcaaggcaaa gcgccatccg ctttgcacct
agttacaggg gataagcgtt cctcaactga 480gcctttaaca acagtgccat
caataagttt aagttaataa tacggcaaat taatgaaaca 540gaaatctact
tcctttaaag aagttacaga gcagaaaatg acaccttaaa caaaccagca
600tttgaatatg gggggttcta ttcagttgat ccggatattc tttgctaata
ttatctccaa 660gtctgacatt tactacgaat taatccatga at
6922394DNACapsicum 2gcaaaagctt aataccatgt gtctccagca ggatgcagat
acagacaact gctgatacta 60ctgatctgag ttattggttg aactggaggt tcttgttttg
tgcaatatgg gtcttgacac 120caacagttgc agcagtgatc attctatgga
agtatgaacg atcagttaat atcatacagg 180aatctgatag taggggggat
tgccagaaaa gttctttgct tttgtatttt gacaaagctt 240ggagaccatg
tgtgaaaaga ataaacccaa tttgtttggc ggctttccga gtttctgcac
300ttgctttact cacactagtg attgtctctg attttgttgt tcatggaggt
gacatatttt 360tctattatac tcagtaagta ttccttttat aatg
3943607DNACapsicummisc_feature(280)..(280)n is a, c, g, or
tmisc_feature(431)..(431)n is a, c, g, or
tmisc_feature(447)..(447)n is a, c, g, or
tmisc_feature(461)..(461)n is a, c, g, or
tmisc_feature(498)..(498)n is a, c, g, or
tmisc_feature(513)..(513)n is a, c, g, or t 3tgtgcacttt ggcagttgag
gcccctcggt cttcataatt tctaacatct gagctgaatc 60taaacgagca taacgcaatg
gagcaactgc aaaacatctt agttgttcat aaccataggg 120ggaaacatat
attactagag aaacagcaaa ctgttttact caggccacta attatagtcc
180ttaccttcat gtatggcact ggtgcatctc tgagacctaa aatgttggcc
aaatttgaaa 240caattagaga tgttcaaaga gttgggtaaa atccaagcgn
aaccacgggg gacttacacg 300tagcataaac agtggactag ctcctgtaca
ttatcggacg agcagccaat ctcatccaac 360agaacaaagt agtgatttgg
cctgctagtt ccctgcattg tgttttctac attatgtagt 420gaaaaaaaga
ngaagataaa cgaaagngtt cctaagagga ntatgccagt gctagcacaa
480agtcatatca aactttantg atcattctac aanagttaca tgcgtagctt
cagagtcttc 540atcagaaaag gtttgctagt aaagtactca caactcttgc
cgcatgtgca cacatgtaga 600agttgtt
60741155DNACapsicummisc_feature(244)..(244)n is a, c, g, or
tmisc_feature(788)..(788)n is a, c, g, or t 4aataaggagc attgaaaatt
gctgctctca aagctcctgg ttatggagat cgaaaaaggc 60aatatcttga tgacattgct
gtccttactg gaggtttgtc cataattaag ttatctcagt 120gttcaatgaa
agctatgttt acttgtatag taattttatt tttgcaatat gcattcaggt
180actgtcatcc gagaggaagt tggtttatac ctggagaagg ctggaagtga
ggtcttaata 240tttncccttt ttcctttttt tttttttttt tttttttttc
tccctccctt tcctctttct 300tcctttcccc ccttttctcc ccttctttct
tttttctttt ttccttccct cccctccctc 360ctcccctccc ttcttcccct
tccctttctc cccccctttt tttcttttcc tctttttttt 420ttcccccccc
cctccttttc ccccccctcc cttctcctct ctcccctctc cttcctcccc
480ccttccccct cctttccctt ttcccccctt tccccctttt tttttctcct
ccccttcctt 540ctccctcccc ctccttcttc ttcctccctc ccccctcctc
cccctttttt tcccctcttc 600cccccctttt cttttccccc tcccccttcc
ctttttttcc ttccccctcc cccccccttc 660tcccccctcc ccttttcccc
ctttttctct ttttcctctt cccccccctc tcttcccttc 720ttcccccctt
ttccttttcc cctttttttt cccccttttt cttttctttc tcctcccctc
780tccctctntt cttccttctc tccccttcct tttcctttct ttttctttcc
