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内蒙古籽用西葫芦病毒病病原鉴定
岳建英1, 韦学锋1, 郑红丽,等1
内蒙古农业大学 园艺与植物保护学院
摘要:
【目的】对内蒙古自治区籽用西葫芦病毒病进行病原鉴定,并确定其分类地位,为该地区籽用西葫芦病毒病的防治提供理论依据。【方法】以采自内蒙古土默特左旗具有典型花叶症状的籽用西葫芦叶片为材料,对其进行sRNA深度测序和RT-PCR检测,并对所得序列进行一致性分析和构建系统发育进化树。【结果】对具有典型花叶症状的籽用西葫芦叶片的sRNA测序结果进行拼接,在数据库中比对共注释到14种病毒,其中西瓜花叶病毒(Watermelon mosaic virus,WMV)、小西葫芦黄化花叶病毒(Zuchhini yellow mosaic virus,ZYMV)和黄瓜花叶病毒(Cucumber mosaic virus,CMV) 所占比例较高,分别为59.48%,20.70%和13.70%,这说明内蒙古自治区籽用西葫芦病毒病为多种病毒混合侵染引起。RT-PCR扩增除了检测到WMV、ZYMV和CMV外,还检测到马蹄莲潜隐病毒(Calla lily latent virus,CLLV)的外壳蛋白特异性条带。系统进化树分析显示,2个籽用西葫芦ZYMV分离物(MK002236和 MK002237)与伊朗蚜虫ZYMV分离物(KU528623)聚合在同一支;籽用西葫芦Z1 样品WMV分离物(MK015646)与意大利柑橘WMV分离物(FJ823122)亲缘关系最近,而Z2样品WMV分离物(MK015645)则与法国甜瓜WMV分离物、法国未知作物WMV分离物、法国西葫芦WMV分离物以及Z1 样品WMV分离物(MK015646)和意大利柑橘WMV分离物形成一个大分支。【结论】内蒙古籽用西葫芦病毒病主要病原为WMV、ZYMV和CMV,自然条件下常发生复合侵染。籽用西葫芦上最初的WMV和ZYMV可能由媒介昆虫或人从其他作物上传播而来,也可能是通过籽用西葫芦种子进口、国内调运而传播到本地的。
关键词:  籽用西葫芦  西葫芦病毒病  病原鉴定  sRNA深度测序  内蒙古
DOI:
分类号:
基金项目:内蒙古农业大学高层次人才引进科研启动项目(NDGCC2016-23)
Identification of viral pathogen of zucchini in Inner Mongolia
YUE Jianying,WEI Xuefeng,ZHENG Hongli,et al
Abstract:
【Objective】This study identified the pathogen of zucchini virus disease in Inner Mongolia and determined its taxonomic status to provide basis for the prevention of zucchini virus disease.【Method】Samples with typical symptoms collected from Tumote Zuoqi,Inner Mongolia were subjected to deep sRNA sequencing and RT PCR detection.The obtained sequences were analyzed for consistency and a phylogenetic tree was constructed.【Result】After splicing the sRNA sequencing results of samples with typical symptoms,a total of 14 viruses were annotated in the database,including Watermelon mosaic virus (WMV),Zucchini yellow mosaic virus (ZYMV) and Cucumber mosaic virus (CMV) with high proportions of 59.48%,20.70% and 13.70%,respectively.In addition,the coat protein specific band of Calla lily latent virus (CLLV) was detected.Phylogenetic tree analysis showed that two isolates of zucchini ZYMV (MK002236 and MK002237) were clustered with the Iranian ZYMV isolate (KU528623).The sample Z1 of the zucchini WMV isolates (MK015646) was closest to the Italian citrus WMV isolates (FJ823122).The sample Z2 of zucchini WMV isolates (MK015645) formed a large branch with the French WMV melon isolates,the French unknown crop WMV isolates,the French zucchini WMV isolates,sample Z1 WMV isolates (MK015646) and Italian citrus WMV isolates.【Conclusion】The main pathogens of zucchini virus disease in Inner Mongolia were WMV,ZYMV and CMV,and mixed infections often occurred under natural conditions.WMV and ZYMV on zucchini may be initially transmitted by insect vector or man made from other crops or due to zucchini seeds imports and domestic transport.
Key words:  Cucurbita pepo  zucchini virus disease  identification of pathogen  sRNA deep sequencing  Inner Mongolia