培養基組成分對高氏柴胡癒合組織增殖與柴胡皂苷含量之影響

Influence of Medium Components on Callus Proliferation and Saikosaponin Content of Bupleurum kaoi Liu, Chao et Chuang

陳威臣(Uei-Chrn Chne) ； 夏奇鈮(Chi-Ni Hsia) ； 葉茂生(Mau-Shing Yeh) ； 曹進義(Chin-Yi Tsao) ； 蔡新聲(Hsin-Sheng Tsay)

《台灣農業研究》 57卷2期 (2008/06) Pp. 119-132

https://doi.org/10.6156/JTAR/2008.05702.04

摘要

本研究利用高氏柴胡(Bupleurum kaoi Liu, Chao et Chuang)葉片癒合組織培養於添加0.2 mg/L 2,4-二氯苯氧乙酸(2,4-dichlorophenoxyacetic acid; 2,4-D)之1/2 MS(Murashige & Skoog 1962)基本培養基(D2培養基)，探討其他培養基組成分的添加對癒合組織增殖與柴胡皂苷(saikosaponins)含量之影響。在測試的培養基組成分中，除細胞分裂素(cytokinins)的添加對癒合組織生長有抑制作用外，其他培養基組成分及其濃度分別爲3%蔗糖、0.9%洋菜(Difco Bacto-agar)、0.5 g/L蛋白腖(peptone)、0.5 g/L水解酪蛋白(casein hydrolysate)與100 mL/L椰子汁(coconut water)對於癒合組織生長均有促進效果。將上述可促進癒合組織生長之影響因子配合不同濃度MS鹽類進行試驗結果顯示，癒合組織之增殖以1/4×MS與1/2×MS鹽類濃度較佳，但其繼代培養間隔時間以不超過6週爲宜；上述MS鹽類濃度試驗所得癒合組織經高壓液相層析(high performance liquid chromatography; HPLC)分析結果顯示，以1×MS鹽類濃度培養之癒合組織具有較高之柴胡皂苷乾重含量。

關鍵字

柴胡 ； 藥用植物 ； 細胞培養 ； 二次代謝物 ； 高壓液相層析

並列摘要

Medium components and their best concentrations of cytokinin, sucrose, Difco Bacto-agar, peptone, casein hydrolysate, coconut water were separately tested for callus growth of Bupleurum kaoi Liu, Chao et Chuang. Saikosaponins production of callus tissues was monitored. A half-strength MS salt medium supplemented with 0.2 mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) was used as a basal medium (D2 medium) for callus growth. The results showed that additional N6-benzyladenine (BA) and N-furfuryladenine (kinetin) inhibited callus growth in all concentration tested in this study. Other than cytokinin, all factors could increase callus growth at a proper range of concentrations. It was concluded that 3% sucrose, 0.9% Difco Bacto-agar, 0.5 g/L peptone, 0.5 g/L casein hydrolysate, and 100 mL/L coconut water had the best effects on callus growth of B. kaoi. Various strengths of MS basal salts in combination with the aforementioned components at the best concentrations were further tested. Among four strengths of MS tested, half-strength MS treatment had the highest fresh and dry weight of callus growth after 4 weeks of culture. However, the highest saikosaponin a, c, d contents examined by high performance liquid chromatography (HPLC) was obtained from callus grown on the full-strength MS basal medium.

並列關鍵字

Bupleuri Radix ； Medicinal plant ； Cell culture ； Secondary metabolite ； High performance liquid chromatography