PLANT-REGENERATION FROM HIGHLY EMBRYOGENIC CALLUS, CELL-SUSPENSION AND PROTOPLAST CULTURES OF TRIFOLIUM-FRAGIFERUM

Using various media, tissue and protoplast cultures plant regeneration systems were developed for Trifolium fragiferum (2n=16). (L.). The be st media for induction of embryogenic cultures were based on Kao (1977 ) or Kao and Michayluk (1975). Somatic embryogenesis was observed in c ultures derived from green leaf mesophyll protoplasts of branching pla nts, somatic embryo protoplasts and cell suspension protoplasts, leafl ets and various explants of immature zygotic embryos. The process of s omatic embryogenesis was maintained for over two years on Murashige an d Skoog's (1962) medium supplemented with 0.5 mg 1(-1) benzyladenine a nd 0.05 mg 1(-1) naphthaleneacetic acid. These long term cultures were capable of regenerating plants that were fertile and produced seeds. These results were compared with those from protoplast, tissue and org an culture of other species of the Trifolium genus.