为探索拟南芥的冷诱导转录因子CBF3和抗寒基因COR15A在"黔白"系列大白菜的转化体系,以"黔白"系列大白菜3个优良自交系作为研究对象,分别用大白菜4d苗龄的带柄子叶、半子叶、下胚轴作为外植体,利用农杆菌介导的方法将同时含有CBF3和COR15A双价抗寒基因的表达载体转入大白菜,就影响大白菜再生和遗传转化的因素进行研究。通过抗生素筛选得到大白菜T0代转基因抗性植株36株,经PCR检测后获得转基因阳性植株6株。经抗寒性初步鉴定,转基因大白菜植株抗寒性超过对照。
To establish"Qianbai"series cabbage regeneration-transforming technology system of cold resistance gene CBF3 and COR15 A,"Qianbai"series 3inbred lines of Chinese cabbage were used as the research object.Using the cotyledons of petiole,half leaf,hypocotyl of 4dage seedlings of Chinese cabbage as explants,and the cotyledon petiole of Zui4 as the receptor of genetic transformation,we transferred the expression vector with bivalent cold resistance gene CBF3 and COR15 Ainto Chinese cabbage by agrobacterium-mediated method.Factors affecting explants regeneration and genetic transformation of Chinese cabbage were also studied.By antibiotic selection,we obtained 6transgenic plants by PCR from 36T0 transgenic plants.Preliminary identification of cold resistance showed that the cold resistance of transgenic cabbage plants is higher than that of control plants.