Pepper hybrid PX-11459057
09699979 ยท 2017-07-11
Assignee
Inventors
Cpc classification
C12N15/8279
CHEMISTRY; METALLURGY
C12N15/8247
CHEMISTRY; METALLURGY
C12N15/8241
CHEMISTRY; METALLURGY
C12N15/8271
CHEMISTRY; METALLURGY
C12N15/8245
CHEMISTRY; METALLURGY
C12N15/8201
CHEMISTRY; METALLURGY
C12N15/8251
CHEMISTRY; METALLURGY
International classification
C12N15/82
CHEMISTRY; METALLURGY
Abstract
The invention provides seed and plants of pepper hybrid PX-11459057 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of pepper hybrid PX-11459057 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.
Claims
1. A pepper plant comprising at least a first set of the chromosomes of pepper line HHA-114-1124, a sample of seed of said line having been deposited under ATCC Accession Number PTA-121393.
2. A seed comprising at least a first set of the chromosomes of pepper line HHA-114-1124, a sample of seed of said line having been deposited under ATCC Accession Number PTA-121393.
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-11459057, a sample of seed of said hybrid PX-11459057 having been deposited under ATCC Accession Number PTA-121689.
8. The seed of claim 6, defined as a seed of pepper hybrid PX-11459057, a sample of seed of said hybrid PX-11459057 having been deposited under ATCC Accession Number PTA-121689.
9. The seed of claim 2, defined as a seed of line HHA-114-1124.
10. A plant part of the plant of claim 1.
11. The plan 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 meristems, cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower, 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 HHA-114-1124, a sample of seed of said line having been deposited under ATCC Accession Number PTA-121393.
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 HHA-114-1124 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-121393; (b) selecting an F1 progeny that comprises the desired trait; (c) backcrossing the selected F1 progeny with a plant of line HHA-114-1124 to produce backcross progeny; (d) selecting backcross progeny comprising the desired trait and the physiological and morphological characteristic of pepper line HHA-114-1124; 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-11459057, or pepper line HHA-114-1124, a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-121689, and ATCC Accession Number PTA-121393, respectively.
21. A plant produced by the method of claim 20.
22. A pepper plant comprising at least a first set of the chromosomes of pepper line HHA-114-1124, a sample of seed of said line having been deposited under ATCC Accession Number PTA-121393, 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. A pepper plant comprising at least a first set of the chromosomes of pepper line HHA-114-1124, a sample of seed of said line having been deposited under ATCC Accession Number PTA-121393, further comprising a single locus conversion.
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-11459057, or pepper line HHA-114-1124 comprising the steps of: (a) crossing a pepper plant of hybrid PX-11459057, or line HHA-114-1124 with itself or a second pepper plant; a sample of seed of said hybrid and line having been deposited under ATCC Accession Number PTA-121689, and ATCC Accession Number PTA-121393, respectively; and (b) allowing seed of a hybrid PX-11459057, or line HHA-114-1124-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-11459057, or HHA-114-1124-derived pepper seed to yield additional hybrid PX-11459057, or line HHA-114-1124-derived pepper seed; (d) growing said additional hybrid PX-11459057, or line HHA-114-1124-derived pepper seed of step (c) to yield additional hybrid PX-11459057, or line HHA-114-1124-derived pepper plants; and (e) repeating the crossing and growing steps of (c) and (d) to generate at least a first further hybrid PX-11459057, or line HHA-114-1124-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 27, further comprising: (f) crossing the further hybrid PX-11459057, or HHA-114-1124-derived pepper plant with a second pepper plant to produce seed of a hybrid progeny plant.
30. A plant part of the plant of claim 7.
31. The plant part of claim 30, further defined as a leaf, an ovule, pollen, a fruit, or a cell.
32. 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.
33. A method of producing a pepper fruit 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.
Description
DETAILED DESCRIPTION OF THE INVENTION
(1) The invention provides methods and compositions relating to plants, seeds and derivatives of pepper hybrid PX-11459057, pepper line HHA-114-1124 and pepper line HHA-114-1030.
(2) Hybrid PX-11459057 is a hot habanero pepper with typical habanero fruit type, typical habanero shape, smell, pungency, and flavor. It has higher yield, mainly due to more fruits per plant, approximately 4 times more than the standard O.P. variety Habanero. This hybrid is grown primarily in the U.S. for harvest as orange fruits for fresh market, with secondary use as a processing pepper for whole pack, dice, salsa, and hot sauce. It exhibits excellent fresh and processed fruit quality due to its orange color and high capsaicin content.
