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利用5'RACE结合一步法RT-PCR分别从山东泰安与临沂冬小麦上克隆了小麦黄花叶病毒(Wheat yellow mosaic virus,WYMV)的全基因组序列。这2个分离物(TADWK和LYJN)基因组间核苷酸一致性分别为97.21%(RNA1)和95.12%(RNA2)。通过对目前已报道的共14个分离物的基因组不同部分的分析,表明5'UTR是WYMV基因组变化幅度最大的区域,而编码区(ORF)及3'UTR的序列一致性较高且变动幅度小。另外,发现LYJN RNA2的5'UTR与已报道的所有分离物RNA2的核苷酸一致率仅为90%左右。综合分析RNA1和RNA2的系统发生树,表明WYMV各基因组片段呈单独进化特征,不同分离物间存在RNA重排。RNA重组分析显示在LYJN RNA1和TADWK RNA2发现了RNA重组。此研究说明WYMV在山东地区存在分离物分化现象,WYMV的5'UTR是基因组中的突变热点。
The whole genome sequence of wheat yellow mosaic virus (WYMV) was cloned from the winter wheat in Taian, Shandong and Linyi using 5 'RACE and one-step RT-PCR respectively. Nucleotide identity between the two isolates (TADWK and LYJN) was 97.21% (RNA1) and 95.12% (RNA2), respectively. The analysis of different genomes of 14 isolates reported at present showed that 5'UTR is the region with the widest variation of WYMV genome and the sequence identity of ORF and 3'UTR is high and the variation range small. In addition, it was found that the nucleotide conformation of the 5'UTR of LYJN RNA2 and all the isolates RNA2 reported was only about 90%. The phylogenetic tree of RNA1 and RNA2 was analyzed comprehensively, which indicated that the WYMV genomic fragments showed individual evolutionary features and RNA rearrangements existed among different isolates. RNA recombinant analysis showed that RNA recombination was found in LYJN RNA1 and TADWK RNA2. This study shows that WYMV exists isolates in Shandong Province. The 5 'UTR of WYMV is a hot spot in the genome.