cctctttctt 840tcccttccct ccccccccct ctttctcctt ccttttccct
cttccttctc tctccttctc 900ttcctcctct ccctcccctc cccctcttct
cccctttttc tccccccccc ttttctcccc 960cccctctcct ccctcctctt
cctctccctt cttcttctcc ccccctctcc cttttttttt 1020cttttccccc
tctttttctt cccttttccc tttctccccc cccccctctt cccctctccc
1080ccccctccct ccctcttctc ctttttcttt tctcccctct ctcccccctc
ccctctcttt 1140ccttttattc ctctc 115551416DNACapsicum 5gtgtcaaaat
ctccatttga gtcttgaaat agttggtctt tgcagagtca aaagttcact 60tcacaagctc
taatactctc tcgactactg taagcaatat taccaagctt cttgtaaaac
120atgaaactgt gaactatcct ctttatacgt agacgtgatg aactttctca
gactaataat 180tggtttaact ggtgttacta tgattagttc ccttctataa
tacatttcag gatcattcct 240aactgatcat tattttttca ggttgtccct
tcaatagcta ttgcttttgt gacatatgag 300caagtgaagg acctattggg
agttgagatc aggatatctg attgatgaag acaggctcca 360atgtttttcc
actgcatacg ataattttgt aggcaaggat agatagagtg ataataacat
420ggcgggttac cccgtctaaa cagagaattt acgattctgt tttagcttgt
gacagaaagt 480ttaggacttt ccagttgtgc ttataattcc ctatgtattt
gcttattcca tattgtcaat 540aagcaaaatt tacaacagtg taatgactaa
tttgaatgtc ttgctatggt tcaagtgaag 600aaaagctgat tgctggttgc
ctttgctaat cctggtctcc attgtacttg tggtcgaatt 660tttctggcgt
cttcaccagt tttcagctct ctttttttca ttggaaaaag ttgaacagtt
720taggtaagaa aaggggtact cactcatgat ctagtgtatt ctgtaatgcc
attttgtatg 780ttctgattaa tatgacgtga atcgtttctt ttaagaaaat
ttgggttgct atgatgtatg 840ctgtctactg aactgcttta tccgctggag
ggctaaacat tttactacct tgcaaagtag 900gtgagtgttc ggtatgtgta
atggatgaaa atcaatgggt ttgagctcga gaatgggcaa 960cacttgacca
gataaattta gactagttga atcaatgatt ttcggatacc agaagcaaaa
1020gaaaaaaaaa gagttggcat ttgcaaaaga gtgatcaatt ctacttctag
tactactctt 1080ttacatgtat gttttcttga aaggtttgaa tgctacacct
caacgatcta tctctcttgc 1140actcacactt gatgcgaaaa aacatactat
gaaaggtagc ttctatggcc ttgctggaaa 1200ccttcttttg gtacatttct
tgctgctagc tttgctggat gttgatttct tcacattttt 1260ctgcaatgcc
tgaatcaaaa gatgaatcat cttgctggta aaaggaaatg aacttttact
1320attaacacaa tcaattagta ttaattttgt gctcctgtcc aaaaaaagaa
tggcatgatg 1380tcaaaacatc taattttcta aggtcctaac tcgtac
141662246DNACapsicum 6ggagaacacc aagcatcagt aattttaaca aataatcgag
tctaaaatat ctggtaataa 60tgatcaagtc cataacacca ctaaaggagc tatgttgtac
ttaatagtag ttaaagtttg 120ggttcatctc acacaggatt gctaatttca
acagttttcc ttgttaatat aaaacctaag 180cgcctcagtc gacatccaac
cagccaaaag gtctgttcat gaatctagac tgcaacttaa 240aagtaactaa
cacatgagca acacaatctt acgctcgcac tcagatcaat caacatgaat
300gagcaatcaa caaacttaag gtactattcc tgtaaaaaaa cagaaataag
cactttttcg 360cttctctttt ttcccttgta aaatgtatac aggaatgcgg
acatatatca tcaatcacat 420gcaaccaatt tcttctttga tagaagttct
aaaatttgta ttttataaaa atgaaagtct 480tgcaccttaa tgatcttaaa
tcactccatt gcctgttaga tgaactccat tttcatgtta 540tgcagatctc
ccagacacat cacaaccatt gaaaaataat caagtttgag tactgtttaa
600acttggtaaa acatagaatg gagcatcagg atatgcacca tgaagtgttg
