| Summary: | 碩士 === 國立臺灣大學 === 農業化學研究所 === 91 === Podophyllum pleianthum Hance has been used for snake bites and healing of wounds in Taiwan. Its secondary metabolite─podophyllotoxin has proven to be an effective anti-tumor agent. Due to slow-growing habit and over-collection, P. pleianthum is listed as an endangered species. The objective of this research is to establish callus and system of suspension cell culture, and to evaluate the content of podophyllotoxin.
Leaf, leafstalk, stem (with shoots tip) and root were used as starting material for callus induction. Our results revealed that B5 medium containing 10 mg/L NAA and 0.1 mg/L Kinetin shows good callus induction rate in dark environment. On the other hand, B5 medium containing 15 mg/L NAA and 0.1 mg/L Kinetin shows good callus induction rate in low light condition. Proliferation rate of cells from leaf and stem (with shoots tip) increased 14.2 and 5.4 fold in modified NP medium containing 10 mg/L NAA.
The extraction of callus induced from the four different explants was prepared by 3 mL of 80 % methanol under 30 min sonication. It was found that podophyllotoxin can only be detected from the extraction of callus induced from root explant. This metabolite showed a maximum absorption at 205 nm and the retention time at 10.10 min, which was identical to authentic compound of podophyllotoxin. The extraction of cells cultured in a liquid modified NP medium had more complex peaks in HPLC chromatogram than that of callus cultured in a soild B5 medium. These results revealed that some other unknown metabolites were simultaneously produced in the suspension cell culture.
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