转基因大豆AtARA6-A001事件采用农杆菌介导大豆遗传转化法获得,其受体为沈农9号,具有耐盐特性,目前已经进入环境释放阶段.为明确该转基因大豆材料的分子特征及其转基因检测方法,推进该转基因事件生物安全评价工作.本研究以转基因大豆AtARA6-A001为研究对象,利用Southern杂交方法及基因组重测序技术鉴定了外源基因的拷贝数及插入位点的位置和方向.同时利用PCR扩增技术获得了外源T-DNA的左右侧翼序列,并基于左右旁侧序列,建立了转AtARA6基因耐盐大豆A001事件的特异性定性PCR检测方法.此方法能特异性检测转基因大豆植株AtARA6-A001根、茎、叶、花和种子样品,并且能够特异性识别转基因大豆事件的身份,这为后续转基因大豆及其产品的检测和监管提供技术支持.
AtARA6 gene from Arabidopsis thaliana was introduced into soybean receptor Shennong 9 by Agrobacterium-mediated transformation of soybean cotyledon nodes,obtaining the salt-tolerant transgenic soybean event AtARA6-A001.To identify the molecular characteristics and setting up a detection method of this transgenic soybean material,we could further accelerate the biosafety evaluation of this transgenic event in the future.In this study,the copy number of exogenous gene of the homozygous transgenic soybean AtARA6-A001 was identified by Southern hybridization as well as the position and direction of the insertion site.Meanwhile,the left and right flanking sequences of exogenous T-DNA were obtained by PCR amplification.Based on these data,a specific qualitative PCR method for detecting AtARA6 gene salt-tolerant soybean A001 event was established.This method has high specificity,and can quickly identify transgenic soybean events,including AtARA6-A001 parents,derivatives,or varieties,as well as plants,tissues,seeds and products for specific detection,achieving effective supervision and management of transgenic soybeans by technical support.
孙星邈;侯云龙;王英哲;关诗宇;邱红梅;张玲
吉林省农业科学院(中国农业科技东北创新中心)大豆研究所,吉林长春 130033吉林省农业科学院(中国农业科技东北创新中心)农业生物技术研究所,吉林长春 130033
大豆;转基因;侧翼序列;重测序;定性PCR检测
soybean;transgene;flanking sequence;resequencing;qualitative PCR detection
《大豆科学》 2024 (001)
29-34 / 6
吉林省农业科技创新工程项目定向委托(CXGC2022DX009);大豆产业技术体系种子生产岗位专家项目(CARS04-P13).
10.11861/j.issn.1000-9841.2024.01.0029
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