山东葫芦科蔬菜上病毒病种类检测及黄瓜花叶病毒分离物的亚组鉴定

摘要

In order to detect and identify the viral causing agent that infecting cucurbit vegetable plants in Shandong,867 cucurbit plant samples of suspected viral diseases from 1 1 vegetable planting areas in Shandong were collected during July to August,2014.9 primer pairs specific to Cucumber mosaic virus(CMV),Cucumber green mottle mosaic virus(CGMMV),Watermelon mosaic virus(WMV),Tobacco mosaic virus(TMV),Zucchini yellow mo-saic virus(ZYMV),Squash mosaic virus(SqMV),Melon yellow spot virus(MYSV),Cucurbit chlorotic yellows virus (CCYV),Prunus necrotic ringspot ilarvirus(PNRSV)and Melon necrotic spot virus(MNSV)were used respectively to perform PCR detection.The results showed that viruses carried rate was 34.8%,10.4%,20.0%,41 .7%,27.0%, 7.8%,2.6%,1 .7%,0 and 0.9% respectively.The natural infection of these viral diseases except PNRSV in Shandong was confirmed,and the incidence of CMV and TMV were higher than others.Two or more than two viruses mixed infections were also very common in field.At the same time,those CMV positive samples from 1 1 regions were selected for subgroup classification.Coat protein (CP)gene and the whole RNA3 nucleotide sequences were determined.Sequence alignment and phylogenetic analyses were performed.The results showed that the CMV iso-lates of cucurbit vegetables in Shandong area were all members of CMV subgroup IB with the highest similarity to a South Korea isolate As(AF013291 ),no other strains were found.%为了检测和鉴定山东地区葫芦科蔬菜上病毒病种类，从山东诸地11个蔬菜种植区采集了867个葫芦科蔬菜疑似感病样品。利用黄瓜花叶病毒、黄瓜绿斑驳花叶病毒、西瓜花叶病毒、烟草花叶病毒、小西葫芦黄花叶病毒、南瓜花叶病毒、甜瓜黄斑病毒、瓜类褪绿黄化病毒、李属坏死环斑病毒及甜瓜坏死斑点病毒等特异性引物对疑似感病样品分别进行 RT-PCR 检测，各病毒检出率分别为34．8％，10．4％，20．0％，41．7％，27．0％，7．8％，2．6％，1．7％，0，0．9％。表明除 PNRSV 外，其他9种病毒在山东各地均有发生，且 CMV 和 TMV 发病率较高，2种或2种以上病毒复合侵染也很普遍。同时为明确山东地区 CMV 分离物的株系分化状况，选取11个地区有代表性的 CMV 阳性样品，测定其外壳蛋白（Coat protein，CP）和 RNA3核苷酸序列并进行序列比对和进化分析，结果显示侵染山东地区葫芦科蔬菜的 CMV 分离物均属于 IB 亚组，与韩国分离物 As（AF013291）相似性最高，未发现其他株系。