# Patent

## Title
Pepper hybrid PX-11487057

## Abstract

The invention provides seed and plants of pepper hybrid PX-11487057 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of pepper hybrid PX-11487057 and the parent lines thereof, and to methods for producing a pepper plant produced by crossing such plants with themselves or with another pepper plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the fruit and gametes of such plants.

## Background
FIELD OF THE INVENTION
The present invention relates to the field of plant breeding and, more specifically, to the development of pepper hybrid PX-11487057 and the inbred pepper lines HSE-114-1108 and HJA-114-1078.
BACKGROUND OF THE INVENTION
The goal of vegetable breeding is to combine various desirable traits in a single variety/hybrid. Such desirable traits may include any trait deemed beneficial by a grower and/or consumer, including greater yield, resistance to insects or disease, tolerance to environmental stress, and nutritional value.
Breeding techniques take advantage of a plant's method of pollination. There are two general methods of pollination: a plant self-pollinates if pollen from one flower is transferred to the same or another flower of the same plant or plant variety. A plant cross-pollinates if pollen comes to it from a flower of a different plant variety.
Plants that have been self-pollinated and selected for type over many generations become homozygous at almost all gene loci and produce a uniform population of true breeding progeny, a homozygous plant. A cross between two such homozygous plants of different genotypes produces a uniform population of hybrid plants that are heterozygous for many gene loci. Conversely, a cross of two plants each heterozygous at a number of loci produces a population of hybrid plants that differ genetically and are not uniform. The resulting non-uniformity makes performance unpredictable.
The development of uniform varieties requires the development of homozygous inbred plants, the crossing of these inbred plants, and the evaluation of the crosses. Pedigree breeding and recurrent selection are examples of breeding methods that have been used to develop inbred plants from breeding populations. Those breeding methods combine the genetic backgrounds from two or more plants or various other broad-based sources into breeding pools from which new lines and hybrids derived therefrom are developed by selfing and selection of desired phenotypes. The new lines and hybrids are evaluated to determine which of those have commercial potential.
SUMMARY OF THE INVENTION
In one aspect, the present invention provides a pepper plant of the hybrid designated PX-11487057, the pepper line HSE-114-1108 or pepper line HJA-114-1078. Also provided are pepper plants having all the physiological and morphological characteristics of such a plant. Parts of these pepper plants are also provided, for example, including pollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.
In another aspect of the invention, a plant of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 comprising an added heritable trait is provided. The heritable trait may comprise a genetic locus that is, for example, a dominant or recessive allele. In one embodiment of the invention, a plant of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 is defined as comprising a single locus conversion. In specific embodiments of the invention, an added genetic locus confers one or more traits such as, for example, herbicide tolerance, insect resistance, disease resistance, and modified carbohydrate metabolism. In further embodiments, the trait may be conferred by a naturally occurring gene introduced into the genome of a line by backcrossing, a natural or induced mutation, or a transgene introduced through genetic transformation techniques into the plant or a progenitor of any previous generation thereof. When introduced through transformation, a genetic locus may comprise one or more genes integrated at a single chromosomal location.
The invention also concerns the seed of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078. The pepper seed of the invention may be provided as an essentially homogeneous population of pepper seed of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078. Essentially homogeneous populations of seed are generally free from substantial numbers of other seed. Therefore, seed of hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 may be defined as forming at least about 97% of the total seed, including at least about 98%, 99% or more of the seed. The seed population may be separately grown to provide an essentially homogeneous population of pepper plants designated PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078.
In yet another aspect of the invention, a tissue culture of regenerable cells of a pepper plant of hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 is provided. The tissue culture will preferably be capable of regenerating pepper plants capable of expressing all of the physiological and morphological characteristics of the starting plant, and of regenerating plants having substantially the same genotype as the starting plant. Examples of some of the physiological and morphological characteristics of the hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 include those traits set forth in the tables herein. The regenerable cells in such tissue cultures may be derived, for example, from embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistils, flowers, seed and stalks. Still further, the present invention provides pepper plants regenerated from a tissue culture of the invention, the plants having all the physiological and morphological characteristics of hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078.
In still yet another aspect of the invention, processes are provided for producing pepper seeds, plants and fruit, which processes generally comprise crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper line HSE-114-1108 or pepper line HJA-114-1078. These processes may be further exemplified as processes for preparing hybrid pepper seed or plants, wherein a first pepper plant is crossed with a second pepper plant of a different, distinct genotype to provide a hybrid that has, as one of its parents, a plant of pepper line HSE-114-1108 or pepper line HJA-114-1078. In these processes, crossing will result in the production of seed. The seed production occurs regardless of whether the seed is collected or not.
In one embodiment of the invention, the first step in “crossing” comprises planting seeds of a first and second parent pepper plant, often in proximity so that pollination will occur for example, mediated by insect vectors. Alternatively, pollen can be transferred manually. Where the plant is self-pollinated, pollination may occur without the need for direct human intervention other than plant cultivation.
A second step may comprise cultivating or growing the seeds of first and second parent pepper plants into plants that bear flowers. A third step may comprise preventing self-pollination of the plants, such as by emasculating the flowers (i.e., killing or removing the pollen).
A fourth step for a hybrid cross may comprise cross-pollination between the first and second parent pepper plants. Yet another step comprises harvesting the seeds from at least one of the parent pepper plants. The harvested seed can be grown to produce a pepper plant or hybrid pepper plant.
The present invention also provides the pepper seeds and plants produced by a process that comprises crossing a first parent pepper plant with a second parent pepper plant, wherein at least one of the first or second parent pepper plants is a plant of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078. In one embodiment of the invention, pepper seed and plants produced by the process are first generation (F1) hybrid pepper seed and plants produced by crossing a plant in accordance with the invention with another, distinct plant. The present invention further contemplates plant parts of such an F1 hybrid pepper plant, and methods of use thereof. Therefore, certain exemplary embodiments of the invention provide an F1 hybrid pepper plant and seed thereof.
In still yet another aspect, the present invention provides a method of producing a plant derived from hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078, the method comprising the steps of: (a) preparing a progeny plant derived from hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078, wherein said preparing comprises crossing a plant of the hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 with a second plant; and (b) crossing the progeny plant with itself or a second plant to produce a seed of a progeny plant of a subsequent generation. In further embodiments, the method may additionally comprise: (c) growing a progeny plant of a subsequent generation from said seed of a progeny plant of a subsequent generation and crossing the progeny plant of a subsequent generation with itself or a second plant; and repeating the steps for an additional 3-10 generations to produce a plant derived from hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078. The plant derived from hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 may be an inbred line, and the aforementioned repeated crossing steps may be defined as comprising sufficient inbreeding to produce the inbred line. In the method, it may be desirable to select particular plants resulting from step (c) for continued crossing according to steps (b) and (c). By selecting plants having one or more desirable traits, a plant derived from hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 is obtained which possesses some of the desirable traits of the line/hybrid as well as potentially other selected traits.
In certain embodiments, the present invention provides a method of producing food or feed comprising: (a) obtaining a plant of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078, wherein the plant has been cultivated to maturity, and (b) collecting at least one pepper from the plant.
In still yet another aspect of the invention, the genetic complement of pepper hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 is provided. The phrase “genetic complement” is used to refer to the aggregate of nucleotide sequences, the expression of which sequences defines the phenotype of, in the present case, a pepper plant, or a cell or tissue of that plant. A genetic complement thus represents the genetic makeup of a cell, tissue or plant, and a hybrid genetic complement represents the genetic make up of a hybrid cell, tissue or plant. The invention thus provides pepper plant cells that have a genetic complement in accordance with the pepper plant cells disclosed herein, and seeds and plants containing such cells.
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. It is understood that hybrid PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 could be identified by any of the many well known techniques such as, for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), 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) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).
In still yet another aspect, the present invention provides hybrid genetic complements, as represented by pepper plant cells, tissues, plants, and seeds, formed by the combination of a haploid genetic complement of a pepper plant of the invention with a haploid genetic complement of a second pepper plant, preferably, another, distinct pepper plant. In another aspect, the present invention provides a pepper plant regenerated from a tissue culture that comprises a hybrid genetic complement of this invention.
Any embodiment discussed herein with respect to one aspect of the invention applies to other aspects of the invention as well, unless specifically noted.
The term “about” is used to indicate that a value includes the standard deviation of the mean for the device or method being employed to determine the value. The use of the term “or” in the claims is used to mean “and/or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. When used in conjunction with the word “comprising” or other open language in the claims, the words “a” and “an” denote “one or more,” unless specifically noted otherwise. The terms “comprise,” “have” and “include” are open-ended linking verbs. Any forms or tenses of one or more of these verbs, such as “comprises,” “comprising,” “has,” “having,” “includes” and “including,” are also open-ended. For example, any method that “comprises,” “has” or “includes” one or more steps is not limited to possessing only those one or more steps and also covers other unlisted steps. Similarly, any plant that “comprises,” “has” or “includes” one or more traits is not limited to possessing only those one or more traits and covers other unlisted traits.
Other objects, features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and any specific examples provided, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.


