Tissue culture and genetic engineering of an important anticancer compound producing plant Veratrum californicum Duran

Veratrum californicum Duran (Liliaceae) is an important monocotyledonous medicinal plant which is the only source of the anticancer compound cyclopamine. The in vitro platform is needed for utilization of Veratrum cells in the production of the important secondary metabolites. Tissue culture, green plant regeneration and genetic engineering of V. californicum were developed. Fine suspension cell lines were established from germinated mature embryos by embloying friable embryogenic calli, AA- and L2-medium as culture media. The suspension cells were cryopreserved successfully and recovered at a high rate. Green plants were regenerated from embryogenic calli maintained on solid medium with 73% regeneration ability (green plants/100 calli) in 27 months old culture. The in vitro plantlets contained the steroid alkaloids cyclopamine and veratramine. In addition, Agrobacterium-mediated and protoplast-based transformation methods were developed. For the first time, the basic tools for the metabolic engineering and biotechnological production of secondary metabolites of V. californicum are now available.