中国农业科学 ›› 2008, Vol. 41 ›› Issue (7): 1969-1974 .doi: 10.3864/j.issn.0578-1752.2008.07.012

• 植物保护 • 上一篇    下一篇

大麦条纹花叶病毒中国株基因组RNA1的全长序列分析

孙现超,邱并生   

  1. 西南大学植物保护学院植物病害检测实验室
  • 收稿日期:2007-04-17 修回日期:1900-01-01 出版日期:2008-07-10 发布日期:2008-07-10

Molecular Character of Barley Stripe Mosaic Virus China Strain(BSMV-CH) RNA1

  1. 西南大学植物保护学院植物病害检测实验室
  • Received:2007-04-17 Revised:1900-01-01 Online:2008-07-10 Published:2008-07-10

摘要: 【目的】获得大麦条纹花叶病毒中国株(BSMV-CH)基因组RNA1的全长cDNA,进行序列分析,明确与国外报道株系间的亲缘关系和分类地位。【方法】以自然侵染BSMV-CH的大麦叶片中提取的总RNA为模板,用RT-PCR方法获得BSMV-CH RNA1的全长cDNA,克隆到载体pMD18并测序,用DNAstar软件将获得的序列与GenBank登录的BSMV RNA1序列进行对比,分析各株系序列RNA1之间的同源性,构建系统发育树分析BSMV-CH与国外株系间的亲缘关系。【结果】BSMV-CH的RNA1全长为3 795 bp(GenBank注册号:AY789693),基因组RNA1 5′端有一个甲基化的帽子结构,3′端有一个PolyA结构和一段239 bp长的类似tRNA的保守序列,BSMV-CH基因组RNA1含有一个长度为3 417 bp的ORF,推测可编码一个1 138个氨基酸的复制酶基因。BSMV-CH的基因组RNA1与国外报道其它株系核苷酸的同源性为95.4%~95.6%。复制酶基因同其它株系核苷酸和氨基酸的同源性分别为94.9%~95.2%和96.5%~96.8%,PolyA的长度比其它株系明显要长。【结论】BSMV-CH和CV42与CV17、ND18、TSE及TSG间的亲缘关系明显低于CV17、ND18、TSG和TSE间的亲缘关系;BSMV-CH与CV42表现相对较近的亲缘关系,但BSMB-CH是不同于CV42的一个BSMV新株系。

关键词: 大麦条纹花叶病毒, 序列分析, 同源性, 亲缘关系

Abstract: [objective] The study conducted to obtain the full-length cDNA of BSMV-CH RNA1, analysis its molecular character and determine its genetic relationship to foreign strains.[method] The full-length cDNA of BSMV-CH RNA1 was obtained by RT-PCR from the total RNA extracted from the barley leaves infected by BSMV-CH, and cloned into pMD18 vector for sequencing. The sequence together with sequences of BSMV RNA1 available in Genbank was used for nucleotide identity analysis and molecular phylogenetic reconstruction. [result] The full-length cDNA of BSMV-CH RNA1 was 3795bp(AY789693) with a 5`cap in the 5`-terminal, a polyA structure and a tRNA-like structure in the 3`terminal. An open reading frame(ORF) was found in BSMV-CH RNA1. The ORF was 3417bp and deduced to encode the replicase with 1138 amino acids. BSMV-CH RNA1 shares 95.4~95.6% nucleotides identity with that of other strains, and the replicase shares 94.9~95.2% and 96.5~96.8% identities with that of other strains in nucleotides and amino acids, respectively. BSMV-CH RNA1 has more polyA in the 3`terminal than that of other stains. [conclusion] The genetic relationship between BSMV-CH or CV42 and other strains is obviously lower than that among other strains. BSMV-CH and CV42 are relatively close to each other. But BSMV-CH is a new strain that is different from CV42.

Key words: Barley stripe mosaic virus, molecular character, identity, genetic relationship