DETAILED DESCRIPTION OF THE INVENTION
The invention provides methods and compositions relating to plants, seeds and derivatives of pepper hybrid PX-11487057, pepper line HSE-114-1108 and pepper line HJA-114-1078.
Pepper hybrid PX-11487057, also known as “PX 11487057”, is a typical fresh market serrano fruit-type, with a cylindrical shape, and typical smell and flavor of a serrano. It has higher yield, mainly due to more fruits per plant, than the standard O.P. variety ‘Hidalgo’. It has higher pungency (2,839 vs 1,121 Scovilles) compared to the standard hybrid ‘Don Vicente’. This hybrid is grown primarily in the U.S. and Mexico for harvest as fresh green fruits. It exhibits excellent fruit quality due to its uniform fruit size, dark green color, lack of surface cracking, and cylindrical shape. The fruits of hybrid PX-11487057 average 9.7 cm long×1.8 cm wide, and weigh an average of 16 g. Immature color is a medium dark green, and fruits mature to red. Fruit pungency was measured by HPLC using fruit grown in Woodland during the summer 2012 growing season at 2,839 Scoville Heat Units (fresh basis). It has somewhat lower yield (765 g/plant, compared to the hybrid ‘Don Vicente’ with 829 g/plant) mainly due to lighter fruits (16.4 vs 18.0 g). Fruit number per plant was similar (46.6 vs. 46.2) Plant height averaged 120 cm, with a semi-spreading plant habit. It matures to green in approximately 85 days after transplanting, compared to 89 days for the standard O.P. variety ‘Hidalgo’. PX-11487057 is resistant to bacterial spot races 1, 2, and 3, (carries Bs2 gene) while ‘Hidalgo’ is susceptible (does not carry Bs2 gene). PX-11487057 is well-adapted to many pepper growing areas of Mexico and the U.S., with excellent fruit quality.
A. Origin and Breeding History of Pepper Hybrid PX-11487057
The parents of hybrid PX-11487057 are HSE-114-1108 and HJA-114-1078. These parents were created as follows:
The hybrid was created by crossing HSE-114-1108 to HJA-114-1078 in the spring of Year 9 crossing block (sown January 6 of Year 9). Seeds were harvested from the female on or around June 25 of Year 9. Hybrid was first tested in Seminis Woodland field during the summer of Year 10.
Parent line HSE-114-1108, also known as “HSE 114-1108”, was developed by backcrossing 2 times the Bs2 gene from Ixtapa into the O.P. serrano cultivar “Hidalgo”. Twenty BC2F1 plants were selfed for two generations, and the resulting BC2F3 families tested with Xanthomonas campestris pv. vesicatoria to determine if they were fixed or not. One plant, designated 0419SEM-568, was selected as being pure and designated HSE-114-1108. Seed of HSE-114-1108 was given to Seminis' Foundation Seed (FS) division in August of Year 8. Subsequent increases of HSE-114-1108 trace back to this plant.
April of Year 9: Planted 100 plants of HSE-114-1108 in the field at CAWO as an observation plot.
Observations during the growing season confirmed HSE-114-1108 is uniform and stable
Parent line HJA-114-1078, also known as “HJA114-1078”, and a subject of U.S. patent application Ser. No. 13/189,318, filed on Jul. 22, 2011, was developed at Seminis's Northern California breeding station at Woodland, Calif. (CAWO) and Seminis's Florida breeding station at Felda (FLFE), and Seminis's Guatemala breeding station at Salama (GUSA). HJA-114-1078 was developed by pure line selection from the cross Durkee Cayenne/Pasilla Bajio, made in Year 1.
The selections from the cross Durkee Cayenne×Pasilla Bajio were made as follows:
November of Year 1: Planted the F1 cross Durkee Cayenne×Pasilla Bajio in the open field at Salama, Guatemala. Selfed 2 individual F1 plants.
July of Year 2: Planted 100 F2 plants in the open field at FLFE. Selected 22 individual F2 plants.
April of Year 3: Planted 22 F3 lines in the field at CAWO. Selected 11 individual F3 plants.
January of Year 4: Planted 11 F4 lines in the field at FLFE. Selected 4 individual F4 plants.
July of Year 4: Planted 4 F5 lines in the field at FLFE. Selected 4 individual F5 plants.
January of Year 5: Planted 4 F6 lines in the greenhouse at CAWO. Selected 2 individual plants and one plant, designated 0413-1618-M, was selected as being pure and designated HJA-114-1078. Seed of HJA-114-1078 was given to Seminis' Foundation Seed (FS) division in October of Year 6. Subsequent increases of HJA-114-1078 trace back to this plant.
January of Year 6: Planted 100 plants of HJA-114-1078 in the field at FLFE as an observation plot.
Observations during the growing season confirmed HJA-114-1078 is uniform and stable.
The parent lines are uniform and stable, as is a hybrid produced therefrom. A small percentage of variants can occur within commercially acceptable limits for almost any characteristic during the course of repeated multiplication. However no variants are expected.
B. Physiological and Morphological Characteristics of Pepper Hybrid PX-11487057, Pepper Line HSE-114-1108 and Pepper Line HJA-114-1078
In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of pepper hybrid PX-11487057 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 1-3.







