河北苹果坏死花叶病毒的发生及近全基因组序列的测定与分析  

Occurrence of apple necrotic mosaic virus in Hebei province and sequence analysis of its whole genome

作　　者：董震杨 李紫腾 胡同乐[1] 王树桐[1] 王亚南[1] 曹克强[1] DONG Zhenyang;LI Ziteng;HU Tongle;WANG Shutong;WANG Yanan;CAO Keqiang(College of Plant Protection,Hebei Agricultural University Baoding 071001,China)

机构地区：[1]河北农业大学植物保护学院,河北保定071001

出　　处：《河北农业大学学报》2023年第1期72-79,共8页Journal of Hebei Agricultural University

基　　金：河北省自然科学基金(C2022204196);河北省重点研发计划(21326506D);国家现代农业(苹果)产业技术体系(CARS-27)。

摘　　要：为明确苹果坏死花叶病毒(Apple necrotic mosaic virus, ApNMV)在河北的发生及其基因组特性,本试验利用RT-PCR技术对河北保定两地ApNMV发病情况进行检测,然后通过高通量测序技术对带毒样本进行序列测定和分析。结果表明,河北农大果园‘红珍珠’‘锦锈红’‘信侬红’3个品种及保定唐县苹果实验基地富士品种ApNMV检出率分别为9.09%、18.18%、100%和65%。ApNMV阳性果树高通量测序结果显示,果树为ApNMV、苹果褪绿叶斑病毒(Apple chlorotic leaf spot virus, ACLSV)、苹果茎沟病毒(Apple stem grooving virus, ASGV)和苹果茎痘病毒(Apple stem pitting virus, ASPV)复合侵染,ApNMV河北分离物(ApNMV-HB)的近全基因组序列包含RNA1~RNA3序列,GenBank登录号分别为OP019626、OP019627和OP019628。RNA1编码120 kD的甲基转移酶/NTP-binding解旋酶(MET/HEL),RNA2编码97 kD的RNA聚合酶(POL),RNA3编码运动蛋白(Movement protein, MP)和外壳蛋白(Coatprotein,CP),4个基因核苷酸和氨基酸序列与代表性分离物相似性分别为89.44%~97.02%、92.14%~98.67%。系统发育关系分析表明,以ApNMV RNA1、RNA2、RNA3(CP和MP)编码的氨基酸为基础构建系统发育树,ApNMV-HB分别与AM95、AM125、XJ、ZZ亲缘关系最近。本研究首次揭示了ApNMV河北分离物的近全基因组序列,丰富了该病毒的基因组序列信息,明确了该分离物与其它分离物的系统发育关系,为进一步了解该病毒的系统发育及遗传进化提供参考。In order to clarify the occurrence and genomic characteristics of apple necrotic mosaic virus(ApNMV) in Hebei, the incidence of ApNMV in two regions of Baoding was detected by RT-PCR, and then the infected sample was sequenced by high-throughput sequencing technology. The results showed that the positive rates of ApNMV were 9.09%, 18.18%, 100% and 65% in the cultivars of ‘Hongzhenzhu’, ‘Jinxiuhong’ and ‘Xinnonghong’ in the Apple Experimental Garden of Agricultural University of Hebei, and ‘Fuji’ in the the Apple Experimental Site of Tangxian County, Baoding. High-throughput sequencing revealed that the apple trees infected by ApNMV were also infected by apple chlorotic leaf spot virus(ACLSV), apple stem grooving virus(ASGV) and apple stem pitting virus(ASPV). The near-complete genome sequence of ApNMV was obtained from the sequencing results. The RNA1~RNA3 GenBank accession numbers were OP019626, OP019627 and OP019628, respectively. The isolate was named ApNMV Hebei isolate(ApNMV-HB). RNA1 encoded a 120 kD methyltransferase/NTP-binding helicase(MET/HEL). RNA2 encoded a 97 kD RNA polymerase(POL). And RNA3 encoded movement protein(MP) and coat protein(CP). The four genes shared sequence similarity with the representative isolates in 89.44%—97.02%of the nucleotide and 92.14%—98.67% of the amino acid. The phylogenetic analysis showed that all representative isolates were clustered in one branch based on the amino acids of ApNMV RNA1, and ApNMV-HB was closest to AM95. All representative isolates were clustered in one branch, except for AM72, based on the amino acids encoded by ApNMV RNA2. ApNMV-HB was closest related to AM125. In the phylogenetic tree based on ApNMV CP, the representative isolates clustered in one branch rooted with AM72, and ApNMV-HB and XJ were relatively close. In the phylogenetic tree based on ApNMV MP, all isolates were divided into 4 branches. ApNMV-HB was clustered with AM75, ApNMV-Qu, AM125 and ZZ in one branch, and was closest to ZZ. We revealed the near-complete genome se

关 键 词：苹果坏死花叶病毒 高通量测序 基因组特性 系统进化 

分 类 号：S432.1[农业科学—植物病理学]