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標題: 建立丹參轉殖毛狀根系統及其二次代謝物生產之研究
Development of hairy root transformation system and its secondary metabolites production of Salvia miltiorrhiza
作者: 林珠鳳
Lin, Jhu-Fong
關鍵字: Salvia miltiorrhiza;丹參;Agrobacterium rhizogenes;hairy root;農桿根群菌;毛狀根
出版社: 農藝學系所
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丹參(Salvia miltiorrhiza Bunge)為唇形科(Labiatae)鼠尾草屬(Salvia)植物,是中國傳統治療心血管疾病的中藥。本研究首先探討影響農桿根群菌(Agrobacterium rhizogenes)感染丹參產生毛狀根之各種因子,用以建立毛狀根轉殖系統,並測試毛狀根培養之最佳條件,以及利用毛狀根生產二次代謝物之可行性。
以N6培養基繼代丹參瓶苗作為轉殖材料,利用葉片、葉柄、莖段及根段等不同部位之培植體進行轉殖,其中以葉片培植體,可得最多之轉殖根數。轉殖試驗中利用葉片培植體以OD600值為1、未經稀釋的農桿根群菌系R1601菌液感染30分鐘,並於感染菌液中添加200 μM acetosyringone,共培養2天後以cefotaxime 500 mg/ L除菌,轉殖所生成的毛狀根數可達最高。
毛狀根的培養以WPM液體培養基添加6 ﹪蔗糖於光照下培養10至12週可達最大生物量。培養於B5液體培養基之二次代謝物最適培養日期依毛狀根系而有所不同,部分毛狀根系(HR-1)其二次代謝物含量在培養10週時最高,而部分毛狀根系(HR-2、HR-3)則在培養12週後仍有增加。丹參毛狀根以含6 ﹪蔗糖B5基本鹽類固態培養基培養,可得較高之二次代謝物含量;光照培養有助於tanshinone ⅡA之生產,暗處理的cryptotanshinone及tanshinoneⅠ含量較高。
其他識別: U0005-2808200612064400
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