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根据番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)基因组序列的复制酶保守区设计1对引物JC1,对石家庄具有典型番茄黄化曲叶病毒症状的番茄样品进行检测,经PCR扩增得到626 bp的片段,序列分析比对表明其为TYLCV的一部分。根据上述序列设计1对特异引物QC,通过PCR分离石家庄的TYLCV全长序列并进行测序。同时利用已报道的DNA-B的通用引物CR01/CR02进行PCR扩增。结果表明石家庄分离物只含有DNA-A组分,全长为2 781 bp(Gen Bank登录号:KF612971),命名为TYLCV-Shijiazhuang。基因组序列比较发现,该分离物与TYLCV-Israel株系相似性为99.0%。基因组序列系统进化分析表明,石家庄病毒分离物与北京分离物TYLCV-Beijing3(Gen Bank登录号:GU983859)、山东分离物TYLCV-SDSG-XC(Gen Bank登录号:KC999851)序列相似性很高,均属于TYLCV-Israel分支。
A pair of primers JC1 was designed according to the replicase conserved region of Tomato yellow leaf curl virus (TYLCV) genome sequence. The tomato samples with typical tomato leaf curl virus symptom in Shijiazhuang were detected by PCR A 626 bp fragment was obtained and sequence analysis revealed that it was a part of TYLCV. According to the above sequences, a pair of specific primer QC was designed and the full-length TYLCV sequence of Shijiazhuang was isolated by PCR and sequenced. At the same time, PCR amplification was carried out by using the commonly used primers CR01 / CR02 of DNA-B. The results showed that the isolates of Shijiazhuang contained only DNA-A component, and the full-length was 2 781 bp (Gen Bank accession number: KF612971), named TYLCV-Shijiazhuang. Genomic sequence comparison revealed that the isolates had a similarity of 99.0% with the TYLCV-Israel line. Phylogenetic analysis showed that the sequences of Shijiazhuang virus isolates were similar to TYLCV-Beijing3 (Gen Bank accession number: GU983859) and TYLCV-SDSG-XC (Genbank accession number: KC999851) Belongs to the TYLCV-Israel branch.