新疆籽用西葫芦花叶病病原的鉴定  

Identification of the Pathogen Causing Mosaic Leaves in Seed Zucchini(Cucurbita pepo)in Xinjiang

作　　者：郝婷丽 陈大群 张燕 杜珍珠 吴勤 刘升学 HAO Ting-Li;CHEN Da-Qun;ZHANG Yan;DU Zhen-Zhu;WU Qin;LIU Sheng-Xue(Analysis and Testing Center,Shihezi University,Shihezi 832003,China)

机构地区：[1]石河子大学分析测试中心,石河子832003

出　　处：《农业生物技术学报》2024年第12期2882-2890,共9页Journal of Agricultural Biotechnology

基　　金：石河子大学分析测试中心大型科研仪器共享平台开放课题(ZZZC2021116)。

摘　　要：籽用西葫芦(Cucurbita pepo)是新疆主要的经济作物之一,病毒危害可导致其产量和品质严重下降,明确病原是进一步防治的基础。本研究从新疆石河子市采集表现花叶症状的籽用西葫芦叶片样品,在电子显微镜下观察到750×11 nm呈弯曲线状的马铃薯Y病毒属(Potyvirus)病毒粒子。提取病毒的双链RNA(double-stranded RNA,dsRNA),采用随机引物对病毒进行RT-PCR,得到弥散条带XHL2-1、XHL2-2和XHL56,在NCBI数据库中Blast搜索比对发现,XHL2-1和XHL56与小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)对应区段的序列相似性分别为92.22%~98.69%和96.21%~99.64%;XHL2-2与西瓜花叶病毒(Water melon mosaic virus,WMV)对应区段的序列相似性在89.38%~98.45%;表明籽用西葫芦花叶病可能是由ZYMV和WMV侵染所致。进一步设计合成ZYMV和WMV的特异性引物,对15个供试样品进行RT-PCR,回收编号为SH2、SH12和SH13的ZYMV特异性扩增片段进行测序分析,结果表明,扩增片段之间的序列相似性超过99.29%,与GenBank中收录的ZYMV的序列相似性为96.09%~99.56%;编号为SH1、SH9和SH11的WMV特异性扩增片段之间的序列相似性超过94.30%,与GenBank中收录的WMV的序列相似性为94.74%~100%;进一步证明新疆籽用西葫芦花叶病是由ZYMV和WMV的复合侵染所致。系统进化树分析表明,侵染新疆籽用西葫芦的ZYMV与来源于东欧、印度等区域的分离物处于同一分支,WMV与来源于中国、法国等区域的WMV分离物亲缘关系较近。本研究明确了ZYMV和WMV的复合侵染是导致新疆籽用西葫芦花叶病害的主要病原,为进一步防治该病害提供理论依据。Seed Zucchini(Cucurbita pepo),a major economic crop in Xinjiang,suffers severe yield and quality losses due to viral infections,and identifying its pathogens is essential for further control measures.In this study,leaf samples displaying mosaic symptoms were collected from Seed Zucchini in Shihezi,Xinjiang.Under an electron microscope,flexuous Potyvirus particles measuring approximately 750×11 nm were observed.The double-stranded RNA(dsRNA)of the virus was extracted,RT-PCR was performed on the virus with random primers and diffuse bands XHL2-1,XHL2-2,and XHL56 were obtained.BLAST searches in the NCBI database revealed that XHL2-1 and XHL56 had sequence similarities of 92.22%~98.69%and 96.21%~99.64%,respectively,with the corresponding regions of Zucchini yellow mosaic virus(ZYMV);XHL2-2 showed a sequence similarity of 89.38%~98.45%with the corresponding region of Watermelon mosaic virus(WMV).These findings indicated that the mosaic disease in Seed Zucchini might be caused by infections of both ZYMV and WMV.To further investigate,specific primers for ZYMV and WMV were designed and synthesized.RT-PCR was conducted on 15 samples,and ZYMV-specific amplification fragments from samples SH2,SH12,and SH13 were sequenced.The sequence similarity among these fragments was over 99.29%,with a similarity of 96.09%~99.56%to ZYMV sequences recorded in GenBank.The WMV-specific amplification fragments from samples SH1,SH9,and SH11 had sequence similarities exceeding 94.30%among themselves,and 94.74%~100%similarity with WMV sequences in GenBank.These results further confirmed that the mosaic disease in Seed Zucchini in Xinjiang was caused by co-infection of ZYMV and WMV.Phylogenetic analysis revealed that the ZYMV infecting Seed Zucchini in Xinjiang clusters with isolates from Eastern Europe and India,while the WMV was closely related to isolates from China,France,and other regions.This study identifies the primary pathogens of mosaic virus disease in Xinjiang's Seed Zucchini as a co-infection by ZYMV and WMV,providing a theoretica

关 键 词：籽用西葫芦(Cucurbita pepo) 花叶病 dsRNA(double-stranded RNA) 鉴定 

分 类 号：S642[农业科学—蔬菜学]