CAJ | 학술논문

本研究从具有典型曲叶病症状的广西靖西烟草病植株上分离到病毒分离物JX-2，全基因组序列测定结果表明，JX-2DNA．A全长2738个核苷酸，共编码6个开放阅读框架（openreadingframes，ORFs），其中病毒链编码AV1（CP）和AV2两个ORFs，互补链编码AC1、AC2、AC3和AC4共4个ORFs。BLAST结果表明，JX-2DNA—A与中国番茄曲叶病毒（Tomato leaf curl Chinavirus，ToLCCNV）各分离物的相似性在93．0％-99．7％之间，其中与ToLCCNV广西番茄分离物ToLCCNV．G32的相似性最高，达99．7％，而与其它双生病毒的同源性均在88．0％以下，表明Jx-2是ToLCCNV的一个分离物。基于Jx-2和已报道的双生病毒属代表种DNA-A全基因组核苷酸序列构建的系统进化树显示，JX-2与ToLCCNV-G32分离物的亲缘关系最近，并与ToLCCNV其它分离物形成一个分支，而与其它10种双生病毒的亲缘关系均相对较远。利用双生病毒卫星DNAβ的特异性引物β01／β02在Jx-2样品中扩增到DNAβ分子（JX-2β），全长为1341个核苷酸，其互补链编码1个ORF（即βC1），并包含一个富含A序列和一个卫星病毒保守序列。序列分析表明，JX-2β与ToLCCNV伴随的DNAβ的相似性在91．0％～96．1％之间，其中与ToLCCNV-G61DNAβ和ToLCCNV—G18DNAβ的相似性最高（96．1％），与其它卫星DNAβ的相似性均低于61．8％。基于JX-2β全基因组核苷酸序列构建的系统进化关系树显示，JX-29与ToLCCNVG61分离物伴随的DNAβ亲缘关系最近，并形成一个独立的分支，再与ToLCCNV其余两个分离物伴随的DNAβ形成一个较大的分支。这是首次报道从烟草中分离到的中国番茄曲叶病毒及其伴随卫星DNA分子的全基因组结构特征。
본연구종구유전형곡협병증상적엄서정서연초병식주상분리도병독분리물JX-2，전기인조서렬측정결과표명，JX-2DNA．A전장2738개핵감산，공편마6개개방열독광가（openreadingframes，ORFs），기중병독련편마AV1（CP）화AV2량개ORFs，호보련편마AC1、AC2、AC3화AC4공4개ORFs。BLAST결과표명，JX-2DNA—A여중국번가곡협병독（Tomato leaf curl Chinavirus，ToLCCNV）각분리물적상사성재93．0％-99．7％지간，기중여ToLCCNV엄서번가분리물ToLCCNV．G32적상사성최고，체99．7％，이여기타쌍생병독적동원성균재88．0％이하，표명Jx-2시ToLCCNV적일개분리물。기우Jx-2화이보도적쌍생병독속대표충DNA-A전기인조핵감산서렬구건적계통진화수현시，JX-2여ToLCCNV-G32분리물적친연관계최근，병여ToLCCNV기타분리물형성일개분지，이여기타10충쌍생병독적친연관계균상대교원。이용쌍생병독위성DNAβ적특이성인물β01／β02재Jx-2양품중확증도DNAβ분자（JX-2β），전장위1341개핵감산，기호보련편마1개ORF（즉βC1），병포함일개부함A서렬화일개위성병독보수서렬。서렬분석표명，JX-2β여ToLCCNV반수적DNAβ적상사성재91．0％～96．1％지간，기중여ToLCCNV-G61DNAβ화ToLCCNV—G18DNAβ적상사성최고（96．1％），여기타위성DNAβ적상사성균저우61．8％。기우JX-2β전기인조핵감산서렬구건적계통진화관계수현시，JX-29여ToLCCNVG61분리물반수적DNAβ친연관계최근，병형성일개독립적분지，재여ToLCCNV기여량개분리물반수적DNAβ형성일개교대적분지。저시수차보도종연초중분리도적중국번가곡협병독급기반수위성DNA분자적전기인조결구특정。
Virus isolate JX-2 was isolated from tobacco showing typical leaf-curled symptoms in Jingxi county of Guangxi Province. The complete nucleotide sequence of JX 2 DNA-A was determined to be 2 738 nucleotides which encoded 6 open reading frames （ORFs） including two template strand-encoding ORFs （AV1 and AV2） and four anti-sense strand-encoding ORFs （AC1, AC2, AC3, and AC4）. The result of BLAST revealed that JX-2 DNA-A showed 93.0%-99.7% sequence identity with those isolates of Tomato leaf curl China virus （ToLCCNV） and shared the highest sequence identity （99.7%） with ToLCCNV isolate G32. Meanwhile less than 88.0% identity was shared with other begomoviruses, which indicated that virus isolate JX-2 infecting tobacco was one of ToLC- CNV isolates. Phylogenetic analysis of JX-2 and other reported begomoviruses DNA-A showed that JX-2 was the most closely related to ToLCCNV isolate G32 and clustered with other ToLCCNV isolates as a separate branch which was apart from other 10 kinds ofbegomoviruses. The fragment of satellite DNA molecule （JX-2β） was amplified with the primers β01 and β02 using JX-2 as template. JX 2β contained 1 341 nueleotides and encoded an ORF （βC1） by anti-sense strand. An adenine-rich region and a satellite conserved region （SCR） were also found in JX-2β. The sequence analysis indicated that JX-2β had 91.0%-96.1% sequence identity with the associated DNA β of ToLCCNV and shared the highest sequence identity with ToLCCNV-G61 DNAβ and ToLCCNV-G18 DNAβ（96.1%）, but got low sequence identity （less than 61.8%） with other reported satellite DNA molecules. Phylogenetic analysis indicated that JX-2β was the most closely related to ToLCCNV-G61 DNAβ and they clustered togeth er as a separate branch, and formed the main branch with other DNAβ associated with ToLCCNV. This was the first report ofgenomic characterization of ToLCCNV and its associated satellite DNA which isolated from tobacco .