Ti和Ri质粒对栝楼双转化的研究

Genetic Transformation of Hairy Roots in Trichosanthes kirilowii Maxim.by Ti and Ri Plasmids

目的 :建立栝楼双转化毛状根培养系统。方法 :①利用根癌农杆菌Agrobacteriumtumefaciens菌株C5 8感染栝楼Trichosantheskirilowii.无菌苗诱导冠瘿瘤 ,然后用发根农杆菌Agrobacteriumrhizogenes菌株 15 834感染其再生植株 ,诱导出毛状根 ;②用纸电泳检测Ti和Ri质粒是否转化成功 ;③运用分光光度计和SDS PAGE检测栝楼组织中所含蛋白。结果 :Ti和Ri质粒转化成功并建立了栝楼双转化毛状根培养系统。结论 :双转化毛状根与一般毛状根相比 ,具有生长更迅速等特点而蛋白含量基本一样 ;因此 。

Objective:To estabish a hairy root culture system by double transfonmation for Trichosathes kirilowii. Method:①Crown galls were induced by direct infection of sterile seedlings of T.kirilonii with Agrobacterium tumefaciens C58, and then the hairy roots were obtained from the regenerated plants by infection with A.rhzogenes 15834;②Transformation of Ti and Ri plasmids was inspected by high-pressure-paper electrophoresis;③ The protein contents in the tissues of T.kirilowii were inspected by spectrophotometer and SDS-PAGE.Result:A hairy root culture system has been established sucessfully by double transformation with Ti and Ri plasmids in T. kirilowii.Conclusion:Compared with the ordinary hairy roots, the double transformed hairy roots grow faster but retain similar protein contents.