Manoj KUNDU, Anil DUBEY, Manish SRIVASTAV, Surendra MALIK

Induction of haploid plants in citrus through gamma-irradiated pollen and ascertainment of ovule age for maximum recovery of haploid plantlets

The present investigation was carried out for the induction of haploid plants in Citrus grandis through in situ parthenogenesis by pollination with gamma-irradiated pollen of C. limetta and C. sinensis, treated with 50, 100, 200, 300, and 400 Gy gamma ray doses followed by in vitro ovule culture. Ovule culture at 50 days after pollination (DAP) was found optimum for the maximum recovery of in vitro-raised plantlets as compared to 35 and 20 DAP. At 50 DAP, irrespective of the pollen parent, plantlet regeneration capacity decreased at 300 and 400 Gy irradiation treatment (0.62% and 0.60%, respectively) with maximum recovery in the nonirradiated control (3.07%). Chromosome counting in actively growing root tips of all the in vitro-raised ovule cultured plantlets revealed that two haploid plants with chromosome number of nine were induced in C. grandis from the ovule culture that was cultured at 50 DAP following pollination with irradiated pollen of C. sinensis at 400 Gy and C. limetta pollen at 300 Gy. However, irradiation doses below 300 Gy were incapable of inducing any haploids in either cross combination. Similarly, all the plantlets regenerated from in vitro embryo culture of mature seeds were found to be diploid in nature, irrespective of the pollen parent and irradiation dose. Molecular analysis of the in vitroraised haploid and diploid plants using SSR markers confirmed the maternal origins of the haploid plants; however, the diploid plants were found zygotic in nature with one allele from the seed parent and the other one from the pollen parent.

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Aleza P, Juarez J, Hernandez M, Pina JA, Ollitrault P, Navarro L (2009). Recovery and characterization of a Citrus clementina Hort. ex Tan. Clemenules haploid plant selected to establish the reference whole Citrus genome sequence. BMC Plant Biol 9: 110.
Bermejo A, Pardo J, Cano A (2011). Influence of gamma irradiation on seedless citrus production: pollen germination and fruit quality. Food Nutri Sci 2: 169-180.
Bouvier L, Zhang YX, Lespinasse Y (1993). Two methods of haploidisation in pear, Pyrus communis L.: greenhouse seedling selection and in situ parthenogenesis induced by irradiated pollen. Theor Appl Genet 87: 229-232.
Chahal GS, Gosal SS (2002). Principles and Procedures of Plant Breeding: Biotechnological and Conventional Approaches. London, UK: Alpha Science International Ltd.
Chai L, Biswas MK, Yi H, Guo W, Deng X (2013). Transferability, polymorphism and effectiveness for genetic mapping of the Pummelo (Citrus grandis Osbeck) EST-SSR markers. Sci Hortic-Amsterdam 155: 85-91.
Chalak L, Legave JM (1997). Effects of pollination by irradiated pollen in Hayward kiwifruit and spontaneous doubling of induced parthenogenetic trihaploids. Sci Hortic-Amsterdam 68: 83-93.
Cicero BM, Wilson RM, Sebastiao MA, Marcos VF, Ildon RN, Douglas WN, Luiz AA, Gomes BB (2005). Inheritance of parthenocarpy in summer squash (Cucurbita pepo L.). Genet Mol Res 4: 39-46.
De Witte K, Keulemans J (1994). Restrictions of the efficiency of haploid plant production in apple cultivar Idared, through parthenogenesis in situ. Euphytica 77: 141-146.
Engvild KC (1985). Pollen irradiation and possible gene transfer in Nicotiana species. Theor Appl Genet 69: 457-461.
Esen A, Soost RK (1971). Unexpected triploids in Citrus: the origin identification, and possible use. J Hered 62: 329-333.
Froelicher Y, Bassene J, Jedidi-Neji E, Dambier D, Morillon R, Bernardini G, Costantino G, Ollitrault P (2007). Induced parthenogenesis in mandarin for haploid production: induction procedures and genetic analysis of plantlets. Plant Cell Rep 26: 937-944.
Germanà MA (1997). Haploidy in Citrus. In: Jain SM, Sopory SK, Veilleux RE, editors. In Vitro Haploid Production in Higher Plants. 5th ed. Dordrecht, the Netherlands: Kluwer Academic Publisher, pp. 195-217.
Germanà MA, Chiancone B (2001). Gynogenetic haploids of Citrus after in vitro pollination with triploid pollen grains. Plant Cell Tiss Org 66: 59-66.