aagcctttct 660gattcaggtt caactttacg agatatttga ttctcatgat
gagtcctggc gttcatcatg 720tcgagggcct tttgatataa agaggccaaa
tgtggcgaga gcccatccaa gttactggag 780tctacatcaa gattggctcc
ctgttctgct tcccaagcct tgaatgcgag gagagttgaa 840cttagatcag
caagagggac agataactgg cgatggaata aattcctaat tcgctggaca
900tgcttctctc tcgactgcat tacaatgaag caaatatcaa cagaaaagat
caactatcaa 960atgtggaaaa gataaaaaca gatcaagtta tcattggaca
ttgtctaata tagaaacacc 1020aatgatacat caatcttctt tacacaaaaa
tacagaacag gctataattg agcgaagttt 1080tagatctaac attctttgat
cttcagcaca gatactaata gtatagacat cgttttgaaa 1140ccaaaacctc
tgacctaaaa ttcaagccac agctcaacta actgaaatta ctaaaaatca
1200atacgcagtt tcttgccatt aactcaacca aataattaaa tagaatattg
tgcaaagtgc 1260aaactcataa cacaagagag aacaactcaa tatgcgtttt
ccttttcatg gtcttcattc 1320aataagggaa gtaaggtttt cccatcattt
aagtttcata acatttcaat ctgctaccag 1380ttgttgaagt atgcttatct
aaaaaggaaa gaagtatgct tgtctaaaag gaactccaat 1440actaagtcat
atcagtctca tgacagtaaa agcaaccaaa gcatttgagg tacacctctt
1500cttcgtatct gttcctgatc tcttgacagt caaacaacga gacgaggtta
acccatttaa 1560gttcttttgg gaaggggtgg gagtgaaaga tgcttcagag
gaacaacaag aagatgcagc 1620agcaactttt taccatccca tgcttaaacc
caaggggtac gctaaaagcc agtaaagtct 1680cagaaaaagt gatttgagag
cactataaga tgaatagtca cgagctccgc aaagacacaa 1740ctcatgatat
agagtactac cttcccaggg aattatattt acgagaattg aacaaagaca
1800tgttacatta tgcctctaaa ttctctcatg cttccaggga agaaaacagg
gacaggaggt 1860gatatacaac ctaacggaaa aagatatatt ccacctttcc
atcatcctgg aggaatagga 1920cggcacaacc ttagaaaaac aagtgctaag
ctaaaaaaag aacggcatct agctctttga 1980caatacgaac aacaacatct
cagtctcaaa ctagatacag ctagctctta ctatatcttt 2040aaaaaggcta
aagtgacaac ggcatgcatc ctaggaaaga aagagaagca ccaagctctt
2100acattggcat cagtttcatc aatagtgagg aaaatagctt gttcaaattc
cctatataat 2160tcccatatcc tgctgccttc agcaacatgc aaaccagtag
caaccagggc acgctcaaag 2220agatttcttg cctttgaaat tccagc
22467725DNACapsicum 7ggctgtagtt ggagatatca tagtgacttg acttgcaaca
tatttcccat gtctcaactt 60accaagcatc caggccaaag aagcagggtc actagactat
tatcaaatta actctagttt 120ttctcagtat acaaagattt catcagagca
tctcctactt tcaataggtc atgaactaga 180ttagagttgc tctcgacgaa
tccataagtg acgccatttt tcatcgggcc gtataacgac 240caaagatctt
gaccaaccga tccaaacaac tcaaagatta ataaatgtat cgttaatctc
300ttcatactcg aggtaccatt tgtacaaaca aaaatcccct tcagtcacgt
ggtttcttct 360tcatacaaat agacaatagt agttataatg caacattgaa
gtacattttg tgctaagatc 420ccattatcct gaaccgatct taccagggca
tttgctcatt gccagagaga cgtagcaata 480aacgtacata caccaacaaa
gtaaacaacc cagctcatgt tatctataaa taaccttatg 540acacaaaaca
tattcatcat catcggctat ttacccacag aattaatatt ctcaaaacag
600attaaagtca tcactgggta tcaccagtca ttcccgccaa agcctaatag
tcgtttgtta 660agcatattga ttacttttca caaccaccta atactacaca
ttcaattaac atcagaaata 720caacc 7258455DNACapsicum 8gcgttttcgc
