发根农杆菌对短叶红豆杉的转化及毛状根中紫杉醇的产生

TRANSFORMATION OF TAXUS BREVIFOLIA BY AGROBACTERIUM RHIZOGENES AND TAXOL PRODUCTION IN HAIRY ROOTS CULTURE

发根农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｒｈｉｚｏｇｅｎｅｓ）Ａ４（ＡＴＣＣ３１７８９）感染短叶红豆杉（Ｔａｘｕｓｂｒｅｖｉｆｏｌｉ－ａ）芽外植体，３０～３５ｄ后可诱导出毛状根，４０ｄ后转化率可达３０％，毛状根经农杆碱单克隆抗体酶联免疫分析，证明已被转化。毛状根生长速度较快，５株毛状根在无激素的Ｂ５液体培养基中悬浮培养２０ｄ，生物量平均增加约９倍，是同等条件下短叶红豆杉愈伤组织液体悬浮培养的２．９倍。经紫杉醇单克隆抗体酶联免疫反应检测，毛状根中含有紫杉醇，其含量为短叶红豆杉愈伤组织的１．３～８．０倍，悬浮培养过程中，毛状根也向培养液中分泌０．０１～０．０３ｍｇ／Ｌ的紫杉醇。

This paper reports the establishment of an efficient transformation system for Taxus brevifolia, using Agrobacterium rhizogenes A4(ATCC31798). Hairy roots are induced from shoot explants after 30～35 d inoculation with the bacteria. Frequency of the explants transformation by A4 is 30% after 40 d. Transformation is confirmed by agropine monoclonal antibody ELISA analysis. The average value of biomass of hairy root cultures suspended in hormone-free B5 medium increases 8.67 times after 20 d culture. It is 2.9 times as much as callus culture from T. brevifolia in the same culture condition. The hairy root culture can synthesize taxol whose content is 1.3～8 times as much as that in the callus from T. brevifolia . Taxol can also be detected in growth medium.