红豆杉(Taxus spp.)为国家一级濒危植物, 含有药用价值高的广谱抗癌成分紫杉醇, 但资源匮乏。为解决紫杉醇药源短缺问题, 该研究建立了红豆杉紫杉烷类离体富集体系。结果表明, 最佳吸收面扩大培养基配方为MS+2 mg·L^-1 6-BA+0.4 mg·L^-1 NAA+0.7 g·L^-1脯氨酸, 扩大率达90%; 高频同步诱导率最高的培养基为DCR+1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA+5 mg·L^-1谷氨酰胺+1 g·L^-1活性炭, 高频同步率达82.2%; 生物量扩增I期效果最佳培养基为MS+0.7 mg·L^-1 6-BA+ 0.07 mg·L^-1 NAA+0.1 mg·L^-1苯丙氨酸+50 mg·L^-1间苯三酚, 芽苗生物量为每株542 mg; 生物量扩增II期效果较好的培养基为MS+0.5 mg·L^-1 6-BA+0.05 mg·L^-1 NAA+0.1 mg·L^-1苯丙氨酸+2 g·L^-1活性炭, 芽苗生物量为每株1 612 mg。富集培养后, 组培材料中紫杉烷类含量远高于天然材料, 紫杉醇含量为天然材料的6.1倍; 巴卡亭III含量为天然材料的8.2倍; 10-去乙酰基巴卡亭III含量为天然材料的68.1倍。该研究首次建立了红豆杉紫杉烷离体富集体系, 突破了紫杉醇药源短缺的瓶颈。

Taxus contains the anti-cancer ingredient paclitaxel, is the top-protected endangered plant in China, and has great medicinal value but scarce resources. To solve the problem of paclitaxel source shortage, in this study, an in vitro culture of a taxane-rich Taxus system was established. The results showed that MS+2 mg·L^-1 6-BA+0.4 mg·L^-1 NAA+0.7 g·L^-1 Pro was the optimal treatment for absorption surface expansion, and the expansion rate reached 90%. The adventitious bud high-frequency synchronous growth induction was in the DCR+1 mg·L^-1 6-BA+0.1 mg·L^-1 NAA+5 mg·L^-1 Glu+1 g·L^-1 AC. The best biomass amplification phase I was in MS+0.7 mg·L^-1 6-BA+0.07 mg·L^-1 NAA+0.1 mg·L^-1 Phe+50 mg·L^-1 PG, in which the biomass reached 542 mg. The optimized biomass amplification phase II was obtained in MS+0.5 mg·L^-1 6-BA+0.05 mg·L^-1 NAA+0.1 mg·L^-1 Phe+2 g·L^-1 AC, in which the biomass reached 1 612 mg. The content of taxanes in tissue culture materials is much higher than that of natural materials. The content of paclitaxel in tissue culture materials was 6.1 times that of natural materials. The contoent of baccatin III in tissue culture materials was 8.2 times that of natural materials. The content of 10-DAB in tissue culture materials was 68.1 times that of natural materials. This study established an in vitro culture system of taxane-rich Taxus, and solved the problem of paclitaxel drug shortage.