Indeterminate and photoperiod-insensitive IIPG-7 and IIPG-11 pigeonpea germplasm

Abstract

The present disclosure relates to two pigeonpea breeding lines designated as IIPG-7 and IIPG-11. The disclosure relates to the seeds of breeding lines IIPG-7 and IIPG-11, to the plants of pigeonpea breeding lines IIPG-7 and IIPG-11, to the plant parts of pigeonpea breeding lines IIPG-7 and IIPG-11, and to methods for producing progeny of pigeonpea breeding lines IIPG-7 and IIPG-11. The disclosure also relates to pigeonpea breeding lines, and plant parts derived from pigeonpea breeding lines IIPG-7 and IIPG-11. The disclosure also relates to methods for producing other pigeonpea cultivars, breeding lines, or plant parts derived from pigeonpea breeding lines IIPG-7 and IIPG-11, and to the pigeonpea plants, varieties, and their parts derived from use of those methods. The disclosure further relates to hybrid / sybrid pigeonpea seeds, plants, and plant parts produced by crossing pigeonpea breeding lines IIPG-7 and IIPG-11 with another pigeonpea cultivar and / or germplasm.

Description

Attorney Docket No. 71900-435893INDETERMINATE AND PHOTOPERIOD-INSENSITIVE IIPG-7AND IIPG-11 PIGEONPEA GERMPLASMCross-Reference to Related Applications

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 779,803, filed March 28, 2025, the entire contents of which are incorporated herein by reference.Federally Sponsored Research

[0002] This invention was made with government support under grant number 7006389 awarded by the United States Department of Agriculture (USDA) National Institute of Food and Agriculture. The government has certain rights in the invention.Background

[0003] Pigeonpea [Cajanus cajan (L.) Mill.] is a diploid (2n = 2x = 22) legume cultivated in tropical and sub-tropical countries in Africa, America, and Asia. This crop has multiple uses, including as a source of protein (20%-25%) in the human diet, as a fodder for fuel and animal feed, as a cover crop fixing nitrogen in soils to improve their structure and fertility, and as medicinal uses. In Puerto Rico, pigeonpea is one of the most important legumes. It is harvested and consumed as immature seed, with a peak of consumption during the Thanksgiving, Christmas, and New Year holidays. The economic contribution of pigeonpea was estimated to be $434,000 in 2017. In the continental United States, pigeonpea has been explored as a forage that can be used as a protein grain for livestock or as a cover crop to reduce soil erosion.

[0004] Pigeonpea may have self- or cross-pollination. The ranges of cross-pollination vary from 3% to 40%, and it is carried out mostly by the bees Apis spp. and Megachile spp. (Hymenoptera: Megachilidae). Also, this legume has papilionaceous flowers or is completely bisexual and zygomorphic. According to the flowering habit, pigeonpea genotypes can be divided into determinate, semi-determinate, and indeterminate. Most cultivated pigeonpeas, and many of their wild relatives [e.g., C. cajanifolius (Haines) Maesen], possess an indeterminate flowering habit. It is believed that genotypes with a determinate flowering habit have been selected or developed in the process of domestication. Also, its photoperiod insensitivity is mostly observed in earlier-maturity genotypes that were developed in pigeonpea breeding programs. The genetics for the indeterminate flowering habit and photoperiod sensitivity have been reported to be inherited by one or multiple dominant genes. The CcTFLI locus, located on chromosome 3, was reported as the candidate gene for flowering habit, while the CCTfamily of genes mostly observedAttorney Docket No. 71900-435893on chromosomes 3, 7, and 11 was reported to be responsible for photoperiod sensitivity in pigeonpea.

[0005] The International Crops Research Institute for the Semi-Arids Tropics (ICRISAT) has made efforts to identify genotypes that are photoperiod insensitive. For instance, more than 10 genotypes (e.g., ICP 1143 Sei, ICP 1146 Sei, ICP 7028 Sei, and ICP 11641 Sei) were reported to have indeterminate flowering habit and photoperiod insensitivity in field evaluations of more than 10,000 genotypes from the pigeonpea worldwide collection conserved at ICRISAT. More recently, researchers tested 36 genotypes, and only ICPL 20325 was photoperiod insensitive and had indeterminate flowering habit. In contrast, and to the best of our knowledge, there are no germplasms or cultivars with both traits that have been released in the Caribbean basin. All local cultivars with indeterminate flowering habit planted in Puerto Rico (e.g., ‘Ariel’, ‘Blanco Yauco’, ‘Guerrero’, ‘Pinto Berrocales’, ‘Pinto Original’, and ‘Super Pinto’) are photoperiod sensitive. This means that, when plants are exposed to long day conditions (>12 h), and generally planted between February and July in Puerto Rico, (USDA plant hardiness zones 12 and 13), they do not initiate flowering until short-day conditions (<12 h) occur. After further evaluation, the time to develop flowers and produce seeds extended up to 208 and 268 days after planting (DAP), respectively. For this reason, indeterminate cultivars are only recommended to be planted during short-day conditions (between October and December in Puerto Rico, zones 12 and 13) where the harvesting period does not exceed 150 DAP.

[0006] There is a need to have indeterminate cultivars that can be planted during long-day conditions to guarantee the harvest of green peas by November through January and year-round. This would contribute to food security and may increase the profits of growers in Puerto Rico because there would be less dependence on imported canned pigeonpeas. Moreover, the use of indeterminate cultivars may have additional agronomic advantages, such us multiple branching, which increases flowering and pod production, and higher seed yields, such as those observed for ‘Ariel’, ‘Guerrero’, ‘ICP 7035’, and ICP 7193 under short day conditions.

[0007] Also, the control of important pests like tobacco budworm [Heliothis virescens (F.) Lepidoptera: Noctuidae] or pod borers (Helicoverpa spp. Lepidoptera: Noctuidae) may be easier in indeterminate genotypes that have long fruiting branches and loose inflorescences than in determinate genotypes that possess clustered racemes. However, indeterminate genotypes are usually harvested later than determinate genotypes. Thus, the plants and seeds may be exposed to attack by bruchids (Callosobruchus spp. Coleoptera: Bruchidae), larvae of Lepidopteran species, infection of fungal species (Aspergillus spp., Botryodiplodia theobromae Pat., Fusarium spp., and Phomopsis spp., and / or adverse environmental effects (e.g., waterlogging) that canAttorney Docket No. 71900-435893significantly reduce seed yield and quality. Furthermore, the costs for multiple pesticide applications, labor, and / or extra agronomic practices may increase if the crop is in the field for extended periods. Currently, the only determinate cultivars that can be planted under long-day conditions in Puerto Rico are ‘Lazaro’ and ‘Isabella’ because they are photoperiod insensitive. Thus, the development of pigeonpea germplasm that have indeterminate flowering habit, are photoperiod insensitive, and are adapted to the conditions of Puerto Rico is important. Our objective was to develop improved breeding lines IIPG-7 (Reg. no. GP-326, PI 704909) and IIPG-11 (Reg. no. GP-327, PI 704910) that can be used directly as cultivars or in breeding programs to incorporate these important traits into pigeonpea cultivars.Brief Summary of the Disclosure

[0008] In one embodiment described herein is a plant, plant part, seed, or plant cell of pigeonpea breeding line IIPG-7, representative seed of said breeding line having been deposited under ATCC accession number PTA-127908.