TABLE 1







Physiological and Morphological Characteristics of Hybrid PX-11487057













Comparison Variety -



Characteristic
PX-11487057
SERRANO CHILI













1.
Species
C. annuum
C. annuum


2.
Maturity (in region of best





adaptability)





days from transplanting until
   47
   75



mature green stage





days from transplanting until
   95
  106



mature red or yellow stage





days from direct seeding until
   86
  114



mature green stage





days from direct seeding until
  134
  145



mature red or yellow stage




3.
Plant





habit
compact
semi-spreading



attitude
upright/erect
upright/erect




(De Cayenne, Doux





très long des Landes,





Piquant d'Algérie)




plant height
67.3 cm
79.9 cm



plant width
54.1 cm
54.7 cm



length of stem from cotyledon
22.1 cm
65.5 cm



to first flower





length of the third internode
102.7 mm
66.3 mm



(from soil surface)





length of stem
long
long




(Lipari, Marconi,





Rouge long ordinaire)




shortened internode (in upper
absent
present



part)
(California wonder, De





Cayenne)




length of internode (on
long




primary side shoots)
(Corno di toro rosso)




stem: hairiness of nodes
medium
very strong




(Doux très long des





Landes, Farnese)




height
tall
very tall




(Century, Orias)




basal branches
few (2-3)
many (4+)



branch flexibility
willowy (Cayenne
rigid




Long Red)




stem strength (breakage
strong
weak



resistance)




4.
Leaf





length of blade
long
short




(Cupido, Dolmy,





Encore, Mazurka,





Monte)




width of blade
medium
narrow




(Albaregia, Balaton,





Danubia, Marconi,





Merit)




width
50.9 mm
36.1 mm



length
98.6 mm
74.3 mm



petiole length
45.9 mm
28.4 mm



color
dark green
dark green



color (RHS color chart)
147A
147A



intensity of green color
dark (Dolmy, Tinto)
dark



mature leaf shape
lanceolate (Diavolo,
lanceolate




Recio)




leaf and stem pubescence
light
heavy



undulation of margin
weak
very weak




(Doux très long des





Landes)




blistering
weak
very weak




(Pusztagold)




profile in cross section
moderately concave
strongly concave




(Doux italien, Favolor)




glossiness
weak
weak




(De Cayenne, Doux





très long des Landes)



5.
Flower





peduncle: attitude
semi-drooping
semi-drooping




(Blondy)




flowers per leaf axil
   1
   1



calyx lobes
   6.5
   6.1



petals
   6.5
   6



diameter
22.4 mm
20.6 mm



corolla color
white
white



corolla throat markings
yellow
yellow



anther color
purple
purple



style length
exceeds stamen
exceeds stamen



self-incompatibility
present
present


6.
Fruit





group
Small Hot (Serrano)
Small Hot (Serrano)



color (before maturity)
green (California
green




Wonder, Lamuyo)




intensity of color (before
dark
dark



maturity)





immature fruit color
dark green
dark green



immature fruit color (RHS
144A
139A



color chart)





attitude/position
drooping/pendent
drooping/pendent




(De Cayenne, Lamuyo)




length
medium
short




(Fehér, Lamuyo)




diameter
medium
narrow




(Doux italien, Corno di





toro)




ratio length/diameter
very large
large




(De Cayenne,





Kusamon, Spadi)




calyx diameter
14.2 mm
12.4 mm



fruit length
97.5 mm
57.1 mm



fruit diameter at calyx
15.5 mm
14.3 mm



attachment





fruit diameter at mid-point
17.6 mm
15.3 mm



flesh thickness at mid-point
3.3 mm
2.2 mm



average number of fruits per
   61.5
   77.7



plant





% large fruits
8.30% (weight range:
32.10% (weight range




18 to 30 g)
4.5 to 13 g)



% medium fruits
71.10% (weight range:
52.30% (weight range




9 to 17 g)
2.5 to 3.5 g)



% small fruits
20.40% (weight range:
15.60% (weight range 1




1 to 8 g)
to 1.5 g)



average fruit weight
11.9 gm
3.7 gm



fruit shape (longitudinal
narrowly triangular
narrowly triangular



section)
(De Cayenne, Demon)




fruit shape (cross section, at
circular
circular



level of placenta)
(Cherry Sweet, Doux





très long des Landes)




sinuation of pericarp at basal
absent or very weak
absent or very weak



part
(Delphin, Kalocsai V-





2, Milord)




sinuation of pericarp
absent or very weak
absent or very weak



excluding basal part
(Delphin, Milord)




texture of surface
smooth or very slightly
smooth or very slightly




wrinkled
wrinkled




(Milord)




color (at maturity)
red
red




(Fehér, Lamuyo)




intensity of color (at maturity)
dark
dark



mature fruit color
red
red



mature fruit color (RHS color
46A
N34A



chart)





glossiness
strong
strong




(Doux italien, Trophy)




stalk cavity
absent
absent




(Corinto, Corno di toro,





Sweet banana, Sucette





de Provence)




pedicel length
41.0 mm
29.9 mm



pedicel thickness
3.6 mm
2.4 mm



pedicel shape
curved
curved



pedicel cavity
absent
absent



stalk: length
long
long




(De Cayenne, Sierra





Nevada, Sweet banana)




stalk: thickness
thin
thin




(Sweet banana)




base shape
rounded
rounded



shape of apex
moderately acute
very acute/pointed



shape
Elongate (Long Thin
Elongate (Long Thin




Cayenne)
Cayenne)



set
scattered
scattered



depth of interloculary grooves
absent
absent




(De Cayenne)




% fruits with one locule
 0%
 0%



% fruits with two locules
20%
10%



% fruits with three locules
80%
80%



% fruits with four locules
 0%
10%



% fruits with five or more
 0%
 0%



locules





average number of locules
   2.8
   3



thickness of flesh
medium
thin




(Fehér, Lamuyo)




calyx: aspect
enveloping (cup-
enveloping (cup-shaped)




shaped)





(De Cayenne, Sweet





banana)




pungency
hot
hot



capsaicin in placenta
present (De Cayenne)
present



mg capsaicin per gram dry
0.95 mg
2.1 mg



fruit





Scoville Units (dry fruit)
15,109
31,327



flavor
strong pepper flavor
strong pepper flavor



glossiness
shiny
shiny


7.
Seed





seed cavity length
83.8 mm
44.3 mm



seed cavity diameter
11.8 mm
10.4 mm



placenta length
67 mm
34.3 mm



number of seeds per fruit
   46.2
   48.3



grams per 1000 seeds
4.1 gm
4 gm



color
yellow
yellow


8.
Anthocyanin coloration of:





seedling hypocotyl
weak
moderate



stem
weak
moderate



coloration of nodes
moderate
strong



intensity of anthocyanin
medium
strong



coloration of nodes
(Clovis, Lamuyo,





Sonar)




leaf
moderate
weak



pedicel
weak
strong



calyx
absent
moderate



anther
present (Lamuyo)
present



fruit anthocyanin coloration
moderate
absent



beginning of flowering (1st
medium
late



flower on 2nd flowering node)
(Lamuyo, Latino)




time of maturity
medium
very late





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.













TABLE 2







Physiological and Morphological Characteristics of Line HSE-114-1108











CHARACTERISTIC
HSE-114-1108
Guajillo TG













1.
Species
C. annuum
C. annuum


2.
Maturity (in region of best





adaptability)





days from transplanting until
 64
 76



mature green stage





days from transplanting until
113
102



mature red or yellow stage





days from direct seeding until
 97
113



mature green stage





days from direct seeding until
145
139



mature red or yellow stage




3.
Plant





habit
semi-spreading
semi-spreading



attitude
upright/erect
semi-upright/semi-




(De Cayenne, Doux très
erect




long des Landes,





Piquant d'Algerie)




plant height
51.8 cm
58 cm



plant width
36.1 cm
52 cm



length of stem from cotyledon
17.3 cm
15 cm



to first flower





length of the third internode
75.2 mm
55 mm



(from soil surface)





length of stem
medium (Belsir,
long




Lamuyo)




shortened internode (in upper
present (Fehér, Kalocsai
absent



part)
601, Kalocsai 702)




number of internodes between
one to three (Fehér)




the first flower and the





shortened internodes





stem: hairiness of nodes
medium (Doux très long
absent or very weak




des Landes, Farnese)




height
medium (HRF)
tall



basal branches
many (4+)
few



branch flexibility
rigid (Yolo Wonder)
willowy



stem strength (breakage
intermediate
strong



resistance)




