The Establishment Of Optimal Culture System And The Pilot-scale Culture Of Adventitious Root Of Tripterygium Wilfordii Hook. F

Tripterygium wilfordii Hook f. is a traditional Chinese medicine plants as well as awoody climber botany resource. Modern pharmacological studies show that the T. wilfordiihave many activities, such as anti-rheumatic, antibacterial, anticancer, anti-HIV. Inagricultural activity aspect we found it was effectively in toxic action, antifeedant anddevelopment inhibition to pests. many research had been widely studied Because it canproduce a series of biological active substances, including triptolide and alkaloids. With thedramatically increase demands in medicine and biorational pesticide, and also the blindexploitation of wild resources, So it’s an effective way through tissue culture to produceuseful metabolites to due with the problem.The adventitious roots lines was established using the adventitious roots after beingrepeated induced from callus. The growth rate and the content of triptolide, wilforine andwilforgine of the adventitious roots was explored by different auxin, elicitors, fermenter andorganic solvent. The main results and conclusions were given as follows:1. Adventitious roots induce and establishment from suspension culture system:Callus can produce adventitious roots on the1/2MS+0.5mg/L IBA, MS+0.5mg/L IBA,MS+1.0mg/L IBA solid medium. Adventitious root induced by IBA auxin grow better,adventitious roots induced from MS+1.0mg/L IBA solid medium grow stout than other solidmedium; MS+1.0mg/L2,4-D+0.5mg/L KT solid medium can induce callus and adventitiousroots, and the growth of adventitious roots was higher than other adventitious roots aftertransferred to the liquid medium.2. The selection of different growth regulator:Through regulating different growth regulator of the growth and secondary metabolitecontent of adventitious roots in the liquid medium, we access to two better training conditions:MS+4.0mg/L NAA+0.5mg/L KT and MS+2.0mg/L IBA.3. The selection of different elicitors:Through regulating different concentration of MeJA, AgNO3and yeast extract to enhance the growth and secondary metabolite content of adventitious roots in the liquidmedium, we access to two better training conditions: In MS+4.0mg/L NAA+0.5mg/LKT+50μmol/L MeJA cultured conditions，the content of the triptolide in adventitious rootwas31.638μg/g,1.31times as root bark; the content of the triptolide in the culture mediumwas337.170μg/bottle; the content of the triptolide in adventitious root and the culturemedium was422.939μg/flask,1.24times as the control. In MS+4.0mg/L NAA+0.5mg/LKT+30μmol/L AgNO3cultured conditions, the content of the triptolide in adventitious rootwas27.368μg/g,1.28times as root bark; the content of the triptolide in the culture mediumwas313.067μg/bottle; the content of the triptolide in adventitious root and the culturemedium was366.240μg/flask,1.15times as the control.4.5L reactor cultured adventitious roots:The secondary metabolites of adventitious roots cultured40days by using5L stirredfermentor showed no significant difference with the secondary metabolites of adventitiousroots cultured in250mL shake flasks.5. The preliminary study of two-phase cultivation about adventitious roots:Through regulating different concentration of DBP and OA to enhance the growth andsecondary metabolite content of adventitious roots in the liquid medium.The growth rate andHPLC results of adventitious roots showed that the treatment of organic solvent decreased thegrowth rate and the content of secondary metabolites, the secondary metabolites were notabsorbed into the organic solvent, other organic solvents need to be screened.