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近年来,水稻黑条矮缩病毒(RBSDV)和南方水稻黑条矮缩病毒(SRBSDV)在安徽各稻区危害日益严重,生产上仅凭症状难以区分2种病毒。本研究建立的RT-PCR方法可以实现一次性检测和鉴定RBSDV和SRBSDV 2种病毒,根据RBSDV和SRBSDV S9保守序列设计3个引物,不仅可以从单独感染RBSDV或SRBSDV的水稻样本中分别扩增出1条大小不同的特异性条带,而且还可以从感染RBSDV和SRBSDV的混合水稻样本中一次性扩增出2条大小不同的特异性条带。从安徽省庐江县、郎溪县、怀宁县和宣州市4个地区水稻田采集表现明显矮缩病症状的水稻样本,RT-PCR检测结果表明,庐江和郎溪水稻样本受到RBSDV侵染,怀宁和宣州水稻样本受到SRBSDV侵染。选取庐江县、郎溪县、怀宁县和宣州市各1个水稻样本,利用RT-PCR扩增出特异性条带,再分别克隆和测序。序列分析表明,庐江、郎溪2个样本的核苷酸序列与RBSDV-Shandong和RBSDV-Zhjr S9部分片段序列相似性高达99.3%～99.8%,且与RBSDV的亲缘关系最近,说明庐江和郎溪样本感染的病毒是RBSDV的2个分离物;怀宁、宣州2个样本的核苷酸序列与SRBSDV-Shangdong和RBSDV-2 S9部分片段序列相似性高达99.5%,且与SRBSDV亲缘关系最近,说明怀宁和宣州样本感染的病毒是SRBSDV的2个分离物。
In recent years, the black streaked dwarf virus (RBSDV) and southern black streaked dwarf virus (SRBSDV) have become more and more serious in the rice paddies in Anhui Province. It is hard to differentiate the two viruses in their production. The RT-PCR method established in this study can detect and identify two kinds of viruses, RBSDV and SRBSDV in one go. Three primers were designed according to the conserved sequences of RBSDV and SRBSDV S9, which can not only be amplified from rice samples infected with RBSDV or SRBSDV 1 specific bands of different size, but also from hybrid rice samples infected with RBSDV and SRBSDV amplified a total of two different sizes of specific bands. The rice samples showing obvious dwarf disease symptoms were collected from paddy fields in Lujiang County, Langxi County, Huaining County and Xuanzhou City in Anhui Province. RT-PCR results showed that RBSDV was infected in Lujiang and Langxi rice samples Ning and Xuanzhou rice samples were infected with SRBSDV. One rice sample from Lujiang County, Langxi County, Huaining County and Xuanzhou City was selected. The specific bands were amplified by RT-PCR and cloned and sequenced respectively. Sequence analysis showed that the nucleotide sequences of Lujiang and Langxi samples were 99.3% -99.8% similar to the RBSDV-Shandong and RBSDV-Zhjr S9 fragment sequences, and their genetic relationship with RBSDV was the closest, indicating that Lujiang and Langxi samples The two viruses isolated from RBSDV were two isolates from Huaining and Xuanzhou. The nucleotide sequence of the two samples was 99.5% similar to that of SRBSDV-Shangdong and RBSDV-2 S9, and the closest relationship with SRBSDV The virus that Huaining and Xuanzhou samples infected was 2 isolates of SRBSDV.