agtttgcagt tgcattatgc gtattctccc aaccttccca agagcgaact 60tccaccttca
gctttaaaga aaatcctttg ctgtttttat cagaagctgc aataagttgc
120ataagctcta gtatgtaaac tctaacatgg ttggggagcc gaaagttttc
agagaactct 180tccaatctct cccaaactgc ctgcctgaca ctctgcagat
gatccaagtc accagctaat 240ttgctcaaag atgacaagta atagtgtaga
cactgatgct cacgggattc gtcagtcaac 300tcctgcaaga tggtcttcaa
tatactaagg tagagatctc gaagattctg gatgctgaaa 360gcttcttttt
taaagtccgc caccgatcca gcttcacgct ggaaatgagc catgacttca
420tcatatatat ctgcaaccgc tcgaaagcaa caacc 4559357DNACapsicum
9gaatatttta aaaatcaagc aagaactcaa ctcaagtatg ttcttccggg gttggatatt
60tcacatgctt gaccagttga catgttgaat tctgtggcat tgtcatctga tttaataaac
120aatatgggtt tctcttctcc atgcccactc ttttgcattt tgtgtacttc
aattcaagga 180tccatctttt gcatcttcag gtcaagacat aggcgggagt
gcaaagttac cactaacacc 240aacaccacct ccgagtggtt tgacatcaaa
tccgttgcca acaactgaac aacaaagaag 300cccgcatgga catgcagttg
caccggccat tgctggtaaa gctaccccta aaccatc
35710515DNACapsicummisc_feature(320)..(320)n is a, c, g, or
tmisc_feature(349)..(349)n is a, c, g, or t 10gcagctgaat tctaaaaaag
agctgagctt gggacaattc aaagagacaa gatcgttaag 60gaagtccaga gcagctgaat
tcacatattt tacctgtgga gcatgtccat tagagtctga 120gacagatgta
agattactta gattatggat ttttttttgt tgctcaatga tgcattgctc
180tcgctctttt tgtgatcttc tctcctcctc caactccatg gcaagcttct
ctcgctctaa 240ttcgtactaa taaatgaaga gtaagcagtc ataagccatg
tcaacacaag ataagatcag 300tcttcttctc ttaacaaaan caagaaaaat
agtaatgata attaccgtna gtaggtcatt 360tctgagtttt agtatctctt
gctcaagcac ctcggaatgt gatccctgtg taggaggttt 420catcccatat
aagatatttt tatgcaactt atgaatatat aaatcctgag agacaagaca
480ctcacctgaa gtttcatgcg tagttccctc aattt 515111905DNACapsicum
11gaagcaatag aaaatattgt ctctttagca aaaaagttac ggggcagatc tggtaaggaa
60attcatgcta ctctaatagt ggttttaata tcacgttcaa accacggatc ccatttaagt
120gttcgcatct gtcaatacct ctctctagat ttaatataaa aagttgttgt
atatgaaaca 180tctgaacata attttcaaga ctggttaatc tgattaaaat
atgccttgtg tccaaaactc 240ctatagcaca ttcatgtttg ataccacaat
ggccaggaat attcttcctt aacgtaatta 300tatcaggttt tcctgcatga
taagggactt gacttctgag gacttcgatg tgatatttgg 360ctcccctcgc
agcataacaa tcgattttgg gggaactcta ggcgttgagt ttgtttagat
420tttaggtttt agtacagcag cgtattgatt ttgattgtca ttggtatttg
ggtacttatt 480aacatgggaa gatccgttga gttactttgt gatggaaggg
atgggtattc tgccgtggga 540gcaacccact ggtggtgttt accgtcatcg
ccgccaatag ttttctaagg ttttatggta 600ttattattga ttccgttcaa
gtaacactca caatagtcat attgctactt aaaaagttaa 660atagtctcta
tgtactaatt tatatgatgc agctaaattc aaaattcaaa tagagatttt
720tgaaatgggg cgaaggaagt gttgaattta tgttcatttc atcaaaacct
ctaatttttt 780gttttcaaaa ttactaaaat ccacataaag aaaagatgtt
ttgactctag tattttgaac 840atttcacata aagttggata attaactggc
tctcgaattg taatttgtta ttagttgcca 900gcaaacaccc tcttccttta
ggacttccct tttcctccta gcagccactg gtaaaaagtt 960attcatatta
ggtgtttcaa tcaagcaaat ggatacatga tatgatattt gtttatattt
