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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。
In order to explore the transformation system of cold-inducible transcription factor CBF3 and cold-tolerance gene COR15A in Chinese white cabbage, three elite inbred lines of Chinese cabbage “Guizhou White” were used as research objects, The Chinese cabbage plants with 4-day-old seedling cotyledons, semi-cotyledons and hypocotyls were used as explants respectively. Agrobacterium tumefaciens-mediated method was used to transfer the expression vector containing both CBF3 and COR15A genes into Chinese cabbage. Factors affecting the regeneration and genetic transformation of Chinese cabbage were studied. 36 Chinese cabbage T0 transgenic plants were obtained by antibiotic screening, and 6 transgenic plants were obtained by PCR assay. The preliminary identification of cold resistance, cold-resistant transgenic cabbage plants more than the control.