| Summary: | 碩士 === 元智大學 === 生物科技與工程研究所 === 106 === Snow lotus, Saussurea involucrata, is a medicinal plant that is known to contain plant phytochemicals of great pharmacological value. Tissue culture propagation of snow lotus with bioactive compounds will provide the substitute for pharmaceutical market demand. Extensive exploitation of the biosynthetic pathways to induce the accumulation of the flavonoids is in need. Snow lotus suspension cell cultures grow faster than the callus and can lead to fast accumulation of flavonoids such as syringin (anti-inflammatory), chlorogenic acid (anti-oxidant), and rutin (anti-cancer). These compounds can be further increased to certain levels through genetic engineering through Agrobacterium mediated-transformation approach. Two antibiotics, hygromycin, kanamycin, and a herbicide Basta with various concentrations were used for the screening of the calli. Following co-culture, about 11.3% resistant calli grew under the selection of 30 mg/L hygromycin. Three putative transformants cell lines by Agrobacterium-mediated transformation were confirmed by genomic PCR. The RT-PCR analysis confirms the gene expression levels of transformants. Three cell lines were chosen for the detection of products and line E13 shows the highest product content of 116 nmol/g FW. Cryopreservation can prevent somatic variation in cells. The cryoprotectant solution, Sol-R3 (E) showed 100 % of growth recovery after 1week storage in liquid nitrogen. The transformants showed the function of ectoine as an osmoprotectant and an antifreeze agent to cells.
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