Somatic embryogenesis of bupleurum falcatum L. Ⅰ. Effects of growth regulator, glutamine and activated charocal

The high frequency of direct somatic embryogenesis (PEDC’s pattern) was observed from leaf explants of Bupleurum falcatum L. The direct embryogenesis from leaf segments of regenerated plantlets showed the highest frequency on the MS media supplemented with 0.5mg/ℓ 2.4-D, while that of spontaneous plantlets, with 1.0mg/ℓ 2, 4-D. The former was more established than the latter. Propagation of embryogenic clump was promoted on the agar medium supplemented with 3mg/ℓ kinetin in presence of 0.4% AC, while suspension culture showed the best reaction in the medium supplemented with 2mg/ℓ kinetin. The supplement of glutamine in presence of 0.4% AC promoted remarkably the development of globular embryo. Addition of AC had a marvelous efficacy on the propagation of embryogenic clumps and normal embryo development. Synchronization of somatic embryogenesis was achieved in suspension culture by the secondary sieving the cell clumps at globular embryo stage. The aggregates of 200-500 μm in size were uniformly matured into normal embryos in the medium without growth regulator.