Study On The Inducement Of Hairy Root From Trichosanthes Bracteata Voigt And The Investigation Of The Inducing Mechanism

Aimed at developing a set of methods efficient in inducing the hairy roots with Agrobacterium rhizogenes from Trichosanthes bracteata Voigt to meet needs for hairy roots for bioreactor culture research and application; and also in order to investigate the roles of explants, Agrobacterium rhizogenes, affecting factors and their magnitude and the underlying mechanism in the transformation, therefore to determine the optimum explants, pre-culture time, tested strain concentration, infection time, co-culture time, affecting factors and their effects working together and to obtain the optimum transforming condition by performing orthogonal test, this project was launched. With cotyledons and stems from Trichosanthes bracteata Voigt plantlets as explants, Agrobacterium rhizogenes R1601 and R1000 as infection agents, transformation was performed using different methods and under various circumstances . The transformed hairy roots were detected for agropine to make sure the transformation was successful.A conclusion was reached as follows: Agrobacterium rhizogenes R1601 was an ideal strain for hairy roots transformation from Trichosanthes bracteata Voigt which exhibited extraordinary transformation rate(60%). OD600=0.7 was the optimum infection concentration for the tested transforming agent, with temperature at 25℃, pH at 6.0, 20min of infection and 48h of co-culture and phytohormone at 1.0mg/L being the most favourable hairy root-inducting pattern. The detection of agropine and trichosathin was conducted and proved available in the resulted hairy roots. R1000 remained the best alternative to R1601 though it had lower transforming capability. The entire transformation consists of three major respects: the explant preparation, culture of agent strains and transformation. Each of the affecting factors work on the process by acting on one or more respects of it.