| 蓝浆果农杆菌介导法基因转化条件研究 |
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| 引用本文： | 刘捷,杨丽娜,章镇,蔡斌华,陶建敏.蓝浆果农杆菌介导法基因转化条件研究[J].江苏农业学报,2008,24(5). |
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| 作者姓名： | 刘捷 杨丽娜 章镇 蔡斌华 陶建敏 |
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| 作者单位： | 南京农业大学园艺学院,江苏,南京,210095 |
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| 基金项目： | 国家农业综合开发科技示范项目，国家高技术研究发展计划(863计划) |
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| 摘 要： | 以蓝浆果品种蓝丰的叶片为转化受体,通过对农杆菌侵染时间、共培养时间、外植体暗培养时间、卡那霉素(Km)选择压以及头孢霉素(Cef)浓度等因素研究,确定转化的最佳试验参数.试验结果:农杆菌侵染5 min,共培养4 d,暗处理21 d时gus基因瞬时表达率最高;在筛选初期添加Km 20 mg/L、Cef 250 mg/L,既能得到不定芽,又达到筛选的目的.利用GUS组织化学染色、PCR扩增方法对蓝浆果的Km抗性植株进行检测,初步证实AtNHX1基因已经整合到2株蓝丰株系基因组中.
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| 关 键 词： | 蓝浆果 农杆菌介导 基因转化 gus基因 PCR检测 |
Agrobacterium-mediated Transformation Factors of Blueberry |
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| LIU Jie,YANG Li-na,ZHANG Zhen,CAI Bin-hua,TAO Jian-min.Agrobacterium-mediated Transformation Factors of Blueberry[J].Jiangsu Journal of Agricultural Sciences,2008,24(5). |
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| Authors: | LIU Jie YANG Li-na ZHANG Zhen CAI Bin-hua TAO Jian-min |
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| Institution: | LIU Jie,YANG Li-na,ZHANG Zhen,CAI Bin-hua,TAO Jian-min(College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China) |
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| Abstract: | Transgenic plants were generated from leaves of bluecrop blueberry by Agrobacterium-mediated transformation with AtNHX1 gene,and the following factors affecting transformation were investigated: dipping time,dark-culture time,co-culture time,concentration of kanamycin(Km),and concentration of cefotaxine(Cef).By analyzing the frequency of gus gene transient-expression and bud regeneration rate,the optimal transformation conditions were worked out: 5 min of leaf dipping,4 d of co-culture and 21 d of dark-cult... |
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| Keywords: | blueberry Agrobacterium mediation gene transformation gus gene PCR analysis |
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