摘要
为明确贵州烟草新病毒病害的病原及其分子特征、传播方式和致病性状,有效防控该病毒病,利用病毒检测试纸条、电镜观察、间接ELISA、RT-PCR和dsRNA技术等方法对病原进行检测和鉴定;通过RT-PCR扩增病原的全基因组序列,借助系统发育树和重组分析解析病原进化关系及可能存在的重组事件;通过生物学接种病原至本氏烟和云烟87,观察是否发病及发病症状。结果表明,在病株汁液中电镜观察到(12~14)nm×(700~900)nm的线性病毒粒子;马铃薯Y病毒(Potato virus Y,PVY)检测试纸条检测结果呈阳性,但PVY特异性引物RT-PCR检测呈阴性;通过dsRNA技术获取病原基因组的随机克隆片段,经测序为辣椒脉斑驳病毒(Chilli veinal mottle virus,ChiVMV)基因组片段;灌根、摩擦、介体均能传播该病毒株系,其侵染本氏烟的典型症状为褪绿、黄化、疱斑、叶缘下卷和植株矮化,侵染云烟87品种的典型症状为斑驳、疱斑和叶缘下卷;该株系的基因组序列全长为9782 nt(登录号:OP378160),与同以烟草为寄主的ChiVMV YN-tobacco在系统发育关系上最为接近;进一步的重组分析显示,该株系可能为重组分离物。综上所述,贵州烟草新病害病原为ChiVMV的一个株系,命名为ChiVMV贵州烟草株系(GZ-tobacco),该株系能够引起烟草典型病毒病症状。
In order to clarify the pathogen and its molecular characteristics,transmission mode and pathogenicity of tobacco new virus disease in Guizhou,and effectively prevent and control the virus disease,the pathogen was detected and identified by virus detection strip,electron microscope observation,indirect ELISA,RT⁃PCR and dsRNA technology.The whole genome sequence of the pathogen was amplified by RT⁃PCR,and the evolutionary relationship of the pathogen and the possible recombination events were analyzed by phylogenetic tree and recombination analysis.Through biological inoculation of the pathogen to N.benthamiana and Yunyan 87,the incidence and symptoms were observed.The results showed that the linear virus particles of(12—14)nm×(700—900)nm were observed in the juice of the diseased plants by electron microscopy.Potato virus Y(PVY)detection test strip was positive,but PVY specific primer RT⁃PCR was negative.The random clone fragments of the pathogen genome were obtained by dsRNA technology and sequenced as the Chilli veinal mottle virus(ChiVMV)genome fragments.The virus strains could be transmitted by root irrigation,friction and mediator.The typical symptoms of N.benthamiana were chlorosis,yellowing,blister spot,leaf margin curling and plant dwarfing.The typical symptoms of Yunyan 87 were mottle,blister spot and leaf margin curling.The genome sequence length of the strain was 9782 nt(accession number:OP378160),which was the closest to ChiVMV YN⁃tobacco in phylogenetic relationship.Further recombination analysis showed that the strain might be a recombinant isolate.In summary,the new pathogen of tobacco disease in Guizhou is a new strain of ChiVMV,named ChiVMV Guizhou tobacco strain(GZ⁃tobacco),which can cause typical symptoms of tobacco virus disease.
作者
张长城
姜军
潘首慧
薛原
张权
李梓阔
汪汉成
史彩华
方守国
章松柏
李熙全
ZHANG Changcheng;JIANG Jun;PAN Shouhui;XUE Yuan;ZHANG Quan;LI Zikuo;WANG Hancheng;SHI Caihua;FANG Shouguo;ZHANG Songbai;LI Xiquan(Key Laboratory of Sustainable Crop Production in the Middle Reaches of the Yangtze River,Ministry of Agriculture and Rural Affairs(Co-construction by Ministry and Province)/Yangtze University,Jingzhou 434025,China;Anshun Branch of Guizhou Tobacco Company,Anshun 561000,China;Guizhou Academy of Tobacco Science,Guiyang 550081,China)
出处
《河南农业科学》
北大核心
2025年第8期102-110,共9页
Journal of Henan Agricultural Sciences
基金
国家自然科学基金项目(31972243)
贵州省烟草公司安顺市公司科技项目(2022520400140040)。
关键词
烟草
辣椒脉斑驳病毒
株系
分子鉴定
生物学特性
序列分析
Tobacco
Chilli veinal mottle virus
Isolate
Molecular identification
Biological characteristics
Sequence analysis
