| 简单高效的根癌农杆菌介导的水稻基因转化方法 |
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| 引用本文： | 李美茹,李洪清.简单高效的根癌农杆菌介导的水稻基因转化方法[J].实验生物学报,2003,36(4):289-294. |
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| 作者姓名： | 李美茹 李洪清 |
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| 作者单位： | [1]中国科学院华南植物研究所，广州510650 [2]华南师范大学生命科学院，广州510631 |
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| 摘 要： | 本文在研究影响农杆菌介导的水稻转化的主要因素基础上，建立了一套简单、高效的水稻转基因系统。将水稻成熟胚来源的愈伤组织用农杆菌EHA101／pHQ9，EHA101／pHQ10，EHA101／pHQT3感染后，筛选抗性愈伤，经分化获得转化株。抗性愈伤的平均得率为约100个愈伤／g愈伤外植体，抗性愈伤的分化频率平均高达85％。转基因植株的GUS染色、Southern杂交结果表明，T-DNA上的外源基因已整合进转基因植物的基因组中。转基因植株T1代对潮霉素的抗性表明，多数转基因株系符合孟德尔分离比3：1。该系统的建立将有助于应用T—DNA标签法和基因打靶法进行水稻功能基因组的研究。
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| 关 键 词： | 根癌农杆菌 水稻 遗传转化 转基因植株 功能基因组 |
A simple and highly efficient Agrobacterium-mediated rice transformation system] |
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| Mei Ru Li,Hong Qing Li.A simple and highly efficient Agrobacterium-mediated rice transformation system][J].Acta Biologiae Experimentalis Sinica,2003,36(4):289-294. |
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| Authors: | Mei Ru Li Hong Qing Li |
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| Institution: | South China Institute of Botany, Chinese Academy of Sciences, Guangzhou 510650. |
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| Abstract: | A simple and highly efficient rice transformation system was established based on the studying of factors influencing the Agrobacterium-mediated rice transformation. Embryogenic calli derived from mature embryos were infected and cocultivated with A. tumefaciens EHA101 harboring binary vectors: pHQ9, pHQ10, pHQT3. The highest transformation frequency was about 100 hygromycin resistance calli per gram of calli explants, and above 85% of these calli could be regenerated into plants. The putative transgenic plants were confirmed by GUS assay and southern-blot analysis. Hygromycin resistance tests indicated that the segregation of transgene in T1 progeny corresponded to the Mendelian ratio, 3:1. This system will benefit the functional genomic study of rice by using T-DNA insertion mutagenesis and gene targeting. |
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