Gadiou Sébastien, Kúdela Otakar, Ripl Jan, Rabenstein Frank, Kundu Jiban K, Glasa Miroslav
Department of Virology, Crop Research Institute, Prague 6-Ruzyně, Czech Republic.
Institute of Virology, Slovak Academy of Sciences, 84505 Bratislava, Slovakia.
Plant Dis. 2009 Nov;93(11):1209-1213. doi: 10.1094/PDIS-93-11-1209.
The tritimovirus Wheat streak mosaic virus (WSMV) is widespread throughout the world and represents a severe threat to cereal crop production. To increase knowledge of genetic diversity of WSMV in Europe, until now scarce, capsid protein (CP) sequences of several Czech, French, Italian, Slovak, and Turkish isolates have been determined. A multiple alignment of CP nucleotide sequences using available WSMV sequences revealed only limited sequence variation among 3 previously sequenced European isolates and the 14 European isolates sequenced in this study. Moreover, these isolates were characterized by an identical 3-nucleotide deletion, resulting in the lack of the Gly codon within the CP region of the polyprotein. The results indicate that this monophyletic group of isolates (designated as WSMV-ΔE) is common and widely dispersed throughout the European continent. The close relationship of WSMV-ΔE isolates implies a single common ancestor and, presumably, subsequent dispersal throughout Europe from a single focus. We developed two simple assays for specific and accurate detection of WSMV-ΔE isolates. First, a conserved ClaI restriction site in the core CP gene sequence unique to WSMV-ΔE isolates was used for restriction fragment length polymorphism analysis of amplified polymerase chain reaction (PCR) products. Second, the conserved and specific codon gap in WSMV-ΔE sequences was used as a target to design specific primers functional in one-step reverse-transcription PCR detection of WSMV-ΔE isolates.
小麦线条花叶病毒(WSMV)属于小麦花叶病毒属,在全球广泛分布,对谷类作物生产构成严重威胁。为了增进对欧洲地区WSMV遗传多样性(目前相关了解较少)的认识,现已测定了来自捷克、法国、意大利、斯洛伐克和土耳其的多个分离株的衣壳蛋白(CP)序列。利用现有的WSMV序列对CP核苷酸序列进行多序列比对分析,结果显示,之前测序的3个欧洲分离株与本研究中测序的14个欧洲分离株之间,序列变异程度有限。此外,这些分离株的特征是存在一个相同的3核苷酸缺失,导致多聚蛋白CP区域内缺乏甘氨酸密码子。结果表明,这一单系类群的分离株(命名为WSMV-ΔE)很常见,在欧洲大陆广泛分布。WSMV-ΔE分离株之间的密切关系意味着它们有一个共同的祖先,并且推测是从一个单一的源头随后扩散到了整个欧洲。我们开发了两种简单的检测方法,用于特异性和准确地检测WSMV-ΔE分离株。第一种方法是利用WSMV-ΔE分离株核心CP基因序列中一个保守的ClaI限制性酶切位点,对扩增的聚合酶链反应(PCR)产物进行限制性片段长度多态性分析。第二种方法是将WSMV-ΔE序列中保守且特异的密码子间隔作为靶点,设计特异性引物,用于一步法逆转录PCR检测WSMV-ΔE分离株。