转基因抗虫玉米LG11中m2cryAb-vip3A融合基因及其蛋白的表达分析  

Expression Analysis of m2cryAb-vip3A Gene in Transgenic Insect-resistant Maize LG11

作　　者：李文兰[1] 崔洁 李冬梅 李海涛 孟昭东[1] 岳润清 Li Wenlan;Cui Jie;Li Dongmei;Li Haitao;Meng Zhaodong;Yue Runqing(Maize Research Institute,Shandong Academy of Agricultural Sciences/National Engineering Laboratory of Wheat and Maize/Key Laboratory of Biology and Genetic Improvement of Maize in Northern Yellow-Huai River Plain,Ministry of Agriculture,Jinan 250100,China;Yantai Rural Economic Management Station,Yantai 264001,China)

机构地区：[1]山东省农业科学院玉米研究所/小麦玉米国家工程研究中心/农业部黄淮海北部玉米生物学与遗传育种重点实验室,山东济南250100 [2]烟台市农村经济经营管理站,山东烟台264001

出　　处：《山东农业科学》2024年第6期25-29,共5页Shandong Agricultural Sciences

基　　金：泰山学者人才工程项目;国家现代农业产业技术体系建设项目(CARS-02-09)

摘　　要：抗虫转基因作物中杀虫蛋白的表达量对抗虫效果至关重要,研究外源Bt基因在作物不同生育时期和不同组织器官中的时空表达模式,对于抗虫转基因玉米产业化后的害虫抗性治理和农业转基因生物安全管理具有重要意义。LG11抗虫转基因玉米中的抗虫基因是山东省农业科学院通过人工合成方法将外源Bt杀虫蛋白Cry1Ab和Vip3Aa的主要结构域组合形成的新型抗虫融合蛋白M2cryAb-vip3A,连续两年对该转化体材料抗虫基因m2cryAb-vip3A的表达模式进行分析,利用实时定量PCR方法从转录水平对LG11苗期的根、茎和叶,吐丝期的根和茎,成熟期的根、叶和籽粒中的m2cryAb-vip3A基因的表达模式进行分析;利用酶联免疫吸附法(ELISA)从翻译水平对苗期的根、茎和叶,吐丝期的根、茎、花丝和花粉,成熟期的根、茎、叶、籽粒和苞叶中的M2cryAb-vip3A蛋白表达量进行测定。结果表明,m2cryAb-vip3A抗虫融合基因及其编码蛋白在抗虫转基因玉米不同生育时期的不同组织器官中表达差异较大,以幼苗期叶片中的表达量最高。该研究结果可为LG11未来的商业化推广和农业转基因生物安全管理提供有用的信息。The expression level of insecticidal protein in insect-resistant transgenic crops is very important to the effect of insect-resistant transgenic crops.The study on the temporal and spatial expression patterns of exogenous Bt gene in different developmental stages and different tissues and organs is of great significance for pest resistance management and agricultural GMO safety management after insect-resistant transgenic maize industrialization in the future.The anti-insect gene in LG11 insect-resistant transgenic maize is a fusion protein M2cryAb-vip3A formed by combining the main domains of Cry1Ab and Vip3Aa proteins by synthetic method.The expression pattern of the anti-insect gene m2cryAb-vip3A in the transformed material was analyzed for two consecutive years.Real-time quantitative PCR was used to analyze the expression patterns of m2cryAb-vip3A gene in roots,stems and leaves at seedling stage,roots and stems at spinning stage,roots,leaves and kernels at maturity stage.The expression of M2cryAb-vip3A protein in roots,stems and leaves at seedling stage,roots,stems,filaments and pollen at spinning stage,roots,stems,leaves,kernels and bract at maturity stage was determined by enzyme-linked immunosorbent assay(ELISA).The results showed that the expression of m2cryAb-vip3A fusion anti-insect gene and its encoded protein were significantly different in different tissues and organs at different developmental stages.From the perspective of development,the expression level of m2cryAb-vip3A fusion anti-insect gene and its encoding protein was the highest in seedling stage,and the expression level of m2cryAb-vip3A fusion anti-insect gene and its encoding protein was the highest in leaves from the perspective of tissues and organs.

关 键 词：m2cryAb-vip3A 表达模式 转基因 玉米 

分 类 号：S513[农业科学—作物学] Q786[生物学—分子生物学]