何首乌毛状根的诱导及其培养

Hairy-Root Induction and Culture of Polygonium Multiflorum Thunb.

利用发根农杆菌LBA940 2和R16 0 1诱导药用植物何首乌产生毛状根。由PCR扩增和Southern杂交证实 ,由LBA940 2诱导的毛状根整合了Ri质粒的TL DNA片段 ;冠瘿碱的高压纸电泳证实 ,由两种菌诱导的毛状根整合了Ri质粒的TR DNA片段。毛状根经抗生素和温度除菌后 ,表现出粗壮、绒毛密集、分枝多 ,且在无激素的培养基上能够快速稳定繁殖的特性。经筛选得到何首乌毛状根优良离体培养系PC9e和PC1e。

Transformed hairy roots of Polygonium multiflorum Thunb.were obtained by the infection of two kinds of Agrobacterium rhizogenes strains LBA9402 and R1601 and an in vitro culture system of the hairy roots was established. It was clearly demonstrated that T-DNA of A.rhizogenes Ri plasmid was integrated into the hairy roots from LBA9402 by the experiments of PCR, Southern hybridization and paper electrophoresis. The transformed roots from R1601 was identified by paper electrophoresis. It was also showed that there were some significant characteristics of hairy roots after eliminating bacteria with higher temperature and antibiotic, such as robustness, more branches and hairs, fast growth capacity, genetic and biochemical stability, and growth in hormone-free media. After cultivation of two months,the optimum culture clones of hairy roots were identified,which were induced with R1601-PC1e and induced with LBA9402-PC9e.