Study On The Tissue Culture And The Accumulation Of The Alkaloid From Fritillaria Unibracteata Hisao Et K.C.Hsia

F.unibracteata Hisao et K.C.Hsia as one of the most genuine medicinal materials in Sichuan province contains various alkaloid and saponins and essential trace elements in human body, the dry bulbs of this plant are used as clearing away heat and eliminatingsputum, relieving cough, anti-inflammation and relieving pain, which has always great concerns on the traditional chinese medicine.As the factors of unique growth environment, longer growth cycle, the yield is lower and the price is so high.However,because of the mankind's excessively exploiting,the wild resources of this plant has turned into an endangered population.The method of tissue culture could solve these problems.Explants from the fresh bulb-scale sections of F.unibracteata were first pretreated by low temperature and added a certain concentration Vc to control moldy and brown.Then the induction rate of the callus and adventitious bud was investigated with different ratios concentration of growth hormone. Explants were in-vitro recultured as the optimized induction condition. To peimine for reference substance,the relational alkaloid's accumulation of the samples in the different growth period could be determined by HPLC.The tissue culture experiment results:The highest induction rate of the adventitious buds reached 73.3%, the highest induction rate of the callus reached 53.3%.The most conditions of culture system could be conducted by the highest induction rate,which was:low temperature pretreatment for 48h, light of 16h per day, MS(pH5.8)+NAA1.2 mg/L +6-BA1.6 mg/L+ Vc0.1 g/L. The accumulation of relational alkaloid in the adventitious buds rised by the time,the highest accumulation was more than double to the original adventitious buds.The best period of accumulation was conducted,was about 32d.The construction of the optimized culture system would be helpful to protect the natural resources of F.unibracteata Hisao et K.C.Hsia and start the new approach of the natural resources; Research on the accumulation in the different growth period,which had importance to study and exploit it's secondary metabolites,and to achieve the modernization of Chinese medicine.