(3) The fruits of PX-11459057 average 6.2 cm long3.5 cm wide, with typical habanero shape and smell. Immature fruit color is green, and fruits mature to orange. Fruit pungency is hot, around 8,000 Scoville Heat Units (fresh basis). It has higher yield of 812 g/plant, compared to the O.P. variety Habanero at 197 g/plant mainly due to more fruits per plant (64 fruits vs. 19 fruits). Fruit weight was not as heavy as the O.P. Habanero (11 g vs 12 g). Plant height averages 85 cm, with a semi-spreading plant habit. It is ready to harvest at the orange stage at approximately 105 days after transplanting, compared to 125 days for the standard O.P. variety Habanero. PX-11459057 is well-adapted to the main habanero growing areas of the U.S. and Mexico, with excellent fresh and processed fruit quality.
(4) Hybrid PX-11459057 is a typical habanero. The fruits average 6.0 cm long4.4 cm wide, with typical habanero shape and smell. Immature color is a medium green, and fruits mature to a dull orange. Fruit pungency was measured by HPLC using fruit grown in Woodland during the summer 2009 growing season at 114,450 Scoville Heat Units (dry basis). It has higher yield (869 g/plant, compared to the hybrid Chichen Itza at 674 g/plant) mainly due to heavier fruit weight (13.8 vs 11.8 g/fruit). Fruit number per plant was similar to the hybrid Chichen Itza (63.2 vs 57.4, respectively). Plant height averaged 95 cm, with a semi-spreading plant habit. It matures to orange in approximately 107 days after transplanting, compared to 126 days for the standard O.P. variety Habanero. PX-11459057 is resistant to Tobamovirus P0, and exhibits excellent tolerance to gemini viruses compared to the standard O.P. variety Habanero.
A. Origin and Breeding History of Pepper Hybrid PX-11459057
(5) The parents of hybrid PX-11459057 are HHA-114-1124 and HHA-114-1030. 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-11459057, Pepper Line HHA-114-1124 and Pepper Line HHA-114-1030
(6) In accordance with one aspect of the present invention, there is provided a plant having the physiological and morphological characteristics of pepper hybrid PX-11459057 and the parent lines thereof. A description of the physiological and morphological characteristics of such plants is presented in Tables 1-3.
(7) TABLE-US-00001 TABLE 1 Physiological and Morphological Characteristics of Hybrid PX-11459057 CHARACTERISTIC PX-11459057 HABANERO 1. Species C. chinense C. chinense 2. Maturity (in region of best adaptability) days from transplanting until 45 95 mature green stage days from transplanting until 91 115 mature red or yellow stage days from direct seeding until 88 138 mature green stage days from direct seeding until 134 158 mature red or yellow stage 3. Plant habit spreading compact attitude upright/erect upright/erect (De Cayenne, Doux trs long des Landes, Piquant d'Algrie) plant height 57.4 cm 64.1 cm plant width 63 cm 64.1 cm length of stem from cotyledon 16.3 cm 11.2 cm to first flower length of the third internode 45 mm 51 mm (from soil surface) length of stem medium (Belsir, short Lamuyo) shortened internode (in upper present present part) (Fehr, Kalocsai 601, Kalocsai 702) number of internodes between more than three more than three the first flower and the (Kalocsai 702) shortened internodes stem: hairiness of nodes absent or very weak weak (Arlequin) height medium (HRF) tall basal branches many (4+) few (2-3) branch flexibility willowy (Cayenne Long willowy Red) stem strength (breakage strong strong resistance) 4. Leaf length of blade short (De Cayenne, medium Szentesi cseresznye) width of blade narrow (De Cayenne, broad Pusztagold, Szentesi cseresznye) width 40 mm 60.6 mm length 68 mm 92.5 mm petiole length 14.3 mm 37.9 mm color dark green medium green color (RHS Color Chart value) 137A 137B intensity of green color dark (Dolmy, Tinto) medium mature leaf shape ovate (Balico, Sonar) broad/elliptic leaf and stem pubescence absent absent undulation of margin medium (Tenor) medium blistering strong (Greygo, PAZ strong pallagi) profile in cross