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对一株进境隔离种植中疑似病毒症状的荷兰百合进行检测、鉴定。采用RT-PCR方法对疑似样品的部分基因序列进行扩增、测序,比对后,建立系统发育树进行亲缘关系分析。结果表明,采用香石竹潜隐病毒属和马铃薯X病毒属(CarlapCar1I/M4T和Potex 5/Potex 1RC)简并引物均可扩增到预期大小的片段(1 257 nts和737 nts)。序列分析显示,扩增片段与车前草花叶病毒(Plantago asiatica mosaic virus,Pl AMV)3’端序列(包含TGBp2、TGBp1、CP,3’-NTS)和部分依赖于RNA的RNA聚合酶基因序列一致性可达79.9%~99.5%和80.5%~99.3%。利用PlAMV的特异性引物(PlAMV-cpup/PlAMV-cpdw)进行检测,结果进一步证实该百合样品含有PlAMV。通过建立系统发育树得出,该病毒分离物与来自荷兰的PlAMV百合分离物优先聚为一簇,表明二者的亲缘关系最近。这是我国口岸首次截获车前草花叶病毒的报道。
To a quarantine plant in the isolation of suspected symptoms of the Netherlands lily for testing, identification. The partial gene sequences of the suspected samples were amplified, sequenced and compared by RT-PCR, and the phylogenetic tree was established for phylogenetic analysis. The results showed that the expected size fragments (1 257 nts and 737 nts) could be amplified using both Degenera carnavirus and degenerate primers of Carvaccine XIV (CarlapCar1I / M4T and Potex 5 / Potex 1RC). Sequence analysis showed that the sequence of the 3 ’end of the amplified fragment and Plantago asiatica mosaic virus (Pl AMV) (including TGBp2, TGBp1, CP, 3’-NTS) and partial RNA- Consistency up to 79.9% ~ 99.5% and 80.5% ~ 99.3%. The detection of PIAMV-specific primers (PlAMV-cpup / PlAMV-cpdw) further confirmed that the lily sample contains PlAMV. By establishing a phylogenetic tree, the isolate of this virus preferentially clustered with the PlAMV lily isolate from the Netherlands, indicating that the two are most closely related. This is the first time in China’s ports intercepted grasshopper mosaic virus reported.