4.
Leaf





length of blade
medium (Atol, Blondy,
medium




Marconi, Merit,





Anthea)




width of blade
medium (Albaregia,
broad




Balaton, Danubia,





Marconi, Merit)




width
35.9 mm
55 mm



length
74.2 mm
110 mm



petiole length
33.8 mm
45 mm



color
dark green
dark green



color (RHS Color Chart value)
146B
139A



intensity of green color
dark (Dolmy, Tinto)
dark



mature leaf shape
lanceolate (Diavolo,
ovate




Recio)




leaf and stem pubescence
moderate
absent



undulation of margin
absent (De Cayenne)
absent



blistering
weak (Pusztagold)
weak



profile in cross section
flat (De Cayenne,
flat




Recio)




glossiness
very weak (Diavolo)
medium


5.
Flower





peduncle: attitude
drooping (Heldor,
drooping




Lamuyo)




flowers per leaf axil
 1
 1



calyx lobes
 5
 5



petals
 5
 6



diameter
18.5 mm
22.5 mm



corolla color
white
white



corolla throat markings
yellow
yellow



anther color
purple
purple



style length
exceeds stamen
exceeds stamen



self-incompatibility
present
present


6.
Fruit





group
Small Hot (Serrano)
Anaheim Chili





(Sandia)



color (before maturity)
green (California
green




wonder, Lamuyo)




intensity of color (before
light
light



maturity)





immature fruit color
light green
light green



immature fruit color (RHS
137B
144A



Color Chart value)





attitude/position
drooping/pendent (De
drooping/pendent




Cayenne, Lamuyo)




length
medium (Fehér,
long




Lamuyo)




diameter
narrow (Doux très long
narrow




des Landes)




ratio length/diameter
medium (Adra, Cherry
large




Sweet, Daniel, Delphin,





Edino)




calyx diameter
10.6 mm
19.2 mm



fruit length
61.6 mm
100.9 mm



fruit diameter at calyx
12.5 mm
22.5 mm



attachment





fruit diameter at mid-point
12.5 mm
25.9 mm



flesh thickness at mid-point
2.2 mm
3 mm



average number of fruits per
140.8
 79



plant





% large fruits
10 (weight range: 7 to
35 (weight range 11 to




10 g)
15 g)



% medium fruits
54 (weight range: 4 to
65 (weight range 6 to




6 g)
10 g)



% small fruits
36 (weight range: 1 to





3 g)




average fruit weight
3.9 gm
8.9 gm



fruit shape (longitudinal
narrowly triangular (De
narrowly triangular



section)
Cayenne, Demon)




fruit shape (cross section, at
circular (Cherry Sweet,
elliptic



level of placenta)
Doux très long des





Landes)




sinuation of pericarp at basal
absent or very weak
weak



part
(Delphin, Kalocsai V-2,





Milord)




sinuation of pericarp excluding
absent or very weak
weak



basal part
(Delphin, Milord)




texture of surface
smooth or very slightly
slightly wrinkled




wrinkled (Milord)




color (at maturity)
red (Fehér, Lamuyo)
red



intensity of color (at maturity)
medium
dark



mature fruit color
red
red



mature fruit color (RHS Color
46B
46A



Chart value)





glossiness
weak (Doux très long
medium/moderate




des Landes)




stalk cavity
absent (Corinto, Corno
absent




di toro, Sweet banana,





Sucette de Provence)




pedicel length
31.2 mm
24.4 mm



pedicel thickness
2.5 mm
8.3 mm



pedicel shape
curved
curved



pedicel cavity
absent
absent



stalk: length
long (De Cayenne,
medium




Sierra Nevada, Sweet





banana)




stalk: thickness
thin (Sweet banana)
medium



base shape
rounded
rounded



shape of apex
very acute/pointed (De
very acute/pointed




Cayenne, Hot chili)




shape
elongate (Long Thin
elongate (Long Thin




Cayenne)
Cayenne)



set
scattered
scattered



depth of interloculary grooves
absent (De Cayenne)
shallow



% fruits with one locule
36.70%
30%



% fruits with two locules
 9.80%
30%



% fruits with three locules
28.30%
40%



% fruits with four locules
25.20%




average number of locules
 1.9
 3



thickness of flesh
thin (Banán, Carré doux
medium




extra hâtif, Dous très





long des Landes)




calyx: aspect
enveloping (cup-
enveloping




shaped) (De Cayenne,





Sweet banana)




pungency
hot
hot



capsaicin in placenta
present (De Cayenne)
present



flavor
mild pepper flavor
strong pepper flavor



glossiness
shiny
moderate


7.
Seed





seed cavity length
55.8 mm
120.7 mm



seed cavity diameter
7.1 mm
20.1 mm



placenta length
38.1 mm
45 mm



number of seeds per fruit
43.2
132



grams per 1000 seeds
5 gm
3.5 gm



color
yellow
yellow


8.
Anthocyanin coloration of:





seedling hypocotyl
absent (Albaregia,
absent




Albena)




stem
moderate
absent



node
moderate
weak



stem: intensity of anthocyanin
medium (Clovis,
very weak



coloration of nodes
Lamuyo, Sonar)




leaf
absent
absent



pedicel
absent
absent



calyx
absent
absent



anther
present (Lamuyo)
present



fruit coloration
absent (Lamuyo)
absent



beginning of flowering (1st
early (Carré doux extra
late



flower on 2nd flowering node)
hâtif, Cupido, Fehér,





Flaviano, Lito, Trophy)




time of maturity
late (Daniel, Doux
late




d'Espagne)





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.













TABLE 3







Physiological and Morphological Characteristics of Line HJA-114-1078











CHARACTERISTIC
HJA-114-1078
Tam Vera Cruz













1.
Species
C. annuum
C. annuum


2.
Maturity (in region of best





adaptability)





Days from transplanting until
 72
 57



mature green stage





Days from transplanting until
138
103



mature red or yellow stage





Days from direct seeding until
 98
 83



mature green stage





Days from direct seeding until
164
129



mature red or yellow stage





Beginning of flowering (1st
late (Daniel, Piquant
early



flower on 2nd flowering node)
d'Algérie, Zingaro)




Time of maturity
Very late (Cancun,
late




California, wonder)



3.
Plant





Habit
semi-spreading
compact



Attitude
semi-upright/semi-
semi-upright/semi-




erect (Clovis, Sonar)
erect



Plant height
35.3 cm
32.2 cm



Plant width
33.8 cm
33 cm



Length of stem from cotyledon
12.8 cm
11.6 cm



to first flower





Length of the third internode
98.2 mm
64.3 mm



(from soil surface)





Length of stem
medium (Belsir,
medium




Lamuyo)




Shortened internode (in upper
present (Fehér, Kalocsai
absent



part)
601, Kalocsai 702)




Number of internodes between
more than three
one to three



the first flower and the
(Kalocsai 702)




shortened internodes (varieties





with shortened internodes)





Stem: hairiness of nodes
absent or very weak
weak




(Arlequin)




Height
medium (HRF)
short



Basal branches
none
few



Branch flexibility
rigid (Yolo Wonder)
willowy



Stem strength (breakage
intermediate
strong



resistance)




4.
Leaf





Length of blade
long (Cupido, Dolmy,
medium




Encore, Mazurka,





Monte)




Width of blade
medium (Albaregia,
medium




Balaton, Danubia,





Marconi, Merit)




