Abstract
This study reports haploid plantlet regeneration through gynogenesis in Citrus clementina Hort. ex Tan., cv. Nules, induced by in vitro pollination with pollen grains of Oroblanco, a triploid cultivar of grapefruit. It indicates that parthenogenesis induced in vitro by triploid pollen can be an alternative method to obtain haploids in monoembryonic cultivars of Citrus. Actually, despite considerable efforts, androgenesis has not been yet successful in many genotypes of Citrus. Pollination and mature stage of pistils was necessary for gynogenic embryo regeneration. Fourteen haploid gynogenic embryos of Nules clementine were obtained. Embryo conversion was high (85.7%) and embryos vigorously germinated producing twelve plantlets. Chromosome counting, performed in the roots of obtained embryos, showed the haploid level (n=x=9). Isozyme analyses confirmed the expected homozygous state of embryos and plantlets.
Similar content being viewed by others
References
Bouvier L, Zhang YX & Lespinasse Y (1993) Two methods of haploidization in pear, Pyrus communis L.: greenhouse seedling selection and in situ parthenogenesis induced by irradiated pollen. Theor. Appl. Genet. 87: 229-232
Castano CI & De Proft MP (1996) Seed set in Cichorium intybus L. by pollination of flowers developed in vitro. Plant Cell, Tissue and Organ Cul. 46: 211-218
Chalak L & Legave JM (1997) Effects of pollination by irradiated pollen in Hayward kiwifruit and spontaneous doubling of induced parthenogenetic trihaploids. Scientia Horticulturae 68: 83-93
Chen Z, Wang H & Liao H (1980) The induction of Citrus pollen plants in artificial media. Acta Genetica Sinica 7: 189-192
Germanà MA (1997) Haploidy in Citrus. In: Jain SM, Sopory SK & Veilleux RE (eds) In Vitro Haploid Production in Higher Plants, Vol. 5 (pp 195-217). Kluwer Academic Publishers
Germanà MA, Crescimanno FG, De Pasquale F & Wang YY (1991) Androgenesis in 5 cultivars of Citrus limon L.Burm. f. Acta Hort 300: 315-324
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
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
Grosser JW, Gmitter FG Jr & Chandler JL (1988) Intergeneric somatic hybrid plants from sexually incompatible woody species: Citrus sinensis and Severina disticha. Theor. Appl. Genet. 75: 397-401
Hidaka T, Yamada Y & Shichijo T (1979) In vitro differentiation of haploid plants by anther culture in Poncirus trifoliata (L.) Raf. Japan J. Breed. 29: 248-254
Hidaka T, Yamada Y & Shichijo T (1981) Plantlet formation from anthers of Citrus aurantium L. Proc. Int. Soc. Citriculture: 153-155
Hofer M & Lespinasse Y (1996) Haploidy in apple. In: Jain SM, Sopory SK & Veilleux RE (eds) In Vitro Haploid Production in Higher Plants, Vol.3 (pp 261-276). Kluwer Academic Publishers
Kasha KJ & Kao KN (1970) High frequency haploid production in barley (Hordeum vulgare L.). Nature (Lond) 225: 874-876
Kobayashi S, Ohgawara T, Saito W, Nakamura Y & Omura M (1997) Production of triploid somatic hybrids in Citrus. J. Japan Soc. Hort. Sci. 66(3-4): 453-458
Lillie RD (1951) Simplification of the manufacture of Schiff reagent for use in histochemical procedures. Stain Tech. 26: 163-165
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-497
Nitsch J P & Nitsch C (1969) Haploid plants from pollen grains. Science 163: 85-87
Oiyama, II & Kobayashi S (1993) Haploid obtained from diploid X triploid crosses of Citrus. J. Japan Soc. Hort. Sci. 62(1): 89-93
Ollitrault P, Allent V & Luro F (1996) Production of haploid plants and embryogenic calli of clementine (Citrus reticulata Blanco) after in situ parthenogenesis induced by irradiated pollen. Proc Int Soc Citriculture, Vol. 2: 913-917
Pandey KK, Przywara L & Sanders PM (1990) Induced parthenogenesis in Kiwifruit (Actinidia deliciosa) through the use of lethally irradiated pollen. Euphytica 51: 1-9
Raquin C (1985) Induction of haploid plants by in vitro culture of Petunia ovaries pollinated with irradiated pollen. Zeitschrift fur Pflanzenzuchtung. 94: 166-169
Rode JC & Dumas de Vaulx (1987) Obtention de plantes haploides de carotte (Daucus carota L) issue de parthénogenèse in situ par du pollen irradié et culture induite in vitro de graines immatures. C. R. Acad. Sci. Paris, Serie III, 305: 225-229
Sauton A & Dumas de Vaulx (1987) Obtention de plantes haploides chez le melon (Cucumis melo L.) par gynogenèse induite par du pollen irradiè. Agronomie, 7: 141-148
Sari N, Abak K, Pitrat M. Rode JC & Dumas de Vaulx R (1994) Induction of parthenogenetic haploid embryos after pollination by irradiated pollen in watermelon. HortScience 29: 1189-1190
Sharma AK & Sharma A (1972) Chromosome techniques. Butterworths, London. University Park Press, Baltimore (p 575)
Snape JW, Sitch LA, Simpson E & Parker BB (1988) Tests for the presence of gametoclonal variation in barley and wheat doubled haploids produced using the Hordeum bulbosum system. Theor. Appl. Genet. 75: 509-513
Soost RK & Cameron JW (1980) 'Oroblanco', a triploid pummelograpefruit hybrid. HortScience 15(5): 667-669
Tilton VR & Russell SH (1983) In vitro pollination and fertilization of soybean Glycine max (L.) Merr. (Leguminosae). In: Pollen: biology and implication for plant breeding. Proc of the symposium, Villa Feltrinelli.: 281-286
Torres AM, Soost RK & Diedenhofen U (1978) Leaf isozymes as genetic markers in Citrus.American J. Bot. 65: 869-881
Tuyl JMvan, Jeu MJ de, De Jeu MJ & Sawhney VK (1997)Methods for overcoming interspecific crossing barriers. In: Shivanna KR (ed) Pollen Biotechnology for Crop Production and Improvement (pp. 273-292)
Wang J, Xia HJ, Zhou C & Yang HY (1997) Establishment of an experimental system for artificial germination and in vitro pollination with de-exined pollen in Nicotiana tabacum. Acta Botanica Sinica 39: 405-410
Wernsman EA, Matzinger DF & Rufty RC (1989) Androgenetic vs. gynogenetic doubled haploids of tobacco. Crop Sci. 29: 1151-1155
Zhang YX & Lespinasse Y (1991) Pollination with gammairradiation pollen and development of fruits, seeds and parthenogenetic plants in apple. Euphytica 54: 101-109
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Germanà, M., Chiancone, B. Gynogenetic haploids of Citrus after in vitro pollination with triploid pollen grains. Plant Cell, Tissue and Organ Culture 66, 59–66 (2001). https://doi.org/10.1023/A:1010627310808
Issue Date:
DOI: https://doi.org/10.1023/A:1010627310808