南方农业学报2025，Vol.56Issue(2)：431-440,10.DOI:10.3969/j.issn.2095-1191.2025.02.008
侵染洋桔梗的烟草曲茎病毒分子鉴定及遗传进化分析
Molecular identification and genetic evolutionary analysis of tobacco curly shoot virus infecting Eustoma grandiflorum(Raf.)Shinners
摘要
Abstract
[Objective]To clarify the pathogen causing Eustoma grandiflorum(Raf.)Shinners symptoms of raised leaf veins and crumpled leaves in Nanning,Guangxi,and to analyzed the genetic evolutionary relationship of virus isolates,which could provide theoretical basis for the prevention and control of viral diseases in flower crops.[Method]A Eustoma grandiflorum(Raf.)Shinners with typical geminivirus symptoms,such as raised leaf veins and crumpled leaves,was used as the study object.The total DNA of Eustoma grandiflorum(Raf.)Shinners leaves was detected by PCR using geminivirus universal degenerative primers(PA/PB).The whole genome sequence of related virus pathogens was ob-tained by designing back-to-back primers(TbCSV-F/TbCSV-R)and gene cloning.The spliced sequences were compared and analyzed by BLASTn,and the whole genome sequence and genetic evolution of related virus pathogens were ana-lyzed by biological softwares SnapGene,SDT and MEGA 7.0.[Result]PA/PB primer was used to amplify one 500 bp specific target band from the samples.BLASTn analysis showed that the nucleotide sequence similarity between the ob-tained sequences and the isolates of tobacco curly shoot virus(TbCSV)registered in GenBank was more than 91%.It was confirmed that the collected Eustoma grandiflorum(Raf.)Shinners plants were infected by Begomovirus virus.β satellite universal primers(β01/β02)could not be used to amplify the bands,confirming that β-satellite molecules were not pre-sent in the leaves of infected Eustoma grandiflorum(Raf.)Shinners.Tbcsv-f/TBCSV-R was used to amplify the total DNA of infected samples by PCR.The obtained product was purified,cloned,sequenced and sequentially splicing to obtain a complete viral genome sequence with a length of 2743 bp.The nucleotide similarity analysis on the obtained sequence in-dicated that the viral isolate was a TbCSV isolate,named TbCSV-GX-YJG.Amino acid homology analysis showed that the nucleotide and amino acid sequence similarity of the positive chain encoding AV1 and AV2 genes with TbCSV-SHJ01(GenBank accession number MW779539.1)was the highest,reaching 100%.The results of evolutionary tree analysis showed that TbCSV-GX-YJG and isolates from Guangxi(TbCSV-SHJ01)and Sichuan(TbCSV-SC740,GenBank acces-sion number MH165181.1)clustered in the same small branch,indicating a close genetic relationship;and isolates from Yunnan(TbCSV-CN,GenBank accession number KM383752.1),Bangladesh(TbCSV-YT8,GenBank accession num-ber HG003650.1),India(TbCSV-WSF1 and TbCSV-TC240,Genbank accession numbers were KM383752.1 and KF551584.0)were in different branches,indicating that TbCSV isolates had certain genetic diversity.[Conclusion]TbCSV of Begomovirus is the viral pathogen that infects Eustoma grandiflorum(Raf.)Shinners displaying raised leaf veins and crumpled leaves in Guangxi.关键词
洋桔梗/烟草曲茎病毒/遗传进化/广西Key words
Eustoma grandiflorum(Raf.)Shinners/tobacco curly shoot virus/genetic and evolutionary/Guangxi分类
农业科技引用本文复制引用
王井园,李战彪,梁小在,谢慧婷,陈锦清,李祐聪,秦碧霞,莫翠萍,蔡健和,朱英芝..侵染洋桔梗的烟草曲茎病毒分子鉴定及遗传进化分析[J].南方农业学报,2025,56(2):431-440,10.基金项目
广西重点研发计划项目(桂科AB23026068) （桂科AB23026068）
广西自然科学基金青年基金项目(2024GXNSFBA010326) （2024GXNSFBA010326）
广西农业科学院基本科研业务专项(桂农科2021YT071,桂农科2024ZX08) Guangxi Key Research and Development Plan Project(Guike AB23026068) （桂农科2021YT071,桂农科2024ZX08）
Youth Project of Guangxi Natural Science Foundation(2024GXNSFBA010326) （2024GXNSFBA010326）
Basic Research Project of Guangxi Academy of Agricul-tural Sciences(Guinongke 2021YT071,Guinongke 2024ZX08) （Guinongke 2021YT071,Guinongke 2024ZX08）