Leaf width
71.7 mm
35.9 mm



Leaf length
66.9 mm
75.8 mm



Petiole length
29.9 mm
24.1 mm



Color
medium green
medium green



RHS Color Chart Value
146B
146A



Intensity of green color
light (Piquant d'Algérie,
medium




Pusztagold)




Mature leaf shape
ovate (Balico, Sonar)
lanceolate



Leaf and stem pubescence
absent
light



Undulation of margin
absent (De Cayenne)
medium



Blistering
absent (Century, Recio,
very strong




Solfiane)




Profile in cross section
flat (De Cayenne,
strongly concave




Recio)




Glossiness
very weak (Diavolo)
medium



Peduncle: attitude
semi-drooping (Blondy)
drooping


5.
Flower





Flowers per leaf axil
 1
 1



Calyx lobes
 6
 6



Petals
 6
 6



Diameter
20.9 mm
24.1 mm



Corolla color
white
white



Corolla throat markings
yellow
yellow



Anther color
purple
purple



Style length
exceeds stamen
exceeds stamen



Self-incompatibility
present
present


6.
Fruit





Group
Cayenne (Cayenne
Jalapeno (Jalapeno)




Long Red)




Color (before maturity)
green (California
green




wonder, Lamuyo)




Intensity of color (before
dark
medium



maturity)





Immature fruit color
dark green
medium green



Immature fruit color
137A
143B



RHS Color Chart value





Attitude/position
drooping/pendent (De
drooping/pendent




Cayenne, Lamuyo)




Length
very long (Arabal,
medium




Como di toro, Marconi)




Diameter
broad (Clovis, Lamuyo)
medium



Ratio length/diameter
very large (De Cayenne,
medium




Kusamon, Spadi)




Calyx diameter
13.1 mm
19 mm



Length
96.2 mm
73 mm



Diameter at calyx attachment
54.5 mm
27.6 mm



Diameter at mid-point
15.8 mm
27 mm



Flesh thickness at mid-point
25.3 mm
4.1 mm



Average number of fruits per
 96
 39.1



plant





% large fruits
9%
13.50%




weight range: 20-29 g
weight range: 30-39 g



% medium fruits
51%
50%




weight range: 10-19 g
weight range: 20-29 g



% small fruits
40%
36.50%




weight range: 1-9 g
weight range: 10-19 g



Average fruit weight
12.8 gm
22 gm



Shape in longitudinal section
narrowly triangular (De
trapezoidal




Cayenne, Demon)




Shape in cross section (at level
circular (Cherry Sweet,
quad-rangular



of placenta)
Doux très long des





Landes)




Sinuation of pericarp at basal
absent or very weak
very weak



part
(Delphin, Kalocsai V-2,





Milord)




Sinuation of pericarp excluding
absent or very weak
medium



basal part
(Delphin, Milord)




Texture of surface
smooth or very slightly
slightly wrinkled




wrinkled (Milord)




Color (at maturity)
red (Fehér, Lamuyo)
red



Intensity of color (at maturity)
dark
medium



Mature fruit color
red
red



Mature fruit color RHS Color
N34A
42A



Chart value





Glossiness
medium/moderate
medium




(Carre doux extra hâtif,





Lamuyo, Sonar)




Stalk cavity
absent (Corinto, Corno
absent




di toro, Sweet banana,





Sucette de Provence)




Pedicel length
38.3 mm
18.9 mm



Pedicel thickness
3 mm
3.9 mm



Pedicel shape
curved
curved



Pedicel cavity
absent
absent



Stalk: length
long (De Cayenne,
medium




Sierra Nevada, Sweet





banana)




Stalk: thickness
medium (Doux italien,
thin




Surpas)




Base shape
rounded
rounded



Shape of apex
rounded (Cherry Sweet)
moderately acute



Shape
elongate (Long Thin
Oblong (Jalapeno)




Cayenne)




Set
concentrated
concentrated



Depth of interloculary grooves
absent (De Cayenne)
absent



Number of locules
predominantly two (De
predominantly two




Cayenne)
(De Cayenne)



% fruits with two locules
73.30%
 7%



% fruits with three locules
26.70%
30%



Average number of locules
 2
 1



Thickness of flesh
medium (Fehér,
medium




Lamuyo)




Calyx: aspect
enveloping (cup-
enveloping




shaped) (De Cayenne,





Sweet banana)




Pungency
hot
hot



Capsaicin in placenta
present (De Cayenne)
present



Flavor
moderate pepper flavor
mild pepper flavor



Glossiness
moderate
moderate


7.
Seed





Seed cavity length
85.6 mm
61 mm



Seed cavity diameter
14.5 mm
17.5 mm



Placenta length
59.2 mm
34.1 mm



Number of seeds per fruit
 23
 80



Grams per 1000 seeds
10 gm
5.7 gm



Color
yellow
yellow


8.
Anthocyanin





Anthocyanin coloration of
absent (Albaregia,
weak



hypocotyl
Albena)




Plant: anthocyanin coloration of
weak
strong



stem





Plant: anthocyanin coloration of
weak
strong



nodes





Stem: intensity of anthocyanin
weak (California
strong



coloration of nodes
wonder, Clio, Doux





d'Espagne, Dous très





long des Landes,





Golden calwonder)




Plant: anthocyanin coloration of
absent
absent



leaf





Plant: anthocyanin coloration of
absent
absent



pedicel





Plant: anthocyanin coloration of
absent
weak



calyx





Flower: anthocyanin coloration
present (Lamuyo)
present



in anther





Fruit: anthocyanin coloration
weak
absent





*These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.






C. Breeding Pepper Plants
One aspect of the current invention concerns methods for producing seed of pepper hybrid PX-11487057 involving crossing pepper lines HSE-114-1108 and HJA-114-1078. Alternatively, in other embodiments of the invention, hybrid PX-11487057, line HSE-114-1108, or line HJA-114-1078 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid PX-11487057 and/or the pepper lines HSE-114-1108 and HJA-114-1078, or can be used to produce plants that are derived from hybrid PX-11487057 and/or the pepper lines HSE-114-1108 and HJA-114-1078. Plants derived from hybrid PX-11487057 and/or the pepper lines HSE-114-1108 and HJA-114-1078 may be used, in certain embodiments, for the development of new pepper varieties.
The development of new varieties using one or more starting varieties is well known in the art. In accordance with the invention, novel varieties may be created by crossing hybrid PX-11487057 followed by multiple generations of breeding according to such well known methods. New varieties may be created by crossing with any second plant. In selecting such a second plant to cross for the purpose of developing novel lines, 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. Once initial crosses have been made, inbreeding and selection take place to produce new varieties. For development of a uniform line, often five or more generations of selfing and selection are involved.
Uniform lines of new varieties may also be developed by way of double-haploids. 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. 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 invention, any of such techniques may be used in connection with a plant of the invention and progeny thereof to achieve a homozygous line.
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 have the characteristic being transferred, but are like the superior parent for most or almost all other loci. The last backcross generation would be selfed to give pure breeding progeny for the trait being transferred.
The plants of the present invention are particularly well suited for the development of new lines based on the elite nature of the genetic background of the plants. In selecting a second plant to cross with PX-11487057 and/or pepper lines HSE-114-1108 and HJA-114-1078 for the purpose of developing novel pepper lines, it will typically be preferred 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 may include, in specific embodiments, high seed yield, high seed germination, seedling vigor, high fruit yield, disease tolerance or resistance, and adaptability for soil and climate conditions. Consumer-driven traits, such as a fruit shape, color, texture, and taste are other examples of traits that may be incorporated into new lines of pepper plants developed by this invention.
D. Performance Characteristics
As described above, hybrid PX-11487057 exhibits desirable traits, as conferred by pepper lines HSE-114-1108 and HJA-114-1078. The performance characteristics of hybrid PX-11487057 and pepper lines HSE-114-1108 and HJA-114-1078 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented below.







