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研究生:蔡苾暉
論文名稱:五味子組織培養之研究
論文名稱(外文):Tissue culture of Schisandra chinensis (TURCZ.)BAILL
指導教授:蔡新聲蔡新聲引用關係陳忠川陳忠川引用關係
學位類別:碩士
校院名稱:中國醫藥學院
系所名稱:中國藥學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:1999
畢業學年度:87
語文別:中文
論文頁數:95
中文關鍵詞:五味子癒合組織組織培養
相關次數:
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中文摘要
五味子(Schisandra chinensis (TURCZ.)BAILL)為木蘭科(Magnnoliaceae)五味子屬(Schizandra MICHX)之多年生落葉木質藤本藥用植物。最早收載於神農本草經,列為上品,列代諸家本草均有記載。自古以來即為中醫常用之收斂固澀藥,主治肺虛喘咳、夢遺滑精等。五味子的有效成分主要為木脂素(lignans),現代藥理研究發現五味子具有保肝、拮抗氧自由基對抗細胞損害及提高免疫力等作用。
近年來因山地的開發及大量採摘,且中醫藥漸受重視之影響,五味子之需求量也相對提高。因此本研究利用組織培養或以懸浮細胞生產五味子之主要二次代謝物,達成大量生產藥用成分schizandrin 及gomisin A的目標,或將有助於改善國內中藥市場對五味子仰賴進口之現況。茲將試驗結果摘錄如下:
1.將五味子種子培養於含1 mg/l GA3 之MS基本鹽類培養基中,可誘導癒合組織產生。將誘導出之癒合組織培養於含2 mg/l NAA,0.5 mg/l BA之MS基本鹽類培養基中,可生產大量的癒合組織供試驗用。
2.含有半量MS基本鹽類添加10﹪椰子汁、1 mg/l NAA、0.5 mg/l BA、1﹪sucrose和1﹪Difco agar,pH值約5.7之固體培養基,於黑暗及25±1℃之恆溫下,是五味子癒合組織生長的最佳條件。
3.在半量MS基本鹽類只添加0.5mg/l BA的培養基中,五味子癒合組織的生長雖然不佳,卻可以產生gomisin A。
4.1﹪蔗糖對五味子癒合組織的生長為最利。但schizandrin的形成則以7﹪的蔗糖和3﹪的葡萄糖較佳。
5.添加水解酪蛋白(250, 500, 750, 1000 mg/l)不利於五味子癒合組織之生長。
6.添加(1, 2, 4 g/l) peptone的培養基有助於schizandrin的生成,但添加(2, 4 g/l) peptone卻不利於五味子癒合組織之生長。
7.添加10﹪椰子汁有預防五味子癒合組織褐化之效果,並有利於schizandrin的生成。
8.培養於含gelrite培養基的癒合組織,均有水晶狀透明化的現象,顯示gelrite對癒合組織不利。但0.3﹪gelrite卻有促進schizandrin生成之作用。
Studies on the Tissue Culture of Schisandra chinensis (TURCZ.) BAILL.
Pi-Hui Tsai
Graduate Institute of Chinese Pharmaceutical Sciences
China Medical College
Taichung, Taiwan, Republic of China
Summary
Schisandra chinensis (TURCZ.) BAILL. (Magnoliaceae), commonly called as "Wu Wei Zi" in Chinese, is one of the most important Chinese medicinal herbs. It grows wild in the northeastern China and Japan (Reid 1986). It was first recorded in Shen-Nong-Pen-Ts''''ao-Chin under upper category. The dried fruits (ripened red berries) and whole plants are used in the traditional Chinese medicine as antihepatotoxic, antiasthmatic, antitussive, antidiabetic, sedative and general tonic (Wagner et al. 1996). It also has antioxidative (Lu and Liu 1992) and antibacterial (Ikeya et al. 1991) activity. The herb mainly contains lignans, monoterpenes and sequiterpenes. In addition, it also contains vitamin C and E, fumaric acid and stigmasterol (Wagner et al. 1996). The pharmaceutical industry is mainly dependent upon wild populations for supplies of the plant material. Due to large-scale and over-harvesting of wild material from forests, S. chinensis is rapidly disappearing.
The main objectives of present study with S. chinensis were: (1) to optimize conditions for rapid proliferation of the seed-derived callus, and (2) to explore the possibility of producing secondary metabolites in large quantity in callus and suspension cell cultures. These objectives were proposed to produce active principles of S. chinensis on commercial scale from callus/suspension cell cultures. The callus induced from mature seeds on MS basal medium supplemented with 3% sucrose and 1.0 mg/l GA3, was used in the present study.
The results are summarizes as follows:
1. Strength MS medium (1/2-strength of MS basal salts supplemented full complements of NaFeEDTA and organic supplements) with 1 mg/l NAA, 0.5 mg/l BA, 10% coconut milk (CM), 1﹪sucrose and 1﹪Difco agar, supported proliferation of seed-derived callus of Schisandra chinensis (TURCZ.) BAILL.
2. Addition of casein hydrolysate (250, 500, 750, 1000 mg/l) in the medium did not support proliferation of callus.
3. Addition of peptone (1, 2, 4 g/l) stimulated production of schizandrin in callus cultures. However, callus production decreased with increase (2 and 4 g/l) in peptone concentration in the medium.
4. Addition of 0.5 mg/l BA without NAA in callus proliferation medium promoted the production of gomisin A, but failed to support callus growth.
5. Higher concentration (7% w/v) of sucrose and 3﹪glucose promoted the production of schizandrin in the callus cultures.
目錄
謝辭 Ⅰ
中文摘要 Ⅱ
英文摘要 Ⅳ
略字解 Ⅵ
第一章 緒 言 1
第二章 總 論
第一節 五味子之本草考察 2
第二節 五味子之藥用植物考察 9
第三節 五味子之生藥學考察 15
第四節 五味子之藥理學考察 20
第五節 植物組織培養學之考察 22
第三章 試驗部分
第一節 材料與方法 40
第二節 結 果 49
第三節 討 論 73
參考文獻 79
附錄 91
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