[1]郭兵福,刘 杰,洪慧龙,等.一种简便大豆原位转基因方法研究[J].大豆科学,2012,31(03):347-352,357.[doi:10.3969/j.issn.1000-9841.2012.03.003]
　GUO Bing-fu,LIU Jie,HONG Hui-long,et al.A Simple Soybean in Planta Transformation Method[J].Soybean Science,2012,31(03):347-352,357.[doi:10.3969/j.issn.1000-9841.2012.03.003]
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一种简便大豆原位转基因方法研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第31卷 期数: 2012年03期 页码: 347-352,357 栏目: 出版日期: 2012-03-25
Title:
A Simple Soybean in Planta Transformation Method
文章编号:
1000-9841（2012）03-0347-06
作者:
郭兵福1; 2; 刘 杰1; 2; 洪慧龙2; 邱丽娟2
1.南昌大学 生命科学与食品工程学院，江西 南昌 330031；
2.中国农业科学院 作物科学研究所/国家农作物基因资源与遗传改良重大科学工程/农业部北京大豆生物学重点实验室，北京100081
Author(s):
GUO Bing-fu1; 2; LIU Jie1; 2; HONG Hui-long2; QIU Li-juan2
1.College of Life Sciences,Nanchang University,Nanchang 330031,Jiangxi;
2.The National Key Facility for Crop Gene Resources and Genetic Improvement(NFCRI)/MOA Key Lab of Soybean Biology(Beijing),Institute of Crop Science,Chinese Academy of Agricultural Sciences,Beijing 100081,China
关键词:
大豆; 根癌农杆菌; 顶芽; 原位转化
Keywords:
Soybean; Agrobacterium; Epicotyl tip; In planta transformation
分类号:
S565.1
DOI:
10.3969/j.issn.1000-9841.2012.03.003
文献标志码:
A
摘要:
以萌发大豆幼苗顶芽为外植体，经纵切及农杆菌侵染后种植到大田中，转化植株用除草剂进行表型鉴定，存活植株进行PCR验证，并分析目的基因EPSPS在T2代转基因植株中的遗传情况；总计获得草铵膦涂抹表型鉴定阳性T0植株75株，草甘膦喷雾鉴定阳性T1植株65个，PCR测序阳性T1植株6个，PCR测序检测转化效率为0.14%；获得T2代PCR阳性植株52个，初步证明目的基因EPSPS能在子代中遗传；该方法能有效解决基因型依赖及再生植株困难等问题，缩短转化周期，为根癌农杆菌阶段的大豆遗传转化体系的优化与改良提供了参考。
Abstract:
In this paper, the terminal bud of germinated soybean seedlings were longitudinal cut and infected by Agrobacterium firstly, and then transplanted the seedlings into field. Eliminated the seedlings without branches on main stem, transformed plants leaves were painted or sprayed with 100 mg·L-1 glufosinate, 3 days later observed leaves reaction and accounted for the resistant plant number. All alive plants were identified by PCR and analyzed the expression of EPSPS gene in T2 generation. There were 75 and 65 plants with resistance to 100 mg·L-1 glufosinate in T0 and T1 generation, respectively; 6 and 52 positive plants in T0 and T1 generation by sequencing the PCR product, respectively; the result showed that EPSPS gene could inherited in offspring. This transgenic method could short transformation cycle, enhance efficacy and provide reference for the optimization and improvement of Agrobacterium transformation.

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备注/Memo

基金项目：转基因生物新品种培育重大专项（2008ZX08004-001）。

第一作者简介：郭兵福（1985-），男，在读硕士，研究方向为大豆转基因。E-mail：gbfhq@163.com。
通讯作者：邱丽娟（1963-），女，研究员，博士生导师，主要从事大豆基因资源挖掘与利用研究工作。E-mail：qiu_lijuan@263.net。

更新日期/Last Update: 2014-08-16