TABLE 4







Performance Characteristics for Hybrid PX-11487057 and Comparative varieties
















Color
Maturity
Yield
Length
Width
Av. wt
% dry
SHU


Variety
mature
(DAT)
g/plt
cm
cm
g/fruit
matter
fresh


















Hidalgo
red
89
438
7.7
1.7
11.1
8.8
1011


Don Vicente
red
85
829
8.7
2.5
18.0
7.7
1121


PX-11487057
red
85
765
9.7
1.8
16.4
10.3
2839









E. Further Embodiments of the Invention
In certain aspects of the invention, plants described herein are provided modified to include at least a first desired heritable trait. Such plants may, in one embodiment, be developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to a genetic locus transferred into the plant via the backcrossing technique. The term single locus converted plant as used herein refers to those pepper plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a variety are recovered in addition to the single locus transferred into the variety via the backcrossing technique. By essentially all of the morphological and physiological characteristics, it is meant that the characteristics of a plant are recovered that are otherwise present when compared in the same environment, other than an occasional variant trait that might arise during backcrossing or direct introduction of a transgene.
Backcrossing methods can be used with the present invention to improve or introduce a characteristic into the present variety. The parental pepper plant which contributes the locus for the desired characteristic is termed the nonrecurrent or donor parent. This terminology refers to the fact that the nonrecurrent parent is used one time in the backcross protocol and therefore does not recur. The parental pepper plant to which the locus or loci from the nonrecurrent parent are transferred is known as the recurrent parent as it is used for several rounds in the backcrossing protocol.
In a typical backcross protocol, the original variety of interest (recurrent parent) is crossed to a second variety (nonrecurrent parent) that carries the single locus of interest to be transferred. The resulting progeny from this cross are then crossed again to the recurrent parent and the process is repeated until a pepper plant is obtained wherein essentially all of the morphological and physiological characteristics of the recurrent parent are recovered in the converted plant, in addition to the single transferred locus from the nonrecurrent parent.
The selection of a suitable recurrent parent is an important step for a successful backcrossing procedure. The goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original variety. To accomplish this, a single locus of the recurrent variety is modified or substituted with the desired locus from the nonrecurrent parent, while retaining essentially all of the rest of the desired genetic, and therefore the desired physiological and morphological constitution of the original variety. The choice of the particular nonrecurrent parent will depend on the purpose of the backcross; one of the major purposes is to add some commercially desirable trait to the plant. The exact backcrossing protocol will depend on the characteristic or trait being altered and the genetic distance between the recurrent and nonrecurrent parents. Although backcrossing methods are simplified when the characteristic being transferred is a dominant allele, a recessive allele, or an additive allele (between recessive and dominant), may also be transferred. In this instance it may be necessary to introduce a test of the progeny to determine if the desired characteristic has been successfully transferred.
In one embodiment, progeny pepper plants of a backcross in which a plant described herein is the recurrent parent comprise (i) the desired trait from the non-recurrent parent and (ii) all of the physiological and morphological characteristics of pepper the recurrent parent as determined at the 5% significance level when grown in the same environmental conditions.
New varieties can also be developed from more than two parents. The technique, known as modified backcrossing, uses different recurrent parents during the backcrossing. Modified backcrossing may be used to replace the original recurrent parent with a variety having certain more desirable characteristics or multiple parents may be used to obtain different desirable characteristics from each.
With the development of molecular markers associated with particular traits, it is possible to add additional traits into an established germ line, such as represented here, with the end result being substantially the same base germplasm with the addition of a new trait or traits. Molecular breeding, as described in Moose and Mumm, 2008 (Plant Physiology, 147: 969-977), for example, and elsewhere, provides a mechanism for integrating single or multiple traits or QTL into an elite line. This molecular breeding-facilitated movement of a trait or traits into an elite line may encompass incorporation of a particular genomic fragment associated with a particular trait of interest into the elite line by the mechanism of identification of the integrated genomic fragment with the use of flanking or associated marker assays. In the embodiment represented here, one, two, three or four genomic loci, for example, may be integrated into an elite line via this methodology. When this elite line containing the additional loci is further crossed with another parental elite line to produce hybrid offspring, it is possible to then incorporate at least eight separate additional loci into the hybrid. These additional loci may confer, for example, such traits as a disease resistance or a fruit quality trait. In one embodiment, each locus may confer a separate trait. In another embodiment, loci may need to be homozygous and exist in each parent line to confer a trait in the hybrid. In yet another embodiment, multiple loci may be combined to confer a single robust phenotype of a desired trait.
Many single locus traits have been identified that are not regularly selected for in the development of a new inbred but that can be improved by backcrossing techniques. Single locus traits may or may not be transgenic; examples of these traits include, but are not limited to, herbicide resistance, resistance to bacterial, fungal, or viral disease, insect resistance, modified fatty acid or carbohydrate metabolism, and altered nutritional quality. These comprise genes generally inherited through the nucleus.
Direct selection may be applied where the single locus acts as a dominant trait. For this selection process, the progeny of the initial cross are assayed for viral resistance and/or the presence of the corresponding gene prior to the backcrossing. Selection eliminates any plants that do not have the desired gene and resistance trait, and only those plants that have the trait are used in the subsequent backcross. This process is then repeated for all additional backcross generations.
Selection of pepper plants for breeding is not necessarily dependent on the phenotype of a plant and instead can be based on genetic investigations. For example, one can utilize a suitable genetic marker which is closely genetically linked to a trait of interest. One of these markers can be used to identify the presence or absence of a trait in the offspring of a particular cross, and can be used in selection of progeny for continued breeding. This technique is commonly referred to as marker assisted selection. Any other type of genetic marker or other assay which is able to identify the relative presence or absence of a trait of interest in a plant can also be useful for breeding purposes. Procedures for marker assisted selection are well known in the art. Such methods will be of particular utility in the case of recessive traits and variable phenotypes, or where conventional assays may be more expensive, time consuming or otherwise disadvantageous. Types of genetic markers which could be used in accordance with the invention include, but are not necessarily limited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), 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) (EP 534 858, specifically incorporated herein by reference in its entirety), and Single Nucleotide Polymorphisms (SNPs) (Wang et al., Science, 280:1077-1082, 1998).
F. Plants Derived by Genetic Engineering
Many useful traits that can be introduced by backcrossing, as well as directly into a plant, are those which are introduced by genetic transformation techniques. Genetic transformation may therefore be used to insert a selected transgene into a plant of the invention or may, alternatively, be used for the preparation of transgenes which can be introduced by backcrossing. Methods for the transformation of plants that are well known to those of skill in the art and applicable to many crop species include, but are not limited to, electroporation, microprojectile bombardment, Agrobacterium-mediated transformation and direct DNA uptake by protoplasts.
To effect transformation by electroporation, one may employ either friable tissues, such as a suspension culture of cells or embryogenic callus or alternatively one may transform immature embryos or other organized tissue directly. In this technique, one would partially degrade the cell walls of the chosen cells by exposing them to pectin-degrading enzymes (pectolyases) or mechanically wound tissues in a controlled manner.
An efficient method for delivering transforming DNA segments to plant cells is microprojectile bombardment. In this method, particles are coated with nucleic acids and delivered into cells by a propelling force. Exemplary particles include those comprised of tungsten, platinum, and preferably, gold. For the bombardment, cells in suspension are concentrated on filters or solid culture medium. Alternatively, immature embryos or other target cells may be arranged on solid culture medium. The cells to be bombarded are positioned at an appropriate distance below the macroprojectile stopping plate.
An illustrative embodiment of a method for delivering DNA into plant cells by acceleration is the Biolistics Particle Delivery System, which can be used to propel particles coated with DNA or cells through a screen, such as a stainless steel or Nytex screen, onto a surface covered with target cells. The screen disperses the particles so that they are not delivered to the recipient cells in large aggregates. Microprojectile bombardment techniques are widely applicable, and may be used to transform virtually any plant species.
Agrobacterium-mediated transfer is another widely applicable system for introducing gene loci into plant cells. An advantage of the technique is that DNA can be introduced into whole plant tissues, thereby bypassing the need for regeneration of an intact plant from a protoplast. Modern Agrobacterium transformation vectors are capable of replication in E. coli as well as Agrobacterium, allowing for convenient manipulations (Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recent technological advances in vectors for Agrobacterium-mediated gene transfer have improved the arrangement of genes and restriction sites in the vectors to facilitate the construction of vectors capable of expressing various polypeptide coding genes. The vectors described have convenient multi-linker regions flanked by a promoter and a polyadenylation site for direct expression of inserted polypeptide coding genes. Additionally, Agrobacterium containing both armed and disarmed Ti genes can be used for transformation.
