| 橡胶树体胚遗传转化中早代转化胚状体的PCR鉴定 |
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| 引用本文： | 李季,黄天带,蔡海滨,华玉伟,黄华孙.橡胶树体胚遗传转化中早代转化胚状体的PCR鉴定[J].热带作物学报,2013,34(5):866-869. |
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| 作者姓名： | 李季 黄天带 蔡海滨 华玉伟 黄华孙 |
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| 作者单位： | 中国热带农业科学院橡胶研究所 农业部橡胶树生物学与遗传资源利用重点实验室 海南儋州571737 |
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| 基金项目： | 国家自然科学基金（No. 30960322）；国家自然科学基金（No. 31200503）；现代农业产业技术体系建设专项（No. NYCYTX-34）。 |
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| 摘 要： | 橡胶树体胚遗传转化体系已成功建立,但抗性胚状体的循环增殖可获得大量抗性胚状体的同时,其后期的继代工作繁重.本文对橡胶树基因组DNA的CTAB提取方法进行了优化,同时筛选出了适合PCR检测的胚块重量.文中建立的胚块DNA的CTAB提取方法及筛选出的1～3mg重的胚块提取的DNA量均可满足早代胚状体PCR分析鉴定,且PCR鉴定结果与胚块的GUS染色结果吻合.该方法可用于橡胶树体胚遗传转化抗性胚状体早代筛选.
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| 关 键 词： | 橡胶树 遗传转化 次生体胚 PCR GUS |
The PCR Analysis of Early Embryos in Hevea Somatic Embryogenesis Transformation |
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| LI Ji,HUANG Tiandai,CAI Haibin,HUA Yuwei and HUANG Huasun.The PCR Analysis of Early Embryos in Hevea Somatic Embryogenesis Transformation[J].Chinese Journal of Tropical Crops,2013,34(5):866-869. |
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| Authors: | LI Ji HUANG Tiandai CAI Haibin HUA Yuwei and HUANG Huasun |
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| Abstract: | Agrobacterium tumefaciens-mediated Hevea somatic embryogenesis transformation system was successfully established. But a large propagation efficiently propagated from the secondary embryogenesis would lead to heavy burden in the following subculture. Thus the analysis of early positive embryo clones is very important. In this paper, the method of DNA extraction was optimized according to the conventional method of CTAB, meanwhile, the suitable weight of embryo used for PCR analysis was chosen. Experiments indicated that the optimized DNA extraction method and the scale of 1~3 mg all could accord with the PCR analysis of early embryos. Moreover, the results of PCR analysis were coincided with GUS histochemical staining. Hence, the method in this paper is available to early embryos screen in somatic embryogenesis transformation. |
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| Keywords: | Rubber tree Genetic transformation Secondary embryo PCR GUS |
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