丹參組織培養苗馴化處理及其植株丹參酮含量分析

In Vitro Plantlets Acclimation and Tanshinones Analysis of Salvia Miltiorrhiza

陳威臣(Uei-Chern Chen) ； 曹進義(Chin-Yi Tsao) ； 夏奇鈮(Chi-Ni Hsia)

《台灣農業研究》 56卷1期 (2007/03) Pp. 21-30

https://doi.org/10.6156/JTAR/2007.05601.03

摘要

本研究探討吲哆丁酸(indole-3-butyric acid; IBA)對丹參(Salvia miltiorrhiza)組培芽體(in vitro shoot)發根之影響，以及培養容器封口透氣資材與馴化條件對組培苗移植存活率關係，並分析成株丹參酮之含量。組培芽體於添加0.5 mg L^(-1) IBA之半量Murashige and Skoog基本鹽類(1/2MS)培養基，利用鋁箔紙封口培養2週後，再以三層藥包紙進行封口置換處理(AF2+DP4)，其發根率可達90%。將組培苗(plantlet)移植於已滅菌之BioMix培養土：蛭石：珍珠石=1:1:1 (v/v)混合介質，再以透明塑膠袋覆蓋植株培養於日夜溫為22℃，10h / 18℃，14h、光照強度約60 μmol m^(-2)s^(-1)之生長箱進行馴化處理，透氣預處理組培苗存活率可達80.1%，較持續以鋁箔紙封口培養前處理組培苗存活率則只有27.8%。此外，經由高效率液相層析儀(high performance liquid chromatography; HPLC)分析丹參各檢品之丹參酮類(tanshinones): tanshinone-I (Tan-I), tanshinone-IIA (Tan-IIA), cryptotanshinone (Crypto)含量結果顯示，組培苗經田間栽植9個月後植株根部（紅根）之Tan-I+Tan-IIA+Crypto總含量約1.85 mg g^(-1) dw，為市售丹參藥材的1.5倍。

關鍵字

丹參 ； 藥用植物 ； 微體繁殖 ； 高壓液相層析 ； 二次代謝物

並列摘要

An efficient micropropagation protocol for producing high quality plantlets that are easy to macclimatize in field was developed for Salvia miltiorrhiza. In vitro grown shoots induced rooting (90%) after culturing on half-strength Murashige and Skoog’s basal medium (1/2MS) supplemented with 0.5 mg L^(-1) indole-3-butyric acid (IBA). Culture vessels were capped with aluminum foil for 2 weeks, and thereafter replaced with 3 layers of dispense paper for 4 weeks (AF2+DP4) were better than controls capped with aluminum foil for 6 weeks (AF6). Rooted shoots were transplanted into plastic pot containing a mixture of BioMix: vermiculite: perlite (1: 1: 1 ratio) (v/v). For acclimation, these pots were covered with transparent plastic bags and kept in a growth chamber under 14 h light at 22℃ and 10 h dark at 18℃ for 4 weeks. At 4 weeks of observation, these conditions resulted in the highest survival rate of 80.1% compared to AF6 treatment (27.8%). To compare, tanshinones contents (tanshinone-I, tanshinone-IIA and cryptotanshinone), the bioactive compounds in Salvia miltiorrhiza were estimated in the aerial parts and roots between in vitro micropropagated plants and market crude drug of S. miltiorrhiza by high performance liquid chromatography (HPLC). The quantity of tanshinones in red roots of in vitro micropropagated plants was measured as 1.85 mg g^(-1) dw about 1.5-fold higher than market crude drug.

並列關鍵字

Danshen ； Medicinal plant ； Micropropagation ； High performance liquid hromatography HPLC ； Secondary metabolite