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利用7种植物病毒的抗体,对采自重庆7个区县的67份南瓜病毒病样品进行了抗原直接包被酶联免疫吸附分析(ACP-ELISA)检测。检测结果表明,在这67份样品中,至少59份感染了所要检测病毒,其中,感染黄瓜花叶病毒(Cucumber mosaic virus,CMV)、烟草花叶病毒(Tobacco mosaic virus,TMV)、番茄花叶病毒(Tomato mosaic virus,ToMV)、芜菁花叶病毒(Turnip mosaic virus,TuMV)、蚕豆萎蔫病毒2号(Broad bean wilt virus 2,BBWV-2)、马铃薯Y病毒(Potato virus Y,PVY)和马铃薯X病毒(Potato virus X,PVX)的样品分别为80.60%、77.61%、74.63%、82.09%、76.12%、83.58%和55.22%。在检测呈阳性的59份样品中有93.22%的样品受到2种以上病毒的复合侵染,而受7种病毒复合侵染的样品高达60.00%,表明在南瓜上病毒复合侵染现象相当严重。进一步对54份受CMV侵染的南瓜样品检测发现,CMV亚组Ⅰ(CMVⅠ)病毒侵染的样品有49份,占90.74%,CMV亚组Ⅱ(CMVⅡ)病毒侵染的样品为11份,占20.37%;其中6份(11.11%)样品检测到CMVⅠ和CMVⅡ的复合侵染。以免疫捕获反转录PCR(IC-RT-PCR)扩增了一个CMVⅠ分离物(CMV-CQ)近全长CP基因并进行序列分析,结果证实其属于CMV亚组Ⅰ型,其核苷酸序列与国内CMVⅠ型赤豆分离物(CMV-RB)的同源性最高,为98.1%。
67 samples of pumpkin virus disease collected from 7 districts and counties in Chongqing were detected by direct enzyme-linked immunosorbent assay (ACP-ELISA) using seven kinds of plant virus antibodies. The results showed that at least 59 of the 67 samples were infected with the virus to be detected, among which Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), tomato mosaic Tomato mosaic virus (ToMV), Turnip mosaic virus (TuMV), Broad bean wilt virus 2 (BBWV-2), Potato virus Y (PVY) and The samples of Potato virus X (PVX) were 80.60%, 77.61%, 74.63%, 82.09%, 76.12%, 83.58% and 55.22%, respectively. Among the 59 positive samples, 93.22% of the samples were infected with more than two viruses and 60.00% of the samples were infected with the seven viruses, indicating that the virus complex infection in pumpkin was quite serious. Further tests on 54 CMV infected pumpkin samples revealed that CMV subunit Ⅰ (CMVⅠ) virus infected 49 samples (90.74%), CMV Ⅱ (CMV Ⅱ) infected 11 samples, Accounting for 20.37%. Among 6 samples (11.11%), CMVⅠand CMVⅡinfection were detected. The full-length CP gene of a CMVⅠ isolate (CMV-CQ) was amplified by immunocapture reverse transcription PCR (IC-RT-PCR) and sequenced. The results showed that it belonged to type Ⅰ of CMV subunit. The nucleotide The sequence has the highest homology with CMV type Ⅰ red bean isolate (CMV-RB), which is 98.1%.