[0009] One aspect of the present disclosure is a breeding line of pigeonpea IIPG-7 plant, plant part, or seed of said breeding line of pigeonpea IIPG-7 described herein. In another aspect is the breeding line of pigeonpea IIPG-7, wherein the breeding line of pigeonpea IIPG-7 has at least one phenotype selected from the group consisting of: a) indeterminate flowering habit, b) photoperiod insensitivity, c) higher accumulative yield per year compared to indeterminate photoperiod-sensitive pigeonpea cultivars, d) shorter time to harvest compared to indeterminate pigeonpea cultivars, e) red and yellow flowers, f) red pods with green speckles, and g) completely green seeds at an immature stage. In one aspect, the breeding line of pigeonpea IIPG-7 reaches harvest maturity at about 89-172 days after planting (DAP).

[0010] In another aspect is a tissue culture comprising at least one cell or protoplasm of the plant or plant part of the breeding line of pigeonpea IIPG-7.

[0011] In another aspect is a germplasm of a breeding line of pigeonpea IIPG-7. In another aspect, the germplasm produces seeds in both long-day and short-day conditions.

[0012] In another aspect is a method for producing a breeding line of pigeonpea IIPG-7 seed, comprising fertilizing the breeding line of pigeonpea IIPG-7 and harvesting the resultant breeding line of pigeonpea IIPG-7 seed. In one aspect, the breeding line of pigeonpea IIPG-7 seed produced by the method described herein results from a seed of breeding line of pigeonpea I IPG-7 seed as a result of self-fertilization. In one aspect, the breeding line of pigeonpea IIPG-7 has a 100-seed weight between 12 and 16 grams (g).Attorney Docket No. 71900-435893

[0013] In one embodiment described herein is a plant, plant part, seed, or plant cell of pigeonpea breeding line I IPG-11 , representative seed of said breeding line having been deposited under ATCC accession number PTA-127909.

[0014] In one aspect is a breeding line of pigeonpea 11 PG-11 plant, plant part, or seed of said breeding line of pigeonpea IIPG-11 described herein. In another aspect is the breeding line of pigeonpea 11 PG-11 , wherein the breeding line of pigeonpea I IPG- 11 has at least one phenotype selected from the group consisting of: a) indeterminate flowering habit, b) photoperiod insensitivity, c) higher accumulative yield per year compared to indeterminate photoperiodsensitive pigeonpea cultivars, d) shorter time to harvest compared to indeterminate pigeonpea cultivars, e) red and yellow flowers, f) green pods with brown speckles, and g) completely green seeds at an immature stage. In one aspect, the breeding line of pigeonpea IIPG-11 reaches harvest maturity at about 88-165 days after planting (DAP).

[0015] In another aspect is a tissue culture comprising at least one cell or protoplasm of the plant or plant part of the breeding line of pigeonpea 11 PG- 11.

[0016] In another aspect is a germplasm of a breeding line of pigeonpea 11 PG- 11. In another aspect, the germplasm produces seeds in both long-day and short-day conditions.

[0017] In another aspect is a method for producing a breeding line of pigeonpea IIPG-11 seed, comprising fertilizing the breeding line of pigeonpea I IPG-11 and harvesting the resultant breeding line of pigeonpea 11 PG-11 seed. In one aspect, the breeding line of pigeonpea 11 PG-11 seed produced by the method described herein results from a seed of breeding line of pigeonpea 11 PG- 11 seed as a result of self-fertilization. In one aspect, the breeding line of pigeonpea I IPG-11 has a 100-seed weight between 13 and 17 grams (g).

[0018] One or more embodiments or aspects may be incorporated in a different embodiment or aspect although not specifically described. That is, all embodiments and aspects can be combined in any way or combination.Brief Description of the Figures

[0019] The above summary, as well as the following detailed description of embodiments of the invention, will be better understood when read in conjunction with the appended drawings. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities shown. In the drawings:

[0020] Figure 1 shows pods and mature seeds of IIPG-7 (Fig. 1A and Fig. 1C) and IIPG-11 (Fig. 1 B and Fig. 1 D) pigeonpea [Cajanus cajan (L.) Mill.] breeding lines. Fig. 1 A shows the pods of pigeonpea breeding line IIPG-7. The pods are largely red with variable green mottling. IfAttorney Docket No. 71900-435893depicted in black and white, the pods will appear as largely dark grey with lighter grey specks. As shown in Fig. 1B are the pods of pigeonpea breeding line IIPG-11 , which is a mix of red and green speckles. If depicted as a black and white figure, the pod will appear as largely light grey with darker grey mottling. Seeds of both lines are green at immature stages and mature seeds of IIPG-7 are tan with reddish spots (Figure 1C), which, in black and white, appear as lighter grey with darker grey spots, in contrast to mature seeds of IIPG-11, which are reddish at harvest (Figure 1D), which, in black and white, appear darker grey..

[0021] Figure 2 depicts pigeonpea [Cajanus cajan (L.) Mill.] breeding line I IPG-11 flowering. The flowers have red color petals on the outside and yellow on the inside. If shown in black and white, the outer petals will therefore appear a darker shade of grey, and the inside, a lighter shade of grey.DETAILED DESCRIPTION OF THE DISCLOSURE

[0022] The present disclosure relates to two novel pigeonpea (Cajanus cajan (L.) Mill.) breeding lines designated as IIPG-7 and IIPG-11, their seeds, plants, plant parts, and hybrids. Also provided herein are methods for producing IIPG-7 and 11 PG-11.

[0023] It is to be understood that the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. As used herein, the terms “including,” “comprising,” or “having” and variations thereof herein are meant to encompass the items listed thereafter and equivalents thereof as well as additional items.

[0024] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise.

[0025] As used herein, the term “plant parts” includes, without limitation, plant protoplasts, plant cell tissue cultures from which multiple plant species including legumes can be regenerated, plant calli, plant clumps, plant cells, embryos, pollen, ovules, pericarp, seed, flowers, florets, heads, spikes, stems, stalks, leaves, roots, root tips, anthers, and the like. When indicating that a plant is crossed or selfed this indicates that any plant part of the plant can be used. For instance, the plant part does not need to be attached to the plant during the crossing or selfing, only the pollen might be used.

[0026] It also is understood that any numerical range recited herein includes all values from the lower value to the upper value. For example, if a concentration range is stated as 1 % to 50%, it is intended that values such as 2% to 40%, 10% to 30%, or 1% to 3%, etc., are expresslyAttorney Docket No. 71900-435893enumerated in this specification. These are only examples of what is specifically intended, and all possible combinations of numerical values between and including the lowest value and the highest value enumerated are to be considered to be expressly stated in this application.

[0027] Pigeonpea breeding lines or cultivars should be homozygous and easily reproducible. Crosses between two different homozygous lines may produce uniform hybrid plants that may be heterozygous for many gene loci, while crosses between heterozygous plants may result in genetically diverse plants that are not uniform. However, plants that are self-pollinated and selected over many generations may become homozygous at almost all genetic loci and thus produce uniform populations of true breeding progeny. The term “homozygous plant” as used herein is a plant with homozygous genes at > 90% or more of its loci.

[0028] The type of breeding or selection methods depends on the mode of plant reproduction, the trait(s) being improved, and the type of variety used commercially (e.g., Fi hybrid variety, pure line variety, etc.). For highly heritable traits, a choice of superior individual plants evaluated at a single location will be effective, whereas for traits with low heritability, selection may be based on mean values obtained from replicated evaluations of families of related plants. Popular selection methods which may be used for the pigeonpea cultivars described herein include pedigree selection, modified pedigree selection, gamete selection, mass selection, backcrossing, and recurrent selection.

