Production of taxanes in callus and suspension cultures of Taxus baccata L.

Abstract

The selection and cloning of cell lines for the establishment of long-term cell cultures of Taxus baccata L. is important for the prospective biotechnological production of taxanes. Callus cultures were established from young stems of adult trees and aseptically grown seedlings. All parts of young seedlings were more suitable for cell proliferation on callus-induction media as compared to young stems of adult trees. The best growing calli successfully achieved an average 8-fold increase in fresh weight after 20 months of culture. The growth characteristics of two seedling-derived cell lines were determined. The best growing calli were used for establishment of suspension cultures. Gamborg's B5 medium supplemented with 3 mg l-1 2,4-dichlorophenoxyacetic acid, 0.5 mg l-1 kinetin and 1.5% polyvinylpyrrolidone, a phenolic-binding agent, was used as agar-solidified and liquid medium. A 20-month-old callus culture of T. baccata (VI/Ha) produced paclitaxel up to 0.0109 ± 0.0037 % of extracted dry weight basis. The content of taxanes was determined by high performance liquid chromatography or a competitive inhibition enzyme immunoassay system (CIEIA). A kinetic study of callus growth and taxane production was performed.