[Isolation and Identification of a Novel Cypovirus from Daphnis nerii].

Zhigao Zhan, Yuzhou Xiao, Zhuorong Liu, Wenchao Zhang, Jinchang Wang, Hongxiu Wang, Limei Guan, Gang Yang, Zhao Huang, Liang Jin
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Abstract

In order to develop a novel effective biological insecticide for controlling oleander hawk moth, a new pathogen was isolated from naturally diseased Daphnis nerii. Based on scanning electron microscopy, full-length amplification of cDNAs (FLAC), and phylogenetic analysis of genome segments 2and 10,the virus was identified as a new type of cypovirus (Da phnis nerii cypovirus [DnCPV]). Electrophoresis analysis showed that DnCPV had a genome comprising 10double-stranded RNA (dsRNA) segments, ranging from 892 to 4160bp.Using FLAC, the cDNAs from the 10 dsRNA segments of the new CPV were cloned and genome segments 2and 10 were sequenced. Sequencing results showed that segment 2 encoded RNA-dependent-RNA-polymerases (RdRps) and segment 10 encoded polyhedrin. These two segments shared conserved terminal sequences of AGUCAAA and AGC at the 5’and 3’ends,respectively.These conserved terminal sequences were not consistent with any of the known CPV types.Phylogenetic analysis of the RdRp and polyhedrin indicated that this CPV was more closely related to CPV type 19 and type 5than other CPV types. Based on the unique conserved terminal sequences and the electrophoresis pattern of the new virus, we tentatively named it DnCPV Nanchang isolate: DnCPV-NC.

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一种新型蛇形病毒的分离鉴定。
为了开发一种新型有效的生物杀虫剂防治夹竹桃飞蛾,从夹竹桃自然病害中分离出一种新的夹竹桃飞蛾病原。通过扫描电镜、cdna全长扩增(FLAC)和基因组第2段和第10段的系统发育分析,确定该病毒为一种新型沙河病毒(Da phnis nerii沙河病毒[DnCPV])。电泳分析显示,DnCPV基因组包含10个双链RNA (dsRNA)片段,长度从892 ~ 4160bp不等。利用FLAC克隆了新CPV 10个dsRNA片段的cdna,并对其基因组2和10段进行了测序。测序结果显示,第2段编码rna依赖rna聚合酶(RdRps),第10段编码多角蛋白(polyhedrin)。这两个片段分别在5′和3′端共享AGUCAAA和AGC的保守末端序列。这些保守的末端序列与任何已知的CPV类型都不一致。RdRp和多面蛋白的系统发育分析表明,该CPV与CPV 19型和CPV 5型的亲缘关系较其他CPV型更为密切。根据新病毒独特的保守末端序列和电泳图谱,我们暂定其为DnCPV南昌分离株:DnCPV- nc。
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