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題 名 | 誘引劑對高氏柴胡懸浮細胞生長與柴胡皂苷含量累積之影響=Influence of Elicitors on Cell Proliferation and Saikosaponin Production of Bupleurum kaoi in Suspension Culture |
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作 者 | 陳威臣; 夏奇鈮; 葉茂生; 曹進義; 蔡新聲; | 書刊名 | 臺灣農業研究 |
卷 期 | 61:2 2012.06[民101.06] |
頁 次 | 頁112-123 |
分類號 | 434.92 |
關鍵詞 | 高氏柴胡; 藥用植物; 懸浮細胞培養; 柴胡皂苷; 誘引劑; Bupleurum kaoi; Medicinal plant; Cell suspension culture; Saikosaponin; Elicitor; |
語 文 | 中文(Chinese) |
中文摘要 | 高氏柴胡(Bupleurum kaoi Liu, Chao et Chuang)懸浮細胞於含有0.2 mg/L二氯苯氧基乙酸(2,4-dichlorophenoxyacetic acid, 2,4-D)之1/2 MS液態培養基中於低光照下以100 rpm轉速培養,細胞之生長在接種後0-2週為緩慢生長期,在2-4週呈現快速生長期,在培養4週後進入生長靜止期。振盪速度試驗結果顯示,80 rpm較100 rpm振盪速率在培養4週後,具有較高之細胞生長量。細胞接種密度試驗結果顯示,以12%密度接種之細胞生長最佳,接種密度較高(24%)或較低(6%)的細胞生長表現均較差,但同樣接種密度(12%)下,以較大培養容器為佳(125-mL vs. 250-mL錐形瓶)。添加誘引劑之試驗結果顯示,在處理2週後,所有誘引劑處理之細胞鮮重皆受到顯著抑制;但在細胞乾重方面,酵母抽出物處理與對照組之乾重相較並無顯著差異,僅茉莉酸與高濃度離層酸處理對細胞乾重有顯著抑制之效果,但三種誘引劑處理對懸浮細胞之柴胡皂苷(saikosaponin a及saikosaponin d)累積均有促進效果,柴胡皂苷含量可提高1.9-2.0倍,產量可提高1.6-1.8倍。進一步施用500 mg/L酵母抽出物作為誘引劑,結果顯示施用2週後柴胡皂苷含量較對照組增加2.1倍,而柴胡皂苷產量可達對照組之2.8倍,顯示在細胞生長後期施用誘引劑並在特定反應時間後收穫細胞,有助於二次代謝產物柴胡皂苷之產生與累積。 |
英文摘要 | Bupleurum kaoi, a native medicinal plant in Taiwan which contains saikosaponins higher than other commercial Bupleurum species, was chosen for producing saikosaponins in vitro. Cell suspension cultures of B. kaoi were cultured in a half-strength MS liquid medium containing 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and used for studying effect of speed of shaker, size of flasks and inoculum density on cell proliferation as well as studying effect of elicitors, yeast extract (YE), abscisic acid (ABA), and jasmonic acid (JA), on saikosaponin production. Results showed that cell proliferation of B. kaoi in suspension cultures was higher using shaker speed at 80 rpm than that of 100 rpm with peak period of cell growth at 3-4 weeks. Cell proliferation of B. kaoi was also affected by inoculum density and flask size, with the optimum inoculum density of 12% by adding 6 mL packed cell volume (PCV) cultures in 250-mL flasks. Three elicitors (YE, ABA, and JA) tested in 3-week-old suspension cultures of B. kaoi were found reducing cell fresh weight and dry weight significantly. However, one week after the treatments of elicitors, saikosaponin contents and its productivity increased 1.9-2.0 folds and 1.6-1.8 folds, respectively. Further study using 500 mg/L YE as elicitor in cultures of B. kaoi for two weeks, similar increasing tendency on saikosaponin contents and its productivity was found with 2.1-fold and 2.8-fold, respectively. This study concludes that adding 500 mg/L YE into cell suspension cultures of B. kaoi after the lag phase of cell cycle for one to two weeks would not reduce cell growth but increase saikosaponin production. |
本系統中英文摘要資訊取自各篇刊載內容。