丹參毛狀根培養建立與其丹參酮產量之研究

Production of Tanshinones of Salvia miltiorrhiza in Hairy Root Culture

陳威臣(Uei-Chern Chen) ； 詹効松(Hsiao-Sung Chan) ； 李秋儀(Choi-Yi Lee) ； 曹進義(Chin-Yi Tsao) ； 劉禎祺(Jen-Chyi Liu) ； 李艷琪(Yahn-Chir Lee) ； 夏奇鈮(Chi-Ni Hsia)

《台灣農業研究》 57卷4期 (2008/12) Pp. 305-316

https://doi.org/10.6156/JTAR/2008.05704.07

摘要

本研究以丹參(Salvia miltiorrhiza Bunge)無菌瓶苗葉片經農桿根群菌(Agrobacterium rhizogenes, R1601)感染所得之毛狀根爲材料，測試毛狀根接種部位、接種量及培養時間在液態培養系統中，對於生質量與其丹參酮類(tanshinones)含量之影響。分別以固態與液態培養根尖與根段培植體之試驗結果顯示，根尖培植體於液態培養條件下有較高生質量與丹參酮類含量。毛狀根接種量試驗以每20mL培養液接種5-20條根尖培植體，結果顯示當培養達8週時，10至20根尖處理組間之乾重與丹參酮類含量的差異並不顯著，其中以10根尖處理效果較佳。毛狀根之錐形瓶液態時序培養結果顯示，培養液中蔗糖與氨態氮含量於培養第3週時分別降至初始濃度之9%與10%，此時毛狀根乾重累積有減緩之現象，推測蔗糖與氨態氮可能爲丹參毛狀根生長限制因子。丹參毛狀根在培養6週後可達最高乾重約爲5.8 g L^(-1)，此時收穫之毛狀根與培養液經高效液相層析法(high performance liquid chromatography; HPLC)分析毛狀根與培養液之結果顯示，其三種主要丹參酮類產量僅約11.4 mg L^(-1)；若繼續培養至第8週則丹參酮類產量可達約64 mg L^(-1)。

關鍵字

丹參 ； 時序培養 ； 氮源 ； 蔗糖 ； 二次代謝物

並列摘要

Leaf explants of Salvia miltiorrhiza were inoculated with Agrobacterium rhizogenes strain R1601 to induce formation of hairy roots. Root cultures were established and maintained in a semi-solid or a liquid B5 medium. They were used to study effect of source of inoculum, size of inoculum, type of media and incubation period on production of root biomass and tanshinones in liquid culture. Results showed that root tips were superior to other root segments as inoculum for production of root biomass and tanshinones in liquid cultures. The treatments of 10, 15 and 20 root tips/20 mL B5 medium significantly increased root growth and production of tanshinones, compared to the treatment of 5 root tips/20 mL B5 medium. Concentration of sucrose and NH4(superscript +) in B5 liquid cultures rapidly reduced to 9% and 10%, respectively, in 3 weeks, resulting in a decrease in biomass accumulation. This indicates that carbohydrate and nitrogen might be important limiting factors for hairy root growth of S. miltiorrhiza. Production of dry weight biomass in liquid B5 medium peaked at 5.8 g L^(-1) after 6 weeks, whereas production of tanshinones peaked at 64 g L^(-1) after 8 weeks.

並列關鍵字

Danshen ； Salvia miltiorrhiza ； Time-course study ； Nitrogen source ； Carbohydrate source ； Secondary metabolite ； Tanshinones ； Hairy root culture