[1]许娜,闫强,崔晓霞,等.发根农杆菌介导的大豆DAD基因沉默嵌合植株的获得[J].大豆科学,2015,34(04):576-581.[doi:10.11861/j.issn.1000-9841.2015.04.0576]
　XU Na,YAN Qiang,CUI Xiao-xia,et al.Agrobacterium-mediated GmDAD1 Gene Silence of Transgenic Hairy Roots in Chimeric Soybean[J].Soybean Science,2015,34(04):576-581.[doi:10.11861/j.issn.1000-9841.2015.04.0576]
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发根农杆菌介导的大豆DAD基因沉默嵌合植株的获得

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第34卷 期数: 2015年04期 页码: 576-581 栏目: 出版日期: 2015-08-25
Title:
Agrobacterium-mediated GmDAD1 Gene Silence of Transgenic Hairy Roots in Chimeric Soybean
作者:
许娜1; 闫强1; 2; 崔晓霞1; 郭娜1; 窦道龙2; 邢邯1
1南京农业大学 大豆研究所/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室/作物遗传与种质创新国家重点实验室，江苏 南京 210095；
2南京农业大学 植物保护学院，江苏 南京 210095
Author(s):
XU Na1;  YAN Qiang1; 2;  CUI Xiao-xia1; GUO Na1;  DOU Dao-long2;  XING Han1
1Soybean Research Institute of Nanjing Agricultural University/National Center for Soybean Improvement /Key Laboratory of Biology and Genetics and Breeding for Soybean, Ministry of Agriculture/State Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095, China; 2Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095, China
关键词:
大豆; 转基因根毛; 嵌合植株; 发根农杆菌; GmDAD
Keywords:
Soybean; Hairy root; Chimeric plant; A rhizogenes; GmDAD
DOI:
10.11861/j.issn.1000-9841.2015.04.0576
文献标志码:
A
摘要:
利用发根农杆菌介导的转化方法，获得了大豆-DAD-基因在根组织中沉默的嵌合植株。同时在pHellsGate12载体基础上插入一个GFP表达盒使得对转基因组织的筛选变得更加简便；通过对荧光组织进行qPCR检测发现转基因组织与对照相比，DAD-基因转录水平降低45%~70%。以上结果表明大豆-DAD-基因在转基因根组织中已被成功沉默，并且利用改造后的pHellsGate12:GFP载体通过荧光可以对转基因组织进行初步快速筛选。
Abstract:
Agrobacterium rhizogenes is able to transform plant genomes and induce the production of hairy roots.In this work, we described the use of Arhizogenes in soybean to rapidly assess gene expression and function. We first cloned the soybean –DAD- gene into pHellsgate12:GFP vector, which was reconstructed by inserted a 35S:GFP:nos expression cassette into SacI digested pHellsGate12, then transformed the DAD-gene fragment into soybean by using this chimeric hairy root transgenic system.The transcription level of DAD in the fluorescence hairy roots were reduced to 45%~70%.The results revealed that the DAD gene was silenced in transgenic hairy roots, and the reconstructed RNAi vector could simplify the progress of positive hairy roots screen.

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备注/Memo

基金项目：国家现代农业产业技术体系（CARS-004-PS10）；转基因生物新品种培育重大专项（2014ZX08004-002）；南京农业大学自然科学重大项目（KYZ201202-3）；长江学者和创新团队发展计划（PCSIRT13073）；江苏省现代作物生产协同创新中心资助。

第一作者简介：许娜（1990-），女，硕士，主要从事大豆遗传育种研究。E-mail:xuna@njau.edu.cn。
通讯作者：邢邯（1963-），男，教授，博导，主要从事大豆育种研究。E-mail:hanx@njau.edu.cn。

更新日期/Last Update: 2015-08-31