Study On Tissue Culture Of Epimedium Wushanense T.S.Ying

Epimedium wushanense T.S. Ying is traditional Chinese medicinal plant, which was recorded in national pharmacopoeia (2010), it has been used2000years of history.The research showed that it had significantly physiologically and pharmaceutical activity. So it has a great value of exploitation.There are not many published paper research on the tissue rapid propagation of E. wushanense. A large number of seedlings will be obtained by using biological technological technology in a short period of time. It is important for the protection of wild plant resources,source of drugs and sustainable ues. In this paper, the propagation by tissue culture were studied, incluing different explants buds inducation and proliferation, callus inducation and differentiation, root inducation of tube plantlets of E. wushanense. The main results were showed as the following:(1) Tender leaves、rachis、buds and embryo of E. Wushanense were optimal for explant primary culture.The best sterilizing method for explants were disinfected with75%ethanol for30s,then treated with0.1%HgCl2, leaves and rachis were soaked for6min, respectively; the buds was soaked for (4+2) min, which could control pollution rate within5%,the survival rate was above75%. The seeds were soaked with0.1%HgCl2for2minutes, the ratio of survival was50%.(2) The induction rate of callus from rachis was higher than that from leaves and embryos,but the callus which were induced from rachis and leaves was hardly differentiated. Additionally,the WPM medium was easier than the MS、B5and N6mediums to callus induction of E. Wushanense,and2,4-D played an important role in callus inducation. The optimal callus induction medium from embryo of E. Wushanense was MS+2,4-D2mg·L-1+IBA2mg·L/-1+NAA0.5mg·L-1,and the optimal callus induction medium from leaves of E. Wushanense was WPM+2,4-D2mg·L-1+6-BA0.5mg·L-1,the ratios of callus induction was79.2%.(3) The optimal medium for the shoot regeneration from embryo derived from in vitro cultured individual was MS+6-BAlmg·L-1+NAA0.5mg·L-1+IBA1mg·L-1,shoot regeneration rate was73.3%,6-BA played an important role in callus inducation. (4) Direct shoot regeneration was achieved by culturing E. Wushanense embryos and buds. Basic medium and6-BA showed the significant effect on the formation of buds,and the optimal medium was WPM+6-BA2.0mg·L-1+NAA0.1mg·L-1+GA30.5mg·L-1;6-BA was important for bud induction from E. Wushanense embryo,and the suitable medium was MS+IBA2mg·L-1+6-BA0.5mg·L-1, the ratios of bud inducation was77.8%; The best bud proliferation medium was MS+6-BA1.0mg·L-1+NAA0.5mg·L-1, and shoot multiplication rate was2.17.(5) The optimal growth of roots medium was1/2MS+IBA0.5mg·L-1, which was added0.05%active carbon.The rooting ratio was90%, and the buds growed heathy and strong.