基因枪法和农杆菌介导的Bt抗虫基因转化芥蓝

发布时间：2018-10-18 11:52

【摘要】：以芥蓝品种小香菇为试材,分别采用基因枪法和农杆菌介导转化法将Bt抗虫基因Cry2Aa2转化到芥蓝中,并对两种转化方法进行了优化和比较。结果表明:基因枪法在轰击距离为6cm、轰击次数为1次时转化效率最高,为0.43%;农杆菌介导转化法以带子叶下胚轴为外植体、预培养2d、侵染10min、共培养2d、抑菌培养5d后转入筛选培养基为最佳参数,转化效率为0.37%。经PCR和PCR-Southern鉴定,基因枪法和农杆菌介导转化法均成功将目的基因导入芥蓝基因组中。经抗虫性表型鉴定,T_0代转基因芥蓝植株对甘蓝夜蛾幼虫的抗性高于非转基因植株。
[Abstract]:The Bt insect-resistant gene Cry2Aa2 was transformed into Brassica oleracea by gene gun method and Agrobacterium tumefaciens-mediated transformation. The two transformation methods were optimized and compared. The results showed that the transformation efficiency was the highest when the bombardment distance was 6 cm and the number of bombardment was 1, and the transformation efficiency was 0.43, and Agrobacterium tumefaciens mediated transformation was based on the hypocotyls of scallop leaves. The optimum parameters were pre-culture for 2 days, infection for 10 minutes, co-culture for 2 days, inhibition of bacteria for 5 days, and then transferred to the screening medium. The transformation efficiency was 0.37%. The results of PCR and PCR-Southern showed that the target gene was successfully introduced into the genome of kale by gunshot method and Agrobacterium tumefaciens-mediated transformation. The resistance of transgenic kale plants in generation 0 to larvae of Spodoptera spp. was higher than that of non-transgenic plants.
【作者单位】： 华南农业大学园艺学院;
【基金】：广东省教育厅特色创新项目(2014KTSCX026) 广东省教育厅青年人才项目(2014KQNCX035) 华南农业大学校长基金项目(K13012)
【分类号】：S635.9
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