Melzer M J, Ogata D Y, Fukuda S K, Shimabuku R, Borth W B, Sether D M, Hu J S
Plant and Environmental Protection Sciences, University of Hawaii, Honolulu.
Agricultural Diagnostic Services Center, University of Hawaii, Honolulu.
Plant Dis. 2010 May;94(5):641. doi: 10.1094/PDIS-94-5-0641B.
Tomato yellow leaf curl disease, caused by the begomovirus Tomato yellow leaf curl virus (TYLCV; family Geminiviridae), is an economically important disease of tomato (Solanum lycopersicum L.) that can be very destructive in tropical and subtropical regions (1). In October 2009, tomato plants showing stunted new growth, interveinal chlorosis, and upward curling of leaf margins were reported by a residential gardener in Wailuku, on the island of Maui. Similar symptoms were observed in approximately 200 tomato plants at a University of Hawaii research farm in Poamoho, on the island of Oahu in November 2009. The similarity between these symptoms and those of tomato yellow leaf curl disease and the presence of whiteflies (Bemisia spp.), the vector of TYLCV, suggested the causal agent was a geminivirus such as TYLCV. Total nucleic acids were extracted from a tomato plant sample from Wailuku and Poamoho and used in a PCR assay with degenerate primers PAR1c715 and PAL1v1978 for geminivirus detection (4). The ~1.5-kbp amplicon expected to be produced from a geminivirus template was generated from the symptomatic tomato plant samples but not from a greenhouse-grown control tomato plant. The amplicons were cloned by the pGEM-T Easy vector (Promega, Madison, WI). Three clones from each sample were sequenced, revealing 97 to 99% nucleotide identity to TYLCV sequences in GenBank and a 98.9% nucleotide identity between the Wailuku (Accession No. GU322424) and Poamoho (Accession No. GU322423) isolates. A multiplex PCR assay for the detection and discrimination between the IL and Mld clades of TYLCV was also performed on these isolates (2). A ~0.8-kbp amplicon was generated from both isolates confirming the presence of TYLCV and their inclusion into the TYLCV-IL clade (2). Seven symptomatic and three asymptomatic tomato plant samples from Poamoho were tested for TYLCV using a squash-blot hybridization assay (3) utilizing a digoxigenin-labeled probe derived from the ~1.5-kbp PCR amplicon. All symptomatic tomato plants and one asymptomatic tomato plant were found to be infected with TYLCV. How the virus entered Hawaii and how long it has been present is unknown. The most plausible route is through infected plant material such as an asymptomatic alternative host rather than viruliferous whiteflies. It appears TYLCV is not a recent introduction into Hawaii since the Wailuku gardener observed similar disease symptoms for a few years before submitting samples for testing. In January 2010, TYLCV was also detected in two commercial tomato farms on Oahu, posing a serious threat to the state's $10 million annual tomato crop. References: (1) H. Czosnek and H. Laterrot. Arch. Virol. 142:1392, 1997. (2) P. Lefeuvre et al. J. Virol. Methods 144:165, 2007. (3) N. Navot et al. Phytopathology 79:562, 1989. (4) M. R. Rojas et al. Plant Dis. 77:340, 1993.
番茄黄化曲叶病由双生病毒科的番茄黄化曲叶病毒(TYLCV)引起,是番茄(Solanum lycopersicum L.)的一种具有重要经济影响的病害,在热带和亚热带地区可能极具破坏性(1)。2009年10月,毛伊岛怀卢库的一位园艺爱好者报告称,番茄植株出现新梢生长受阻、脉间黄化以及叶缘向上卷曲的症状。2009年11月,在瓦胡岛波莫霍的夏威夷大学研究农场的约200株番茄植株上也观察到了类似症状。这些症状与番茄黄化曲叶病的症状相似,且存在TYLCV的传播媒介粉虱(烟粉虱属),这表明致病因子是一种双生病毒,如TYLCV。从怀卢库和波莫霍的番茄植株样本中提取了总核酸,并用于使用简并引物PAR1c715和PAL1v1978进行双生病毒检测的PCR分析(4)。从有症状的番茄植株样本中产生了预期由双生病毒模板产生的约1.5kbp扩增子,但从温室种植的对照番茄植株中未产生。扩增子通过pGEM-T Easy载体(Promega,麦迪逊,威斯康星州)进行克隆。对每个样本的三个克隆进行测序,结果显示与GenBank中TYLCV序列的核苷酸同一性为97%至99%,怀卢库分离株(登录号GU322424)和波莫霍分离株(登录号GU322423)之间的核苷酸同一性为98.9%。还对这些分离株进行了用于检测和区分TYLCV的IL和Mld分支的多重PCR分析(2)。从两个分离株中均产生了约0.8kbp的扩增子,证实了TYLCV的存在以及它们属于TYLCV-IL分支(2)。使用源自约1.5kbp PCR扩增子的地高辛标记探针,通过压片杂交分析(3)对波莫霍的七个有症状和三个无症状番茄植株样本进行了TYLCV检测。发现所有有症状的番茄植株和一株无症状番茄植株感染了TYLCV。病毒如何进入夏威夷以及它已存在多长时间尚不清楚。最合理的途径是通过受感染的植物材料,如无症状的替代寄主,而不是带毒粉虱。由于怀卢库的园艺爱好者在提交样本进行检测前几年就观察到了类似的病害症状,因此TYLCV似乎不是最近才传入夏威夷的。2010年1月,在瓦胡岛的两个商业番茄农场也检测到了TYLCV,这对该州每年价值1000万美元的番茄作物构成了严重威胁。参考文献:(1)H. Czosnek和H. Laterrot。Arch. Virol. 142:1392,1997。(2)P. Lefeuvre等人。J. Virol. Methods 144:165,2007。(3)N. Navot等人。Phytopathology 79:562,1989。(4)M. R. Rojas等人。Plant Dis. 77:340,1993。