| RT-PCR快速检测3种烟草病毒技术 |
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| 引用本文： | 谷瑞华,王进忠,文思远,王升启,张爱环,魏艳敏.RT-PCR快速检测3种烟草病毒技术[J].华北农学报,2007,22(B08):21-23. |
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| 作者姓名： | 谷瑞华 王进忠 文思远 王升启 张爱环 魏艳敏 |
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| 作者单位： | [1]北京农学院植物科学技术系,北京102206 [2]军事医学科学院放射医学研究所,北京100850 |
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| 基金项目： | 北京市教育委员会项目（KM200610020007）;北京市自然基金（5043026） |
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| 摘 要： | 为了快速提取植物叶片的总RNA,并对烟草病毒进行检测,对提取及检测方法进行了研究。结果表明：以黄瓜叶片、白肋烟叶片为试材,用TRIzol试剂盒在1 h内提取到纯度高、完整性好的总RNA。设计烟草花叶病毒、黄瓜花叶病毒和马铃薯X病毒的特异性引物,RT-PCR检测,从感病叶片中分别扩增出313,326,392 bp的目的片断,而健康叶片中无此扩增带。构建这3种病毒重组质粒,对核苷酸序列与GeneBank中报道的进行比较,同源性分别为99.36%,97.55%,98.21%。用这种方法快速、可靠,为烟草病毒的检测提供科学依据。
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| 关 键 词： | 烟草花叶病毒 黄瓜花叶病毒 马铃薯X病毒 RT-PCR |
| 文章编号： | 1000-7091（2007）增刊-0021-03 |
| 修稿时间： | 2007-04-05 |
A Rapid RT-PCR Detection for the Three Kinds of Viruses in Tobacco |
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| GU Rui-hua, WANG Jin-zhong, WEN Si-yuan, WANG Sheng-qi, ZHANG Ai-huan, WEI Yan-min.A Rapid RT-PCR Detection for the Three Kinds of Viruses in Tobacco[J].Acta Agriculturae Boreali-Sinica,2007,22(B08):21-23. |
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| Authors: | GU Rui-hua WANG Jin-zhong WEN Si-yuan WANG Sheng-qi ZHANG Ai-huan WEI Yan-min |
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| Institution: | 1. Department of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, China; 2. Beijing Institute of Radiation Medicine, Academy of Military Medical Science, Beijing 100850, China |
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| Abstract: | In order to extract the total RNA from the plant leaf and to detect the viruses of tabacoo rapidly, a extraction method and detection method were studied. The TRIzol Reagent box was used to extract the total RNA from the leaves cucumber and N. tabacum cv. White burly. The total RNA with the high purity and integrality was obtained in 1 h. Based on the genome conservative sequence of tabacoo mosaic virus, cucumber mosaic virus and potato X virus, special primers were designed respectively. With the RT-PCR, the target fragments about 313,326 and 392 bp were ampilfied separately from the infected sample, but not health sample. There recombinant plasmids were constructed and their sequences were compared severally with these three kinds of virus nucleotide sequences in GeneBank. The homology achieve separately 99.36%, 97.55 %, 98.21%. These methods were rapid and reliable. It supplied scientific base for the detection to viruses of tabacoo. |
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| Keywords: | Tabacoo mosaic virus Cucumber mosaic virus Potato X virus RT-PCR |
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