Wu J B, Dai F M, Zhou X P
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, P. R. China.
Institute of Plant Protection, Shanghai Academy for Agricultural Sciences, Shanghai 201106, P. R. China.
Plant Dis. 2006 Oct;90(10):1359. doi: 10.1094/PD-90-1359C.
Tomato yellow leaf curl virus (TYLCV) is a devastating pathogen of tomato that causes significant yield losses in many tropical and subtropical regions (2). In China, however, there has as yet been no report of this virus, although other begomoviruses have been reported infecting tomato (1,3). A yellow mosaic disease was observed on tomato with 90% disease incidence during March 2006 in fields of Sunqiao, Shanghai Province, China. Triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) tests indicated that tomato plants were not infected by Tomato mosaic virus or Cucumber mosaic virus. Tomato plants were found to be infested with Bemisia tabaci, suggesting a begomovirus etiology. The disease agent was transmitted to tomato by whiteflies and produced yellow mosaic and stunting symptoms that were identical to those observed in the field. Total DNA was isolated from eight collected leaf samples. Polymerase chain reaction (PCR) was performed with begomovirus degenerate primers PA and PB (3), and an amplicon of the expected size (~500 bp) was obtained in all eight samples but not from healthy leaf samples. The PCR products from two samples (SH1 and SH2) were cloned and sequenced. All residues in the sequences were confirmed by comparison of duplicate clones. Alignment of the sequences showed that they shared 97.4% nucleotide sequence identity (GenBank Accession No. AM282874-75), suggesting that they were infected by an identical virus. Overlapping primers Full/F (5'-AGCCCAATACATTGGGCC ACGA-3') and Full/R (5'-CGTAAGTTTCCTCAACGGACTGC-3') were then designed to amplify the full length DNA-A of SH2. The sequence was determined to be 2,781 nucleotides long (GenBank Accession No. AM282874). A comparison with other begomoviruses shows SH2 DNA-A has the highest nucleotide sequence identity (99.8%) with TYLCV isolate Tosa from Japan (GenBank Accession No. AB192966). The above results indicate that the virus associated with yellow mosaic disease of tomato in Shanghai is an isolate of TYLCV. To our knowledge, this is the first report of TYLCV in China and the first report of a begomovirus in Shanghai. References: (1) X. F. Cui et al. J. Virol. 78:13966, 2004. (2) E. Moriones and J. Navas-Castillo. Virus Res. 71:123, 2000. (3) Z. H. Li et al. Arch. Virol. 149:1721, 2004.
番茄黄化曲叶病毒(TYLCV)是番茄的一种毁灭性病原体,在许多热带和亚热带地区会导致严重的产量损失(2)。然而,在中国,尽管已有其他双生病毒感染番茄的报道(1,3),但尚未有关于该病毒的报道。2006年3月,在中国上海孙桥的田间观察到一种番茄黄花叶病,发病率达90%。三抗体夹心酶联免疫吸附测定(TAS - ELISA)测试表明,番茄植株未感染番茄花叶病毒或黄瓜花叶病毒。发现番茄植株上有烟粉虱侵害,提示病因可能是双生病毒。该病原体通过粉虱传播给番茄,并产生与田间观察到的相同的黄花叶和矮化症状。从采集的8个叶片样本中提取了总DNA。用双生病毒简并引物PA和PB进行聚合酶链反应(PCR)(3),在所有8个样本中均获得了预期大小(约500 bp)的扩增子,而健康叶片样本中未获得。对两个样本(SH1和SH2)的PCR产物进行克隆和测序。通过重复克隆的比较确认了序列中的所有残基。序列比对表明,它们的核苷酸序列同一性为97.4%(GenBank登录号AM282874 - 75),表明它们感染了同一种病毒。然后设计重叠引物Full/F(5'-AGCCCAATACATTGGGCC ACGA-3')和Full/R(5'-CGTAAGTTTCCTCAACGGACTGC-3')来扩增SH2的全长DNA - A。确定该序列长度为2781个核苷酸(GenBank登录号AM282874)。与其他双生病毒比较显示,SH2 DNA - A与来自日本的TYLCV分离株Tosa的核苷酸序列同一性最高(99.8%)(GenBank登录号AB192966)。上述结果表明,与上海番茄黄花叶病相关的病毒是TYLCV的一个分离株。据我们所知,这是TYLCV在中国的首次报道,也是双生病毒在上海的首次报道。参考文献:(1)X. F. Cui等人,《病毒学杂志》78:13966,2004年。(2)E. Moriones和J. Navas - Castillo,《病毒研究》71:123,2000年。(3)Z. H. Li等人,《病毒学档案》149:1721,2004年。