Abstract
Interspecific somatic hybrid plants were obtained by symmetrical electrofusion of mesophyll protoplasts of Medicago sativa with callus protoplasts of Medicago arborea. Somatic hybrid calli were picked manually from semi-solid culture medium after they were identified by their dual color in fluorescent light. Twelve putative hybrid calli were selected and one of them regenerated plants. The morphogenesis of the somatic hybrid calli was induced by the synthetic growth regulator 1,2 benzisoxazole-3-acetic acid. Somatic hybrid plants showed intensive genome rearrangements, as evidenced by isozyme and RFLP analysis. The morphology of somatic hybrid plants was in general intermediate between the parents. The production of hybrids by protoplast fusion between sexually incompatible Medicago species is related to the in vitro respon siveness of the parental protoplasts. The possibility of using somatic hybrid plants in alfalfa breeding is discussed.
Similar content being viewed by others
References
Arcioni S, Damiani F, Pezzotti M, Lupotto E (1990) Alfalfa, Lucerne (Medicago spp.). In: Bajaj YPS (ed) Biotechnology in arricultureture and forestry vol 10, Legumes and oilseed crops I. Springer, Berlin Heidelberg New York Tokyo, pp 197–241
Arcioni S, Damiani F, Pupilli F (1994) Somatic hybridization in the genus Medicago In: Bajaj YPS (ed) Biotechnology in agriculture and forestry vol 27. Somatic hybridization in crop improvement I. Springer, Berlin Heidelberg New York Tokyo, pp 145–164
Arcioni S, Mariani A, Damiani F, Pupilli F (1995) Somatic hybridization and embryo rescue for the introduction of wild germ plasm. In: McKersie BD, Brown DCW (eds) Biotechnology and the improvement of forage legumes. CAB international, Wallingford Oxford, UK (in press)
Branca C, Ricci A, Torelli A, Bassi M (1993) Effect of 1,2 benzisoxazole-3-acetic acid on plant regeneration from protoplasts of Nicotiana tabacum. Plant Cell Rep 12:121–124
Bingham ET (1979) Maximizing heterozygosity in autopolyploids. In: Lewis WH (ed) Polyploidy. Biological relevance. Plenum Press, New York London, pp 471–489
Corleto A, Venezian ME, Magini L, Eroli A, Cordella S (1980) Prove di adattamento e produzione di arbusti da pascolo in diverse località della Puglia e della Basilicata. Rivista di Agron 14:42–49
Dumbier MW, Bingham ET (1975) Maximum heterozygosity in alfalfa: results using double haploid-derived autotetraploids. Crop Sci 15:527–531
Elgin JH Jr, Ostazeky SA (1982) Evaluation of selected alfalfa cultivars and related Medicago species for resistance to race 1 and 2 anthracnose. Crop Sci 22:39–42
Frearson EM, Power JB, Cocking EC (1973) Isolation, culture and regeneration of Petunia leaf protoplasts. Dev Biol 33:130–137
Isaac I (1957) Wilt lucerne caused by species of Verticillium. Ann Biol 45:550–558
Kao KN, Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at very low population density in liquid media. Planta 126:105–110
Lesins KA, Lesins I (1979) Genus Medicago (Leguminosae): a taxogenetic study. W.Junk, The Hague Boston London
Maliga P, Lazar G, Joo F, Nagy AH, Menczel L (1977) Restoration of morphogenetic potential in Nicotiana by somatic hybridization. Mol Gen Genet 157:291–296
Mariani A (1968) Impiego dell'incrocio interspecifico nel miglioramento genetico dell'erba medica. Genet Agr 22:35–51
Mariani A, Arcioni S, Veronesi F (1978) Cytological analysis and electrophoresis patterns of seeds proteins in Medicago sativa, Medicago glutinosa and their hybrids. Genet Agr 32:21–39
Mariotti D, Arcioni S, Pezzotti M (1984) Regeneration of Medicago arborea L. plants from tissue and protoplast cultures of different organ origin. Plant Sci Lett 37:149–156
McEvans A (1966) The breeding potential of wild perennial Medicago spp. Acta Agric 16:46–49
Mendis MH, Power JB, Davey MR (1991) Somatic hybrids of the forage legumes Medicago sativa L. and Medicago falcata L. J Exp Bot 42:1565–1573
Michaud R, Lehman WF, Rumbaugh MD (1988) World distribution and historical development. In: Hanson AA, Barnes DK, Hill RR Jr. (eds) Alfalfa and alfalfa improvement. Agronomy monograph vol 29, American Society of Agronomy, Crop Science Society of America, Soil Science Society of America. Madison, Wisconsin, USA, pp 25–91
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Piccirilli M, Pupilli F, Arcioni S (1988) Lotus tenuis Wald and Kit. In-vitro conditions for plant regeneration from protoplasts and callus of various explants. Plant Sci 55: 77–82
Phillips GC, Collins GB (1979) In vitro tissue culture of selected legumes and plant regeneration from callus culture of red clover. Crop Sci 19:59–64
Pupilli F, Damiani F, Arcioni S (1991) Protoplast fusion in the genus Medicago and isoenzyme analysis of parental and somatic hybrid cell lines. Plant Breed 106:122–131
Pupilli F, Scarpa GM, Damiani F, Arcioni S (1992) Interspecific somatic hybrid plants in the genus Medicago developed through protoplast fusion. Theor Appl Genet 84:792–797
Pupilli F, Businelli S, Caceres ME, Damiani F, Arcioni S (1995) Molecular, cytological and morpho-agronomical characterization of hexaploid somatic hybrids in Medicago. Theor Appl Genet 90:347–355
Quiros CF, Bauchan GR (1988) The genus Medicago and the origin of the Medicago sativa complex. In: Hanson AA, Barnes DK, Hill RR Jr. (eds) Alfalfa and alfalfa improvement. Agronomy monograph vol 29, American Society of Agronomy, Crop Science Society of America, Soil Science Society of America. Madison, Wisconsin, USA, pp 25–91
Renfro BL, Sprague EW (1959) Reaction of Medicago species to eight alfalfa pathogens. Agron J 51:481–483
SAS user's guide (1988) Release 6.03 edition. SAS Institute Incorporated, Cary, North Carolina
Téoulé E (1983) Hybridation somatique entre Medicago sativa L. et Medicago falcata L. Cr Acad Sci Paris 297:13–16
Uchimiya H, Murashige T (1974) Evaluations of parameters in the isolation of viable protoplasts from cultured tobacco cells. Plant Physiol 54:936–944
Author information
Authors and Affiliations
Additional information
Communicated by F. Salamini
Rights and permissions
About this article
Cite this article
Nenz, E., Pupilli, F., Damiani, F. et al. Somatic hybrid plants between the forage legumes Medicago sativa L. and Medicago arborea L.. Theoret. Appl. Genetics 93, 183–189 (1996). https://doi.org/10.1007/BF00225744
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00225744