不同大豆基因型对大豆遗传转化效率的影响及外源T-DNA插入分析  

Effects of Different Soybean Genotypes on the Transformation Efficiency of Soybean and Analysis of the T-DNA Insertions in the Soybean Genome

作　　者：杨静[1] 邢国杰[1] 杜茜[2] 隋丽[2] 郭东全[1] 牛陆[1] 杨向东[1] 

机构地区：[1]吉林省农业科学院农业生物技术研究所/吉林省农业生物技术重点实验室,吉林长春130033 [2]吉林省农业科学院植物保护研究所,吉林公主岭136100

出　　处：《大豆科学》2016年第4期562-567,共6页Soybean Science

基　　金：国家转基因生物新品种培育重大专项(2016ZX08004-004);吉林省科技发展计划项目(20150204011NY);吉林省农业科技创新工程项目

摘　　要：大豆通常被认为是较难转化的豆科作物之一,大豆基因型是影响大豆转化效率的重要因素。采用农杆菌介导大豆子叶节遗传转化方法,对2个国外大豆品种和9个国内品种进行转化研究,对转化再生植株进行BAR试纸条和PCR检测。结果表明：不同大豆品种再生率及转化效率存在显著差异。国外品种Jack和Bert,南方大豆品种华春6号和东北大豆品种沈农9号具有较高的再生率和转化效率,转化效率分别达到6.45%、3.80%、3.24%和2.86%。对来源于沈农9号的PCR阳性植株进行Southern杂交检测,结果表明：外源基因已导入受体大豆品种,该受体品种实际转化效率为2.14%。对外源T-DNA插入结构进一步分析表明：低拷贝（1-2个）T-DNA插入比例为75%。本研究筛选了4个转化效率较高的大豆品种,为开展高效大豆遗传转化研究提供依据;同时,作为大豆主栽品种,利用华春6号和沈农9号作为受体品种开展转基因研究,对于加快转基因大豆新品种培育研究也具有重要的参考价值。Soybean is usually considered as a recalcitrant leguminous crop to Agrobacterium-mediated transformation. Soybean genotypes exert significant effects on the transformation efficiency. In this study, two abroad soybean cultivars and 9 domestic cultivated soybeans were employed for Agrobacterium-mediated transformation. PCR and LibertyLink strip analysis on the regenerated plants showed that different soybean genotypes exhibited significant differences in both regeneration and transforma- tion rates. Among them, Jack, Bert, Huachun 6 and Shennong 9 showed higher transformation efficiency than other 7 cuhivars with 6.45%, 3.80%, 3.24% and ：2. 86% of transformation rate. Southern blot analysis was also carried out for the PCR-positive plants derived from Shennong 9. The results showed the integration of T-DNA in the soybean genome with 2. 14% of transformation rate obtained for the recipient cultivar. Further analysis showed that 75% of the transgenic plants tested contained low copies（1-2） T-DNA insertions. Taken together, in this study, four soybean cuhivars with high transformation rates were selected for Agrobacterium-mediated transformation, Huachun 6 and Shennong 9 could be further utilized for development of transgenic soybean cultivars.

关 键 词：大豆基因型 农杆菌介导转化 转化效率 T-DNA插入 

分 类 号：S565.1[农业科学—作物学]