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Characterization of recurrent somatic embryogenesis of alfalfa on auxin-free medium
Authors:Wayne A Parrott  Matthew A Bailey
Institution:(1) Department of Agronomy, University of Georgia, 30602 Athens, GA, USA
Abstract:Callus cultures from 300 genotypes of alfalfa (Medicago sativa L.) were initiated from leaf, petiole, and internode explants placed on Blaydes medium containing 10.74 mgrM agr-naphthaleneacetic acid, 11.42 mgrM indole-3-acetic acid, and 9.29 mgrM kinetin. Five genotypes produced somatic embryos. Upon transfer of these embryos to growth regulator-free Murashige and Skoog medium with B5 vitamins, new somatic embryos repeatedly formed directly on older somatic embryos without an intervening callus phase in a cycle lasting about 30 days. These cultures have been maintained for two years, during which time their embryogenic capacity has remained stable. New embryogenic cultures could be started repeatedly from these genotypes. The elimination of sugars from the medium could stop recurrent embryogenesis. Glucose, maltose, and fructose stimulated recurrent embryogenesis more effectively than sucrose. Sucrose was superior to lactose, while sorbitol and mannitol did not stimulate recurrent somatic embryogenesis. The absence of nicotinic acid in the medium, as long as sucrose was present, was lethal to embryos of three of the five tested genotypes. The ability of this system to propagate embryos exponentially offers potential for development of new gene transfer systems and application to artificial seed technology.Abbreviations NAA agr-naphthaleneacetic acid - RSE recurrent somatic embryogenesis
Keywords:carbohydrate source  lucerne  nicotinic acid
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