[0029] Plant breeding methods may also include analysis and characterization of plant genome using well known lab techniques, including, but not limited to, starch gel electrophoresis, isozyme electrophoresis, restriction fragment length polymorphism (RFLPs), randomly amplified polymorphic DNAs (RAPDs), DNA amplification fingerprinting (DAF), simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). Molecular markers associated with alleles of interest may also be used to select for plants that contain those alleles during crossing or breeding programs.

[0030] Molecular markers may be used during the breeding process for the selection of qualitative and / or quantitative traits. For example, markers closely linked to alleles or markers containing sequences within the actual alleles of interest can be used to select plants that contain the alleles of interest during a crossing or backcrossing breeding program. The markers may also be used to select for the genome of the recurrent parent and against the markers of the donor parent. Using this procedure can minimize the amount of genome from the donor parent that remains in the selected plants. It can also be used to reduce the number of crosses back to the recurrent parent needed in a backcrossing program.DefinitionsAttorney Docket No. 71900-435893

[0031] The following definitions are meant to clarify, but not limit, the terms defined. If a particular term used herein is not specifically defined, such term should not be considered indefinite. Rather, terms are used within their accepted meanings.Definitions of plant characteristics.

[0032] In the description and examples that follow, a number of terms are used. To provide a clear and consistent understanding of the specification and claims, including the scope to be given such terms, the following definitions are provided.

[0033] Allele: Any of one or more alternative forms of a genetic 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.

[0034] Backcrossing: A process in which a breeder repeatedly crosses hybrid progeny, for example a first generation hybrid (Fi), 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.

[0035] Breeding lines: A group of identical pure-breeding diploid or polyploid organism, distinguished from other individuals of the same species by some unique genotype of phenotype.

[0036] Bulk: When F2 and subsequent generations are harvested in mass or as bulks to raise the next generation.

[0037] Cotyledon : A cotyledon is a type of seed leaf. The cotyledon contains the food storage tissues of the seed.

[0038] Crossing: The mating of two parent plants.

[0039] Cross-pollination: Fertilization by the union of two gametes from different plants.

[0040] Crop-performance: Used synonymously with plant performance and refers to of how well a plant grows under a set of environmental conditions and cultivation practices. Crop performance can be measured by any metric associates with a crop's productivity (e.g., yield), appearance and / or robustness (e.g., color, morphology, height, biomass, maturation rate), product quality (e.g., seed protein content, seed oil content, seed carbohydrate content, etc.), cost of goods sold (e.g., the cost of creating a seed, plant, or plant product in a commercial, research, or industrial setting) and / or a plant's resistance / tolerance to disease (e.g., a response associated with deliberate or spontaneous infection by a pathogen), pests, microbes, fungi, and / or environmental stress (e.g., drought, flooding, low nitrogen or other soil nutrients, wind, hail, temperature, day length, etc.). Crop performance can also be measured by determining a crop's commercial value and / or by determining the likelihood that a particular inbred, hybrid, or variety will become a commercial product, and / or by determining the likelihood that the offspring of anAttorney Docket No. 71900-435893inbred, hybrid, or variety will become a commercial product. Crop performance can be a quantity (e.g., the volume or weight of seed or other plant product measured in liters or grams) or some other metric assigned to some aspect of a plant that can be represented on a scale (i.e., assigning a 1 to 10 value to a plant based on its disease tolerance).

[0041] Cultivar: A plant that has been selectively bred or developed by humans for desired characteristics, such as size, color, taste, or disease resistance, and can be reproduced true to type through various propagation methods.

[0042] Days after planting (DAP): Alternatively, “days to maturity” (DAM). The average number of days from the time a seed is sown or a seedling is transplanted to the first harvest. Environmental conditions like temperature, soil type, and sunlight play a role in plant growth and therefore, maturity.

[0043] Day-neutral: Plants that form flowers regardless of day length.

[0044] Determinate: Plants where the apical bud develops into the flower. The sequence of the inflorescence production is basipetal, and the flowers occur more or less in the same plane. The parent cultivar ‘ICPL 86012’ has determinate flowering habit and it is day neutral.

[0045] Diploid: A cell or organism having two sets of chromosomes.

[0046] Dry seed weight: The weight of seeds after all moisture has been removed. This metric is used to assess seed size, quality, and other characteristics.

[0047] Embryo: The embryo is the small plant contained within a mature seed.

[0048] 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 or a gene editing conferring male sterility.

[0049] Enzymes: Molecules which can act as catalysts in biological reactions.

[0050] Fi Hybrid: The first generation progeny of the cross of two non-isogenic plants. Similarly, an “P3” symbol, for example, denotes a generation resulting from the selfing of the F2generation along with selection for type and rogueing of off types. The “F” number is a term commonly used in genetics and designates the number of the filial generation. The “F3” generation denotes the offspring resulting from the selfing or self-mating of members of the generation having the next lower “F” number, that is, the “F2” generation.

[0051] Genotype: The genetic constitution of a cell or organism.

[0052] Haploid: A cell or organism having one set of the two sets of chromosomes in a diploid.

[0053] Harvest maturity: The stage of development when a crop is ready to be harvested, ensuring optimal quality, flavor, and shelf life for produce.Attorney Docket No. 71900-435893

[0054] Hybrid: An offspring of two animals or plants of different subspecies, breeds, varieties, species, or genera.

[0055] Indeterminate: Pigeonpea breeding lines of the present disclosure have the growth characteristic of being indeterminate plants. In this case, the inflorescence is large with a vegetative apical bud resulting in continuous growth. The flowers occur in axillar racemes spread over a considerable length of the stem. The parent cultivar ‘Guerrero’ and all cultivars with indeterminate flowering habit (e.g., ‘Ariel’, ‘Blanco Yauco’, ‘Kaki’, ‘Pinto Berrocales', ‘Super Pinto’, among others) are also indeterminate plants. Unfortunately, these cultivars have sensitivity to the photoperiod.

[0056] Linkage: A phenomenon wherein alleles on the same chromosome tend to segregate together more often than expected by chance if their transmission was independent.

[0057] Long-day: A long-day plant requires a short night to flower. These plants bloom when they receive more than 12 hours of light. Herein, long-day conditions mean from February to July in Puerto Rico, USDA Plant hardiness zone 12 and 13.

[0058] 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.

[0059] Pedigree: A record of ancestry or purity of breed.

[0060] Phenotype: The detectable characteristics of a cell or organism, which characteristics are the manifestation of gene expression.

[0061] Photoperiod: Photoperiod refers to the duration of light and darkness within a 24-hour period, and it is an environmental signal for plants influencing processes like flowering, dormancy, and seasonal adaptation.

[0062] Photoperiod insensitive: A plant’s ability to flower or develop regardless of the length of daylight and darkness. Also known as day-neutrality or day neutral plants.

[0063] Photoperiod sensitive: Also refers as delay to flower under long-days as compared to short-days. The growth and development of a plant, especially flowering, is influenced by the length of daylight and darkness in a 24-hour period.

[0064] Plant: A plant refers to a whole plant, any part thereof, or a cell or tissue culture derived from a plant, comprising any of whole plants, plant components or organs (e.g., leaves, stems, roots, etc.), plant tissues, seeds, embryos, pollen, plant cells, protoplasts and / or progeny of the same. A plant cell is a biological cell of a plant, taken from a plant or derived through culture of a cell taken from a plant.Attorney Docket No. 71900-435893

[0065] Plant height: Plant height is taken from the top of the soil to the top node of the plant and is measured in centimeters, meters, or feet.

