Hairy Roots Inducing Of Tripterygium Wilfordii Hook. F

Tripterygium wilfordii Hook f, woody climber, is a traditional Chinese medicine plants as well as an important resource to produce biorational pesticides. Modern pharmacological studies show that the T. wilfordii have many activities, such as anti-rheumatic, antibacterial, anticancer, anti-HIV. In agricultural activity aspect we found it is effective in toxic action, antifeedant and development inhibition to pests. With the dramatically increasing demands in medicine and biorational pesticide, and also the blind exploitation of wild resources, it brings a lot pressures to the environment. Hairy roots are growing fast, not subject to the effects of season and without harmful substance pollution compared to conventional artificial cultivation, so they get more attention in recent years. In this study, we use Agrobacterium rhizogenes ATCC15834and A4to infect T. wilfordii to obtain hairy roots. The main results were as follows:1. Hairy roots of T. wilfordii are obtained with infecton by A. rhizogenes ATCC15834and A4on the midia of MS and1/2MS. The inductivity is the highest (80.36%) with the group of A4+1/2MS, the second is the group of A4+MS (72.73%), the lowest is the group of ATCC15834+1/2MS (46.30%). It indicates that A. rhizogenes ATCC15834and A4both have the capacity to induce hairy roots of T. wilfordii, but T. wilfordii is more sensitive to A. rhizogenes A4, and it is better-adapted to the media1/2MS.2. The content of triptolide, wilforgine and wilforine in T. wilfordii hairy roots are39.98、1100.20and234.61μg/g DW, with22.72,3.04and16.38times higher than that of callus respectively. The accumulation of triptolide in adventitious roots is the highest, the second one is the hairy roots, the last one is the callus, and the content in dventitious roots and hairy roots are both higher than that of naturally growing roots. Wilforgine contents in hairy root can achieve the same level with that of naturally growing root, while the content of wilforine is lower than naturally growing roots.3. The optimistic culture conditions of hairy roots on the solid medium was examined. Orthogonal test of4factors, which were concentration of sugar, macroelement content of the medium, inoculum size and pH of the medium respectively, and was designed with3levels each. The biomass of hairy roots cultured was measured and20g/L sugar,4.09g/L macroelement content of the medium (1650mg/L NH4NO3,1900mg/L KNO3,370mg/L MgSO4·7H2O,170mg/L KH2PO4) with pH of5.5and4g/L inoculum size resulted in the maximum biomass (14.71g).