[1]杨静,邢国杰,杜茜,等.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[J].大豆科学,2016,35(04):562-567.[doi:10.11861/j.issn.1000-9841.2016.04.0562]
　YANG Jing,XING Guo-jie,DU Qian,et al.Effects of Different Soybean Genotypes on the Transformation Efficiency of Soybeanand Analysis of the T-DNA Insertions in the Soybean Genome[J].Soybean Science,2016,35(04):562-567.[doi:10.11861/j.issn.1000-9841.2016.04.0562]
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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

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年04期 页码: 562-567 栏目: 出版日期: 2016-07-20
Title:
Effects of Different Soybean Genotypes on the Transformation Efficiency of Soybeanand 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
( 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)
Author(s):
YANG Jing1; XING Guo-jie1; DU Qian2; SUI Li2; GUO Dong-quan1; NIU Lu1; YANG Xiang-dong1
1. Agro-Biotechnology Institute，Jilin Academy of Agricultural Sciences /Jilin Provincial Key Laboratory of Agricultural Biotechnology，Changchun130033，China; 2. Institute of Plant Protection，Jilin Academy of Agricultural Sciences，Gongzhuling 136100，China
关键词:
大豆基因型;  农杆菌介导转化;  转化效率;  T-DNA 插入
Keywords:
Soybean ［Glycine max ( L. ) Merr. ］ genotypes;  Agrobacterium-mediated transformation;  Transformationrate;  T-DNA insertions
分类号:
S565. 1
DOI:
10.11861/j.issn.1000-9841.2016.04.0562
文献标志码:
A
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。
Abstract:
Soybean is usually considered as a recalcitrant leguminous crop to Agrobacterium-mediated transformation. Soybeangenotypes exert significant effects on the transformation efficiency. In this study，two abroad soybean cultivars and 9 domesticcultivated soybeans were employed for Agrobacterium-mediated transformation. PCＲ and LibertyLink strip analysis on the regeneratedplants showed that different soybean genotypes exhibited significant differences in both regeneration and transformationrates. Among them，Jack，Bert，Huachun 6 and Shennong 9 showed higher transformation efficiency than other 7 cultivarswith 6. 45%，3. 80%，3. 24% and 2. 86% of transformation rate. Southern blot analysis was also carried out for the PCＲ-positiveplants derived from Shennong 9. The results showed the integration of T-DNA in the soybean genome with 2. 14% oftransformation rate obtained for the recipient cultivar. Further analysis showed that 75% of the transgenic plants tested containedlow copies( 1-2) T-DNA insertions. Taken together，in this study，four soybean cultivars with high transformation rateswere selected for Agrobacterium-mediated transformation，Huachun 6 and Shennong 9 could be further utilized for developmentof transgenic soybean cultivars.

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备注/Memo

基金项目: 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。第一作者简介: 杨静( 1982-) ，女，硕士，助理研究员，主要从事大豆转基因育种研究。E-mail: jinggyang@126. com。通讯作者: 杨向东( 1976-) ，男，博士，副研究员，主要从事大豆生物技术育种研究。E-mail: xdyang020918@126. com。

更新日期/Last Update: 2016-08-22