1020acataattat tacttaatca cagttaattt atgcattttt tactttatta
attaatacaa 1080acatcgatat gactaaatat ttatcgatac cacacagtca
tacattttat cagtttgaaa 1140atttactcaa attaataata ggccaaaagg
aaaaggtaaa attgatggat tttgaggcaa 1200atggctaggg cctcttctta
atataccttg attgtactat tcaactctgt gtaacttttt 1260tgaaagaaaa
aaaacggtaa gtttccctat gaattcaata ttgttttttt aaaagaaata
1320cattatattt cccaaaaaaa atcaggagga aaataaatta ctaaaatagt
tctttcataa 1380ttaatttgaa gttcatatgc atataaccag agggaggaga
ggggggaggg gataggttgg 1440tcgagggggc tcatcagaac cttcttagat
agaaaattac cttgtttgta tatgtttaaa 1500ttatttttta tgtgaaactc
ctttgataag tatcagacat aataagcttc ttaatttgtt 1560caataactat
atatattata agtcacatag gctcttcaat cagtacataa taacattata
1620tatatgattt gaataccatc caaatcaaac tatagattat aacataattc
ttattaagca 1680attaatacac ttttaaggat aatcattgtg cctaggtgat
ggtccagtag aaattggaac 1740tccttttggt ggcaatggaa ccaattattc
ttgtgaacga aattctttct ttcaatcgtt 1800gtatttagaa attggttctg
agaattttga gattggttca gaaaatttcg aaactggctc 1860tgaaaatttg
aaactaggtt caaaaatatt tcgaaacttc tgaga 1905122405DNACapsicum
12gagttactat ttcttcccta aatagtcaaa atgatatctt tgggatgttc ggttattcct
60gtatgtgtta cttaaaagca ccttaagttt gccattaatt tctcacaact tcttggagaa
120tattgtaaat tgtttcatat gattttaggt gttgatggaa gcattattat
tgcagatggg 180aaaccgcgac cttgttttca ctgctttttg gagggatttg
cttcgctgga aggttcacat 240tgttttctca cttttgctga ttcatgttcc
gcttttgtta ttatcatgat tagtaacccc 300ttgttaagaa ccaatgacta
caagtatgct ttgcttttaa aaatagtttg atccatgaag 360gtttagtgca
tcttgtaaga gaaggttgag ggaaattgtc tacaaaatag atgctctgac
420atgcttccat attcatatga agaatggtct aatttcaaat tgaactcttt
gctctgtgac 480cttcctctgt gctttatggc tccttaattc tttgttacat
ctgaaaaact atactcatca 540aaagtaaaac gaaatataaa catctgaaga
aaaacttgtt ataaccccca actttttctt 600ccctattcaa agataggatg
ccatctgctt cactaaatac tcacattgtg tgtgaaattc 660atttcatgta
aatggatttg ctcttttcaa ctgttggtaa agtaaaacgt tgctaaataa
720tggcaaagca atatcttgtg ccctggatat tttatactcc acagttaacc
taatcgttgt 780tctcttccac atggaagaac aattcctttc ctttttgaaa
aacaatacat gattttcatc 840tttagaaaat tgcgaaagta acacattttc
taattttgtt tagccagcga actattggag 900cagttaacgc tgtccttgtg
tttggaatca tcgcttcttt cacagctctt gtggtaattt 960ccattcttga
ttcagtagaa aaaaactgct attattatct ttaggataat gtagctgttt
1020tacattttac ttttagggtg cagtattgat attcatcttt gtggacagta
attttcgtat 1080ttcactgaaa ttggtgccat cgtatctgac tcatgccaat
atatgcctca attggtgaca 1140ccaacatagg atagctgccg cctgaattat
ttagatggcc tggagtctgg ggacttttaa 1200tgttagatgt tgcatgggat
ggtaaatgct tcctgccaac aggtttggcc agtggcgggt 1260ccaaaaatta
taatggtaga tatgaaaaaa gtgaattact tcacatagta ctacatgcat
1320attagattgg tagatgatta tttcattcat ttgaactttt taatgctcgg
agatgcttag 1380ctttgtttct tgttatttat gccatagaca tgatatttct
tactttccat gagtgcagac 1440cttttctgat gaatttcttc gaaaggacat
taatagaatt tctatttgag atattggatg 1500atttagtaaa tggttcgtcc