section flat (De Cayenne, moderately concave Recio) glossiness medium (Alby, Eolo) strong 5. Flower peduncle: attitude semi-drooping (Blondy) semi-drooping flowers per leaf axil 2 2.2 calyx lobes 6 5.8 petals 5.8 6.2 diameter 13.8 mm 15.4 mm corolla color white white corolla throat markings white white style length exceeds stamen same as stamen self-incompatibility present absent 6. Fruit group Habanero (Scotch Habanero (Scotch Bonnet) Bonnet) color (before maturity) green (California green wonder, Lamuyo) intensity of color (before medium dark maturity) immature fruit color medium green dark green immature fruit color (RHS 144A 143A Color Chart value) attitude/position drooping/pendent (De drooping/pendent Cayenne, Lamuyo) length medium (Fehr, short Lamuyo) diameter medium (Doux italien, medium Corno di toro) ratio length/diameter medium (Adra, Cherry very small Sweet, Daniel, Delphin, Edino) calyx diameter 11.8 mm 12 mm fruit length 51.5 mm 39.6 mm fruit diameter at calyx 18.7 mm 19.9 mm attachment fruit diameter at mid-point 25.8 mm 28.7 mm flesh thickness at mid-point 1.3 mm 1.5 mm average number of fruits per 49.5 26.6 plant % large fruits 4.30 (weight range: 11 35.70 (weight range: 11 to 20) to 20) % medium fruits 41.20 (weight range: 6 58.30 (weight range: 6 to 10) to 10) % small fruits 54.50 (weight range: 1 6 (weight range: 1 to 5) to 5) average fruit weight 5.8 gm 6.7 gm fruit shape (longitudinal moderately triangular cordate/heart-shaped section) (Fehr, Marconi) fruit shape (cross section, at quadrangular circular level of placenta) sinuation of pericarp at basal strong (Alfa) medium part sinuation of pericarp excluding strong (De Cayenne, strong basal part Doux italien) texture of surface strongly wrinkled/rough slightly wrinkled (Sierra Nevada) color (at maturity) orange (Ariane) orange intensity of color (at maturity) medium medium mature fruit color orange orange mature fruit color (RHS Color 24B N25B Chart value) glossiness strong (Doux italien, very strong/shiny Trophy) stalk cavity absent (Corinto, Corno absent di toro, Sweet banana, Sucette de Provence) pedicel length 33 mm 31.9 mm pedicel thickness 1.4 mm 1.6 mm pedicel shape curved curved pedicel cavity absent absent stalk: length long (De Cayenne, long Sierra Nevada, Sweet banana) stalk: thickness thin (Sweet banana) thin base shape rounded rounded shape of apex moderately acute very acute/pointed shape Oblate (Sunnybrook) Oblate (Sunnybrook) set scattered scattered depth of interloculary grooves deep (Majister, Surpas) medium number of locules predominantly four or predominantly four or more (Palio, PAZ more szentesi) % fruits with one locule 0% 0% % fruits with two locules 0% 0% % fruits with three locules 0% 6.70% % fruits with four locules 93.30% 86.70% % fruits with five locules 6.70% 6.70% average number of locules 4.1 4 thickness of flesh very thin (De Cayenne, thin Macska srga, Petit marseillais, Recio) calyx: aspect enveloping (cup- enveloping (cup- shaped) (De Cayenne, shaped) Sweet banana) pungency hot hot capsaicin in placenta present (De Cayenne) present mg Capsaicin per gram dry fruit 8.7 mg 10.9 mg Scoville Units (dry fruit) 136,915 165,336 flavor strong pepper flavor mild pepper flavor glossiness shiny shiny 7. Seed seed cavity length 45.6 mm 29.9 mm seed cavity diameter 21.8 mm 23.6 mm placenta length 19.8 mm 16.2 mm number of seeds per fruit 66.8 166 grams per 1000 seeds 3 gm 3 gm color yellow yellow 8. Anthocyanin coloration of: seedling hypocotyl moderate moderate stem weak weak node weak weak stem: intensity of anthocyanin very weak weak coloration of nodes leaf absent absent pedicel absent absent calyx absent absent anther present (Lamuyo) present fruit coloration absent (Lamuyo) absent beginning of flowering (1.sup.st late late flower on 2.sup.nd flowering node) (Daniel, Piquant d'Algrie, Zingaro) time of maturity medium (Lamuyo, very late Latino, Sonar) *These are typical values. Values may vary due to environment. Other values that are substantially equivalent are also within the scope of the invention.
