山东烟草番茄黄化曲叶病毒的分子检测和序列分析

Molecular Detection and Sequence Analysis of Tomato Yellow Leaf Curl Virus Infecting Tobacco in Shandong Province

2012年7月从山东泰安烟草种植区采集疑似感染番茄黄化曲叶病毒的烟草样品。利用双生病毒的通用引物PA/PB对样品进行扩增和测序,所得序列经NCBI BLAST比对证明,该感病烟草的病毒分离物为番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)。随后对TYLCV进行全基因组序列扩增和测序(GenBank登录号JX856172),经分子比对和进化分析发现,其与山东泰安番茄分离物TYLCV(JF414236)同源性最高为99%,并与山东其他地区和天津、北京等地报道的TYLCV同属一个大的进化分支,均属于以色列株系(TYLCV-IL)。本研究首次明确了在自然条件下烟草受到TYLCV的侵染。

In July of 2012, the diseased tobacco samples were collected from the tobacco growing areas near the tomato plantations in which tomato yellow leaf curl virus disease and whitefly Bemisia tabaci occurred very seriously. The DNA of the samples were tested by PCR using degenerate primers （PA/PB） of geminivirus. The fragments of expected size （0.5 kb） were obtained and sequenced. The blast result showed the causal agent of the disease was a tomato yellow leaf curl virus isolate （TYLCV-SDTT）. Then complete genome sequence of the isolate was amplified. Analysis of similarity between the obtained complete genome sequence and those deposited in the GenBank database was performed. The results shows that TYLCV-SDTT （Gerfl3ank accession JX856172） shared homology of 99% with TYLCV tomato isolate （JF414236） in Shandong province, and belonged to the same large branch with other TYLCV isolates belonging to Israel strain of TYLCV （TYLCV-IL）. This is the first report that Tomato yellow leaf curl virus infects tobacco under natural conditions in China.