| 利用农杆菌浸种法建立白三叶草遗传转化体系的研究 |
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| 引用本文： | 王丹,吴燕民,刘水,吕慧,雷江丽. 利用农杆菌浸种法建立白三叶草遗传转化体系的研究[J]. 中国农业科技导报, 2009, 11(1): 96-101 |
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| 作者姓名： | 王丹 吴燕民 刘水 吕慧 雷江丽 |
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| 作者单位： | （1．深圳市园林科学研究所, 广东 深圳 518003； ,2.中国农业科学院生物技术研究所, 北京 100081； 3.深圳市铁汉园林绿化有限公司, 广东 深圳 518034） |
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| 基金项目： | 深圳市科技与信息局项目 |
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| 摘 要： | 以白三叶萌动种子作为转化受体,利用npt-Ⅱ筛选基因对影响农杆菌介导的遗传转化外部因素进行深入
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| 关 键 词： | 白三叶；农杆菌浸种法；遗传转化 |
| 收稿时间： | 2008-10-06 |
| 修稿时间： | 2008-11-28 |
Studies on the Establishment of Genetic Transformation System of White Clover(Trifolium repens L.)by Soaking Seeds with Agrobacterium tumefaciens |
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| WANG Dan,WU Yan-min,LIU Shui,LV Hui,LEI Jiang-li. Studies on the Establishment of Genetic Transformation System of White Clover(Trifolium repens L.)by Soaking Seeds with Agrobacterium tumefaciens[J]. Journal of Agricultural Science and Technology, 2009, 11(1): 96-101 |
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| Authors: | WANG Dan WU Yan-min LIU Shui LV Hui LEI Jiang-li |
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| Affiliation: | （1.Shenzhen Institute of Landscape Science, Guangdong Shenzhen 518003|2.Biotechnology Research Institute, ;Chinese Academy of Agricultural Sciences, Beijing 100081|3. Shenzhen Techand Landscape Co., Ltd., ;Guangdong Shenzhen 518034, China） |
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| Abstract: | The germinating seeds of white clover were used as transformation receptor, and npt-Ⅱgene was chosen as a selection gene. External factors that affect the efficiency ofgenetic transformation were studied and optimized. A high-frequency transformation systemof white clover had been established by adopting Agrobacterium tumefaciens soaked seedstransformation technology. The seeds of white clover germinated for 1 hour, then addingAgrobacterium with concentration of 0.9(OD600) under 5 min+30 s vacuum treatment. After24 hours infection in oscillating culture and 1 day co-cultivation, the transferredplants were screened by 50 mg/L kanamycin. This transformation system was so effective,that positive transformation rate reached 45.5%. The transgenic plants were also examinedby PCR analysis and GUS staining. It was proved that npt-Ⅱ and GUS gene had beenintegrated into the genome. |
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| Keywords: | white clover soaking seeds with Agrobacterium tumefaciens genetic transformation |
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