Abstract
Embryogenic protoplasts of Dancy tangerine (Citrus reticulata Blanco) were X-ray irradiated at three doses and electrofused with iodoacetic acid-treated embryogenic protoplasts of Page tangelo (C. reticulata Blanco × C. paradisi Macf.). Shoots could regenerate only from the fusion combination with the lowest irradiation dose, but were recalcitrant to rooting. In vitro grafting was applied to obtain complete plants. Chromosome examination showed that the plants contained mainly diploid and aneuploid cells, together with few tetraploid cells, indicating that they were mixoploids. Random amplified polymorphic DNA analyses with 10-mer arbitrary primers confirmed the plants as true somatic hybrids. This is the first report on regeneration of mixoploid hybrid plants via protoplast asymmetric fusion in Citrus. Negative effects of ionizing irradiation on regeneration of embryoids and plantlets and possible agronomic interest of the mixoploid plants are also discussed.
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Bates, G.W., 1990. Asymmetric hybridization between Nicotiana tabacum and N. repanda by donor recipient protoplast fusion: Transfer of TMV resistance. Theor Appl Genet 80: 481–487.
Deng, X.X., S.Y. Xiao, Z.A. Deng & W.C. Zhang, 1993. Interspecific somatic hybrid of Ichang papeda with Valencia sweet orange. Chinese J Biotech 9: 103–107.
Derks, F.H.M., J.C. Hakkert, W.H.J. Verbeek & C.M. Colijn-Hooymans, 1992. Genome composition of asymmetric hybrids in relation to the phylogenetic distance between the parents, nucleus-chloroplast interaction. Theor Appl Genet 84: 930–940.
Dudits, D., E. Maroy, T. Praznovszky, Z. Olah, J. Gyorgyey & R. Cella, 1987. Transfer of resistance traits from carrot into tobacco by asymmetric somatic hybridization: regeneration of fertile plants. Proc Natl Acad Sci USA 84: 8434–8438.
Feher, A., J. Preiszner, Z. Litkey, Gy. Csanadi & D. Dudits, 1992. Characterization of chromosome instability in interspecific somatic hybrids obtained by X-ray fusion between potato (Solanum tuberosum L.) and S. brevidens Phil. Theor Appl Genet 84: 880–890.
Forsberg, J., U. Lagercrantz, & K. Glimelius, 1998. Comparison of UV light, X-ray and restriction enzyme treatment as tools in production of asymmetric somatic hybrids between Brassica napus and Arabidopsis thaliana. Theor Appl Genet 96: 1178–1185.
Grosser, J.W. & F.G. Gmitter Jr, 1990. Protoplast fusion and citrus improvement. Plant Breed Rev 8: 339–374.
Grosser, J.W., J. Jiang, F.A.A. Mouraofo, E.S. Louzada, K. Baergen, J.L. Chandler & F.G. Gmitter Jr, 1998a. Somatic hybridization, an integral component of citrus cultivar improvement. I. Scion improvement. Hort Sci 33: 1057–1059.
Grosser, J.W., J. Jiang, E.S. Louzada, J.L. Chandler & F.G. Gmitter Jr, 1998b. Somatic hybridization, an integral component of citrus cultivar improvement. II. Rootstock improvement. Hort Sci 33: 1060–1061.
Grosser, J.W., P. Ollitrault & O. Olivares-Fuster, 2000. Somatic hybridization in citrus: an effective tool to facilitate variety improvement. In Vitro Cell Dev Biol Plant 36: 434–449.
Hinnidaels, S., L. Bariller, A. Mouras, V. Sidorov, J. Del-Favero, J. Veuskens, I. Negrutiu & M. Jacobs, 1991. Highly asymmetric intergeneric nuclear hybrids between Nicotiana and Petunia: evidence for recombination and translocation events in somatic hybrid plants after 'gamma-fusion'. Theor Appl Genet 82: 609–614.
Liu, J.H. & X.X. Deng, 1999. Production of hybrid calluses via protoplast asymmetric fusion between Microcitrus papuana and Newhall navel orange. Plant Cell Tiss and Organ Cult 59: 81–87.
Liu, J.H. & X.X. Deng, 2000. Regeneration and characterization of plants derived from protoplast asymmetric fusion in Citrus. Acta Bot Sin 42: 1144–1149.
Liu, J.H., C.G. Hu & X.X. Deng, 2000. Production of intergeneric somatic hybrid plants via protoplast electrofusion in Citrus (in Chinese). Acta Bio Exp Sin 33: 325–332.
Liu, J.H., C.G. Hu & X.X. Deng, 1999. Regeneration of diploid intergeneric somatic hybrid plants between Microcitrus and Citrus via electrofusion. Acta Bot Sin 41: 1177–1182.
