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采用分子手段鉴定了安徽淮北番茄曲叶病的病原。从安徽淮北番茄温室采集番茄叶片样本,用CTAB法从番茄叶片中提取总DNA。利用简并引物PA、PB进行PCR扩增,从大部分样本中扩增出双生病毒500 bp特异性条带。将扩增片段克隆并测序,该片段序列全长541 bp,与我国报道的番茄黄化曲叶病毒(TYLCV)各个分离物序列相似性均很高,达到95.9%~99.4%,可以初步明确侵染安徽淮北番茄的双生病毒为番茄黄化曲叶病毒(TYL-CV)。安徽淮北TYLCV 500 bp片段序列与山东TYLCV相似性最高,达99.4%,而且从构建的系统关系树也可以看出,安徽淮北TYLCV与山东TYLCV的亲缘关系最近,推测安徽淮北番茄黄化曲叶病毒(TYLCV)可能来源于山东。
The pathogen of tomato leaf curl disease in Huaibei of Anhui Province was identified by molecular method. The tomato leaf samples were collected from the tomato greenhouse in Huaibei, Anhui Province, and the total DNA was extracted from the tomato leaf by CTAB method. Degenerate primers PA and PB were used for PCR amplification, and a 500 bp specific band of geminivirus was amplified from most samples. The cloned fragment was cloned and sequenced. The full length of this fragment was 541 bp, which was highly homologous to all isolates of TYLCV reported in China, reaching 95.9% ~ 99.4% The twin virus that infected the Huaibei tomato in Anhui Province was TYL-CV. The 500 bp fragment of TYLCV from Huaibei in Anhui Province has the highest similarity with TYLCV in Shandong province, reaching 99.4%. And from the constructed phylogenetic tree, it can be seen that the genetic relationship between TYLCV and TYLCV in Huaibei, Anhui Province is the closest, suggesting that Anhui Huaibei tomato yellow leaf curl virus (TYLCV) may come from Shandong.