[0066] Plant parts: Plant parts (or a pigeonpea plant, or a part thereof) includes but is not limited to protoplasts, cells, leaves, stems, roots, root tips, anthers, pistils, seed, grain, embryo, pollen, ovules, cotyledon, hypocotyl, pod, flower, shoot, tissue, petiole, cells, meristematic cells, stipules, vine, tendril, and the like.

[0067] Pod: It consists of the shell or wall (pericarp) and the seeds.

[0068] Progeny: Progeny includes an Fi plant produced from the cross of two plants where at least one plant includes pigeonpea cultivar and progeny further includes, but is not limited to, subsequent F2, F3, F4, F5, F6, F7, F8, F9, F and Fngenerational crosses with the recurrent parental line.

[0069] Self-pollination: The transfer of pollen from the anther to the stigma of the same plant.

[0070] Short-day: Also known as a ‘long-night’ plant. Short day plants form flowers only when day length is less than about 12 hours. Pigeonpea is typically a short-day plant, requiring a daylight length of 12.5 hours to initiate flowering and seed production. Herein, short-day conditions mean from October to December in Puerto Rico, zones 12 and 13. In contrast to the pigeonpea cultivars of the present disclosure, the parent cultivar ‘Guerrero’ and all cultivars with indeterminate flowering habit (e.g. ‘Ariel’, ‘Blanco Yauco’, ‘Kaki’, ‘Pinto Berrocales’, ‘Super Pinto’, among others) that have sensitivity to the photoperiod, and are recommended to be planted only under short-day conditions.

[0071] Sybrid population: This breeding method has been designed in such a way that portions of both additive and nonadditive genetic variances are exploited for enhancing the productivity of the often-cross-pollinated crops. The primary breeding tool in this technology is the insect-aided cross-pollination with no male-sterility system involved.

[0072]

[0073] 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.

[0074] Varietal: A naturally occurring genetic variation within a species, distinguished by specific characteristics.Typical pigeonpea characteristics.Typical Uses

[0075] Alternative names: Pigeon pea, Angola pea, Congo pea, dhal, no-eye pea, gungo pea, and red gram.Attorney Docket No. 71900-435893

[0076] Alternative scientific names: Cajanus indicus Spreng., Cajanus flavus DC., Cytisus cajanus L.

[0077] Cajanus cajan is grown as a pulse crop (crop harvested for dry seed) or eaten green as a vegetable. The grain is popularly consumed in India, Asia, Africa, and the Caribbean basin. India is the largest importer and producer, where seed is sold as dhal (dry split pea). Unprocessed seed should not be consumed by humans or livestock. The seed contains tannin and trypsin inhibitors (trypsin inhibitors are removed through cooking).

[0078] The protein content in split seeds is similar to soybean and ranges from 21-28% (Phatak et al., 1993). It is also widely used as a good source of dietary vitamins and minerals.

[0079] Cajanus cajan makes an excellent, high-protein forage for livestock. Crude protein ranges from 28-36% (Phatak et al., 1993). In Florida, plants yielded 3.1 tons / acre of biomass (Duke, 1983). Livestock may browse foliage, but damage to the branches may result, so continuous grazing should be avoided. Palatability has been reported to increase with age of the plant (Cook et al., 2005).

[0080] Cajanus cajan can be grown as a forage intercropped with sorghum and / or millet. The deep taproot of C. cajan draws water from deeper soil depths than most legumes, so will not interfere with the water uptake of other crops and grasses. It is not generally seeded as forage with other legumes like cowpea [Vigna unguiculata (L.) Walp], but mainly with grasses (Cook et al., 2005).

[0081] As a green manure it can fix about 62 lb. N / acre up to the time when pods are produced (Phatak et al., 1993).

[0082] Cajanus cajan has been used as a trap crop for Heliothis spp. (moth pests), in affected cotton (Mullen et al., 2003). A trap crop is a form of companion planting used to attract pests away from nearby crops. Plantings are also used as live fences and windbreaks in many regions.Typical characteristics.

[0083] In general, C. cajan is an erect perennial, warm season crop that is widely grown in the tropics and subtropics. It is a shrub that can grow to 12 ft tall (>3.5 m) under long-day conditions, but usually only reaches 3 to 6 ft (1 -2 m).

[0084] The ribbed stem grows upright and is covered in short, soft hairs (pubescent) and is woody at the base. Its deep tap root is fast-growing. The pubescent, stalked leaflets are 2 to 4 inches long (5-10 cm) and % to 11 / 2 inches wide (2-4 cm), with minute resinous glands underneath.Attorney Docket No. 71900-435893

[0085] The bi-laterally symmetrical, bell-shaped blooms are yellow or yellow and red, may be in pairs, and grow in the angle between stem and leaf on an unbranched inflorescence. The upper two lobes are united and two-toothed, and the lower lip is smaller and three-toothed. The lower bracts fall off early.

[0086] The two-valve, pointed seedpods are produced in clusters, and mottled red. They are 2 to 3.5 inches (5-9 cm) long,1Z> inch (12 mm) wide, flat, covered with soft hairs, and taper to a sharp point. The seeds can be green, red, or black at immature stages. The round or oval seeds may be red, black, light beige to dark brown or tan with red mottling at maturity. The 2 to 9-seeded pods do not shatter in the field.

[0087] Cajanus cajan has a deep taproot system that grows large, cylindrical nodules. This nodulation can be initiated by a variety of rhizobial strains (Allen and Allen, 1981).

[0088] The distribution of C. cajan is an old-world food crop that grows best in the tropics and subtropics but has also been successfully grown in the Southern United States, Hawaii, and Puerto Rico. It is uncertain whether C. cajan is native to Africa (Allen and Allen, 1981) or India (Duke, 1983). Nevertheless, the plant has been in cultivation in parts of Africa and India for thousands of years.

[0089] The habitat of C. cajan grows is typically in forests in its native range. It can be found in warmer, dryer regions of the tropics and subtropics as well as more temperate regions.

[0090] Cajanus cajan is adapted to a wide range of soil types. It grows best in well-drained soils and will not survive waterlogged conditions. It can be grown in a pH range of 4.5-8.4 (Cook et al., 2005). Cajanus cajan grows best under hot conditions (65-86 °F) and can grow in temperatures greater than 95 °F (Cook et al., 2005). Mullen et al. (2003) found that a soil temperature of 64 °F or greater is required at time of planting, as lower temperatures will lengthen the period of establishment. Nevertheless, it is a hardy plant that can grow at temperatures as low as 41 °F (Phatak et al., 1993) and has been successfully grown in warmer temperate regions such as North Carolina (Duke, 1983). Frost will defoliate the plant.

[0091] Cajanus cajan is drought resistant and can survive under very dry conditions because of its deep root system. It has been found to grow throughout a six-month dry season (Cook et al., 2005); however, flowering will be delayed and seed yields will decrease under long periods of drought (Mullen et al., 2003). It is less adapted to humid, wet conditions.

