生物技术通报 ›› 2020, Vol. 36 ›› Issue (10): 80-87.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0479

• 研究报告 • 上一篇    下一篇

稻曲病菌菌株GZ-14-07中携带的全病毒基因组特征研究

张婷婷1,3(), 滕丽1,2, 蔡小姚1, 龙慧1, 李升愉1, 刘红美1()   

  1. 1.贵州医科大学 生物与医学工程重点实验室,贵阳 550000
    2.贵州医科大学 基础医学院,贵阳 550000
    3.贵州医科大学 环境污染与疾病监控省部共建教育部重点实验室,贵阳 550000
  • 收稿日期:2020-04-23 出版日期:2020-10-26 发布日期:2020-11-02
  • 作者简介:张婷婷,女,博士,副教授,研究方向:微生物;E-mail:ztt-gd@163.com
  • 基金资助:
    贵州省科技计划项目(黔科合基础[2017]1153);贵阳市科技计划项目(筑科合同[2017]5-24);贵州省大学生创新创业训练计划项目(201510660016)

Genomic Characterization of a Totiviridae from the Ustilaginoidea virens Strain GZ-14-07

ZHANG Ting-ting1,3(), TENG Li1,2, CAI Xiao-yao1, LONG Hui1, LI Sheng-yu1, LIU Hong-mei1()   

  1. 1. Key Laboratory of Biology and Medical Engineering,Guizhou Medical University,Guiyang 550000
    2. School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550000
    3. Key Laboratory of Environmental Pollution Monitoring and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang 550000
  • Received:2020-04-23 Published:2020-10-26 Online:2020-11-02

摘要:

稻曲病菌是引起真菌类病害水稻稻曲病的病原。从稻曲病菌菌株GZ-14-07中分离到一株真菌病毒,命名为Ustilaginoidea virens RNA virus 6(UvRV6),对其进行了序列分析。通过CF-11纤维素粉法提取菌株GZ-14-07中的真菌病毒UvRV6,采用随机引物扩增法获得UvRV6的全长基因组序列,使用DNAMAN、BlastP和MEGA等软件对UvRV6的全长序列进行拼接、比对和系统进化树分析。研究发现UvRV6全基因组长度为5 043个碱基,GC为48.8%,UvRV6基因组序列中存在两个开放阅读框(ORF1和ORF2),ORF1长2 280个碱基,编码760个氨基酸(79.6 kD),ORF2长2 385个碱基,编码830个氨基酸(91.5 kD)。ORF1和ORF2编码的蛋白,分别与全病毒科维多利亚属真菌病毒成员的衣壳蛋白和依赖于RNA的RNA聚合酶显著出较高的相似性。另外,UvRV6的ORF1和ORF2之间有一个AUGA序列,在AUGA序列上游存在一段能形成一个H型的假节序列,这两个序列元件的存在使UvRV6两个开放阅读框(ORF1和ORF2)能通过翻译终止后又重新启动翻译的机制,融合成一个大的开放阅读框。最后,通过UvRV6和其他全病毒科成员的依赖于RNA的RNA聚合酶蛋白序列构建的系统进化树分析得出,UvRV6与属于全病毒科维多利亚属的成员聚集成一簇。明确了UvRV6属于全病毒科维多利亚属的新成员。

关键词: 稻曲病菌, 真菌病毒, dsRNA, 全病毒科, 维多利亚属

Abstract:

Ustilaginoidea virens is a pathogen that causes fungi-like disease Ustilaginoidea oryzae. A mycovirus isolated from strain GZ-14-07 of U. virens was designated as Ustilaginoidea virens RNA virus 6(UvRV6)and sequenced. Mycovirus UvRV6 was extracted from the strain GZ-14-07 using CF-11 cellulose powder method and its full-length genomic sequence was obtained by random-primer amplification method. Sequence analysis,alignment and phylogenetic tree of the UvRV6 were performed using DNAMAN,BlastP and MEGA software. The complete genome sequence of the UvRV6 was determined to be 5 043 bp in length,with a GC content of 48.8%,and contained two open reading frames(ORF1 and ORF2). ORF1 had a length of 2 280 bp and encoded a 760-amino-acid protein(79.6 kD),and ORF2 was 2 385 bp long and encoded a 830-amino-acid protein(91.5 kD). ORF1- and ORF2-encoded protein had a high percentage of sequence similarity to the capsid protein and the RNA-dependent RNA polymerases(RdRps)of genus Victorivirus in the family Totiviridae,respectively. Moreover,there was an AUGA between ORF1 and ORF2,and a sequence forming H-type pseudo-knot existed in the upstream of the AUGA. These 2 sequence elements allowed ORF1 and ORF2 to be merged as a bigger ORF via a mechanism of re-initiating translation after translation ended. A phylogenetic analysis based on the alignment of RdRps of UvRV6 and other totiviruses showed the UvRV6 was clustered with the members of Victorivirus in the family Totiviridae. Therefore,UvRV6 is confirmed as a new member of Victorivirus in the family Totiviridae.

Key words: Ustilaginoidea virens, mycovirus, dsRNA, Totiviridae, Victorivirus