Taxane production in hairy roots of Taxus x media var. Hicksii carrying taxadiene synthase gene

Paclitaxel is the active agent of Taxol® (Bristol-Myers Squibb), one of the most effective anticancer plant – derived drug showing activity against different cancers. Moreover paclitaxel has shown great promise as a locally delivered antirestenotic agent and also seems to be an effective agent for the treatment of Alzheimer's disease and other neurodegradive diseases [1,2]. A very promising approach for the production of paclitaxel and related taxanes, without forest harvestation, is provided by plant cell suspension cultures and its production has been recently successfully scaled-up [3]. Two lines of hairy roots were obtained as a result of infection of 10-year old plantlets cultivated in vitro on solid DCR medium [4]. Two bacterial strains were used: Agrobacterium rhizogenes and modified A. tumefaciens with Ri plasmid. These both bacterial strains carried taxadiene synthase gene. Taxadiene synthase leads cyclization of GGPP to taxa 4(5), 11(12)-diene. This is first committed step of paclitaxel biosynthetic pathway. The hairy roots were cultivated in liquid hormone-free DCR-Medium [5] in dark. Medium supplementation with precursor L-phenylalanie and/or elicitor methyl jasmonate are underway to test the growth of roots and taxane accumulation in roots and post-culture media.

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