番木瓜环斑病毒海南南瓜分离物全基因组序列分析  

Complete genomic sequence of an Papaya ringspot virus isolate from pumpkin in Hainan

作　　者：车海彦[1] 曹学仁[1] 罗大全[1] CHE Haiyan;CAO Xueren;LUO Daquan(Environment and Plant Protection Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou,Hainan 571101,China)

机构地区：[1]中国热带农业科学院环境与植物保护研究所,海南海口571101

出　　处：《西北农林科技大学学报（自然科学版）》2019年第3期38-43,51,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家公益性行业(农业)科研专项(201303028);2018年海南省重大科技计划项目(zdkj201813)

摘　　要：【目的】研究海南南瓜上番木瓜环斑病毒(Papaya ringspot virus,PRSV)的全基因组特征及系统进化情况,为南瓜病毒病的综合防控提供依据。【方法】通过DAS-ELISA和RT-PCR等方法,检测采自海南澄迈桥头镇疑似感染PRSV的南瓜叶片中是否存在PRSV,采用分段扩增测序方法拼接获得PRSV-HnPumpkin基因组,利用NC-BI中的BLAST工具、DNAStar和MEGA 6.06软件进行序列分析及系统关系树构建。【结果】DAS-ELISA和RT-PCR检测结果表明,采自海南澄迈桥头镇疑似感染PRSV的南瓜叶片中存在PRSV(PRSV-HnPumpkin)。PRSV-HnPumpkin基因组全长为10 327nt,具有典型的PRSV基因组结构特征,含有1个开放阅读框,编码1个含有3 345个氨基酸的多聚蛋白。PRSV-HnPumpkin与已报道的其他PRSV分离物核苷酸和氨基酸之间的相似性分别为80.6%～95.1%和90.7%～97.9%。系统关系分析表明,PRSV-HnPumpkin与亚洲分离物处于同一组中,且与我国台湾分离物亲缘关系较近。【结论】获得了PRSV-HnPumpkin全序列,但其进化来源有待进一步探讨。【Objective】The study clarified the genome characteristics of Papaya ringspot virus(PRSV)infecting pumpkin in Hainan and its phylogenetic relationship with reported PRSV isolates to provide basis for the comprehensive prevention of pumpkin virus disease.【Method】PRSV was detected from suspected virus-infected pumpkin leaf samples by DAS-ELISA and RT-PCR methods.The complete genomic sequence of PRSV-Hnpumpkin isolate was determined by assembling six fragments amplified from the sample by RT-PCR.Sequence was analyzed by BLAST and DNAStar.Neighbor-joining phylogram was constructed by MEGA 6.06.【Result】DAS-ELISA and RT-PCR confirmed the presence of PRSV(PRSV-HnPumpkin)in the suspected virus-infected pumpkin leaf samples from Qiaotou,Chengmai county in Hainan.Its genome contained 10 327 nt with typical characteristics of previously reported PRSV isolates.The genome contained one ORFs,coding for one polyprotein with 3 345 amino acids.The identities of nucleotide and amino acids with other isolates in GenBank were 80.6%-95.1%and 90.7%-97.9%,respectively.Phylogram analysis revealed that PRSV-HnPumpkin and Asian isolates were in the same group,and it shared closer relationship with the isolates from Taiwan,China.【Conclusion】The complete sequence of PRSV-HnPumpkin was obtained,and further investigation is needed to identify its evolution source.

关 键 词：番木瓜环斑病毒 南瓜病毒病 基因序列分析 

分 类 号：S432.4.1[农业科学—植物病理学]