The Anther And Protoplast Culture In Dioscorea Zingiberensis

Dioscorea zingiberensis C.H.Wight, a native medicinal resource plant in China, contain the highest level of diosgenin in the rhizome comparing with other speicies in the world. After acclimated cultivation, its diosgenin content has been declined year and year. Biotechnology combining routine breeding is an expected efficient pathway to solve the problem. In this study, Whole plantlets were regenerated from anther culture, some of them were proved to be haploid; embryogenesis calli were induced and well suspension culture system was established and be used in protoplast isolation and culture. which offered a new approach for further breeding to enhance diosgenin content.The main results are as follows: 1. Anther culture and plantlets regenerated(1) The screening of basic culture medium: Anther at the later uni-nucleate stage was cultured in W14,B5, MS,N6 basic culture medium, calculate rate of inducement displayed that average inducement rate is 2.15%,0.50%,0.18%,0.10%, it showed that W14 basic culture medium suit for induce calli.(2) the influence of genotypes: the induction rate of 4 genotypes were different when cultured in W14+2,4-D2.0 mg/L+KT2.0 mg/L+NAA0.5 mg/L+60g/L maltose, the rate of induction was highest in Henan Nanyang genotype and reach to 5.20%, and the rate of induction was only 0.80% in Hubei Wudang.(3) The influence of 2,4-D and KT in induction of calli: The rate of calli induction from anther of Hubei Wudang increased with increasing the concentration of 2,4-D, when culture medium with 2 mg/L 2,4-D, highest induction rate(1.00%) was obtained; The rate of calli induction decreased with increasing the concentration of KT; So suitable hormone was 2,4-D 2 mg/L. However, the response of different genotypes to 2 hormone was significant different.(4) Differentiation of calli: two-step culture was used, Firstly, calli were transferred to shooting culture medium, Secondly transferred to rooting culture medium after shooting up. Shooting culture medium was MS basic culture medium containing BA 2.0...