Studies On Transformation Of Medical Resource Plant Dioscorea Zingiberensis

Dioscorea zingiberensis C.H.Wright is a native species in China, which is an important perennial, medical resource plant. The diosgenin in its rhizome is the important material for steroid hormonal medicines. In all Dioscorea species, the concent of diosgenin in wild Dioscorea zingiberensis is the highest, and reported to be 16.15%. Nowadays, there are some problems on Dioscorea zingiberensis breeding. First, wild Dioscorea zingiberensis resources were reduced for the excessively herborized and utilization. Second, the contention of diosgenin was decreased in the domestic plants. Third, the production of rhizome and the contention of diosgenin were not steady in tetraploid of Dioscorea zingiberensis. Genetic engineering is a new pathway to improve the breeding of Dioscorea zingiberensis.First, it has established a high frequency regeneration system of Dioscorea zingiberensis in this experiment. The most appropriate Murashige-koog (MS) medium with different concentration of 6-BA and NAA was determined, which could induce axillary buds, callus, differention and roots growth. rFCA-RRM1 and rFCA-RRM2 have the function of increasing biological-production. This study accomplish to gain the transformant of Dioscorea zingiberensis by Agrobaterium-mediated and gene gun, respectively. The Agrobacterium tumefaciens EHA105 with rFCA-RRM1 were transformated into Dioscorea zingiberensis callus and discussed effect of concentration of Agrobacterium,infected time,co-culture time,the selection pressure.The resistant callus became whole plants by regeneration, radication, strength and transplant, then to be the transgenic plants. The PCR results showed that transgenic plants contained HPT and target gene. It proved the successful transmutations. The diosgenin was extracted from wild plants'rhizome and cullus, and detected the contention by HPLC.