转epsps-G6基因棉花的Southern杂交及遗传特性分析  

Analysis of Southern Blot and Genetic Character in Transgenic Cotton Expressing epsps-G6 Gene

作　　者：燕树锋[1,2] 许蒙蒙 祝水金[2] 郭新海 铁双贵[1] YAN Shufeng;XU Mengmeng;ZHU Shuijin;GUO Xinhai;TIE Shuanggui(Cereal Crops Research Institute,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Agronomy Department,College of Agriculture and Biotechnology,Zhejiang University,Hangzhou 310085,China)

机构地区：[1]河南省农业科学院粮食作物研究所,河南郑州450002 [2]浙江大学农业与生物技术学院农学系,浙江杭州310085

出　　处：《华北农学报》2018年第3期19-23,共5页Acta Agriculturae Boreali-Sinica

基　　金：转基因生物新品种培育重大专项(2014ZX08005-005)

摘　　要：为鉴定epsps-G6基因成功导入棉花基因组并分析转基因抗草甘膦棉花的拷贝数和遗传特性,以花粉管通道法获得的8个转epsps-G6基因抗草甘膦棉花株系(G6-1、G6-4、G6-5、G6-7、G6-11、G6-19、G6-20、G6-25)为材料,通过Southern点杂交、双酶切和单酶切的Southern杂交后进行分析,结果表明,外源epsps-G6基因已成功整合到棉花基因组上,且其中4个转化株系含有单拷贝插入的epsps-G6基因。选用其中3个单拷贝插入、自交纯合且草甘膦抗性强的株系G6-1、G6-5、G6-19,分别与转基因遗传背景材料(中棉所49)和陆地棉标准系(TM-1)进行正反交,遗传分析结果表明,这3个转基因株系均符合3∶1的孟德尔分离规律,不受细胞质效应的影响。In order to identify the epsps-G6 gene successfully introduced into the cotton genome and analyze the copy number and genetic characteristics of the transgenic glyphosate-resistant cotton,eight transformation events（ G6-1 ,G6-4,G6-5,G6-7,G6-11,G6-19,G6-20,G6-25）obtained by the pollen tube pathway method were used as materials. The results of southern Dot Blotting,double and single digestion enzyme Southern hybridization indicated that the exogenous epsps-G6 gene had been successfully integrated into the cotton genome and that four of the transformants contained a single copy of epsps-G6 gene. Three homozygous lines（G6-1,G6-5,G6-19）by self-pollination,with a single copy insertions and high glyphosate resistance,were selected to reciprocal cross with genetic background cultivar CCRI-49 and genetic standard line TM-1. The results of genetic analysis suggested that the three transformation events were inconsistent with the Mendelian separation rule of 3 ∶ 1,which was not influenced by the cytoplasmic effect.

关 键 词：棉花 epsps-G6基因 草甘膦 除草剂 花粉管通道法 

分 类 号：S562.03[农业科学—作物学]