[0092] To establish the plant, typically seeds are sown 1.5 inches deep on 1 to 3 foot rows at 8 to 10 lb. per acre. The seeds are drilled in a weed-free seedbed or transplanted as seedlings on 6 foot rows if grown as an intercrop. Seedlings usually emerge within 3 weeks but will grow slowly.Attorney Docket No. 71900-435893

[0093] For successful pod development, it is important to grow the plant under full sunlight and never under waterlogged conditions. Fertilization is not recommended as the plant can grow well in soils with low phosphorus levels (Phatak et al., 1993) and has shown little response to fertilizers (Duke, 1983).

[0094] Cajanus cajanus can be managed as an annual shrub or a perennial plant. As a perennial plant it has been successfully used in alley cropping systems with cereals and legumes. Under good management, the plant can live up to five years. As the plant ages, the stem will become woodier, and leaf regeneration will decline.

[0095] Harvesting should be done after first frost, as leaf drop will make harvesting easier. Harvesting methods are similar to those for soybean and can be done with a combine.

[0096] The plant will not survive heavy, continuous grazing or heavy cutting, but may be pruned. Weeds can be cultivated in the inter-rows. It cannot survive fire.

[0097] Potential pests and problems for cultivation include Heliothis spp. which can significantly decrease yields. Other pests include cutworms (Noctudidae), thrips (Thysanoptera), mirids (Miridae), and green stink bugs [(e.g., Nezara viridula L. (Heteroptera:Pentatomidae)]. The seeds are eaten by pod borers (e.g., Helicoverpa spp., and Heliothis spp.) in the field and by bruchids (Callosobruchus spp. Coleoptera: Bruchidae) during storage (Cook et al., 2005). Also, C. cajan can be affected in the field by Fusarium wilt (caused by Fusarium udum Butler) or other Fusarium spp. (Ravikumara et al., 2022), and Phytophthora blight (Mishra et al., 2025); and by Aspergillus spp., Fusarium spp. on seed in the storage (Bankole et al., 1995) which are important fungal pathogens.

[0098] The time required to reach maturity can vary greatly due to seed variety, temperature, and photoperiod. Cajanus cajan is a short-day plant, requiring a daylight length of 12.5 hours to initiate flowering and seed production (Cook et al., 2005). Cajanus cajan requires 65-80 days to flower and at least 40 additional days to create mature seeds (Mullen et al., 2003). It is 60% selfpollinated (Cook et al., 2005), but varieties grown within 2-3 miles may have cross-pollination (Mullen et al., 2003). The plant is a prolific seed producer. Yields of mature seed pods can be from 0.25 to 1 ton / acre (Mullen et al., 2003). There are roughly 7,000 to 8,000 seeds per pound (Cook et al., 2005; USDA-NRCS, 2012). The seed pods can withstand weather damage (Mullen et al., 2003). After the first year of growth, seed production declines in subsequent years (Duke, 1983).ExamplesExample 1: Methods of developing pigeonpea cultivars.Attomey Docket No. 71900-435893Locations for breeding and experimental trials

[0099] Breeding and evaluation trials were conducted in Isabela and Lajas Research Substations at the University of Puerto Rico from 2019 to 2024. Isabela is located to the northwest of Puerto Rico at 126 masl. Isabela has temperatures ranging from 20 °C to 31 °C, mean annual precipitation of 1592 mm, and average relative humidity of 70%. This location has Oxisol soils (Goto series, very fine, kaolinitic, isohyperthermic Typic Eutrustox). Lajas is located to the southwest of the island at 9 masl and has soils belonging to the Mollisol and Vertisol orders, San Anton (fine-loamy, mixed, superactive, isohyperthermic Cumulic Haplustoll) and Fraternidad (fine, smectitic, isohyperthermic Typic Haplustert) series, respectively. It has temperatures between 19°C and 33°C, annual precipitation of 1143 mm, and average relative humidity of 80%. Pigeonpea plantings under short-day conditions (<12 h) are mostly conducted from October to December to increase seed of all type of pigeonpea genotypes (i.e., photoperiod sensitive and insensitive) in both locations. In contrast, plantings under long-day conditions (>12 h) are mostly carried out from February to July. Puerto Rico is USDA Plant hardiness zones 12 and 13.Germplasm development

[0100] IPG-7 and IIPG-11 breeding lines were developed from the bi-parental cross ‘ICPL 860127‘Guerrero’ conducted by emasculation and pollinations at the Isabela Research Substation, University of Puerto Rico, in February 2019. ICPL 86012 was selected because it is photoperiod insensitive, has early flowering (<85 DAP) and early maturity (<146 DAP), and has seed yields between 300 and 2400 kg ha-1in Puerto Rico. ICPL 86012 has a determinate flowering habit, and plant height does not exceed 2 m even under long-day conditions. Guerrero was selected because it is an indeterminate cultivar developed in Puerto Rico that has a beige seed (19 g per 100 mature seeds), which is the most common market class in the island. Plant height varied from 1.0 m (short-day conditions) to >3.0 m (long-day conditions); flowering time and harvest maturity can extend up to 97 and 157 DAP, respectively; and seed yield can reach up to 1800 kg ha-1under short-day conditions. Guerrero is photoperiod sensitive.

[0101] Twenty-four Fi seeds of the ICPL 86012 / Guerrero cross were planted in May 2019 in Isabela. Four rows with six plants per row, with 0.6 m between plants and 0.9 m between rows, were used. The seeds were sown at a depth of 3 cm, and over-head irrigation was provided to promote optimum vegetative and reproductive growth. All 24 hybrids were indeterminate and photoperiod sensitive, with both dominant traits derived from Guerrero. The FI:2seed was harvested December 2019 through January 2020. Sixty F2seeds derived from each Fi (n = 1440 plants) were planted by hand in single rows with the distance and conditions mentioned above inAttorney Docket No. 71900-435893February 2020 in Isabela. A total of -200 F23 plants with indeterminate flowering habit and photoperiod insensitivity were selected in August 2020. Fifty seeds from the F3to F5progenies were sown in single rows using two different spacings in Isabela. During the short-day conditions, plants were established at 0.3 m by 0.9 m between plants and rows, and for long-day conditions plants were established at 0.6 m by 0.9 m between plants and rows. These populations were evaluated in five continuous cycles of planting from October 2020 to January 2021, February to August 2021, and October 2021 to January 2022, in Isabela. Ten plants with indeterminate flowering habit that were photoperiod insensitive, had green seed at immature stage, and had 100-mature seed with a weight >13 g were selected and advanced by the pedigree method in each planting. The F56 derived from two plants that possessed all the traits of interest were selected for yield trials in Isabela and Lajas. Also, the Fe y were used to evaluate the agronomic performance of both genotypes compared with indeterminate cultivars under short-day conditions in both locations. Ultimately, the Fysseed from the two genotypes was harvested separately in bulk to develop IIPG-7 and IIPG-11 breeding lines.Field trials an experimental design

[0102] IIPG-7 and IIPG-11 breeding lines, along with their parents (i.e., ICPL 86012 and Guerrero), and determinate photoperiod-insensitive cultivars Isabela and Lazaro, were evaluated at the Isabela and Lajas Research Substations at the University of Puerto Rico in March 2022 (Table 1). Likewise, all these genotypes and indeterminate photoperiod-sensitive cultivars Ariel, Blanco Yauco, Kaki, IGP 7035, Pinto Berrocales, and Super Pinto were evaluated in November 2022 and February 2023 in both locations and in May 2023 and October 2024 in Isabela. Drip and overhead irrigation were used in Lajas and Isabela, respectively. Also, plants were grown under the weather conditions and type of soils described in Locations for breeding and experimental trials for both locations.