(8) TABLE-US-00002 TABLE 2 Physiological and Morphological Characteristics of Line HHA-114-1124 CHARACTERISTIC HHA-114-1124 HABANERO 1. Species C. chinense C. chinense 2. Maturity (in region of best adaptability) days from transplanting until 88 88 mature green stage days from transplanting until 116 114 mature red or yellow stage days from direct seeding until 139 139 mature green stage days from direct seeding until 175 165 mature red or yellow stage 3. Plant habit compact compact attitude upright/erect upright/erect (De Cayenne, Doux trs long des Landes, Piquant d'Algrie) plant height 47.5 cn 60.5 cm plant width 57.8 cm 60.5 cm length of stem from cotyledon 10.5 cm 14.4 cm to first flower length of the third internode 40.8 mm 39.8 mm (from soil surface) length of stem medium (Belsir, medium Lamuyo) shortened internode (in upper present present part) (Fehr, Kalocsai 601, Kalocsai 702) number of internodes between one to three (Fehr) one to three the first flower and the shortened internodes stem: hairiness of nodes weak weak (Andevalo, Clovis) height medium (HRF) medium basal branches many (4+) many (4+) branch flexibility willowy (Cayenne Long willowy Red) stem strength (breakage strong strong resistance) 4. Leaf length of blade medium (Atol, Blondy, long Marconi, Merit, Anthea) width of blade broad (California broad wonder, Golden calwonder, Seifor, Solario) width 60.4 mm 68.4 mm length 105.5 mm 115.4 mm petiole length 21.3 mm 52.0 mm color dark green dark green color (RHS Color Chart value) 146A 146A intensity of green color dark (Dolmy, Tinto) dark mature leaf shape lanceolate (Diavolo, ovate Recio) leaf and stem pubescence light light undulation of margin strong (Sucette de strong Provence, Tosca) blistering very strong (Florian) strong profile in cross section moderately concave moderately concave (Doux italien, Favolor) glossiness strong (Andevalo, strong Floridor) 5. Flower peduncle: attitude semi-drooping (Blondy) semi-drooping flowers per leaf axil 2.6 3 calyx lobes 6.3 6 petals 6 6.2 diameter 15.1 mm 16.5 mm corolla color white white corolla throat markings yellow yellow anther color purple purple style length less than stamen same as stamen self-incompatibility absent absent 6. Fruit group Habanero (Scotch Habanero (Scotch Bonnet) Bonnet) 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 144A Color Chart value) attitude/position drooping/pendent (De drooping/pendent Cayenne, Lamuyo) length short (Delphin, Petit short carr doux) diameter medium (Doux italien, medium Corno di toro) ratio length/diameter medium (Adra, Cherry medium Sweet, Daniel, Delphin, Edino) calyx diameter 12.6 mm 12.4 mm fruit length 44.6 mm 46.4 mm fruit diameter at calyx 26.8 mm 23.3 mm attachment fruit diameter at mid-point 27.9 mm 31.2 mm flesh thickness at mid-point 1.4 mm 1.7 mm average number of fruits per 52.2 47 plant % large fruits 33 (weight range: 11 23.60 (weight range: 13.5 to 20) to 20) % medium fruits 51.50 (weight range: 6 61.40 (weight range: 7 to 10) to 12.5) % small fruits 15 (weight range: 1 28 (weight range: 1 to 5) to 6) average fruit weight 8.0 gm 8.9 gm fruit shape (longitudinal trapezoidal (Delta, moderately triangular section) Piperade) fruit shape (cross section, at circular (Cherry Sweet, circular level of placenta) Doux trs long des Landes) sinuation of pericarp at basal weak (Donat) weak part sinuation of pericarp excluding weak (Clovis, Sonar) weak basal part texture of surface strongly wrinkled/rough strongly wrinkled/rough (Sierra Nevada) color (at maturity) orange (Ariane) orange intensity of color (at maturity) medium medium mature fruit color orange orange mature fruit color (RHS Color N25B N25B Chart value) glossiness very strong/shiny very strong/shiny (Floridor, Kappy) stalk cavity absent (Corinto, Corno absent di toro, Sweet banana, Sucette de Provence) pedicel length 33.5 mm 33.4 mm pedicel thickness 2.0 mm 1.7 mm pedicel shape curved curved pedicel cavity present present stalk: length long (De Cayenne, long Sierra Nevada, Sweet banana) stalk: thickness thin (Sweet banana) thin base shape cupped rounded shape of apex moderately acute moderately acute shape Conical (Pimento) Oblate (Sunnybrook) set scattered scattered depth of interloculary grooves deep (Majister, Surpas) deep number of locules predominantly three predominantly four or (Century) more % fruits with one locule 0% 0% % fruits with two locules 0% 0% % fruits with three locules 66.70% 6.70% % fruits with four locules 23% 70% % fruits with five locules 10% 23.40% average number of locules 3.4 4.2 thickness of flesh thin thin (Bann, Carr doux extra htif, Dous trs long des Landes) calyx: aspect non-enveloping/saucer- non-enveloping/saucer- shaped shaped (Lamuyo, Sonar) pungency hot hot capsaicin in placenta present (De Cayenne) present mg Capsaicin per gram dry fruit 10.8 mg 12.9 mg Scoville Units (dry fruit) 172,287 160,356 flavor strong pepper flavor strong pepper flavor glossiness shiny shiny 7. Seed seed cavity length 25.7 mm 36.6 mm seed cavity diameter 20.4 mm 25.5 mm placenta length 19.2 mm 17.7 mm number of seeds per fruit 26.6 113.5 grams per 1000 seeds 3.5 gm 3.9 gm color yellow yellow 8. Anthocyanin coloration of: seedling hypocotyl strong (Lamuyo) strong stem moderate weak node weak moderate stem: intensity of anthocyanin medium (Clovis, medium coloration of nodes Lamuyo, Sonar) leaf strong strong pedicel absent absent calyx absent absent anther present (Lamuyo) present fruit coloration absent (Lamuyo) absent beginning of flowering (1.sup.st late medium flower on 2.sup.nd flowering node) (Daniel, Piquant d'Algrie, Zingaro) time of maturity very late (Cancun, early California wonder) *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
