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Study on the adventitious root culture of Tripterygium wilfordii under shaking culture conditions

Li Yan;Zhao Lei;Wen Peng-Fei;Ge Fu-Hua;Zhang Xing;College of Life Science,Northwest A & F University;Shaanxi Province Technology and Engineering Center of Biopesticide;  
Tripterygium wilfordii Hook.f.,adventitious roots were used in vitro to study the effects of inoculation density,liquid medium volume per flask,step-by-step amplification,defoaming agent,macroporous adsorption(MA) resin type,and concentrations of triptolide,wilforgine,and wilforine in roots and medium on adventitious root growth after shake-flask culture.Results showed that an inoculation density of 15 g/L was suitable for the subculture of adventitious roots and accumulation of secondary metabolite.The medium utilization rate was the highest in a 250-m L flask with 100 m L of liquid medium.Adventitious root growth declined slightly withthe increase in flask volume.In a 5-L bottle,the adventitious root growth and triptolide,wilforgine,and wilforine concentrations were 91.6%,91.8%,91.7%,and 96.9%,respectively,compared to the control,XAD-7 resin markedly enhanced adventitious root growth and secondary metabolite content among the six kinds of MA resin.The adventitious root growth and triptolide,wilforgine,and wilforine content increased 1.2-,2.9-,2.4-,and2.2-fold,respectively,compared to the control when XAD-7 resin was at 0.5 g/bottle.Adventitious root growth and secondary metabolite content all decreased at differing degrees when the defoaming agent was added to the medium.Adventitious root growth and triptolide,wilforgine,and wilforine contents were 85%,78%,64%,and 87%,respectively,compared to the control when LX-603 was added to the medium;however,foam was markedly inhibited during the culturing process.This study demonstrated that step-by-step amplification was a feasible approach for adventitious root culture.Furthermore,this research provides the basis for the scale-up culture of T.wilfordii adventitious roots using a bioreactor.
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