| 农杆菌介导的大豆子叶节遗传转化技术流程及操作要点 |
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| 引用本文： | 赵晓雯,吴芳芳,狄少康,柏锡,纪巍,李勇,才华,朱延明.农杆菌介导的大豆子叶节遗传转化技术流程及操作要点[J].大豆科学,2011,30(3):362-368. |
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| 作者姓名： | 赵晓雯 吴芳芳 狄少康 柏锡 纪巍 李勇 才华 朱延明 |
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| 作者单位： | 东北农业大学生命科学学院; |
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| 基金项目： | 国家高技术研究发展计划(“863”计划)资助项目(2008AA10Z153); 转基因生物新品种培育科技重大专项资助项目(2008ZX08004-002) |
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| 摘 要： | 大豆遗传转化效率低,是大豆转基因育种亟待解决的重要问题。以大豆子叶节为外植体采用农杆菌介导法,系统地研究了大豆子叶节遗传转化各技术环节的操作要点,包括工程菌液制备、无菌苗获得及外植体制备、农杆菌的侵染及共培养、不定芽诱导、不定芽伸长、根诱导、驯化与移栽、抗性植株的获得及转基因植株的检测等,建立了一套较稳定、高效的大豆子叶节遗传转化体系。并利用该体系,将野生大豆耐盐碱相关基因A1对大豆品种绥农28、合丰50、合丰55、东农50进行遗传转化,系统地探讨了大豆基因型、影响T-DNA的转移效率的各环节、筛选剂浓度及不同基因型转化效率等几个重要因素。
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| 关 键 词： | 大豆 遗传转化 农杆菌介导 子叶节 技术流程 |
Technique flow and Key Operation Points of Agrobacterium-mediated Genetic Transformation of Soybean Cotyledonary Node |
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| ZHAO Xiao-wen,WU Fang-fang,DI Shao-kang,BAI Xi,JI Wei,LI Yong,CAI Hua,ZHU Yan-ming.Technique flow and Key Operation Points of Agrobacterium-mediated Genetic Transformation of Soybean Cotyledonary Node[J].Soybean Science,2011,30(3):362-368. |
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| Authors: | ZHAO Xiao-wen WU Fang-fang DI Shao-kang BAI Xi JI Wei LI Yong CAI Hua ZHU Yan-ming |
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| Institution: | ZHAO Xiao-wen,WU Fang-fang,DI Shao-kang,BAI Xi,JI Wei,LI Yong,CAI Hua,ZHU Yan-ming(College of Life Science,Northeast Agicultural University,Harbin 150030,Heilongjiang,China) |
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| Abstract: | The low soybean genetic transformation frequency was one of the difficult problems that needed be resolved in soybean transgenic breeding.In this study,the soybean cotyledonary node was used as explants,and the technique flow and key operation points of Agrobacterium-mediated transformation method,including Agrobacterium preparation,explant and aseptic seedling preparation,infection and co-cultivation,adventitious bud induction,shoot elongation,regeneration of transgenic plantlets,plant habituation,obtainin... |
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| Keywords: | Soybean Genetic transformation Agrobacterium-mediated Cotyledonary node Technique flow |
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