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齿兰环斑病毒(odontoglossum ringspot virus,ORSV)是严重危害兰科植物的主要病毒之一。本研究根据NCBI基因库中ORSV外壳蛋白基因设计特异性引物,应用RT-PCR方法对ORSV进行检测研究。本试验所使用的植物总RNA提取方法,可以提取出完整且纯度高的总RNA,能够满足RT-PCR的要求;本研究所建立起的ORSV RT-PCR检测体系,可以从感病的指示植物及染病蝴蝶兰中扩增出与预期的332 bp大小完全一致的扩增产物,而健康的指示植物无此扩增产物。对此片段的测序研究也证明了RT-PCR的检测。利用BLAST及DNAMAN 6.0.3.99软件将我们检测的蝴蝶兰齿兰环斑病毒与来自中国海南、台湾以及新加坡的病毒分离物进行同源性分析,其同源性均达98%以上。
Odontoglossum ringspot virus (ORSV) is one of the major viruses that seriously damage orchid plants. In this study, specific primers were designed according to ORSV coat protein gene in NCBI gene bank, and RT-PCR method was used to detect ORSV. The total RNA extracted by this experiment can extract complete and high purity total RNA, which can meet the requirement of RT-PCR. The ORSV RT-PCR detection system established in this study can be used to detect the susceptible plant And infected phalaenopsis amplified exactly the same size as the expected 332 bp amplification products, while healthy indicator plants did not have this amplification product. Sequencing studies of this fragment also demonstrated the detection of RT-PCR. Homology analysis of phalaenopsis ornithine spot blot detected by BLAST and DNAMAN 6.0.3.99 software with the virus isolates from Hainan, Taiwan and Singapore showed that the homology of the tested virus was over 98%.