摘要
目的：比较Ｒｉ质粒转化的掌叶大黄发根与愈伤组织、悬浮细胞以及掌叶大黄生药中游离蒽醌成分的含量。方法：用发根农杆菌Ａｇｒｏｂａｃｔｅｒｉｕｍｒｈｉｚｏｇｅｎｅｓ９４０２野生型菌株感染掌叶大黄外植体以诱导发根，建立发根离体培养系，用Ｓｏｕｔｈｅｒｎｂｌｏｔ鉴定发根；分别诱导掌叶大黄愈伤组织和悬浮细胞；用高效液相色谱法测定发根、愈伤组织、悬浮培养细胞以及掌叶大黄生药中芦荟大黄素等游离蒽醌化合物的质量含量。结果：从子叶柄上诱导出了发根，并建立了培养体系；Ｓｏｕｔｈｅｒｎ杂交结果证明Ｒｉ质粒ＴＤＮＡ转移并整合到植物细胞基因组中；用叶片作外植体诱导出了愈伤组织；悬浮培养细胞用叶片、叶柄外植体诱导的愈伤经液体培养获得；掌叶大黄发根中各种游离蒽醌化合物含量普遍高于愈伤组织，但低于生药和悬浮培养细胞。结论：用掌叶大黄发根培养物生产蒽醌化合物是一条值得探索的新途径。
Objective: To compare the contents of free anthraquinones in Ri plasmid transformed roots, calli, suspension cells and wild roots of Rheum palmatum L.. Methods: Inducing hairy roots by infecting explants of Rheum palmatum L. using wild Agrobacterium rhizogenes 9402 strain, identifying transformed roots by Southern blot with rol C gene as probe. Inducing calli and suspension cells of Rheum palmatum L., respectively. Quantitatively determining the contents of six free anthraquinones——aloe emodin, rhein, chrysophanol, emodin, physcion, 8 O methylchrysophanol in hairy roots, calli, suspension cells wild roots of Rheum palmatum L. by high performance liquid chromatography(HPLC) technique. Results: The hairy roots were induced and the culture system was established. Result of Southern blot indicated that T DNA of Ri plasmid was transferred into the hairy root cells. The calli were induced from leaf fragments and the suspension cells were received by liquid culturing of the calli induced from leaf and stem fragments.The determination result showed that the content of each free anthraquinone in hairy roots was generally higher than that in calli, but lower than that in suspension cells or wild rhubarb roots. Conclusion: Production of anthraquinones by hairy root cultures is a valuable approach
出处
《北京医科大学学报》
CAS
CSCD
1998年第6期500-502,524,共4页
Journal of Peking University(Health Sciences)
基金
国家自然科学基金
国家教委博士点基金资助
关键词
大黄
发根
愈伤组织
DNA印迹法
蒽醌类
Rheum /chem Hairy roots ☆ Calli ☆ Southern blot Anthraquinones/anal