In those plant strains where Agrobacterium-mediated transformation is efficient, it is the method of choice because of the facile and defined nature of the gene locus transfer. The use of Agrobacterium-mediated plant integrating vectors to introduce DNA into plant cells is well known in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S. Pat. No. 5,563,055).
Transformation of plant protoplasts also can be achieved using methods based on calcium phosphate precipitation, polyethylene glycol treatment, electroporation, and combinations of these treatments (see, e.g., Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al., Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature, 312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986; Marcotte et al., Nature, 335:454, 1988). Transformation of plants and expression of foreign genetic elements is exemplified in Choi et al. (Plant Cell Rep., 13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet., 107:462-469, 2003).
A number of promoters have utility for plant gene expression for any gene of interest including but not limited to selectable markers, scoreable markers, genes for pest tolerance, disease resistance, nutritional enhancements and any other gene of agronomic interest. Examples of constitutive promoters useful for plant gene expression include, but are not limited to, the cauliflower mosaic virus (CaMV) P-35S promoter, which confers constitutive, high-level expression in most plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985), including in monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990; Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemly duplicated version of the CaMV 35S promoter, the enhanced 35S promoter (P-e35S); 1 the nopaline synthase promoter (An et al., Plant Physiol., 88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell, 1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as described in U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter (P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem; the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform virus promoter; a commelina yellow mottle virus promoter; and other plant DNA virus promoters known to express in plant cells.
A variety of plant gene promoters that are regulated in response to environmental, hormonal, chemical, and/or developmental signals can also be used for expression of an operably linked gene in plant cells, including promoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., Plant Cell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell, 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson et al., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid (Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wun1, Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such as methyl jasmonate, salicylic acid, or Safener. It may also be advantageous to employ organ-specific promoters (e.g., Roshal et al., EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988; Bustos et al., Plant Cell, 1:839, 1989).
Exemplary nucleic acids which may be introduced to plants of this invention include, for example, DNA sequences or genes from another species, or even genes or sequences which originate with or are present in the same species, but are incorporated into recipient cells by genetic engineering methods rather than classical reproduction or breeding techniques. However, the term “exogenous” is also intended to refer to genes that are not normally present in the cell being transformed, or perhaps simply not present in the form, structure, etc., as found in the transforming DNA segment or gene, or genes which are normally present and that one desires to express in a manner that differs from the natural expression pattern, e.g., to over-express. Thus, the term “exogenous” gene or DNA is intended to refer to any gene or DNA segment that is introduced into a recipient cell, regardless of whether a similar gene may already be present in such a cell. The type of DNA included in the exogenous DNA can include DNA which is already present in the plant cell, DNA from another plant, DNA from a different organism, or a DNA generated externally, such as a DNA sequence containing an antisense message of a gene, or a DNA sequence encoding a synthetic or modified version of a gene.
Many hundreds if not thousands of different genes are known and could potentially be introduced into a pepper plant according to the invention. Non-limiting examples of particular genes and corresponding phenotypes one may choose to introduce into a pepper plant include one or more genes for insect tolerance, such as a Bacillus thuringiensis (B.t.) gene, pest tolerance such as genes for fungal disease control, herbicide tolerance such as genes conferring glyphosate tolerance, and genes for quality improvements such as yield, nutritional enhancements, environmental or stress tolerances, or any desirable changes in plant physiology, growth, development, morphology or plant product(s). For example, structural genes would include any gene that confers insect tolerance including but not limited to a Bacillus insect control protein gene as described in WO 99/31248, herein incorporated by reference in its entirety, U.S. Pat. No. 5,689,052, herein incorporated by reference in its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, herein incorporated by reference in their entirety. In another embodiment, the structural gene can confer tolerance to the herbicide glyphosate as conferred by genes including, but not limited to Agrobacterium strain CP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat. No. 5,633,435, herein incorporated by reference in its entirety, or glyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No. 5,463,175, herein incorporated by reference in its entirety.
Alternatively, the DNA coding sequences can affect these phenotypes by encoding a non-translatable RNA molecule that causes the targeted inhibition of expression of an endogenous gene, for example via antisense- or cosuppression-mediated mechanisms (see, for example, Bird et al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be a catalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desired endogenous mRNA product (see for example, Gibson and Shillito, Mol. Biotech., 7:125, 1997). Thus, any gene which produces a protein or mRNA which expresses a phenotype or morphology change of interest is useful for the practice of the present invention.
G. Definitions
In the description and tables herein, a number of terms are used. In order to provide a clear and consistent understanding of the specification and claims, the following definitions are provided:
Allele: Any of one or more alternative forms of a gene locus, all of which alleles relate to one trait or characteristic. In a diploid cell or organism, the two alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.
Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F1), back to one of the parents of the hybrid progeny. Backcrossing can be used to introduce one or more single locus conversions from one genetic background into another.
Crossing: The mating of two parent plants.
Cross-pollination: Fertilization by the union of two gametes from different plants.
Diploid: A cell or organism having two sets of chromosomes.
Emasculate: The removal of plant male sex organs or the inactivation of the organs with a cytoplasmic or nuclear genetic factor or a chemical agent conferring male sterility.
Enzymes: Molecules which can act as catalysts in biological reactions.
F1 Hybrid: The first generation progeny of the cross of two nonisogenic plants.
Genotype: The genetic constitution of a cell or organism.
Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.
Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
Marker: A readily detectable phenotype, preferably inherited in codominant fashion (both alleles at a locus in a diploid heterozygote are readily detectable), with no environmental variance component, i.e., heritability of 1.
Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.
Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer to genetic loci that control to some degree numerically representable traits that are usually continuously distributed.
Resistance: As used herein, the terms “resistance” and “tolerance” are used interchangeably to describe plants that show no symptoms to a specified biotic pest, pathogen, abiotic influence or environmental condition. These terms are also used to describe plants showing some symptoms but that are still able to produce marketable product with an acceptable yield. Some plants that are referred to as resistant or tolerant are only so in the sense that they may still produce a crop, even though the plants are stunted and the yield is reduced.
Regeneration: The development of a plant from tissue culture.
Royal Horticultural Society (RHS) color chart value: The RHS color chart is a standardized reference which allows accurate identification of any color. A color's designation on the chart describes its hue, brightness and saturation. A color is precisely named by the RHS color chart by identifying the group name, sheet number and letter, e.g., Yellow-Orange Group 19A or Red Group 41B.
Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.
Single Locus Converted (Conversion) Plant: Plants which are developed by a plant breeding technique called backcrossing, wherein essentially all of the morphological and physiological characteristics of a pepper variety are recovered in addition to the characteristics of the single locus transferred into the variety via the backcrossing technique and/or by genetic transformation.
Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.
Tissue Culture: A composition comprising isolated cells of the same or a different type or a collection of such cells organized into parts of a plant.
Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a pepper plant by transformation.
H. Deposit Information
A deposit of pepper hybrid PX-11487057 and inbred parent lines HSE-114-1108 and HJA-114-1078, disclosed above and recited in the claims, has been made with the American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The dates of deposits were Oct. 23, 2013, Oct. 23, 2013, and Jun. 20, 2011, respectively. The accession numbers for those deposited seeds of pepper hybrid PX-11487057 and inbred parent lines HSE-114-1108 and HJA-114-1078 are ATCC Accession No. PTA-120665, ATCC Accession No. PTA-120666, and ATCC Accession No. PTA-11946, respectively. Upon issuance of a patent, all restrictions upon the deposits will be removed, and the deposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for a period of 30 years, or 5 years after the last request, or for the effective life of the patent, whichever is longer, and will be replaced if necessary during that period.
Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be obvious that certain changes and modifications may be practiced within the scope of the invention, as limited only by the scope of the appended claims.
All references cited herein are hereby expressly incorporated herein by reference.

