利用pmi作为筛选基因对辣椒进行Bt基因转化的研究  

Transformation of Bt gene into Hot Pepper using pmi gene as Selectable Marker

作　　者：黄皓[1] 雷建军[1] 曹必好[1] 陈长明[1] 陈清华[1] 陈国菊[1] 

机构地区：[1]华南农业大学园艺学院,广州510642

出　　处：《辣椒杂志》2013年第2期23-28,共6页Journal of China Capsicum

基　　金：国家863项目(2009AA10Z104);广东省攻关项目(2010A020102001;2011A020102004);广州市科技局项目(2011Y2-0001511B)

摘　　要：根据Cry2Aa2的保守基因序列设计引物,从而扩增出完整的Cry2Aa2序列,连接到含有磷酸甘露糖异构酶基因(pmi)的表达载体pCAMBIA1301-PMI上,采用农杆菌介导的方法转化辣椒,利用甘露糖筛选体系对辣椒转化体进行筛选,对转基因植株进行分子生物学检测和抗虫试验,72h后统计抗虫情况。结果表明:获得了含有Cry2Aa2转基因植株;食用转Bt基因辣椒叶片和果实的斜纹夜蛾幼虫的校正死亡率均高于阴性对照,且大多表现出僵化和厌食的状态,对转基因的辣椒伤害较小,说明Bt基因成功导入辣椒中。In this study, primers were designed to amplify Cry2Aa2 from T -vector containing Cry2Aa2, according to the sequence of Cry2Aa2 gene submitted in the Genebank. The target fragments were linked to pCAMBIA1301-PMI and introduced into hot pepper by Agrobacterium mediated transformation,and the PMI/ Man system was adapted in the Agrobacterium-mediated transformation. Resistance to insects was also analyzed in the transgenic plants 72 h after the resistant test. The results showed that the transgenic pepper plants were obtained with Cry2Aa2, the adjusted mortality of insects on transgenic plants was higher than that of normal plants, and the insects on transgenic plants grew slowly and did little damage to the leaves and fruits, which indicated that the Bt gene had integrated with the pepper genome successfully.

关 键 词：辣椒 Cry2Aa2基因 甘露糖 遗传转化 

分 类 号：S641.3[农业科学—蔬菜学]