Isakeit T, Idris A M, Sunter G, Black M C, Brown J K
Department of Plant Pathology and Microbiology, Texas A&M University, College Station.
Department of Plant Sciences, University of Arizona, Tucson.
Plant Dis. 2007 Apr;91(4):466. doi: 10.1094/PDIS-91-4-0466A.
Tomato yellow leaf curl virus (TYLCV), a monopartite virus in the genus Begomovirus (family, Geminiviridae) from the Middle East, is one of the most damaging whitefly-transmitted viruses of tomato (Lycopersicon esculentum) worldwide. TYLCV was first identified in the United States in 1997 in Florida (4), and most recently, in the Pacific Coast states of Mexico where fresh market tomatoes are grown for the U.S. market (1). During September 2006, tomatoes grown from transplants in Waller County, TX exhibited shortened internodes, stunting and puckering of leaflets, green vein banding, and diffuse chlorosis. The disease incidence in two fields (4 ha total) was 95% and yield was substantially reduced. Many of the transplants were symptomatic at planting. The transplants originated from two facilities in Hidalgo County, TX. Both facilities had experienced heavy infestations of the whitefly, Bemisia tabaci (Genn.), during transplant production. At the same time, transplants produced in Uvalde and Bexar counties, TX, where whitefly infestations were also prevalent, had similar virus symptoms. Total DNA was extracted from the leaves of symptomatic tomato plants from 10 samples from these four counties and amplified by PCR (2). DNA samples from Waller, Hidalgo, and Uvalde counties were cloned, and a partial fragment of the viral coat protein gene (core Cp) was sequenced. BLAST analysis of the core Cp sequences of each sample confirmed the presence of TYLCV. No other begomovirus was detected, and all attempts to amplify a bipartite begomovirus by PCR using degenerate DNA-B specific primers (3) were unsuccessful. The full-length TYLCV DNA was amplified from three samples using the rolling circle amplification method as described (1), cloned, and the sequences were determined. The three sequences shared 99.6 to 100% nt identity and so only one sequence was deposited in the NCBI GenBank database (Accession No. EF110890) (1). Analysis of the complete genome nucleotide sequence corroborated TYLCV identity predicted by core Cp analysis that was 98.1% identical with TYLCV from Egypt (GenBank Accession No. AY594174) and Spain (GenBank Accession No. AJ489258), 97.6% with TYLCV from Mexico (GenBank Accession No. DQ631892), and 96.5% with TYLCV-Is (GenBank Accession No. X15656). Additionally, a Southern blot with TYLCV as the probe detected replicating (double-stranded) TYLCV DNA in all samples consisting of three plants from Uvalde County and 21 plants from Bexar County. To our knowledge, this is the first report of TYLCV in Texas that occurred in two transplant production areas approximately 400 km apart. Transplants produced in Uvalde and Bexar counties were planted there, while Hidalgo County transplants were shipped outside of the usual range of the whitefly. Hidalgo County has a subtropical climate, which can allow overwintering of TYLCV and the whitefly vector, allowing the establishment and spread of this virus in the future. References: (1) J. K. Brown and A. M. Idris. Plant Dis. 90:1360, 2006. (2) J. K. Brown et al. Arch. Virol. 146:1581, 2001. (3) A. M. Idris and J. K. Brown. Phytopathology 88:648, 1998. (4) J. E. Polston et al. Plant Dis. 83:984, 1999.
番茄黄化曲叶病毒(TYLCV)是一种来自中东的双生病毒科菜豆金色花叶病毒属的单分体病毒,是全球番茄(Lycopersicon esculentum)最具危害性的粉虱传播病毒之一。TYLCV于1997年在美国佛罗里达州首次被发现(4),最近在墨西哥太平洋沿岸各州也有发现,这些地区种植的新鲜市场番茄供应美国市场(1)。2006年9月,得克萨斯州沃勒县种植的移栽番茄出现节间缩短、小叶发育不良和起皱、绿色叶脉带以及弥漫性黄化现象。两块田地(总计4公顷)的发病率为95%,产量大幅下降。许多移栽苗在种植时就表现出症状。这些移栽苗来自得克萨斯州伊达尔戈县的两家育苗场。在移栽苗生产期间,两家育苗场都遭受了烟粉虱(Bemisia tabaci (Genn.))的严重侵害。与此同时,得克萨斯州乌瓦尔德县和贝克斯县生产的移栽苗也出现了类似的病毒症状,这两个县的粉虱侵害也很普遍。从这四个县的10个样本中出现症状的番茄植株叶片中提取总DNA,并通过PCR进行扩增(2)。对来自沃勒县、伊达尔戈县和乌瓦尔德县的DNA样本进行克隆,并对病毒外壳蛋白基因的部分片段(核心Cp)进行测序。对每个样本的核心Cp序列进行BLAST分析,证实存在TYLCV。未检测到其他菜豆金色花叶病毒,并且使用简并DNA - B特异性引物通过PCR扩增双分体菜豆金色花叶病毒的所有尝试(3)均未成功。使用所述的滚环扩增方法从三个样本中扩增出全长TYLCV DNA,进行克隆并确定序列。这三个序列的核苷酸同一性为99.6%至100%,因此仅将一个序列存入NCBI GenBank数据库(登录号EF110890)(1)。对完整基因组核苷酸序列的分析证实了通过核心Cp分析预测的TYLCV同一性,其与来自埃及(GenBank登录号AY594174)和西班牙(GenBank登录号AJ489258)的TYLCV同一性为98.1%,与来自墨西哥(GenBank登录号DQ631892)的TYLCV同一性为97.6%,与TYLCV - Is(GenBank登录号X15656)同一性为96.5%。此外,以TYLCV为探针的Southern杂交检测到在所有样本中均存在复制型(双链)TYLCV DNA,这些样本包括来自乌瓦尔德县的三株植物和来自贝克斯县的21株植物。据我们所知,这是得克萨斯州TYLCV的首次报道,该病毒出现在相距约400公里的两个移栽苗生产区。在乌瓦尔德县和贝克斯县生产的移栽苗在当地种植,而伊达尔戈县的移栽苗则运往粉虱通常分布范围之外的地区。伊达尔戈县具有亚热带气候,这可能使TYLCV和粉虱传播媒介得以越冬,从而使该病毒在未来得以定殖和传播。参考文献:(1)J.K. Brown和A.M. Idris。植物病害。90:1360,2006。(2)J.K. Brown等人。病毒学档案。146:1581,2001。(3)A.M. Idris和J.K. Brown。植物病理学。88:648,1998。(4)J.E. Polston等人。植物病害。83:984,1999。