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利用农杆菌介导法成功地将RiT-DNA转入药用植物盾叶薯蓣 ,产生了毛状根 ,经分子信标探针检测农杆菌Ri质粒上的T DNA已整合进植物基因组中。研究建立了毛状根大量快速繁殖技术 ,基本技术要求为 :1/ 2MS液体培养基 ,2 8℃培养温度 ,35 0lux弱光条件下有利于毛状根的增殖培养 ,提高生物量。HPLC测定结果显示 ,转基因获得的毛状根其薯蓣皂甙元的含量分别是微块茎、愈伤组织和植物体合成量的 5 . 6 8倍、6 . 12倍和 2 . 6 8倍。
The Agrobacterium-mediated method successfully transferred RiT-DNA into the medicinal plant Dioscorea zingiberensis to produce hairy roots. The T DNA of Agrobacterium Ri plasmid was detected by molecular beacon probe and integrated into plant genome. The establishment of a large number of rapid propagation of hairy root technology, the basic technical requirements: 1 / 2MS liquid medium, 28 ℃ incubation temperature, 35 0lux low light conditions are conducive to the proliferation of hairy roots culture to improve biomass. The results of HPLC showed that the contents of diosgenin in hairy roots obtained from transgenic plants were respectively 5.86, 6.12 and 2.68 times of those of microtuber, callus and plant.