侵染湖北圆叶牵牛的甘薯曲叶病毒分子鉴定及遗传进化分析

罗婉笛; 王鹏; 莫翠萍; 蔡健和; 章松柏; 李战彪

doi:10.16768/j.issn.1004-874X.2022.10.011

侵染湖北圆叶牵牛的甘薯曲叶病毒分子鉴定及遗传进化分析

Molecular Identification and Genetic and Evolutionary Analysis of Sweet Potato Leaf Curl Virus Infecting Ipomoea purpurea (L.) in Hubei

摘要

摘要:
目的明确湖北省荆州市1株表现黄脉的圆叶牵牛的病毒病原，为该类病害的防控提供理论依据。
方法从湖北省荆州市采集1株表现黄脉、卷叶的圆叶牵牛植株叶片，采用双生病毒简并引物PA、PB进行PCR检测，通过分段克隆、核苷酸序列比对分析和进化树构建等方法对获得的全长基因组序列进行比对及遗传进化分析。
结果 PCR检测结果显示，采集的样品中可扩增一条大小为363 bp的目的靶标序列，证实所采集的圆叶牵牛植株受到双生病毒的侵染；通过分段克隆的方法从阳性样品中拼接获得一条全长为2 827 bp的全基因组序列，核苷酸相似性比较发现，该序列与GenBank已登录序列的甘薯曲叶病毒（Sweet Potato Leaf Curl Virus，SPLCV）各分离物的相似性均在91% 以上，其中与湖南分离物（GenBank登录号：KY783941）和江苏分离物（GenBank登录号：FJ176701）的核苷酸序列相似性分别为97.52% 和96.81%，根据国际病毒分类委员会对菜豆金色黄花叶病毒属病毒种核苷酸相似性 > 91% 为同一病毒的分类标准，确定侵染圆叶牵牛的双生病毒为SPLCV的一个分离物。进化树分析发现，该分离物（GenBank登录号：OM287440）与中国及多个亚洲分离物处于一个大分支，特别是与湖南分离物处于同一小分支，说明该研究获得的分离物与湖南分离物具有较近的亲缘关系。
结论侵染湖北圆叶牵牛的病原为SPLCV，而该病毒可能通过种苗或烟粉虱经湖南传入。该研究为SPLCV侵染湖北牵牛花的首次报道。

Abstract:
Objective The study is conducted to identify the viral pathogen of Ipomoea purpurea (L.) with yellow vein in Jingzhou, Hubei Province, and the results will provide theoretical basis for the effective prevention and control of this disease.
Method One I. purpurea (L.) sample with yellow vein and curly leaves was collected from Jingzhou, Hubei Province. The sample was detected by PCR with a pair of begomovirus degenerate primers PA and PB. Gene cloning, nucleotide sequence comparison and phylogentic tree construction were applied for the comparison and genetic and evolutionary analysis of the obtained full-length genome sequence.
Result PCR results showed that a PCR amplicon with an expect size of 363 bp were amplified from the samples, which confirmed that the collected I. purpurea (L.) were infected by begomoviruses. A full-length viral genome sequence of 2 827 bp was obtained from the positive samples by segmenting cloning. Nucleotide similarity comparison showed that the sequence in this study had more than 91% nucleotide identities with other isolates of sweet potato leaf curl virus (SPLCV) available in GenBank, and it had 97.52% and 96.81% nucleotide identities with Hunan isolate (GenBank accession number: KY783941) and Jiangsu isolate (GenBank accession number: FJ176701), respectively. According to the standards of the International Committee on Taxonomy of Viruses for the classification of begomoviruses, begomoviruses with nucleotide identities of over 91% were classified as the same virus, and the begomovirus infecting I. purpurea (L.) was identified as SPLCV. Phylogenetic tree analysis showed that the SPLCV Hubei isolate (GenBank Accession number: OM287440) were clustered in a big branch with the SPLCV isolates from China and other Asian areas, and clustered with Hunan isolate in a small branch, indicating their close evolutionary relationship.
Conclusion SPLCV is the pathogen that infects I. purpurea (L.) in Hubei, and the disease is probably transmitted from Hunan through seedlings or Bemisia tabaci. This is the first report of SPLCV infecting I. purpurea (L.) in Hubei.

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