Germanà MA, Chiancone B (2003). Improvement of Citrus clementina Hort. ex Tan. microspore-derived embryoid induction and regeneration. Plant Cell Rep 22: 181-187.
Germanà MA, Crescimanno FG, De Pasquale F, Ying WY (1992). Androgenesis in 5 cultivars of Citrus limon L. Burm. f.. Acta Hortic 300: 315-324.
Germanà MA, Reforgiato-Recupero G (1997). Haploid embryos regeneration from anther culture of Mapo tangelo (Citrus deliciosa × C. paradisi). Adv Hort Sci 11: 147-152.
Germanà MA, Wang YY, Barbagallo MG, Iannolino G, Crescimanno FG (1994). Recovery of haploid and diploid plantlets from anther culture of Citrus clementina Hort. ex Tan. and Citrus reticulata Blanco. J Hort Sci 69: 473-480.
Grosser JW, Ollitrault P, Olivares-Fuster O (2000). Somatic hybridization in citrus: an effective tool to facilitate variety improvement. In Vitro Cell Dev-Pl 36: 434-449.
Grouh MSH, Vahdati K, Lotfi M (2011). Production of haploids in Persian walnut through parthenogenesis induced by gammairradiated pollen. J Amer Soc Hort Sci 136: 198-204.
Hidaka T, Yamada Y, Shichijo T (1979). In vitro differentiation of haploid plants by anther culture in Poncirus trifoliata (L.) Raf. Jpn J Breed 29: 248-254.
Höfer M, Grafe C (2003). Induction of double haploids in sweet cherry (Purnus avium L.). Euphytica 130: 191-197.
Höfer M, Lespinasse Y (1996). Haploidy in apple. In: Jain SM, Sopory SK, Veilleux RE, editors. In Vitro Haploid Production in Higher Plants, 3: Important Selected Plants. Dordrecht, the Netherlands: Kluwer Academic Publishers, pp. 261-276.
James DJ, Passey AJ, Mackenzie KAD, Menhinick EC (1985). The effects of pollen irradiation on the development of the post fertilisation ovule of apple (Malus Pumila Mill.). In: Chapman GPC, Mantell JH, Davids RW, editors. Experimental Manipulation of Ovule Tissues. New York, NY, USA: Longman, pp. 210-239.
Karasawa K (1971). On the occurrence of haploid seedlings in Citrus natsudaidai Hayata. Bulletin of the Sakushin Gakuin Junior College for Women 1: 1-2.
Katoh N, Hagimori M, Iwai S (1993). Production of haploid plants of melon by pseudofertilized ovule culture. Plant Tissue Culture Lett 10: 60-66.
Kobayashi S, Ohgawara T, Saito W, Nakamura Y, Omura M (1997). Production of triploid somatic hybrids in Citrus. J Jpn Soc Hort Sci 66:453-458.
Kundu M, Dubey A, Mallik SK (2016). Effect of gamma ray irradiation doses on pollen viability and in vitro germination in Citrus. Indian J Agr Sci 86: 106-109.
Kundu M, Dubey A, Srivastav M, Mallik S, Singh B (2014). Effect of gamma ray irradiation and cryopreservation on pollen stainability, in vitro germination, and fruit set in Citrus. Turk J Biol 38: 1-9.
Kurtar ES (2003). An investigation on parthenocarpy in some summer squash (Cucurbita pepo L.) cultivars. Pak J Agron 2: 209-213.
Kurtar ES (2009). Influence of gamma irradiation on pollen viability, germination ability, and fruit and seed-set of pumpkin and winter squash. Afr J Biotechnol 8: 6918-6926.
Kurtar ES, Sarı N, Abak K (2002). Obtention of haploid embryos and plants through irradiated pollen technique in squash (Cucurbita pepo L.). Euphytica 127: 335-344.
Murashige T, Tucker DPH (1969). Growth factor requirements of Citrus tissue culture. First International Citrus Symposium 3: 1155-1161.
Murray MG, Thompson WF (1980). Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8: 4321-4326.
Musial K, Przywara L (1998). Influence of irradiated pollen on embryo and endosperm development in kiwifruit. Ann Bot-London 82: 747-756.
Musial K, Przywara L (1999). Endosperm response to pollen irradiation in kiwifruit. Sex Plant Reprod 12: 110-117.
Naess SK, Swartz JH, Bauchan GR (1998). Ploidy reduction in blackberry. Euphytica 99: 57-73.
Nepi M, Pacini E (1993). Pollination, pollen viability and pistil receptivity in Cucurbita pepo L. Ann Bot-London 72: 527-536.
