西洋参冠瘿组织培养及其人参皂苷Re和人参皂苷Rg_1的产生

Study on the Culture of Crown Gall from Panax quinquefolium and the Prouction of Its Secondary Metabolites——Ginsenosides Re and Rg_1

考察了培养基组成、培养时间、接种量、pH值、肌醇浓度等对冠瘿组织生长及其人参皂苷含量的影响 ;用HPLC检测了冠瘿组织中人参皂苷Re和人参皂苷Rg1 的含量。高压纸层析电泳证实 ,根癌农杆菌Ti质粒上的T DNA片段已整合进入植物细胞核基因组中。在考察的 6种培养基中 ,White培养基最适合人参皂苷Rg1 的累积(0 0 95 % ) ,MS培养基最适合人参皂苷Re的累积 (0 194 % )。以MS为基本培养基培养 36d、32d时人参皂苷Re和人参皂苷Rg1 累积含量最高 (分别为 0 14 7%和 0 0 6 1% ) ;接种量为 4g、2g (FW flask) ,有利于人参皂苷Re和人参皂苷Rg1的累积 ;培养基pH 5 8时人参皂苷Re含量最高 (0 184 % ) ,培养基pH 5 6时人参皂苷Rg1 累积量最高 (0 0 5 4 % ) ;肌醇浓度为 0 0 5g L时 ,能促进人参皂苷Re合成 (0 182 % ) ,浓度为 0 30g L时 ,有利于人参皂苷Rg1 累积 (0 0 5 5 % )。

It was clearly demonstrated that T DNA of Agrobacterium tumefeciens Ti plasmid was integrated into the cells of crown gall in our experiment. This paper reported the influences of some kinds of physical chemistry factors on the growth of crown gall of Panax quinquefolium and the production of its main active compounds —— ginsenoside Re and ginsensoside Rg 1. The results showed that White medium was the best one for ginsensoside Rg 1 accumulation(0 095%) among the six media, but ginsensoside Re accumulation(0 194%) was the highest on the MS medium; The highest contents of ginsensoside Re(0 147%) and ginsensoside Rg 1 (0.061%) were on the culture 36d and 32d after innoculum respectively; The optimum pH was 5 6 for ginsensoside Rg 1 synthesis(0 054%), and 5 8 for ginsensoside Re synthesis(0 184%); The contents of ginsensoside Re and ginsensoside Rg 1 was the highest in the inoculum of 4g and 2g/flask(FW) respectively. The result also indicated that the concentration of inositol in 0 05g/L could obviously promote ginsensoside Re synthesis(0 182%), and in 0 30g/L for ginsensoside Rg 1(0 055%).