Effects of Ultraviolet-b(UV-B) Radiation and Ultrasonic Treatment on the Content of Three Secondary Metabolites in Tripterygium wilfordii Hook.f. Cell Suspension Cultures
Abstract：A method for producing triptolide and alkaloid by Tripterygium. wilfordii Hook.f. suspension culture had been founded, but the secondary metabolites content is low that produced by this method, so the terpenoids secondary metabolites content in T. wilfordii Hook.f. suspension culture system were detected and analyzed by using ultraviolet-b(UV-B) radiation and ultrasonic treatment in this study. UV-B radiation and ultrasonic treatment were conducted to the cell suspension culture system of T. wilfordii at different culture period and treated time. To get the optimal treatment condition, HPLC(high performance liquid chromatography) was used to determine the content of triptolide, wilforgine and wilforine in suspension cell and culture medium of T. wilfordii. The results showed that UV-B radiation and ultrasonic treatment could improve the biomass of suspension cell and the content of the secondary metabolites in T. wilfordii cell suspension culture. After UV-B radiation and ultrasonic treatment, the biomass of suspension cell of T. wilfordii was about 1~2 times higher than that of control. After UV-B radiation for 90 min or ultrasonic treatment for 40 s at 13 d in the cell suspension cultivate period of T. wilfordii, the content of secondary metabolites production in the suspension cell of T. wilfordii reached to the highest of 5.185, 39.747, 42.189 μg/L and 5.185, 40.080, 45.472 μg/L, respectively. It was about 1~3 times higher than that of control. Ultrasonic treatment could promote the secretion of the secondary metabolites of T. wilfordii into the cell culture medium. After the detection found that the conversion rate of triptolide(93.7%), wilforgine(86.6%) and wilforine(88.1%) reached to the highest when treated by the ultrasonic treatment for 40 s respectively. Compared with the conversion rate of control, those conversion rate increased 1~3 times. The results provide foundation for solving the shortage of natural resource and production of triptolide, wilforgine and wilforine in T. wilfordii by using T. wilfordii cell suspension cultivation.
