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›› 2017, Vol. 39 ›› Issue (6): 429-436.

• 论文与报告 • 上一篇    下一篇

闽南地区斜带石斑鱼(Epinephelus coioides ) 神经坏死病毒的基因型与分子进化研究

朱志煌1,林克冰1,周宸2,吴建绍1,葛辉1,郑乐云1,黄种持1   

  1. 1. 福建省水产研究所
    2. 福建水产研究所
  • 收稿日期:2017-11-06 修回日期:2017-11-20 出版日期:2017-12-25 发布日期:2018-01-02
  • 通讯作者: 朱志煌
  • 基金资助:
    厦门南方海洋研究中心项目;福建省海洋高新产业发展专项;福建省海洋生物增养殖与高值化利用重点实验室开放课题基金

The genotyping and molecular evolution analysis of nervous neurosis virus (NNV) in infected Epinephelus coioides in southern Fujian

  • Received:2017-11-06 Revised:2017-11-20 Online:2017-12-25 Published:2018-01-02
  • Contact: zhihuang Zhu

摘要: 本研究以闽南地区具有典型神经坏死病症状的斜带石斑鱼为材料,采用RT-PCR法对其进行病毒检测,对检测到的阳性序列进行双向测序和构建系统发育树,对病毒颗粒进行分离纯化,并通过分子进化模型分析病毒衣壳蛋白基因所受的选择压力。结果显示,采集到的6个石斑鱼样品均呈NNV 阳性,系统树分析发现6个样品的PCR扩增片段均为RGNNV基因型序列,表明闽南地区感染石斑鱼的神经坏死病毒主要为RGNNV基因型病毒;通过PEG法对病毒进行分离提純,获得直径为25~28 nm、呈二十面立体对称结构的无囊膜病毒 颗粒;分子进化分析显示NNV外壳蛋白基因经历了纯化选择,表明病毒在进化过程中没有 出现遗传变异并以相对恒定保守的速率进化。

关键词: 斜带石斑鱼, 基因型, 神经坏死病毒, 分子进化, Epinephelus coioides, genotype, NNV, molecular evolution

Abstract: In this study,we used the nervous neurosis virus ( NNV ) test standard of Office International Des Epizooties (OIE) to detect the viral nervous necrosis of Epinephelus coioides collected from southern Fujian. All the six samples were detected to be infected with NNV. The positive clones were separately sequenced from both forward and reverse directions with vector primers,and the sequences were used to construct the phylogenetic trees. The virus particles were isolated and purified by PEG method. All the sequence information of positive clones was utilized to analyze the selection pressure of viral coat protein gene. The results showed that the collected six samples were infected with RGNNV,indicating that the infectious NNV of grouper was mainly RGNNV in southern Fujian. The vims was isolated and purified to obtain a kind oi icosahedral vims paricles with a diameter of 25~28 nm. Molecular evolution analysis showed that the NNV coat protein gene experienced the purification selection,indicating that the virus retained very low genetic variation.

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