CHERRY VIRUS-A - CDNA CLONING OF DSRNA, NUCLEOTIDE-SEQUENCE ANALYSIS AND SEROLOGY REVEAL A NEW PLANT CAPILLOVIRUS IN SWEET CHERRY
CHERRY VIRUS-A - CDNA CLONING OF DSRNA, NUCLEOTIDE-SEQUENCE ANALYSIS AND SEROLOGY REVEAL A NEW PLANT CAPILLOVIRUS IN SWEET CHERRY
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DOI：
10.1099/0022-1317-76-8-2015
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发表时间：
1995-08-01
影响因子：
3.8
通讯作者：
JELKMANN, W
中科院分区：
文献类型：
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作者：
JELKMANN, W
从cDNA克隆中获得了一个新鉴定的樱桃病毒属成员樱桃病毒A（CVA）的核苷酸序列（7383个核苷酸）。该cDNA是从感染小樱桃病毒（LCV）的植物组织中提取的dsRNA产生的。少量的LCV dsRNA用作模板核酸，并能够构建文库，出乎意料地，其中7.5%的重组质粒对CVA具有特异性。CVA的基因组结构类似于苹果茎沟病毒（ASGV），苹果茎沟病毒是毛细病毒属的模式成员，并且由266 kDa的多聚蛋白（ORF 1）、位于ORF 1内的52 kDa的ORF 2和poly（A）尾组成。266 kDa的ORF 1包含复制相关蛋白的所有元件，并且与“辛德毕斯样”病毒具有高度的同一性。ORF编码C-末端区域的外壳蛋白（CP）。52 kDa的ORF 2与capillo病毒和pneumonia病毒的推定病毒细胞间运动蛋白具有高度的同一性。在免疫印迹分析中鉴定CP，估计其分子量为24 kDa。通过在大肠杆菌中表达抗原作为融合蛋白获得抗血清。CVA CP与其他毛细血管和支气管炎病毒的相应蛋白质之间存在显著的序列同一性。但与ASGV、苹果褪绿叶斑病毒（ACLSV）、苹果茎痘病毒（ASPV）和樱桃斑驳叶病毒（CMLV）的抗血清进行免疫印迹分析时，均未发现交叉反应。用免疫电镜（ISEM）鉴定甜樱桃提取物中弯曲丝状CVA病毒粒子，并用融合蛋白的抗血清进行修饰。通过ISEM、免疫印迹分析和与dsRNA的杂交，在三种不同疾病状态的樱桃来源中鉴定了CVA。CVA与樱桃中目前描述的任何疾病都没有密切关系，但它具有毛细病毒的所有特性。建议将CVA列为毛细病毒属的一个新成员。
The nucleotide sequence (7383 nucleotides) of a newly identified member of the genus Capillovirus, cherry virus A (CVA), was obtained from cDNA clones. The cDNA was generated from dsRNA extracted from plant tissue infected with little cherry virus (LCV). Small amounts of LCV dsRNA served as template nucleic acid and enabled the construction of a library of which, unexpectedly, 7.5% of the recombinant plasmids were specific for CVA. The genome organization of CVA resembles that of apple stem grooving virus (ASGV), the type member of the genus Capillovirus and is composed of a 266 kDa polyprotein (ORF1), a 52 kDa ORF2 located within ORF1 and a poly(A) tail. The 266 kDa ORF1 contains all the elements of a replication-related protein and has high identity with 'Sindbis-like' viruses. The ORF encodes the coat protein (CP) in the C-terminal region. The 52 kDa ORF2 has high identities with the putative viral cell-to-cell movement proteins of capillo- and trichoviruses. The CP was identified in immunoblot analysis and estimated to have a molecular mass of 24 kDa. Antiserum was obtained by expression of antigens as fusion proteins in Escherichia coli. There is significant sequence identity between CVA CP and the corresponding proteins of other capillo- and trichoviruses. However, no serological cross-reaction was obtained in immunoblot analysis with ASGV, apple chlorotic leafspot trichovirus (ACLSV), apple stem pitting virus (ASPV) and cherry mottle leaf virus (CMLV) antisera. Flexuous filamentous CVA virions were identified in extracts of sweet cherry by immunosorbent electron microscopy (ISEM) and decorated with the antiserum to the fusion protein. CVA was identified in three cherry sources of different disease status by ISEM, immunoblot analysis and hybridization to dsRNA. CVA is not closely related to any of the currently described diseases in cherry but it has all the properties of a capillovirus. It is suggested that CVA should be classified as a new member of the genus Capillovirus.