【作者】 王莉；

【导师】 于荣敏；

【作者基本信息】 沈阳药科大学 ， 微生物与生化药学， 2001， 硕士

【摘要】 本实验首次利用发根农杆菌LBA9402和R1601诱导药用植物何首乌产生毛状根。冠瘿碱的高压纸电泳证实，两种菌诱导的毛状根都整合了Ri质粒的T_R-DNA片段；PCR扩增和Southern杂交试验结果证明，LBA9402诱导的毛状根整合了Ri质粒的T_L-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、黑色生长点逐渐增多等特点，且在无激素的培养基上能够快速稳定繁殖。通过基本培养基的筛选，确立了毛状根生长的最佳培养基为MS培养基。TLC定性试验证实毛状根中含有与原植物相同的蒽醌类成分，如大黄素、大黄酚、大黄酸；HPLC定量分析结果显示毛状根培养物中大黄酸的含量为原植物的2.85倍。更多还原

【Abstract】 Transformed hairy roots of Polygonium multzflorum Thunb. were obtained by the infection of two kinds of Agrobacterium rhizogenes strains LBA9402 and Rl601 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 strain R1601 was identified by paper electrophoresis. It was also showed that there were some significant characteristics of hairy roots after eliminating bacteria, such as more branches and black growth tips, fast growth, genetic and biochemical stability, and growth in hormone-free media. As a result, they showed a 11.03-fold increase in biomass in MS medium after 40-days of incubation. The compounds of anthraquinones 5 THESIS FOR MASTER DEGREE ABSTRACT (emodin , chrysophanol and rhein) from cultivated hairy roots were identified by means of HPLC and TLC, and the content of rhein was 2.85 folds higher in hairy roots than that of natural plants.更多还原