In vitro techniques and genetic engineering of Veratrum californicum Duran

• Main Authors: Ritala, Anneli, Rischer, Heiko, Ma, R., Oksman-Caldentey, Kirsi-Marja
• Format: Other Non-Article Part of Journal/Newspaper
• Language: English
• Published: VTT Technical Research Centre of Finland 2007
• Subjects: Arctic; Duran
• Online Access: https://cris.vtt.fi/en/publications/363e5402-3564-4d66-8ac2-a6b8da8160b9; https://publications.vtt.fi/pdf/symposiums/2007/S249.pdf

The genus Veratrum is distributed throughout the northern temperate and Arctic regions of Europe, Asia and North America. Veratrum californicum Duran (Melanthiaceae) is an important monocotyledonous medicinal plant which is the only source of the anticancer compound cyclopamine. In order to be able to utilize Veratrum cells in the production of important secondary metabolites, an in vitro platform is needed. For that purpose, tissue culture, green plant regeneration and genetic engineering of Veratrum californicum were developed. Fine suspension cell lines were established from germinated mature embryos by transferring friable embryogenic calli to AA- and L2-medium. The suspension cells were cryopreserved successfully and recovered at a high rate. In addition, a highly embryogenic and regenerating cell line was established and kept on L2-medium without additional growth regulators. Very good growth was obtained by employing the temporary immersion system of RITA -bioreactors. The highly regenerating cell line and in vitro plantlets contained steroid alkaloids cyclopamine and veratramine. Furthermore, transformation methods were developed for Veratrum californicum. Transgenic cell lines were produced by Agrobacterium tumefaciens and A. rhizogenes -mediated deliveries. Also direct gene transfer techniques such as particle bombardment and protoplast-based techniques were applied. The basic tools for the metabolic engineering and biotechnological production of secondary metabolites of Veratrum californicum are now available.