臺灣白芷組織培養之研究Ⅰ．癒合組織之誘導及培養基成分之探討

Studies on Tissue Culture of Angelica Dahurica Var Formosana Ⅰ. Callus Induction and Medium Evaluation

張文德(Wen-Der Chang) ； 陳忠川(Chung-Chuan Chen) ； 張永勳(Yuan-Shiun Chang) ； 蔡新聲(Hsin-Sheng Tsay)

《中華農業研究》 42卷3期 (1993/09) Pp. 253-264

https://doi.org/10.29951/JARC.199309.0004

摘要

利用組織培養技術生產吾人所需之二次代謝產物，最近受到世界各國研究人員的重視。植物細胞不但生長迅速，二次代謝產物粹取容易，而且有些植物細胞培養所得二次代謝產物之量，更高於原植物體，所以組織及細胞懸浮培養技術，目前已成為製藥工業重要之製藥技術。本研究以臺灣白芷進行癒合組織的誘導及建立細胞懸浮培養，並從培養細胞中抽取有效成分imperatorin，期能經由本研究的探討，為臺灣製藥工程界關發出生產imperatorin之新方法，進而推廣到其他重要藥材有效成分之大量生產，以利於臺灣省產藥材之開發應用。結果發現將臺灣白芷幼嫩之葉柄，經消毒後接種於含有1mg/12,4-D、0.5mg/1 kinetin、3%蔗糖及0.9% agar, pH=5.7之MS基本鹽類固體培養基中，進行暗培養30天後，可由葉柄邊綠長出質地鬆軟，白色微黃之癒合組織；繼代培養則以稍低濃度的2, 4-D(0.25-0.5mg/l)配合0.5mg/1 kinetin、3%蔗醣及1.2% agar之MS基本鹽類固體培養基較佳。將癒合組織切碎後投入上述繼代培養的液體培養基中，於100rpm轉速，光照約200 lux的條件下培養，可建立一分散狀況良好的懸浮培養細胞。Imperatorin成份測定結果顯示，市場品杭白芷根最高，野生品臺灣白芷之根次之，而以野生品臺灣白芷之莖葉、癒合組織及懸浮細胞的含量最少，顯示白芷不同部位之器官imperatorin之含量差異極大。

關鍵字

白芷 ； 癒合組織 ； 懸浮細胞 ； 二次代謝產物

並列摘要

The production of secondary metabolites from suspension cell culture system is one of the most promising methods in medicinal industry. The cells from callus or suspension cultures not only grow faster and the products produced can be extracted more easily than cells from the intact plants, but also the quantities of the products produced are much higher than those of the field-grown plants. The tissue or suspension cell culture techniques have made industrial production of some natural products possible.
Bai-Zhi (Angelica dahurica) was first recorded in Shen-Nung-Pen-Ts'ao-Ching under middlior category and was also recorded in the successive Pen-ts'aos of the descending dynasties. It has been used for the cure of headache. Recently it has also been used for treating a skin disease, Yin-Hsieh Ping with good healing effect. The component imperatorun of Bai-Zhi has been suggested as the major active ingredient for healing the skin disease. The purpose of this study was to establish the tissue culture system including callus induction from petiole and to establish the suspension culture for the production of imperatorin. It was found that the soft and light-yellowish callus from petiole of A. dahurica could be induced on solid medium containing MS basal salts supplemented with 1 mg/l 2,4-D, 0.5 mg/l kinetin and 3% sucrose. The callus could be maintained by successive culture on medium containing MS basal salts, 0.25-2 mg/l 2,4-D, 0.5 mg/l kinetin, and 3% sucrose, solidified with 1.2% agar at pH 5.7. For developing a finely dispersed cell suspensoin culture, the callus was cut and cultured on a medium containing 0.25-0.5 mg/l 2,4-D, 0.5 mg/l kinetin, 3% sucrose and pH 5.2 and shaking at 100 rpm. Analyses by TLC and HPLC indicated that both callus and suspension cells contained imperatorin and byak-angelicin. Although the content of imperatorin in callus and suspension cell was lower than market products and wild species.

並列關鍵字

Angelica dahurica ； Callus ； Suspension culture ； Secondary metabolites