## Claims

1. A pepper plant comprising at least a first set of the chromosomes of pepper line HSE-114-1108, a sample of seed of said line having been deposited under ATCC Accession Number PTA-120666.

2. A seed comprising at least a first set of the chromosomes of pepper line HSE-114-1108, a sample of seed of said line having been deposited under ATCC Accession Number PTA-120666.

3. The plant of claim 1, which is an inbred.

4. The plant of claim 1, which is a hybrid.

5. The seed of claim 2, which is an inbred.

6. The seed of claim 2, which is a hybrid.

7. The plant of claim 4, wherein the hybrid plant is pepper hybrid PX-11487057, a sample of seed of said hybrid PX-11487057 having been deposited under ATCC Accession Number PTA-120665.

8. The seed of claim 6, defined as a seed of pepper hybrid PX-11487057, a sample of seed of said hybrid PX-11487057 having been deposited under ATCC Accession Number PTA-120665.

9. The seed of claim 2, defined as a seed of line HSE-114-1108.

10. A plant part of the plant of claim 1.

11. The plant part of claim 10, further defined as a leaf, an ovule, pollen, a fruit, or a cell.

12. A pepper plant having all the physiological and morphological characteristics of the pepper plant of claim 7.

13. A tissue culture of regenerable cells of the plant of claim 1.

14. The tissue culture according to claim 13, comprising cells or protoplasts from a plant part selected from the group consisting of embryos, meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, seed and stalks.

15. A pepper plant regenerated from the tissue culture of claim 13, wherein said plant otherwise comprises all of the morphological and physiological characteristics of the pepper plant comprising at least a first set of the chromosomes of pepper line HSE-114-1108, a sample of seed of said line having been deposited under ATCC Accession Number PTA-120666.

16. A method of vegetatively propagating the pepper plant of claim 1 comprising the steps of:
(a) collecting tissue capable of being propagated from the plant according to claim 1;
(b) cultivating said tissue to obtain proliferated shoots; and
(c) rooting said proliferated shoots to obtain rooted plantlets.

17. The method of claim 16, further comprising growing at least a first pepper plant from said rooted plantlets.

18. A method of introducing a desired trait into a pepper line comprising:
(a) crossing a plant of line HSE-114-1108 with a second pepper plant that comprises a desired trait to produce F1 progeny, a sample of seed of said line having been deposited under ATCC Accession Number PTA-120666;
(b) selecting an F1 progeny that comprises the desired trait;
(c) backcrossing the selected F1 progeny with a plant of line HSE-114-1108 to produce backcross progeny;
(d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristic of pepper line HSE-114-1108; and
(e) repeating steps (c) and (d) three or more times to produce selected fourth or higher backcross progeny that comprise the desired trait.

19. A pepper plant produced by the method of claim 18.

20. A method of producing a pepper plant comprising an added trait, the method comprising introducing a transgene conferring the trait into a plant of pepper hybrid PX-11487057, or pepper line HSE-114-1108, a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-120665, and ATCC Accession Number PTA-120666, respectively.

21. A plant produced by the method of claim 20.

22. The plant of claim 1, further comprising a transgene.

23. The plant of claim 22, wherein the transgene confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.

24. The plant of claim 1, further comprising a single locus conversion, wherein said plant otherwise comprises essentially all of the morphological and physiological characteristics of the pepper plant comprising at least a first set of the chromosomes of pepper line HSE-114-1108, a sample of seed of said line having been deposited under ATCC Accession Number PTA-120666.

25. The plant of claim 24, wherein the single locus conversion confers a trait selected from the group consisting of male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, modified fatty acid metabolism, environmental stress tolerance, modified carbohydrate metabolism and modified protein metabolism.

26. A method for producing a seed of a pepper plant derived from at least one of pepper hybrid PX-11487057, or pepper line HSE-114-1108 comprising the steps of:
(a) crossing a pepper plant of hybrid PX-11487057, or line HSE-114-1108 with itself or a second pepper plant; a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-120665, and ATCC Accession Number PTA-120666, respectively; and
(b) allowing seed of a hybrid PX-11487057, or line HSE-114-1108-derived pepper plant to form.

27. The method of claim 26, further comprising the steps of:
(c) selfing a plant grown from said hybrid PX-11487057, or HSE-114-1108-derived pepper seed to yield additional hybrid PX-11487057, or line HSE-114-1108-derived pepper seed;
(d) growing said additional hybrid PX-11487057, or line HSE-114-1108-derived pepper seed of step (c) to yield additional hybrid PX-11487057, or line HSE-114-1108-derived pepper plants; and
(e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid PX-11487057, or line HSE-114-1108-derived pepper plant.

28. The method of claim 26, wherein the second pepper plant is of an inbred pepper line.

29. The method of claim 26, comprising crossing line HSE-114-1108 with line HJA-114-1078, a sample of seed of said lines having been deposited under ATCC Accession Number PTA-120666, and ATCC Accession Number PTA-11946, respectively.

30. The method of claim 27, further comprising:
(f) crossing the further hybrid PX-11487057, or HSE-114-1108-derived pepper plant with a second pepper plant to produce seed of a hybrid progeny plant.

31. A plant part of the plant of claim 7.

32. The plant part of claim 31, further defined as a leaf, an ovule, pollen, a fruit, or a cell.

33. A method of producing a pepper seed comprising crossing the plant of claim 1 with itself or a second pepper plant and allowing seed to form.

34. A method of producing a pepper comprising:
(a) obtaining the plant according to claim 1, wherein the plant has been cultivated to maturity; and
(b) collecting a pepper from the plant.

35. A method of producing a plant of pepper hybrid PX-11487057 comprising a single locus conversion, the method comprising crossing a plant of line HJA-114-1078 with a plant of line HSE-114-1108, wherein at least one of said lines further comprises a single locus conversion and otherwise comprises essentially all of the morphological and physiological characteristics of line HJA-114-1078 or line HSE-114-1108, a sample of seed of said lines having been deposited under ATCC Accession No. PTA-11946 and ATCC Accession Number PTA-120666, respectively.