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油桃茎痘相关病毒中国分离物基因组的分子特征分析
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引用本文:杨丽娟,许云霄,周俊,李世访,卢美光.油桃茎痘相关病毒中国分离物基因组的分子特征分析.植物保护学报,2020,47(1):143-149
DOI:10.13802/j.cnki.zwbhxb.2020.2019126
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作者单位E-mail
杨丽娟 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193  
许云霄 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193  
周俊 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193  
李世访 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193  
卢美光 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193 mglu@ippcaas.cn 
中文摘要:为明确我国油桃茎痘相关病毒(nectarine stem-pitting-associated virus,NSPaV)基因组的分子特征,利用RT-PCR和RACE技术对NSPaV中国分离物NSPaV-T04基因组进行克隆,采用最大似然法对得到的NSPaV基因组序列和GenBank中的5条NSPaV基因组序列构建系统发育树,应用RDP软件对NSPaV基因组序列进行重组分析。结果表明:中国分离物NSPaV-T04基因组序列全长为4 991 nt,包括4个开放阅读框(open reading frame,ORF),其中ORF1与ORF2共同编码1个RdRpP1-P2融合蛋白,ORF3编码1个CP,ORF5与ORF3共同编码CP通读蛋白。系统发育树和序列比较分析结果显示,中国分离物NSPaV-T04(MN095353)与美国分离物NSPaV/12P42(KT273410)的亲缘关系最近,核苷酸序列同源性最高,为96.4%;NSPaV-T04的RdRp P1变异较大,与GenBank中5条NSPaV基因组核苷酸序列的同源性为90.5%~96.1%,CP较为保守,核苷酸序列的同源性为96.6%~98.7%。重组分析结果显示,中国分离物NSPaV-T04为鉴定的一个重组体(韩国分离物SK)的亲本序列,表明中国分离物NSPaV-T04可能是一个实际的重组体。
中文关键词:油桃茎痘相关病毒  基因组  分子特征
 
Molecular characterization of nectarine stem-pitting-associated virus from China
Author NameAffiliationE-mail
YANG Lijuan State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China  
XU Yunxiao State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China  
ZHOU Jun State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China  
LI Shifang State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China  
LU Meiguang State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China mglu@ippcaas.cn 
Abstract:In order to molecularly characterize the nectarine stem-pitting-associated virus (NSPaV) from China, the complete nucleotide sequence of an NSPaV isolate T04 (named NSPaV-T04) from a sample of nectarine collected from China was determined by using RT-PCR and RACE methods. Phylogenetic analysis of NSPaV was conducted by using maximum likelihood and recombination analysis of NSPaV was performed by using RDP software. The results showed that the complete genome of NSPaV-T04 comprised 4 991 nucleotides (nt) and contained four open reading frames (ORF), which was similar to several NSPaV isolates deposited in GenBank. ORF1 and ORF2 encoded the RNA-dependent RNA polymerase P1, P2 fusion protein (RdRp P1-P2), and ORF3 encoded the coat protein (CP). ORF3 and ORF5 were identified to encode the CP read-through fusion protein. Phylogenetic analysis showed that isolate NSPaV-T04 (MN095353) was most closely related to isolate NSPaV/12P42 (KT273410) from America. Analysis of the nucleotide and amino acid sequences showed that the complete nucleotide sequence of NSPaV-T04 had the highest similarity (96.4% nt identity) with isolate NSPaV/12P42. Compared with the five NSPaV genome sequences available in GenBank, the NSPaVT04 sequence in ORF1 (RdRp P1) was more variable compared with other domains, sharing 90.5%-96.1% nt identity. The NSPaV-T04 CP sequence was more conserved, with 96.6%-98.7% nt similarity. The recombination analysis of all available genome sequences showed that the genome of a Korean isolate SK might be a recombinant. However, NSPaV-T04 might also be an actual recombinant, based on only one identified parental sequence in triplet in recombination analysis.
keywords:nectarine stem-pitting-associated virus  genome  molecular characterization
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