The Establishment Of The Clone Rapid Propagation Regenerate Individual And The Studys On The Hormone Autotrophy Culture's Mechanism Of Morphogenesis Of Fritillaria Ussuriensis Maxim

Taking bulbs of fritillaria ussuriensis maxim as explants, sieve the optimal explant and medium . and set up high performance regeneration system,discuss the relations between different method in vitro conservation and the rate of survivorship, determine the type of differentiation in cells by paraffin section,analysis the regulations between physiological and biochemistry aspects and morphogensis of hormone autotrophy culture , In order to control the secondary metabolism and ground the foundation for mass production.The optimal basic medium of bud inducement from bulbuds and little stem is MS+NAA0.5mg/L+BA1.5mg/L, the optimal mediums of calli inducement are MS+2.4-D2.0mg/L and MS+2.4-D1.5mg/L,pH=6.0,light,Immerse the bulbuds with GA500mg/L to break dormancy ,and the regeneration circle is 40-50 days,we gain the hormone autotrophy type culture by changing the medium with half reducing the concentration of hormone and then culture with MS free of hormone, during the organogenesis course,the content of free amino acid is to go up and then go down,but the opposite condition is in the content of soluble sugar.The buds have the highest content of free amino acid is 10.90% and the calli has the highest contents of soluble sugar is 6.49% , the amylase activity is higher in buds, the change trend of α-amylase activity is similarly with the whole amylase activity's and it may play an important role in starch degradation. The zymogram of buds is complex which has many line and deep dying , it indicate that many enzyme have different activities and function during the culture period.Fritillaria culture and the crude drug have the same alkaloids spots. And the contents of total alkaloid in adventitious roots is higher than that in calli and buds.Using cryopreservation to conserve, the course is : add 5%DMSO+5%glycerol—>stay 2h in -20°C→conserve in ultra colder→defrosting in 38°C water bath→ wash with MS medium→subculture in fresh MS medium. Also add mannitol and PP333 can conserve fritillaria ussuriensis maxim culture in short period.