Murashige, T. & D.P.H. Tucker, 1969. Growth factor requirements of citrus tissue culture. In: H.D. Chapman (Ed.), Proc. 1st Citrus Symp. Vol 3. Univ of Cali, Riverside, pp. 1155–1161.
Oberwalder, B., B. Ruoß, L. Schilde-Rentschler, V. Hemleben & H. Ninnemann, 1997. Asymmetric fusion between wild and cultivated species of potato (Solanum spp.) - Detection of asymmetric hybrid and genome elimination. Theor Appl Genet 94: 1104–1112.
Ohgawara, T., S. Kobayashi, E. Ohgawara, H. Uchimiya & S. Ishii, 1985. Somatic hybrid plants obtained by protoplast fusion between Citrus sinensis and Poncirus trifoliata. Theor Appl Genet 71: 1–4.
Oiyama, I., S. Kobayashi, K. Yoshinaga, T. Ohgawara & S. Ishii, 1991. Use of pollen from a somatic hybrid between Citrus and Poncirus in the production of triploid. Hort Sci 26: 1082.
Rasmussen, J.O., S. Waara & O.S. Rasmussen, 1997. Regeneration and analysis of interspecific asymmetric potato-Solanum ssp hybrid plants selected by micromanipulation or fluorescenceactivated cell sorting (FACS). Theor Appl Genet 95: 41–47.
Rasmussen, J.O., A. Lossl & O.S. Rasmussen, 2000. Analysis of the plastome and chondriome origin in plants regenerated after asymmetric Solanum ssp. Protoplast fusions. Theor Appl Genet 101: 336–343.
Samoylov, Y.M. & K. Sink, 1996. The role of irradiation dose and DNA content of somatic hybrid calli in producing asymmetric plants between an interspecific tomato hybrid and eggplant. Theor Appl Genet 92: 850–857.
Shi, Y.Z., W.W. Guo & X.X. Deng, 1998. Establishment of RAPD system and identification of citrus somatic hybrid (in Chinese). Acta Hort Sin 25: 105–110.
Spangenberg, G., M.P. Valles, Z.Y. Wang, P. Montavon, J. Nagel & I. Potrykus, 1994. Asymmetric somatic hybridization between tall fescue (Festuca arundinaceae Schreb.) and irradiated Italian ryegrass (Lolium multiforum Lam.) protoplasts. Theor Appl Genet 88: 509–519.
Tian, D. & R.J. Rose, 1999. Asymmetric somatic hybridization between the annual legumes Medicago truncatula and Medicago scutellata. Plant cell Rep 18: 989–996.
Vardi, A., P. Arzee-Gonen, A. Frydman-Shani, S. Bleichman & E. Galun, 1989. Protoplast-fusion mediated transfer of organelles from Microcitrus into Citrus and regeneration of novel alloplasmic trees. Theor Appl Genet 78: 741–747.
Vardi, A., A. Breiman & E. Galun, 1987. Citrus cybrids: production by donor-recipient protoplast - fusion and verification by mitochondrial-DNA restriction profiles. Theor Appl Genet 75: 51–58.
Vlahova, M., S. Hinnisdaels, F. Frulleux, M. Claeys, A. Atanassov & M. Jacobs, 1997. UV irradiation as a tool for obtaining asymmetric somatic hybrids between Nicotiana plumbaginifolia and Lycopersicon esculentum. Theor Appl Genet 94: 184–191.
Varotto, S., E. Nenz, M. Lucchin & P. Parrini, 2001. Production of asymmetric somatic hybrid plants between Cichorium intybus L. and Helianthus annuus L. Theor Appl Genet 102: 950–956.
Wijbrandi, J., A. Posthuma, J.M. Kok, R. Rijken, J.G.M. Vos & M. Koornneef, 1990. Asymmetric somatic hybrids between Lycopersicon esculentum and irradiated Lycopersicon peruvianum. 1: Cytogenetics and morphology. Theor Appl Genet 80: 305–312.
Xiao, S.Y., F.G. Gmitter Jr, J.W. Grosser & S. Huang, 1995. RAPD analysis: a rapid method to identify citrus somatic hybrids (in Chinese). Hereditas 17: 40–42.
Xu, Y.S., M. Murto, R. Dunckley, M.G.K. Jone & E. Pehu, 1993. Production of asymmetric hybrids between Solanum tuberosum and irradiated S. brevidens. Theor Appl Genet 85: 729–734.
Yamamoto, T., Y. Nakajima & K. Oeda, 2000. Morphological changes in homeotic cytoplasmic male-sterile carrots combined with fertile cytoplasm by asymmetrical cell fusion. Plant cell Rep 19: 363–370.
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Liu, J., Deng, X. Regeneration and analysis of citrus interspecific mixoploid hybrid plants from asymmetric somatic hybridization. Euphytica 125, 13–20 (2002). https://doi.org/10.1023/A:1015748411654
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DOI: https://doi.org/10.1023/A:1015748411654