药用植物商陆毛状根培养系统的建立

目的:建立药用植物商陆(Phytolacca acinosa Roxb.)的毛状根离体培养系统.方法:分别用发根农杆菌A4、R1600、C58C1 3个菌株感染商陆的子叶外植体,获得毛状根,并筛选了优质株系;考察了影响毛状根生长的因素;测定了毛状根的生长曲线;利用PCR和高压纸电泳法对毛状根进行tDNA转化的检测.结果:利用发根农杆菌A4、R1600、C58C1 3种菌株成功地从商陆中诱导出毛状根;以水解酪蛋白为氮源、以麦芽糖为碳源最有利于商陆毛状根的生长;毛状根的生长曲线呈"S"形,24 d时质量达到最大(增长18倍);PCR及高压纸电泳检测结果表明Ri质粒的tDNA已整合进毛状根中.结论:成功建立了商陆毛状根离体培养系统,为进一步进行药用成分的工业化生产和引入外源基因改良性状奠定了基础.

Establishment of Phytolacca acinosa Roxb hairy root in vitro culturing system

Objective:To establish a culture system for hairy root of Phytolacca acinosa Roxb. Methods: The cotyledon explants of Phytolacca acinosa were infected with Agrobacterium rhizogenes strains A4, R1600 and C58C1 and the hairy root culture clones were obtained; the better lines were selected. The growth curve was surveyed and extrinsic factors affecting the growth of hairy roots were investigated. The transformation of Ri tDNA was examined through PCR and high voltage paper electrophoresis. Results: The hairy root was obtained from Phytolacca acinosa. The results of PCR and high voltage paper electrophoresis confirmed the transformation of tDNA from Ri plasmid to the hairy root. Conclusion: The acquisition of hairy root of Phytolacca acinosa lays a foundation for the mass production of active components and introduction of foreign gene.