(9) One aspect of the current invention concerns methods for producing seed of pepper hybrid PX-11459057 involving crossing pepper lines HHA-114-1124 and HHA-114-1030. Alternatively, in other embodiments of the invention, hybrid PX-11459057, line HHA-114-1124, or line HHA-114-1030 may be crossed with itself or with any second plant. Such methods can be used for propagation of hybrid PX-11459057 and/or the pepper lines HHA-114-1124 and HHA-114-1030, or can be used to produce plants that are derived from hybrid PX-11459057 and/or the pepper lines HHA-114-1124 and HHA-114-1030. Plants derived from hybrid PX-11459057 and/or the pepper lines HHA-114-1124 and HHA-114-1030 may be used, in certain embodiments, for the development of new pepper varieties.
(10) 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-11459057 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.
(11) 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.
(12) 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.
(13) 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-11459057 and/or pepper lines HHA-114-1124 and HHA-114-1030 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
(14) As described above, hybrid PX-11459057 exhibits desirable traits, as conferred by pepper lines HHA-114-1124 and HHA-114-1030. The performance characteristics of hybrid PX-11459057 and pepper lines HHA-114-1124 and HHA-114-1030 were the subject of an objective analysis of the performance traits relative to other varieties. The results of the analysis are presented in Table 3.
(15) TABLE-US-00003 TABLE 3 Performance data for Hybrid PX-11459057 and comparative varieties. Color Maturity Yld Len Wid Av. wt % dry SHU Variety mature (DAT) g/plt cm cm g/fruit matter Dry basis PX-11459057 orange 105 869 6.0 4.4 13.8 10.6 114,450 Chichen Itza orange 108 674 5.4 3.7 11.8 10.9 214,727 Caribbean Red red 133 204 5 3.2 8.0 11.3 264,832
E. Further Embodiments of the Invention
(16) 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.
(17) 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.
(18) 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.
(19) 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.
(20) 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.
(21) 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.
(22) 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.
(23) 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.
(24) 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.
(25) 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
(26) 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.
(27) 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.
(28) 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.
(29) 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.
(30) 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.
(31) 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).
(32) 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).
(33) 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.
(34) 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., wunl, 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).
(35) 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.
(36) 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.
(37) 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
(38) 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:
(39) 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.
(40) Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (F.sub.1), 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.
(41) Crossing: The mating of two parent plants.
(42) Cross-pollination: Fertilization by the union of two gametes from different plants.
(43) Diploid: A cell or organism having two sets of chromosomes.
(44) 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.
(45) Enzymes: Molecules which can act as catalysts in biological reactions.
(46) F.sub.1 Hybrid: The first generation progeny of the cross of two nonisogenic plants.
(47) Genotype: The genetic constitution of a cell or organism.
(48) Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.
(49) Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.
(50) 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.
(51) Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.
(52) 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.
(53) 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.
(54) Regeneration: The development of a plant from tissue culture.
(55) 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.
(56) Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.
(57) 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.
(58) Substantially Equivalent: A characteristic that, when compared, does not show a statistically significant difference (e.g., p=0.05) from the mean.
(59) 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.
(60) Transgene: A genetic locus comprising a sequence which has been introduced into the genome of a pepper plant by transformation.
H. Deposit Information
(61) A deposit of pepper hybrid PX-11459057 and inbred parent line HHA-114-1124, 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 deposit were Oct. 30, 2014 and Jul. 16, 2014, respectively. The accession numbers for those deposited seeds of pepper hybrid PX-11459057 and inbred parent line HHA-114-1124 are ATCC Accession No. PTA-121689 and ATCC Accession No. PTA-121393, 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.
(62) 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.
(63) All references cited herein are hereby expressly incorporated herein by reference.