抗草甘膦转基因玉米AG16分子特征和抗性鉴定  

Molecular characterization and efficacy evaluation of a transgenic corn event AG16

作　　者：刘苗苗 程家慧 林海燕[1] 沈志成[1] 王志勇[2] 林朝阳[1] 

机构地区：[1]浙江大学植物保护系,昆虫科学研究所,浙江杭州310058 [2]海南大学农学院,海南海口570228

出　　处：《草业科学》2017年第9期1830-1837,共8页Pratacultural Science

基　　金：农业部转基因专项--安全转基因技术研究(2016ZX08010-003)

摘　　要：抗草甘膦转基因玉米(Zea mays)能有效降低杂草防治成本,具有重要的应用前景。浙江大学转基因抗虫植物和生物安全实验室前期通过农杆菌介导法,以玉米Hi-Ⅱ品种为受体导入新型抗草甘膦基因G10evo,获得不同的抗草甘膦玉米转化系。在此基础上,筛选了具有良好草甘膦抗性的转基因玉米转化系AG16。本研究利用PCR、Western杂交、Southern杂交、ELISA等方法对AG16进行分子特征检测,并对AG16的草甘膦抗性水平进行鉴定。结果表明,G10evo在AG16中为单拷贝插入;G10evo蛋白在AG16的根、茎和叶组织中表达;ELISA分析表明,草甘膦喷洒前,嫩茎中G10evo蛋白的表达量达到9.975μg·g-1。温室中草甘膦抗性测定结果显示,AG16能抗4~8倍田间浓度的草甘膦,远高于草甘膦在实际生产中的使用量。因此,AG16草甘膦抗性水平达到生产需求,具有产业应用潜力,为培育具有自主知识产权的抗草甘膦玉米提供了种质资源。Glyphosate-resistant transgenic maize can significantly improve the efficiency of weed control. In pre- vious studies, the newly glyphosate-tolerant gene GlOevo was integrated into the genome of maize named Hi-II by Agrobacteriurn-mediated transformation, and different glyphosate-tolerant maize events were obtained. In the study, transgenic maize AG16 was selected for molecular characterization by PCR, Western blot, Southern blot and ELISA. Southern blot analysis using probe for GlOevo showed that T-DNA was integrated in a single site in the maize genome. Western blot analysis showed that G10evo protein was expressed in leaf, stem and root of AG16. ELISA assay showed that the expression level of G10evo protein in the young stem reached 9.975 μg·g^-1 before glyphosate spraying. Glyphosate spray assay in the greenhouse showed that the AG16 is tolerant to glyphosate at concentrations up to 4 times of recommended field application level. Therefore, the transgenic maize AG16 has the potential of field application, which is expected to become completely independent research and independent intellectual property rights of domestic glyphosate-resistant corn.

关 键 词：G10evo基因 PCR检测 SOUTHERN杂交 Western杂交 蛋白定量 抗性鉴定 杂草防治 

分 类 号：S513[农业科学—作物学]