A way of production podophyllotoxin by root culture of Podophyllum hexandrum Royle (P. hexandrum) was established. In the basis of aseptic plantlets induced from mature embryo of P. hexandrum seed, the rhizome of the plantlets were excised to 2.0 cm and then cultured on the Murashige and Skooge medium (MS) supplemented with different concentrations of growth regulators (IBA: 3-Indolebutyric acid, GA3: Gibberellic acid, Hy: Hydroquinone) and activated charcoal (AC). As the cultures development, the Hy could promote the plantlets growth compared to IBA and GA3, and the length of rhizome reached by 11.59 cm under the concentration of Hy 2.0 mg/L after 40 days; Supplementation of the medium with AC could significantly hasten the plantlet growth and development compared to other treatments with the length and dry weight of rhizome 13.60 cm and 0.33 g under the concentrations of Hy 2.0 mg/L and AC 0.5 g/L after 40 days; MS + sucrose 30.0 g/L + agar 5.0 g/L + Hy 5.0 mg/L + AC 0.5 mg/L (pH=5.8) was optimum for both biomass and accumulation of podophyllotoxin in root culture of P. hexandrum, the content of podophyllotoxin in the root and cultured medium reached 151.50 μg/g and 121.22 μg/g with the accumulation index of podophyllotoxin e0.4487 and e0.3329 respectively after 50 days. This indicated that production podophyllotoxin by root culture of P. hexandrum was feasible and had important application prospect.
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