[0103] A randomized complete block design with three replications was used in each planting. The experimental plot consisted of 40 plants of the above genotypes planted in four 6-m-long rows, with a spacing of 0.9 m between rows and 0.6 m between plants for long-day conditions (i.e., March 2022 and February and May 2023). In contrast, the 40 plants were planted in four 3-m-long rows, and each seed was separated by 0.3 m instead of 0.6 m for short-day conditions (October 2022 and 2024) in Isabela and Lajas.Qualitative and quantitative traits evaluation and statistical analysis

[0104] Data for the number of days to the initiation of flowering, flower color, type of flowering habit (determinate or indeterminate), and plant height were collected at reproductive stages in theAttorney Docket No. 71900-435893plantings on October 2022 at both locations. Plant height was measured from the soil level to the top of the primary inflorescence when pod and seed set had been completed in all genotypes. In contrast, plant heights for the indeterminate photoperiod-sensitive genotypes were collected at 180 DAP under long-day conditions in 2022 and 2023. Also, data for the number of days to harvest were collected when at least 50% of IIPG-7 and I IPG-11 breeding lines and the checks reached harvest maturity in the field. The seed yield was estimated from 20 plants harvested from the two central rows per each genotype. Likewise, data on pod size and phenotypic appearance of pods and immature / mature seeds were collected from 10 plants taken randomly in each experimental plot. The weight of 100 dry seeds (-16% moisture) was recorded from all trials with the exception of the plantings carried out in March 2022.

[0105] Data on plant height, days to flowering and harvest, seed yield, and weight of 100 dryseed were analyzed using SAS 9.4 PROC GLM in each planting in Isabela and Lajas. Fisher’s least significant difference at P < 0.05 was calculated to compare means of these quantitative traits between pigeonpea genotypes.Example 2: Characteristics of Pigeonpea cultivars.Morphological traits and agronomic performance

[0106] Each of IIPG-7 and IIPG-11 breeding lines have indeterminate flowering habit, and plant height varied according to the date of planting. For instance, values <1.2 m were observed for IIPG-7 and 11 PG-11 in the plantings of October 2022 and February 2023 at Isabela and Lajas (Table 1). In contrast, taller plants (2.0-2.6 m) for both BL were observed in March 2022 in both locations and in May 2023 at Isabela (Table 1). However, IIPG-7 and I IPG-11 breeding lines did not reach mean values >2.8 m compared with those observed for the cultivars Ariel, Blanco Yauco, Guerrero, and Super Pinto, where higher vegetative biomass accumulated under long-day conditions.Table 1. Mean values for plant height of IIPG-7 and IIPG-11 pigeonpea [Cajanus cajan (L.) Mill.] breeding lines, their parents, and cultivars checks evaluated at the University of Puerto Rico Isabela and Lajas Research Substations in 2022 and 2023.Isabela LajasGenotype Mar-22 Oct-22 Feb-23 May-23 Mar-22 Oct-22 Feb-23 Mean New breeding linesIIPG-7 2.4 1.1 1.2 2.5 2.0 0.8 1.0 1.6Attorney Docket No. 71900-435893IIPG-11 2.3 1.0 1.1 2.6 - 0.9 1.0 1.5 ParentsGuerrero32.4 1.7 2.9 4.1 2.3 1.6 3.0 2.6 ICP 0.6 1.11.5 0.6 0.8 1.7 1.4 0.886012bCultivars c necks insensitive to the photoperiodbIsabella 1.6 0.7 0.9 1.8 1.6 0.8 0.9 1.2 Lazaro 1.7 0.9 1.1 1.9 1.6 0.9 1.2 1.3 Cultivars checks sensitive to the photoperiodaAriel _c 1.4 3.0 3.8 - 1.4 3.1 2.5 Blanco 2.9 2.6 - 1.6 2.9 3.8 - 1.7YaucoICP 7035 - 1.6 2.8 3.7 - 1.6 3.0 2.5 Kaki - 1.2 2.8 3.8 - 1.1 2.7 2.3 Pinto 2.7 2.5 - 1.4 2.9 4.0 - 1.4BerrocalesSuper 3.0 2.7 - 1.6 3.2 4.0 - 1.6PintoMean 2.0 1.2 2.1 3.1 1.8 1.2 2.1 LSDQ 05 0.1 0.1 0.1 0.1 0.1 0.1 0.1aPlant height was recorc ed at 180 c ays after p anting for genotypes sensitive to the photoperiod.bPlant height was measured from the soil level to the top of the primary inflorescence when pod and seed set had been completed for genotypes insensitive to the photoperiod.cGenotypes that were not tested in March 2022 plantings.

[0107] IIPG-7 and IIPG-11 breeding lines initiated flowering between 42 and 92 DAP and reached harvest maturity at 88-172 DAP compared with cultivars Ariel, Blanco Yauco, Guerrero, Kaki, Pinto Berrocales, and Super Pinto, which initiated flowering between 87 and 109 DAP and reached maturity at 132-172 DAP under short-day conditions in both locations (Tables 2 and 3).All indeterminate check cultivars and photoperiod sensitivity did not initiate flowering at least at 180 DAP, and therefore seed yield was not evaluated under long-day conditions. Similar results were reported for photoperiod-sensitive cultivars Cortada, Guerrero, ICP 7035, Pinto Berrocales, and Super Pinto. Contrarily, IIPG-7 and 11 PG-11 initiated flowering and reached maturity betweenAttorney Docket No. 71900-43589388-92 DAP and 141-144 DAP in March 2022, 76-79 DAP and 122-128 DAP in February 2023, and 77-88 DAP and 165-172 DAP in May 2023, respectively in Isabela. However, 11 PG-7 and IIPG-11 breeding lines did not exceed the 76 and 120 DAP for flowering and harvest time, respectively, in Lajas in March 2022 and February 2023 (Tables 2 and 3). This was expected because the drier conditions of Lajas allowed an earlier harvesting for photoperiod-insensitive pigeonpea genotypes under long-day conditions.Table 2. Mean values for flowering time of IIPG-7 and IIPG-11 pigeonpea [Cajanus cajan (L.) Mill.] breeding lines, their parents, and cultivars checks evaluated at the University of Puerto Rico Isabela and Lajas Research Substations in 2022, 2023, and 2024.Isabela Lajas Genotype Mar- 22 Oct-22 Feb-23 May-23 Oct-24 Mar-22 Oct-22 Feb-23 Mean New breeding linesIIPG-7 92 77 79 88 76 75 84 44 77 IIPG-11 88 73 76 77 78 - 83 42 74ParentsGuerrero NFa91 NF NF 100 NF 109 NF 100 ICP 86012 85 76 80 65 78 82 78 45 74Cultivars checks insensitive to the photoperiodIsabella 86 77 77 68 75 84 79 45 74 Lazaro 89 87 91 76 84 85 90 58 83Cultivars checks sensitive to the photoperiodAriel _b 93 NF NF 97 - 92 NF 94 Blanco - 94 NF NF 94 - 102 NF 97 YaucoICP 7035 - 91 NF NF - - 107 NF 99 Kaki - 87 NF NF - - 86 NF 87 Pinto- 91 NF NF - - 95 NF 93 BerrocalesSuper- 92 NF NF - - 96 NF 94 PintoMean 88 86 81 75 85 82 92 472.2 5.LSDOO5 4 2.2 11.7 4.1 6.4 4.7 3.2aNot flowering at 180 days after planting.bGenotypes that were not tested in March 2022 and October 2024 plantings.Attorney Docket No. 71900-435893Table 3. Mean values for harvesting time of IIPG-7 and IIPG-11 pigeonpea [Cajanus cajan (L.) Mill.] breeding lines, their parents, and cultivars checks evaluated at the University of Puerto Rico Isabela and Lajas Research Substations in 2022, 2023, and 2024.Isabela Lajas Genotype Mar-22 Oct-22 Feb-23 May-23 Oct-24 Mar-22 Oct-22 Feb-23 Mean New breeding linesIIPG-7 144 125 128 172 128 119 127 89 129 IIPG-11 141 125 122 165 127 - 121 88 127 ParentsGuerrero NEa153 NE NE 172 NE 153 NE 159 ICP 86012 144 124 123 116 123 112 119 87 119Cultivars checks insensitive to the ohotoperiodIsabella 142 126 123 124 128 110 120 103 122 Lazaro 160 134 184 193 145 128 134 117 149Cultivars checks sensitive to the photoperiodAriel _b 146 NE NE 164 - 138 NE 149 Blanco- 146 NE NE 152 - 145 NE 148 YaucoICP 7035 - 147 NE NE - - 154 NE 151 Kaki - 145 NE NE - - 132 NE 139 Pinto- 147 NE NE - - 138 NE 143 BerrocalesSuper- 148 NE NE - - 140 NE 144 PintoMean 146 139 136 154 142 117 135 97 LSDQ 05 2.8 4.6 12.2 10.2 3.5 6.9 4.0 16.3aNot evaluated because the plants did not produce pods and seed set under long-day conditions.bGenotypes that were not tested in March 2022 and October 2024 plantings.

