Studies On Tissue Culture Of F.ussurensis Maxim To Producing Alkaloid

This thesis, for the first time, aiming at producing alkaloid, deals with tissue culture of F.ussurensis Maxim, which is on the brink of extinction and only exists in Changbai mountains in China.Comparing the cultured tissues with cultivated bulb shows that the total content of alkaloid in regenerative bulb, adventitious root and callus is higher than that of cultivated bulb about 0.0665%, 0.0475% and 0.1624%, respectively. The content of adventitious bud is lower than that of cultivated bulb about 0.0150%.To evaluate many factors which affect the production of alkaloid in regenerative bulb, we use three main norms: growth rate^ rate of dry/fresh(%) and content of alkaloid(%).In the process of bulb culture, the accumulation of dry matter nearly reaches the highest level in 60 days after inoculation, which is suitable for harvesting. Liquid medium is worse than solid medium for accumulating alkaloid, though it is benefit for the differentiation of the little bulbs. The experiments for basic medium selection indicate that LS medium was the most suitable medium which could produce the highest yield of alkaloid among four types of media. 1.0mg.L"!BA+0.5mg.L"INAA is better than other three combinations. By different sucrose concentrations, the highest growth rate and yield of alkaloid is in 5%.Sugar is a suitable substitue for sucrose. It could not only reduce the cost of medium about 30%, but also increase the yield of alkaloid by about 42.77%. Ribavirin of different concentrations were used in medium. The results indicate that it has significant activity on bulb growth by optimized concentration and time affecting, culturing 60 days in Smg.L"1 ribavirin is the optimized combination. The addition of L-tyrosine is favorable to the growth of bulb, but not to the accumulation of alkaloid. Using boiling water to substitue distilled water reduces the yield of alkaloid by about 17.77%.Among the 7 different combinations of regulators for inducing callus from slices of regenerative bulb, O.Smg.L^NAA is the best. But for producing alkaloid, 1.0mg.L"1NAA+0.5mg.L"1KT is the optimum of all. There is significant difference between light and dark culture of callus, light culture is better than dark.In conclusion,it is possible to produce alkaloid by tissue culture,but need further research.