天蓝苜蓿原生质体培养再生植株

Protoplasts isolated from suspension cell lumps of Medicago lupulina L. started to divide after 2 days in K8p culture medium containing 0.1～2.0 mg/L of 2,4 D, with a maximum division frequency of 38.35%. After 5 weeks of culture, the protoplast derived cell lumps were transferred to liquid/solid double layer media for microcallus regeneration, with a maximum frequency of 0.58%.The whole plants were regenerated from protoplast derived calli via somatic embryogenesis and organogenesis.In somatic embryogenesis,the embryoids were induced on MS and W 14 media with rather wide range (1.0～20.0 mg/L) of 2,4 D concentration. The highest induction frequency of embryoids was 71.0%. In organogenesis,the differentiation media containing lower concentration of 6 BA (0.5～0.7 mg/L) were suitable for adventitious bud formation.The highest frequency of adventitious bud formation from calli was 27.8%. The mature protoplast regenerated plants were obtained 3 months after transplanting the plantlets into soil.