大麦条纹花叶病毒中国株基因组RNA1的全长序列分析.docVIP

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1，邱并生2 （1西南大学植物保护学院植物病害检测实验室，重庆 400716；2中国科学院微生物研究所，北京 100101） 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 Sequence Analysis of Barley Stripe Mosaic Virus China Strain (BSMV-CH) RNA1 SUN Xian-chao1, QIU Bing-sheng2 (1Laboratory of Plant Disease Detection, College of Plant Protection, Southwest University, Chongqing 400716; 2Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101) Abstract: 【Objective】T study was conducted to obtain the full-length cDNA of BSMV-CH RNA1, analyze its sequence and determine its phylogenetic relationship with 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 were used for nucleotide identity analysis and molecular phylogenetic reconstruction. 【Result】The full-length cDNA of BSMV-CH RNA1 was 3 795 bp (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 3 417 bp and deduced to encode the replicase with 1 138 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%