Clonal propagation of Trifolium Pratense,T. Resupinatum and T. Subterraneum by direct somatic embryogenesis on cultured immature embryos

Direct somatic embryogenesis on immature zygotic embryos in vitro has been confirmed for Trifolium pratense and extended to T. resupinatum and T. subterraneum. For all species direct embryo cloning can be achieved on an appropriate basal medium supplemented with 1gl^–1 yeast extract and 0.05 mgl^–1 BAP. Basal medium/sucrose formulation, level of yeast extract and level of BAP affected the nature of in vitro responses. In particular, for T. pratense and T. subterraneum lowering of the yeast extract level suppressed embryoid initiation, and raising of the BAP level stimulated formation of nodular morphogenic callus. For T. resupinatum alteration of the basal medium/sucrose formulation changed the tissue site of embryoid initiation from hypocotyl to cotyledons or both. Control of embryoid initiation is briefly discussed.Abbreviation BAP 6-benzylaminopurine