| 利用根癌农杆菌介导转化大豆成熟种子胚尖获得转基因植株 |
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| 引用本文： | 刘海坤,卫志明.利用根癌农杆菌介导转化大豆成熟种子胚尖获得转基因植株[J].植物生理与分子生物学学报,2004,30(6):631-636. |
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| 作者姓名： | 刘海坤 卫志明 |
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| 作者单位： | 中国科学院上海生命科学研究院植物生理生态研究所,植物分子遗传国家重点实验室,中国科学院研究生院,上海,200032 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 利用根癌土壤农杆菌EHA105/pCAMBIA2301对来自大豆成熟种子的胚尖外植体进行遗传转化,并对农杆菌侵染时间长短以及乙酰丁香酮(AS)浓度等影响转化频率的条件进行了探讨.发现浸染时间以20 h为佳,乙酰丁香酮最佳浓度为200 umo1/L,并探讨了恢复培养的重要性.分别从3个大豆品种合丰35、合丰39、东农42得到了转基因植株,GUS染色及Southern杂交结果证明外源基因整合到大豆基因组中,获得转基因大豆的频率达6.4%～12.1%.
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| 关 键 词： | 大豆 根癌农杆菌 转化 胚尖 |
| 修稿时间： | 2004年5月28日 |
Transgenic Soybean Obtained with Agrobacterium tumefaciens-mediated Transformation of Embryonic Tip of Soybean Mature Seeds |
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| Hai-Kun Liu,Zhi-Ming Wei.Transgenic Soybean Obtained with Agrobacterium tumefaciens-mediated Transformation of Embryonic Tip of Soybean Mature Seeds[J].Journal Of Plant Physiology and Molecular Biology,2004,30(6):631-636. |
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| Authors: | Hai-Kun Liu Zhi-Ming Wei |
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| Institution: | National Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. |
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| Abstract: | Regenerated embryonic tips were inoculated with Agrobacterium tumefaciens strain EHA105, which contains binary vector pCAMBIA2301, and cultured for 20 h. Our results showed that the T-DNA transfer efficiency reached up to 63.3% (Table 1) and the transformation efficiency reached up to 6.4%-12.1% (Table 2). The effect of infection time on T-DNA delivery into soybean embryonic tips was determined (Table 1). We also discuss the effects of days of co-cultivation to transient expression and the effects of different AS concentrations to transient expression of gus gene (Figs. 1,2). These data indicate that the embryonic tip regeneration system can be used for efficient, effective Agrobacterium-mediated transformation. |
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| Keywords: | soybean Agrobacterium tumefaciens transformation embryonic tips |
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