Abstract
White latex in shoots of an oil plant Euphorbia tirucalli contains abundant secondary metabolites such as phytosterols. Using this organism, we have obtained more than 20 cDNA clones related with sterol metabolism by EST analysis and RT-PCR cloning, established plant regeneration by successive cultures on LS medium containing 0.02mg/l thidiazulon and then on hormone-free LS media, and revealed sensitivity of the callus for G418 and hygromycin B. In order to develop transgenic E. tirucalli with higher amount of phytosterols, we have attempted Agrobacterium-mediated introduction of binary vectors for 35S::squalene synthase gene, 35S::squalene epoxydase gene, and beta-amyrin synthase gene RNAi, and are checking corresponding transcripts by RT-PCR. This work was partly performed as one of the technology development projects of the "Green Biotechnology Program" in New Energy and Industrial Technology Development Organization (NEDO).