[0108] IIPG-7 and IIPG-11 have flowers with red color petals on the outside and yellow inside. Pods of IIPG-7 have red color with small green marks (Figure 1A), whereas pods of IIPG-11 are green with brown marks (Figure 1 B) with an average size of 6.5 by 0.7 cm. Seeds of both BL are green at immature stages and mature seeds of IIPG-7 are tan with reddish spots (Figure 1C), unlike mature seeds of IIPG-11, which are reddish at harvest (Figure 1D). In general, IIPG-7 and IIPG-11 BL had average 100-seed weights of 14 and 15 g, respectively (Table 4).Attorney Docket No. 71900-435893Table 4. Mean values for 100-seed weight of IIPG-7 and IIPG-11 pigeonpea [Cajanus cajan (L.) Mill.] breeding lines, their parents, and cultivars checks evaluated at the University of Puerto Rico Isabela and Lajas Research Substations in 2022, 2023, and 2024.Isabela La asGenotype Oct-22 Feb-23 May-23 Oct-24 Oct-22 Feb-23 MeanNew breeding linesIIPG-7 13.7 14.1 13.8 14.8 13.8 14.5 14.1IIPG-11 14.3 15.6 14.9 15.1 14.9 15.5 15.1 ParentsGuerrero 17.1 NEaNE 18.7 16.6 NE 17.5ICPL 86012 12.0 10.7 10.1 11.9 10.7 11.9 11.2 Cultivars checks insensitive to the photoperiodIsabella 14.2 13.9 12.3 13.0 12.7 14.8 13.5 Lazaro 17.8 15.5 14.3 17.4 15.7 15.1 16.0 Cultivars checks sensitive to the photoperiodAriel 19.3 NE NE 19.3 16.7 NE 18.4 Blanco Yauco 19.6 NE NE 19.1 17.7 NE 18.8ICP 7035 18.9 NE NE _b 19.3 NE 19.1 Kaki 21.6 NE NE - 17.9 NE 19.8 Pinto17.8 NE NE - 16.9 NE 17.4 BerrocalesSuper Pinto 15.0 NE NE - 15.4 NE 15.2 Mean 17.0 14.0 13.1 16.2 15.7 14.41.4 1.4 1.0 1.3 1.9 1.0LSDOO5aNot evaluated because plants did not produce seed under long-day conditions.bGenotypes that were not tested in October 2024 planting.

[0109] IIPG-7 and IIPG-11 breeding lines had lower seed yields (720-1080 kg ha-1) than indeterminate and photoperiod-sensitive cultivars Ariel, Blanco Yauco, ICP 7035, and Pinto Berrocales, which reached values >1500 kg ha-1under short-day conditions either at Isabela or Lajas (Table 5). In contrast, IIPG-7 and IIPG-11 had seed yields between 1500 and 2400 kg ha-1under long-day conditions. Furthermore, IIPG-7 and IIPG-11 had significantly higher yields than Lazaro in Lajas in the February 2023 planting (Table 5). Seed yields decreased drastically for IIPG-7 (626 kg ha-1) and IIPG-11 (766 kg ha-1) and even cultivars ICPL 86012, Isabela, and Lazaro (<500 kg ha-1) in May 2023 in Isabela (Table 5). This might be associated with the higherAttorney Docket No. 71900-435893rainfall that occurred in July (113 mm), August (156 mm), and September (138 mm), which may have caused flower and pod abortion. Similar conditions caused seed yield reductions in over65% of pigeonpea genotypes.Table 5. Mean values for seed yield of IIPG-7 and IIPG-11 pigeonpea [Cajanus cajan (L.) Mill.] breeding lines, their parents, and cultivars checks evaluated at the University of Puerto Rico Isabela and Lajas Research Substations in 2022, 2023, and 2024.Isabela La as Genotype Mar-22 Oct-22 Feb-23 May-23 Oct-24 Mar-22 Oct-22 Feb-23 Mean New breeding linesIIPG-7 2449 1010 1541 626 1079 1900 818 1964 1423 IIPG-11 2268 721 1597 766 898 - 909 1949 1301 ParentsGuerrero NEa1419 NE NE 1847 NE 1409 NE 1558 ICPL1711 559 1026 510 773 2440 842 1246 1138 86012Cultivars checks insensitive to the photoperiodIsabella 2793 777 1703 436 1122 2235 950 1685 1463 Lazaro 1975 1221 1283 75 1775 1224 1244 1098 1237 Cultivars checks sensitive to the photoperiodAriel _b 1728 NE NE 2539 - 1094 NE 1787 Blanco- 1564 NE NE 1269 - 1398 NE 1410 YaucoICP 7035 - 1621 NE NE - - 1020 NE 1321 Kaki - 1228 NE NE - - 1426 NE 1327 Pinto- 1265 NE NE - - 1578 NE 1422 BerrocalesSuper Pinto - 1264 NE NE - - 1374 NE 1319 Mean 2239 1198 1430 483 1413 1950 1172 1588 LSDQ05 760.1 543.2 1422.9 348.5 733.6 390.6 523.9 759.2aNot evaluated because the plants did not produce pods and seed set under long-day conditions.bGenotypes that were not tested in March 2022 plantings.

