[1]王全伟,张海玲,白晶,等.农杆菌介导的大豆植株整体转化[J].大豆科学,2008,27(02):190-193,198.[doi:10.11861/j.issn.1000-9841.2008.02.0190]
　WANG Quan-wei,ZHANG Hai-ling,BAI Jing,et al.Agrobacterium-Mediated Plant Transformation of Soybean[J].Soybean Science,2008,27(02):190-193,198.[doi:10.11861/j.issn.1000-9841.2008.02.0190]
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农杆菌介导的大豆植株整体转化

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第27卷 期数: 2008年02期 页码: 190-193,198 栏目: 出版日期: 2008-04-30
Title:
Agrobacterium-Mediated Plant Transformation of Soybean
文章编号:
1000-9841(2008)02-0190-04
作者:
王全伟; 张海玲; 白晶; 徐香玲
哈尔滨师范大学生命科学与技术学院，黑龙江 哈尔滨，150025
Author(s):
WANG Quan-wei; ZHANG Hai-ling; BAI Jing; XU Xiang-ling
Life Science and Technology Institute，Harbin Normal University，Harbin 150025，Heilongjiang，China
关键词:
大豆; 农杆菌; 整体植株转化
Keywords:
Soybean; Agrobacterium; Plant transformation
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2008.02.0190
文献标志码:
A
摘要:
农杆菌介导的植物整体转化法主要以植物的分生组织和生殖器官作为外源基因导入的受体，通过真空渗透法、浸蘸法及注射法等方法使农杆菌与受体材料接触，以完成可遗传细胞的转化，通过抗生素筛选和分子检测鉴定转基因植株后代。以大豆幼苗的顶端生长点和叶腋生长点为靶点，通过农杆菌介导的整体转化法——注射法进行转录因子DREB1C基因的遗传转化，最终获得6株T0代PCR阳性转基因植株，转化率为9.2%。T1代植株的PCR、Southern blot和RT-PCR鉴定结果表明获得1株阳性植株，DREB1C基因以单拷贝形式整合于大豆基因组中，在200 mmolL-1NaCl胁迫条件下在转录水平得到成功表达。该方法克服了大豆组织培养再生难、转化率低的缺点，是一种高频、简单、快捷的非组培遗传转化途径。
Abstract:
Plant meristem and reproductive organs are often taken as the receptor of exogenous gene in Agrobacterium Mediated plant Transformation.Agrobacterium can interact with the receptor and complete genetic transformation by vacuum infiltration，floral dip，injection and so on.Then transgenic plants can be detected by the screening of antibiotics and molecular tests.In this study，the gene DREB1C encoding a transcription factor was transformed into the apical and axils growing points of soybean seedling through Agrobacterium-mediated plant genetic transformation．Six positive lines were obtained through PCR assay in the T0?generation transgenic lines.The frequency of transformation is 9.2%.The T1 generation transgenic lines have been tested through PCR，Southern blot and RT-PCR and one shows positive.The results indicate that the DREB1C gene has been integrated into the genome of soybean and has a single copy．The gene transcripts successfully under 200 mmolL-1NaCl stress．So this non-tissue culture transformation method is simple and it improves the frequency of transformation efficiently.

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备注/Memo

基金项目：哈尔滨师范大学科研基金资助项目（KM2007-03）。

作者简介：王全伟（1978-），女，讲师，博士研究生，研究方向为基因工程。E-mail：wqw125@126.com。

更新日期/Last Update: 2014-10-10