The Effects Of Precursors And Fungal Elicitors For The Secondary Metabolites Production In Tripterygiun Wilfordii Hook. F Suspension Cell Cultures

Tripterygium wilfordii Hook.F(Celastraceae,Tripterygium) is a wild climber. Modernchemical and pharmacological studies show that the T. wilfordii have many activities:diterpenes; triterpenes; alkaloids et al.Clinical researching find it has mang efficacy:anti-rheumatic, antibacterial, anticancer, anti-HIV,and it’s effective to inhibition pests.But thecontent of activities is low in plant. The output of T. wilfordii is too exiguous to appease thedemands in medicine and biorational pesticide. The effect of precursor and endophytic fungielicitor feeding on the production of triptolide, wilforgine and wilforine was studied withsuspension cell cultures of T. wilfordii. The main results were as follows:1. Precursors supplementation increased the accumulation of secondary metabolites, andwith negative effect on cell growth. Different precursors and concentration has differentstimulation:(1) The triptolide contents increased up to1.29-folds by feeding0.1mmol/Lnicotinic acid.(2) Both of the three precursors can improve wilforgine accumulation. Thewilforgine contents increased up to5.58-folds by feeding1.5mmol/L isoleucine.With thetreatment of isoleucine, asparagic acid, nicotinic acid, the wilforgine was up to218.14μg/flask,154.42μg/flask and183.49μg/flask respectively.(3) wilforine reach to28.79μg/flaskand11.08μg/flask when treat with isoleucine and asparagic acid respectively. No visibleeffort was found in nicotinic acid treatment.2. Elicitation of15-day-old cell cultures by F4-3elicitor caused the highest wilforgineand total alkaloids production.3. F4-3、F4-31and F4-20elicitor can improve the secondary metabolites production butrepress the cell growth. Different fungal mycelium homogenates and concentration stimulatedifferent kinds of secondary metabolites accumulation:(1)35mg/L F4-31elicitor promote thewilforgine to127.06μg/flask,3.1-folds than control, and3.2-folds,1.83-folds higher whentreat with F4-31, F4-20, the content was127.88μg/flask and74.21μg/flask respectively.(3)The triptolide contents increased up to2.54-folds by feeding with nicotinic acid.4. Exposed to a same fungal elicitor F4-3, several different cell lines generated thedifferent responses regarding as secondary metabolites production. It’s highly increased thetriptolide, wilforgine, wilforgine to6-folds，5-folds，6.2-folds well1.1-folds，2.5-folds， 1.8-folds and1.2-folds，2.4-folds，1.4-folds than control when use root and stem as cellcultures.