[0110] The benefits of photoperiod insensitivity for pigeonpea breeding:Attorney Docket No. 71900-435893

[0111] IIPG-7 and IIPG-11 breeding lines can be planted under long-day conditions. This characteristic can be incorporated into determinate and indeterminate cultivars that are photoperiod sensitive but possess desirable agronomic and seed traits. For instance, backcrosses of IIPG-7 and I IPG-11 with Ariel, Blanco Yauco, and Guerrero may help to develop cultivars with beige / cream seed with a bigger size (>16 g per 100 mature seeds). Likewise, a new set of crosses combining IIPG-7 and I IPG-11 breeding lines with multiple parents that possess a commercial seed size and color, and higher yield (e.g., Ariel, Blanco Yauco, Super Pinto, Pinto Original, and Pinto Berrocales) may help to introgress all these characteristics in new populations. Gamete selection or recurrent selection in combination with pedigree and bulk breeding methods may be recommended for the selection of multiple traits. The pedigree selection would be used to select plants with photoperiod insensitivity in earlier generations (e.g., F2 and F3) under long-day conditions. With this, homozygous genotypes to this trait could be achieved rapidly. Also, because a low frequency of plants that flower is expected, the space and labor should not be a problem to carry out progeny tests. The bulk method should be used in later generations (F5-F7) to select photoperiod-insensitive genotypes that have a higher agronomic performance and commercial seed size and color, especially if labor and space conditions are limited. Furthermore, crosses between IIPG-7 breeding line with determinate cultivars Isabella or Lazaro can help to develop cultivars with seed for commercial purposes. Pedigree in combination with bulk breeding methods may be an appropriate approach because the dominant trait, indeterminate flowering habit, can be selected and advanced in each generation.

[0112] Applicant has deposited 2500 seeds of pigeonpea breeding line IIPG-7 and 2500 seeds of pigeonpea breeding line IIPG-11 with the American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, VA 20110, USA, as ATCC Deposit No. PTA-127908 for IIPG-7 and ATCC Deposit No. PTA-127909 for I IPG-11. The seeds deposited with the ATCC on May 14, 2025, are from seed stock maintained by Diego Viteri at the University of Puerto Rico since prior to the filing date of this disclosure. Access to this seed will be available during the pendency of this application to the Commissioner of Patents and Trademarks and persons determined by the Commissioner to be entitled thereto upon request. Upon allowance of any claims in the application, Applicant will make the deposit available to the public pursuant to 37 C.F.R. §1.808. This deposit of the two pigeonpea breeding lines will be maintained in the ATCC depository, which is a public depository, for a period of 30 years, or 5 years after the most recent request, or for the enforceable life of the patent, whichever is longer, and will be replaced if it becomes nonviable during that period. Additionally, Applicant has or will satisfy all of the requirements of 37 C.F.R §1.801-1.809, including providing an indication of the viability of theAttorney Docket No. 71900-435893sample upon deposit. Applicant has no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce. Applicant does not waive any infringement of rights granted under this patent or under the Plant Variety Protection Act (7 USC §2321 et seq.). Unauthorized seed multiplication is prohibited.

[0113] All publications, patents, and patent applications cited in this specification are incorporated herein by reference for the teaching to which such citation is used.

[0114] The present disclosure enables one of skill in the relevant art to make and use the inventions provided herein in accordance with multiple and varied embodiments. Various alterations, modifications, and improvements of the present disclosure that readily occur to those skilled in the art, including certain alterations, modifications, substitutions, and improvements are also part of this disclosure. Accordingly, the foregoing description are by way of example to illustrate the discoveries provided herein. Furthermore, the foregoing Description and Examples are exemplary of the present invention and not limiting thereof. The scope of the invention is therefore set out in the appended claims.

[0115] Although specific embodiments of the present disclosure are herein illustrated and described in detail, the disclosure is not limited thereto. The above detailed descriptions are provided as exemplary of the present disclosure and should not be construed as constituting any limitation of the disclosure. Modifications will be obvious to those skilled in the art, and all modifications that do not depart from the spirit of the disclosure are intended to be included with the scope of the appended claims.

Claims

Attorney Docket No. 71900-435893That which is claimed is:

1. A plant or a seed of a breeding line of pigeonpea [Cajanus cajan (L.) Mill.], 11 PG-7, wherein a representative sample of seed of said varietal of pigeonpea I IPG-7 has been deposited under ATCC Accession number PTA-127908.

2. A breeding line of pigeonpea I IPG-7 plant, or a part thereof, of the plant or seed of claim 1, wherein the plant or plant part comprises at least one cell of the breeding line of pigeonpea I IPG-7.

3. A tissue culture comprising at least one cell or protoplast of the plant or plant part of claim 2.

4. A breeding line of pigeonpea 11 PG-7, wherein the cultivar of pigeonpea 11 PG-7 has at least one phenotype selected from the group consisting of:a) indeterminate flowering habit,b) photoperiod insensitivity,c) higher accumulative yield per year compared to indeterminate photoperiodsensitive pigeonpea cultivars,d) shorter time to harvest compared to indeterminate pigeonpea cultivars, e) red and yellow flowers,f) red pods with green speckles, andg) completely green seeds at an immature stage.

5. The breeding line of pigeonpea 11 PG-7, wherein the breeding line reaches harvest maturity at about 89-172 days after planting (DAP).

6. A germplasm of a breeding line of pigeonpea I IPG-7.

7. The germplasm of claim 6, wherein the germplasm produces seeds in both long-day and short-day conditions.

8. A method for producing a breeding line of pigeonpea I IPG-7 seed, wherein the method comprises fertilizing the breeding line of pigeonpea IIPG-7 and harvesting the resultant breeding line of pigeonpea IIPG-7 seed.

9. A breeding line of pigeonpea IIPG-7 seed produced by the method of claim 8, wherein the resulting breeding line of pigeonpea IIPG-7 seed is an offspring or the product of selffertilization.

10. The breeding line of pigeonpea IIPG-7 seed of claim 9, wherein the 100-seed weighs between 12 and 16 g.Attorney Docket No. 71900-43589311. A plant or a seed of a breeding line of pigeonpea [Cajanus cajan (L.) Mill.], IIPG-11, wherein a representative sample of seed of said breeding line of pigeonpea IIPG-11 has been deposited under ATCC Accession number PTA-127909.

12. A breeding line of pigeonpea 11 PG- 11 plant, or a part thereof, of the plant or seed of claim 1, wherein the plant or plant part comprises at least one cell of the breeding line of pigeonpea I IPG-11.

13. A tissue culture comprising at least one cell or protoplast of the plant or plant part of claim 2.

14. A breeding line of pigeonpea IIPG-11 , wherein the breeding line of pigeonpea plant I IPG- 11 has at least one phenotype selected from the group consisting of:a) indeterminate flowering habit,b) photoperiod insensitivity,c) higher accumulative yield per year compared to indeterminate photoperiod-sensitive pigeonpea cultivars,d) shorter time to harvest compared to indeterminate pigeonpea cultivars,e) red and yellow flowers,f) green pods with red speckles, andg) completely green seeds at an immature stage.

15. The breeding line of pigeonpea IIPG-11 , wherein the breeding line of pigeonpea IIPG-11 reaches harvest maturity at about 88-165 days after planting (DAP).

16. A germplasm of a breeding line of pigeonpea IIPG-11.

17. The germplasm of claim 6, wherein the germplasm produces seeds in both long-day and short-day conditions.

18. A method for producing a breeding line of pigeonpea IIPG-11 seed, wherein the method comprises fertilizing the breeding line of pigeonpea IIPG-11 and harvesting the resultant breeding line of pigeonpea IIPG-11 seed.

19. A breeding line of pigeonpea IIPG-11 seed produced by the method of claim 8, wherein the resulting breeding line of pigeonpea IIPG-11 seed is an offspring or the product of selffertilization.Attorney Docket No. 71900-43589320. The breeding line of pigeonpea I IPG-11 seed of claim 9, wherein the 100-seed weighs between 13 and 17 g.

Images