侵染中国河北大豆的大豆花叶病毒的鉴定和全基因组序列分析

Identification and complete genome sequence analysis of soybean mosaic virus from soybean in Hebei,China

本研究利用小RNA深度测序技术在河北卢龙大豆叶片上检测到6株大豆花叶病毒(soybean mosaic virus,SMV),命名为SMV-Gm1~SMV-Gm6。根据小RNA深度测序结果和参考基因组序列设计引物克隆了SMV河北分离物的基因组序列。测序结果经拼接后获得了6个SMV基因组全长序列,大小分别为9588 nt(SMV-Gm1、SMV-Gm3和SMV-Gm5)和9584 nt(SMV-Gm2、SMV-Gm4和SMV-Gm6)。开放阅读框位于基因组第132位至第9332位核苷酸,编码一个多聚蛋白(分子量约为350 kD)。BLAST比对和系统发育分析发现,SMV-Gm1、SMV-Gm3和SMV-Gm5与江苏SMV分离物(登录号:MH919386)的基因组核苷酸序列相似性最高,为98.06%~98.07%,且遗传距离较近,并与江苏、浙江和山西SMV分离物聚为一小簇;SMV-Gm2、SMV-Gm4和SMV-Gm6与韩国SMV分离物(登录号:FJ640954)的基因组核苷酸序列相似性最高,为98.48%~98.51%,且遗传距离较近。

In this study,six isolates of soybean mosaic virus(labelled SMV-Gm1 to SMV-Gm6)were detected from soybean in Lulong,Hebei province using small RNA deep sequencing technology.Based on the sequencing results and a reference genome sequence,specific primers were designed to clone the genome sequences of these SMV isolates.Following assembly,six full-length SMV genome sequences were obtained,with sizes of 9588 nt for SMV-Gm1,SMV-Gm3,and SMV-Gm5,and 9584 nt for SMV-Gm2,SMV-Gm4,and SMV-Gm6.The open reading frame is located between 132nd to 9332nd nucleotides in each genome,encoding a polyprotein with a molecular weight of approximately 350 kD.BLAST alignment and phylogenetic analysis showed that SMV-Gm1,SMV-Gm3,and SMV-Gm5 showed the highest sequence similarity(98.06%to 98.07%)and relatively close genetic distance with the SMV isolate from Jiangsu province(accession number:MH919386),clustering with SMV isolates from Jiangsu,Zhejiang and Shanxi provinces.Similarly,the nucleotide sequences of SMV-Gm2,SMV-Gm4,and SMV-Gm6 exhibited the highest similarity(98.48%to 98.51%)with the SMV isolate from South Korea(accession number:FJ640954),indicating a close genetic relationship.