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利用斑点免疫杂交法(Dot-ELISA)和RT-PCR法对重庆地区发生的辣椒病毒病的病原进行了鉴定和优势病毒分析。利用5种常见蔬菜病毒的血清对采自重庆8个区县的152份辣椒病毒病样品进行Dot-ELISA检测,结果表明,共有118份辣椒样品检测呈阳性,其中,黄瓜花叶病毒(Cucumber mosaic virus,CMV)的侵染最普遍,总检出率高达57.89%;番茄花叶病毒(Tomato mosaic virus,To MV)的总检出率最低,为22.52%;烟草花叶病毒(Tobacco mosaic virus,TMV)、芜菁花叶病毒(Turnip mosaic virus,Tu MV)和蚕豆萎蔫病毒2号(Broad bean wilt virus 2,BBWV-2)的总检出率分别为30.26%、36.18%和24.34%。在118份阳性样品中,有66.10%的样品受两种及以上病毒复合侵染,其中33.33%的样品受到CMV和Tu MV复合侵染,有26.27%的样品受3种病毒复合侵染。设计5种病毒的特异性引物,随机选取16份阳性样品进行RT-PCR扩增,克隆测序结果表明扩增片段确实是5种病毒的相应序列,且RT-PCR与ELISA检测结果基本吻合。检测结果表明CMV是重庆地区辣椒上的优势病毒种类,且该地区辣椒上病毒复合侵染现象普遍。本文首次报道了To MV和Tu MV侵染辣椒。
Dot immunosorbent assay (Dot-ELISA) and RT-PCR were used to identify the pathogen of pepper virus disease in Chongqing and to analyze the dominant virus. Dot-ELISA was used to detect 152 pepper virus samples collected from 8 districts and counties in Chongqing. The results showed that a total of 118 samples of pepper samples were positive. Cucumber mosaic virus (Cucumber mosaic virus) (CMV) was the most common, with a total detection rate of 57.89%. Tomato mosaic virus (To MV) had the lowest detection rate of 22.52%. Tobacco mosaic virus TMV, Tu MV and BBWV-2 were 30.26%, 36.18% and 24.34%, respectively. Among 118 positive samples, 66.10% of the samples were infected by two or more viruses, of which 33.33% were infected by CMV and Tu MV and 26.27% were infected by three viruses. Five specific primers were designed and 16 positive samples were randomly selected for RT-PCR amplification. Cloning and sequencing results showed that the amplified fragment was indeed the corresponding sequence of five viruses, and the results of RT-PCR and ELISA were basically consistent. The results showed that CMV was the predominant virus species in pepper in Chongqing area, and the virus complex infection in pepper in the area was common. This paper reports for the first time that To MV and Tu MV infest peppers.