aacaggtttg cttaatggaa gttcctgaag ctacatctgg 1560tgttacatgt
aaaagaagtg ataacttcaa atgggccact taataggaac gttatgttaa
1620taatggttat tggctaagat gagcttctta atagatatgg tctgctttat
ttgtcctttg 1680tacgtgctat atgcatgaca aaaaggattc aatacattcc
catacatctc gtgaagacca 1740acaaaccaag gttctaagta atgcacatta
aaagaatgtt cattgagata cttgcatgag 1800atggtaaatg ttcatgtcaa
caggttttcc taatggcagg tcgtgaggca tatgagtatt 1860acattggtag
atgattattg gtgcatttgc ggttcttaat gcttatactt gatcccagtt
1920atttgatgtt tatatacgct atacagataa aggatttaat ataatctcat
acatcatgcc 1980aagaccagaa aacaagtgat gtttcaaata acaaatttaa
atattcaatg gcaaatttgt 2040gccaatatta atctatatct tggaaacaca
tgcgtcaagc ttcatgtaat gcactggaaa 2100tggttgagag agaaatctta
atatctcatc ttttacgctt aaggttgttg ccagtggaga 2160cttgcattgg
gaggctcttc tgaaagctaa ttttgaagct gttactcaaa gtatacccat
2220aatcgcactt tcttttgttt atcaggtaac aagtgttcct gatcaactca
tatagtcgcc 2280ctgggattgt gcatctaagt taatcacttg cttttcaatc
gcagaatgtc gtacctgttc 2340tctgcacaaa tcttgaaggg gacctgtcaa
aagtaaggtg attatcagta tcttgcttgc 2400attac
240513696DNACapsicummisc_feature(80)..(80)n is a, c, g, or
tmisc_feature(260)..(260)n is a, c, g, or
tmisc_feature(283)..(283)n is a, c, g, or
tmisc_feature(341)..(341)n is a, c, g, or
tmisc_feature(479)..(479)n is a, c, g, or
tmisc_feature(557)..(557)n is a, c, g, or t 13tttctaagcc agagaatgtc
ctcttgattg ctccttcact ctcattccac tctacaaggt 60gggattcacc ttctttactn
gtaccacaag agaaaagtct gaaagaagat cagaaaagtt 120aaaaatgact
agatataaaa tcaaatgcaa acaagtttgt attgcttctc aaaaaagttc
180gtcttctttc agacatcaat cataaaatcg cgtaggcctc tttacaattc
atttttctac 240cttcaataag caccattaan tcattcacta ttgctgccat
atnttttatc ctggcacctt 300ttgagtataa aactcaccct aatgagtcct
cctccaacgg ngtcatattt tgccttaggg 360agtgtgcata actttagccc
ttttttattt tttggccaac ggtatgtttg atagagtgcc 420aaagaaggtt
gttagagaga gcacactttt atttatttaa aaatgttttt caacacacna
480tctcaagtcc gtcttaatct tttttcacga atcaagcgca tggaaattct
ctttgatagt 540gagtgactca agacctntgc taccatgttg aagggtatga
tcatctcatc caacaactta 600agctagacag aaacttttat atatttaatc
atgtcgttca acaaagaaac cgaaattaga 660gctaatagga ggaagaccac
tgataaaatc acaaac 696141314DNACapsicum 14aactgtaata tcattctcct
ttatggtatc tttttcatca tctacaattt gagttggtga 60aactggcaat gatagtactg
attcaatttc tttatcttgc gatgatgttt gtattattaa 120tggagttggt
ggtggagatg gtaatgacaa
ctttggtagt gacgttattg gtggttccgg 180ggctggtagt gttactatat
ctgtttgcat ttctgatgct ctctccctca aaggcttccc 240ctgatgatct
aattgtggaa ttctacgttt gctttttctt gcacaatttc taactgcttt
300cgaatgtata ccaagttctt ctccaatcgt agttctgatt actcttttat
ctcttgtctc 360agatgaagaa ggatcctgga tgttcttgtt cccagaagaa
acattcaagt aggacgatgt 420tctttcttct ttgcctttgc gtacaaagta
agatgcactc cattcttgta cctatttgta 480acataatata attaagtagt
taaaagtgta ttctagatct ctttaataac ttacgctttt 540agatgaaatg
