首页|利用小RNA深度测序技术鉴定西瓜病毒病病原

利用小RNA深度测序技术鉴定西瓜病毒病病原

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西瓜是一种重要的园艺植物,近年来饱受病毒病危害.为明确西瓜病毒病害病原,对采自山西太谷区表现花叶、皱缩等症状的西瓜叶片进行小RNA深度测序,同时利用RT-PCR的方法并结合生物信息学的手段,综合分析造成西瓜花叶、皱缩症状的病毒病原.结果表明,表现皱缩、花叶症状的西瓜样品被瓜类蚜传黄化病毒(cucurbit aphid-borne yellows virus,CABYV)、甜瓜潜隐病毒(cucumis melo cryptic virus,CmCV)、西瓜病毒 A(watermelon virus A,WVA)、西瓜皱叶病毒 2 号(watermelon crinkle leaf-associated virus 2,WCLaV2)和黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)5种病毒复合侵染.利用RT-PCR的方法分别扩增获得5种病毒的外壳蛋白(coat protein,CP)基因序列,进一步通过序列一致率分析和系统进化分析发现,本研究获得的西瓜CABYV分离物CABYV-SXJZ(GenBank登录号:OP957280)核苷酸序列与中国南瓜CABYV分离物Inner Mongolia(GenBank登录号:EU262627)的核苷酸序列一致率最高,达到100％;西瓜WVA分离物WVA-SXJZ(GenBank登录号:OP957281)核苷酸序列与同样来自中国瓠瓜WVA分离物WVA-Huizhou(Gen-Bank 登录号:MK292710)和西瓜WVA分离物WVA-KF15(GenBank登录号:KY363796)核苷酸序列一致率最高,分别达到93.6％％和99.9％;WCLaV2分离物WCLaV2-SXJZ(GenBank登录号:OP957282)核苷酸序列与WCLaV2巴西西瓜分离物Ju-01(GenBank登录号:LC636075)核苷酸序列一致率最高,达到99.6％;本研究首次从西瓜上获得的CmCV分离物Cm-CV-SXJZ(GenBank登录号:OP957283)核苷酸序列与中国甜瓜分离物CmCV-HLJ(GenBank登录号:MH479773)的核苷酸一致率高达99.9％;本研究获得的CGMMV分离物CGMMV-SXJZ(GenBank登录号:OP957284)核苷酸序列与CGMMV分离物 GDLZ(GenBank 登录号:MK933286)、CG038(GenBank 登录号:MH271443)、CGMMV-pXTl(GenBank 登录号:KY753929)、eWT(GenBank 登录号:KY753928)、C284R(GenBank 登录号:KY753927)、CGMMV-XG(GenBank 登录号:KP868654)、JD2(GenBank登录号:KM873785)和Anhui(GenBank登录号:KT236095)核苷酸序列一致率最高,均达到99.8％.
Identification of the Citrullus lanatus infecting viruses by small RNA deep sequen-cing
Citrullus lanatus is an important horticultural plant.Viral diseases on C.lanatus have become more and more serious in recent years.In order to identify the viruses that infect watermelons,the small RNA deep sequencing was used to analyze samples with mosaic and shrinking symptoms from Taigu District,Shanxi Province.RT-PCR and bioinformatics methods were used to analyze the pathogens.The results showed that the watermelon samples exhibiting mosaic and shrinking symptoms were infected by five viruses including cucurbit aphid-borne yellows virus(CABYV),cucurbit melo cryptic virus(CmCV),watermelon virus A(WVA),watermelon crinkle leaf-associated virus 2(WCLaV2)and cucumber green mottle mosaic virus(CGMMV).The coat protein(CP)sequences of five viruses were amplified by RT-PCR,and further analyzed by sequence identity analysis and phylogenetic analysis.It was found that the nucleotide sequence of CABYV-SXJZ(Gen-Bank No:OP957280)obtained in this study had the highest identity with that of CABYV-Inner Mongolia(GenBank No:EU262627),reaching 100% .The nucleotide sequence of watermelon WVA isolate WVA-SXJZ(GenBank No:OP957281)has the highest identity with that of watermelon WVA isolate WVA-Huizhou(GenBank No:MK292710)and watermelon WVA isolate WVA-KF15(GenBank No:KY363796),which are also from China,reaching 93.6% and 99.9% ,respectively.The nucleotide sequence of WCLaV2 isolate WCLaV2-SXJZ(GenBank No:OP957282)has the highest identity with that of WCLaV2 Brazilian watermelon isolate Ju-01(GenBank No:LC636075),reaching 99.6% .The nucleotide sequence of CmCV isolate CmCV-SXJZ(GenBank No:OP957283)obtained from watermelon for the first time in this study has 99.9% identity to that of Chinese melon isolate CmCV-HLJ(GenBank No:MH479773).The nu-cleotide sequence of CGMMV isolate CGMMV-SXJZ(GenBank No:OP957284)obtained in this study has the highest identity with CGMMV isolate GDLZ(GenBank No:MK933286),CG038(GenBank No:MH271443),CGMMV-pXT1(GenBank No:KY753929),eWT(GenBank No:KY753928),C284R(GenBank No:KY753927),CGMMV-XG(GenBank No:KP868654),JD2(GenBank No:KM873785)and Anhui(GenBank No:KT236095),reaching 99.8% .

Citrullus lanatusviruscucumber green mottle mosaic viruswatermelon crinkle leaf-associated virus 2cucurbit aphid-borne yellows viruscucumis melo cryptic viruswatermelon virus A

杜江、马振男、王晨燕、崔丽艳、王德富、牛颜冰

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山西农业大学生命科学学院,太谷 030801

山西农业大学草业学院,太谷 030801

西瓜 病毒 黄瓜绿斑驳花叶病毒 西瓜皱叶病毒2号 瓜类蚜传黄化病毒 甜瓜潜隐病毒 西瓜病毒A

山西省高等学校科技创新项目山西农业大学科技创新项目山西省基础研究计划(自由探索类)现代农业产业技术体系建设项目

2021L1722020BQ5120210302124676CARS-21

2024

10.13926/j.cnki.apps.001317

植物病理学报
中国植物病理学会

植物病理学报

CSTPCD北大核心
影响因子：0.865
ISSN：0412-0914
年,卷(期)：2024.54(1)
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