Oiyama I, Kobayashi S (1993). Haploids obtained from diploid × triploid crosses of citrus. J Jpn Soc Hortic Sci 62: 89-93.
Ollitrault P, Dambier D, Sudahono Mademba-Sy F, Vanel F, Luro F, Aubert B (1997). Biotechnology for triploid mandarin breeding. In: Fifth International Congress of Citrus Nurserymen; Montpellier, France.
Ollitrault P, Dambier D, Vanel F, Froelicher Y (2000). Creation of triploid Citrus hybrids by electrofusion of haploid and diploid protoplasts. Acta Hort 535: 191-197.
Pandey KK (1974). Overcoming interspecific pollen incompatibility through the use of ionising radiation. Heredity 33: 279-287.
Pandey KK (1978). Gametic gene transfer in Nicotiana by means of irradiated pollen. Genetica 49: 53-69.
Pandey KK, Phung M (1982). Hertwig effect in plants: induced parthenogenesis through the use of irradiated pollen. Theor Appl Genet 62: 295-300.
Pandey KK, Przywara L, Sanders PM (1990). Induced parthenogenesis in kiwifruit (Actinidia deliciosa) through the use of lethally irradiated pollen. Euphytica 51: 1-9.
Peixe A, Campos MD, Cavaleiro C, Barroso J, Pais MS (2000). Gammairradiated pollen induces the formation of 2n endosperm and abnormal embryo development in European plum (Prunus domestica L., cv. Rainha Cláudia Verde). Sci Horic-Amsterdam 86: 267-278.
Raquin C (1985). Induction of haploid plants by in vitro culture of petunia ovaries pollinated with irradiated pollen. Zeitschrift Pflanzenzuchtung 94: 166-169.
Raquin C, Cornu A, Farcy E, Maizonnier D, Pelletier G, Vedel F (1989). Nucleus substitution between Petunia species using gamma ray induced androgenesis. Theor Appl Genet 78: 337-341.
Rode JC, Dumas de Vaulx R (1987). Obtention de plantes haploides de carotte (Daucus carota L.) issues de parthenogenese induite in situ par du pollen irradieet culture in vitro de graines immatures. CR Acad Sci-Paris 305: 225-229 (in French).
Samanta D (2009). Studies on genetic diversity and varietal identification in mango (Mangifera indica L.) using ISSR marker. PhD, Indian Agricultural Research Institute, New Delhi, India.
Sanders PM, Przywara L, Pandey KK (1991). A further investigation of sub-lethal pollen irradiation in Nicotiana (Solanaceae). Pol Bot Stud 2: 177-182.
Sareefa M (2009). DNA finger printing of mango (Mangifera indica L.) using molecular markers. PhD, Indian Agricultural Research Institute, New Delhi, India.
Sari N, Abak K, Pitrat M, Rode JC, Dumas de Vaulx R (1994). Induction of parthenogenetic haploid embryos after pollination by irradiated pollen in water melon. HortScience 29: 1189-1190.
Sauton A, Dumas de Vaulx R (1987). Obtention de plantes haploides chez le melon (Cucumis melo L.) par gynogénèse induite par du pollen irradié. Agronomie 7:141-148 (in French).
Scarano MT, Tusa N, Abbate L, Lucretti S, Nardi L, Ferrante S (2003). Flow cytometry, SSR and modified AFLP markers for the identification of zygotic plantlets in backcrosses between Femminello lemon cybrids (2n and 4n) and a diploid clone of Femminello lemon (Citrus limon L. Burm. F.) tolerant to mal secco disease. Plant Sci 164: 1009-1017.
Toolapong P, Komatsu H, Iwamasa M (1996). Triploids and haploid progenies derived from small seeds of Banpeiyu pummelo, crossed with Ruby Red grapefruit. J Jpn Soc Hortic Sci 65: 255-260.
Yahata M, Yasuda K, Nagasawa K, Harusaki S, Komatsu H, Kunitake H (2010). Production of haploid plant of Banpeiyu pummelo [Citrus maxima (Burm.) Merr.] by pollination with soft X-rayirradiated pollen. J Jpn Soc Hortic Sci 79: 239-245.
Yang HJ (1968). Fertilization and development of embryo on Satsuma orange (Citrus unshiu Marc.) and Natsudaidai (C. natsudaidai Hayata). J Jpn Soc Hortic Sci 37: 102-108.
Zhang YX, Lespinasse Y (1991). Pollination with gamma irradiated pollen and development of fruits, seeds and parthenogenetic plants in apple. Turk J Bot 54: 101-109.