atcacaaact tcaataatac cttgatagct aaatttgata tagaaccgga
600acttgttgaa aacgctgata ggttgctatc ttttccatga gtattactaa
actctgctaa 660cagtttttgt cgattaatct ctttggacga atcagtaagt
cctttgtaat atggacttct 720tggagagtat ctcttttctt cttgaatcac
atttcttttg ttgaattcct tcacattatt 780agctgttaag ctcctcgagt
tctttgaaga cgaacttccc tttcttccct catcgacttt 840tttagcccat
gttggtttca ctgagtcagc tgaatatgtt gatctgactg tagggggtat
900tatgagtgaa ttttgagttt ctgatatcct gcagggaaca gattttgaac
gtgggatcga 960cattgaagag ctgagattga aagttgctgt tttttttttt
ttccttttgt aggatattat 1020aggatcaata gagaactatg gaatttaaca
caattaatat gtttaatgtg aatgggaaaa 1080aaagagtttc ggttatttta
ccattttgtt gttgttgttg ttgttactgt ttccatatct 1140gttaccatgt
ttcttcactg tcgcttttcc tgtttgtact ttctttgatt tgctctactc
1200gagtcgtggg tcctccaaga acagtctgta aggtctgcgt acgctctacc
ttctcccgac 1260ctcacttgtg aaatttcatc gaatgtgttg ttgtaatgtg
aatggaaaag aagc 131415747DNACapsicum 15gaagaagctg ccagagctta
tgatgttgaa gctcggagga tcagaggcaa taaagctaag 60gtaaactttc ctgatgaagc
tccagttcct gccccaaggc aagctgctaa ggtaaatcct 120gagaaggttc
tttctgatga gagttccaac ccagttctgt ccgacaccgc gttaatgaat
180ttgactgatg gatgttgcaa caacctgggc ttttttgaag agaaaacgaa
aaaactgtat 240ggctataaag ttttgcgcac tactactgta aatatgggac
ctaactcata tgtccctcca 300cctgctgctg gtgtttactt caattctgac
caaggaagta actcatttga cccctccgac 360tttggttggg gagaaacgtg
ttccaggact ccagaaatat catctgttct gtcagctgct 420atagaatgtg
atgaaactca atttgttgaa ggtgccgatg tggaggtgaa cctaaaatct
480tgttccaaca atttggtgcc cgatgatgga aacactgcta acaagccacc
tgaagaattt 540ccagcttttg aatcccagat gaagttcttt cacaccccat
atccggaagc aaattgggat 600gtatcagttg atgctttcct taacgcagat
gcatctcagg gtggcgaaaa tgctgtggac 660ctttggtcca ttgatgaact
ttcttcttta atgagaggtg tctactaagc taagaatgcg 720ctatgtgaat
aaggcttcac atgagtt 74716833DNACapsicum 16gattggaaat atatgaagaa
ggggaattac agtggcaatt caagggaatc atatcaatcc 60ttcactaata atcaatctca
tggaagattt gatagatatg gacatcaggc tgctggtcaa 120tccaataatc
agttttcagc tcataacttt actgggagtc aatcaggaga ggatacaggt
180ttagcagttt catctagcag tcaagggagc aaggacatca acaatgctat
gatagccaag 240gcacaaggct tctcagatga agagtacgta cagacaaatc
atgtcattac taaataaaga 300tgcacatgat tccaaacagg tcaacatgtc
aggcatagtt acttgtttat caacacatgt 360tccacatgaa tgggtaatag
attctggtgc cacataccat gttgcagcac ataaaggtgt 420cttctcacac
tgtcataaga tatttacaca gaaaatgata tggtgaatct tcctactgga
480gccaaggcag atatctcaca tgttggagaa gcacagatta taaatgatga
aattgttaga 540gatgtgatgt ttgtccctga cttcaaagct caatctattg
tcagtgtcaa aaatgactaa 600agaactttca tgctttgtat cattctatcc
cgatttctgt gtatttcagg accttcacac 660tggcaaggtg aaggggattg
gtaaggagaa aagaggatta tacatcctta agaaggtctg 720tggattgaat
gatatacatg aaggaagttc tcaaaagttg ctggttgcag aagtagatat
780gcaagactgc aacttatgcc ataggaggct tggacaccct tcttcttagg ttc
833
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