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1h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313,326,392bp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。
关键词:
烟草花叶病毒,
黄瓜花叶病毒,
马铃薯X病毒,
RT-PCR
Abstract: In order to extract the total RNA from the plant leaf and to detect the viruses of tabacoo rapidly, a extractionmethod and detection method were studied.The TRIzol Reagent box was used to extract the total RNA from the leavescucumber and N . tabacum cv. White burly .The total RNA with the high purity and integrality was obtained in 1 h. Based onthe genome conservative sequence of tabacoo mosaic virus, cucumber mosaic virus and potato X virus, special primers weredesigned respect ively.With the RT-PCR, the target fragments about 313, 326 and 392 bp were ampilfied separately from theinfected sample, but not health sample.There recombinant plasmids were constructed and their sequences were comparedseverally 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 scient ific base for the detection to viruses of tabacoo.
Key words:
Tabacoo mosaic virus,
Cucumber mosaic virus,
Potato X virus,
RT-PCR
中图分类号:
GURui-hua, WANGJin-zhong, WENSi-yuan, WANGSheng-qi, ZHANGAi-huan, WEIYan-min. A Rapid RT-PCR Detection for the Three Kinds of Viruses in Tobacco[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.7668/hbnxb.2007.S1.006.