[1]王萍,高世庆,郭永来,等.利用农杆菌介导将抗逆相关基因GmDREB导入大豆的研究[J].大豆科学,2008,27(01):47-51.[doi:10.11861/j.issn.1000-9841.2008.01.0047]
　WANG Ping,GAO Shi-qing,GUO Yong-lai,et al.Transformation of Stress Resistance Related Gene GmDREB into Soybean via Agrobacterium-mediation[J].Soybean Science,2008,27(01):47-51.[doi:10.11861/j.issn.1000-9841.2008.01.0047]
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利用农杆菌介导将抗逆相关基因GmDREB导入大豆的研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第27卷 期数: 2008年01期 页码: 47-51 栏目: 出版日期: 2008-02-25
Title:
Transformation of Stress Resistance Related Gene GmDREB into Soybean via Agrobacterium-mediation
作者:
王萍1; 高世庆2; 郭永来3 ; 程宪国4; 王罡5; 马有志2; 季静5
1淮海工学院海洋学院，江苏连云港 222005；
2中国农业科学院作物科学研究所，北京 100081；
3长春龙洋大豆生物蛋白股份有限公司，吉林长春 130041；
4中国农业科学院资源与农业区划研究所，北京 100081；
5天津大学农业与生物工程学院，天津 300072
Author(s):
WANG Ping1; GAO Shi-qing2; GUO Yong-lai3; CHENG Xian-guo4; WANG Gang5; MA You-zhi2; and JI Jing5
3Changchun Longyang Soybean Biologic Albumen Co.Ltd，Changchun 130062，Jilin；
4 Institute of Agricultural Resources and Regional Planning，Chinese Academy of Agricultural Sciences，Beijing 100081;
5 Agriculture and biology engineering college，Tianjin University，Tianjin 300072，China
关键词:
大豆; 农杆菌; GmDREB基因; 未成熟子叶
Keywords:
Soybean（Glycine maxL.）; Agrobacterium; GmDREB gene; Immature cotyledon
DOI:
10.11861/j.issn.1000-9841.2008.01.0047
文献标志码:
A
摘要:
转基因技术用于作物改良是分子育种技术的重要内容之一，分子育种技术与传统育种技术相结合方法来改良大豆品种的性状已展现出良好的应用前景。以大豆未成熟子叶为外植体，研究了5个基因型体细胞胚发生率以及未成熟子叶对PPT的基础抗性。用农杆菌介导法将含有GmDREB基因的prdGm-200质粒转化大豆未成熟子叶，并探讨了影响农杆菌大豆遗传转化的主要因素。结果表明，5个大豆基因型的未成熟子叶体细胞胚胎发生率存在显著差异，东农40和吉林35的胚胎发生率显著高于黑农41、九9568、九9313。东农40和吉林35未成熟子叶对PPT很敏感，PPT适合筛选浓度为2~3 mg L-1。农杆菌EHA101菌液浓度、浸染时间对转化效率均有影响，农杆菌菌液浓度为OD600=0.5~0.7、浸染时间10~20 min有利于转化。经PPT筛选，PCR、PCR~Southern检测得到抗性体细胞胚和抗性植株，初步证明GmDREB基因转入到大豆中。
Abstract:
Plant genetic transformation techniques have been important to improve crop varieties.Combined with traditional breeding process，molecular breeding techniques have showed bright prospects for improving agronomic characters in soybean.The somatic embryogenesis and basic resistance to PPT in immature cotyledon with 5 genotypes of soybean were tested.Plasmid prdGm-200 containing GmDREB gene was transformed into immature cotyledon of soybean via Agrobacterium-mediation.The main factors affecting Agrobacterium-mediated transformation were also investigated.The result showed that the somatic embryogenesis of immature cotyledon were different among 5 genotypes in soybean.The somatic embryogenesis in Dongnog 42 and Jilin 35 were higher than those in Heinong 41，Jiu 9568 and Jiu 9313.The immature cotyledon of Dongnog 42 and Jilin 35 were sensitive to PPT.The screening concentration of PPT was appropriate with 2－3 mg L-1.Transformation efficiency was also affected by concentration of Agrobacterium-EHA101 and inoculation time.OD 0.5-0.7 and 10-20 minutes of inoculation were helpful to transformation.The resistant somatic embryos and plantlets were identified after PPT screening，PCR and PCR~Southern detection.It was demonstrated that the GmDREB gene had been transformed into soybean.

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备注/Memo

基金项目：教育部大豆生物学重点实验室资助项目（SB06A07）；国家植物转基因研究与产业化专项（JY03-A-18）

作者简介：王萍（1957-），女，教授，博士，主要从事生物技术与植物转基因的研究。Tel：051885895427;Fax： 0 518-85895429; E-mail：y_pwang@yahoo.com.cn

更新日期/Last Update: 2014-10-09