河南省番茄黄化曲叶病毒的分子鉴定及序列分析

Molecular Identification and Sequence Analysis of Tomato Yellow Leaf Curl Virus from Henan Province

从河南省9个地市蔬菜大田内采集到49份表现番茄黄化曲叶病毒病(tomato yellow leaf curl disease,TYLCD)典型黄化及曲叶症状的番茄样品,为了明确引起TYLCD的病原,利用双生病毒简并引物PA/PB对49份番茄样品进行PCR检测。结果表明,从32份样品中扩增得到约500bp的特异性片段,阳性检出率为65.3%。随机选择番茄HNAY3、HNKF2、HNXY1、HNLY1、HNNY1、HNXC2、HNXX1及HNSQ1样品进行DNA-A全基因组扩增及克隆测序,结果表明,DNA-A全长均为2 781nt,具有典型的双生病毒科(Geminiviridae)菜豆金色花叶病毒属(Begomovirus)病毒的基因组结构特征,共编码6个ORFs。聚类分析表明,这8个分离物与我国已报道的番茄黄化曲叶病毒(tomato yellow leaf curl virus,TYLCV)各分离物亲缘关系较近。这些结果表明,引起河南省大田番茄上TYLCD的病原是TYLCV。

Using PCR,49 tomato samples displaying tomato yellow leaf curl disease（TYLCD）symptoms collected from nine cities in Henan province were detected with degenerate primers PA/PB for geminiviruses in order to clear the pathogen of TYLCD.The results showed that a specific fragment of about 500 bp was amplified from 32 samples.The positive detection rate was 65.3%.DNA-A whole genome was amplified,cloned and sequenced from randomly selected HNAY3,HNKF2,HNXY1,HNLY1,HNNY1,HNXC2,HNXX1 and HNSQ1 samples.The length of DNA-A from all samples was 2 781 nt,coding six ORFs,which showed typical virus genome structure of the genus Begomovirus of Geminiviridae.The eight isolates had closer genetic relationship with tomato yellow leaf curl virus（TYLCV）isolates reported previously in China.These results suggest that the pathogen of TYLCD emerging in Henan province is TYLCV.