抗除草剂棉花GV-2的分子特征和遗传稳定性分析  

Molecular Characterization and Inheritance Stability Analysis of Herbicide-resistant Cotton GV-2

作　　者：陆国清[1,2] 马彩霞 孙国清 郭惠明[1] 程红梅[1] LU Guoqing;MA Caixia;SUN Guoqing;GUO Huiming;CHENG Hongmei(Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Biotechnology Research Institute,Tianjin Academy of Agricultural Sciences,Tianjin 300381,China;College of Agriculture,Shanxi Agricultural University,Shanxi Jinzhong 030801,China)

机构地区：[1]中国农业科学院生物技术研究所,北京100081 [2]天津市农业科学院生物技术研究所,天津300381 [3]山西农业大学农学院,山西晋中030801

出　　处：《中国农业科技导报》2023年第1期42-49,共8页Journal of Agricultural Science and Technology

基　　金：国家转基因生物新品种培育重大专项(2020ZX08009-05B)。

摘　　要：分子特征和遗传稳定性是我国转基因植物安全评价流程所需考察的重要数据。利用基因组测序技术对抗除草剂棉花GV-2的T-DNA插入位点、拷贝数及侧翼序列进行解析,并采用实时荧光定量PCR(quantitative real time polymerase chain reaction,qRT-PCR)、Southern杂交、胶体金试纸及ELISA(enzyme linked immunosorbent assay)方法对连续3代(T3~T5)GV-2转化体中目的基因的表达的稳定性进行验证。结果表明,转化体GV-2中T-DNA以单拷贝形式插入到陆地棉A06染色体2 092 124~2 092 194 bp之间,造成棉花基因组中70 bp DNA的缺失。转化体特异性PCR及Sanger测序结果进一步验证了插入位点的正确性。此外,目的基因及其表达蛋白在不同世代转化体中均可稳定遗传,为转基因棉花GV-2转化体的安全评价提供了有效的支撑。Molecular characterization and inheritance stability are important data for safety evaluation of genetic modified crop in China.The whole genome sequencing was used to analyze the T-DNA insertion site,copy number and flanking sequence of the herbicide-resistant transgenic cotton GV-2.The stability of target gene in three successive generations(T3~T5)GV-2 were verified by qRT-PCR(quantitative real time polymerase chain reaction),Southern hybridization,colloidal gold dipstick and ELISA(enzyme linked immunosorbent assay).The results showed that the T-DNA in GV-2 was inserted in the form of single copy,and the insertion site was located in 2092124~2092194 bp on chromosome A06 of upland cotton,resulting in the deletion of 70 bp DNA in the cotton genome.The insertion site and flanking sequence were further confirmed by the specific PCR of the transformant and Sanger sequencing.In addition,the target gene and its expressed protein,as well as the target character could be inherited stably in different generations of transformants.The results provided effective support for safety evaluation of cotton GV-2 transformant.

关 键 词：转基因棉花 抗除草剂 G10evo 分子特征 遗传稳定性 

分 类 号：S562[农业科学—作物学]