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多米尼加共和国加工番茄中番茄褪绿斑点病毒的首次报道。

First Report of Tomato chlorotic spot virus in Processing Tomatoes in the Dominican Republic.

作者信息

Batuman O, Rojas M R, Almanzar A, Gilbertson R L

机构信息

Department of Plant Pathology, University of California-Davis, CA 95616.

Transagricola, S. A. Av. Duarte 269 Navarrete, Santiago, Dominican Republic Apartado 713.

出版信息

Plant Dis. 2014 Feb;98(2):286. doi: 10.1094/PDIS-07-13-0685-PDN.

Abstract

Processing tomatoes (Solanum lycopersicum) are an important industry in the Dominican Republic. In November 2012, symptoms typical of tospovirus infection (bronzing, chlorosis, and necrosis of leaves) appeared in numerous processing tomato fields in the North (>50% incidence in some fields) and a few fields in the South (<1% incidence). Plants in affected fields had large populations of thrips on leaves and in flowers. Symptomatic leaves from four fields in the North (Guayubin, Juan Gomez, Hatillo Palma, and Navarrete) and one field in the South (Azua) were positive for infection by Tomato spotted wilt virus (TSWV) when tested with AgDia immunostrips. However, RT-PCR tests of these samples with a TSWV N gene primer pair (1) were negative, whereas the expected size 590 and 777 bp fragments were amplified with N gene primers for Groundnut ringspot virus (GRSV, 2) and Tomato chlorotic spot virus (TCSV; NF5'ATGTCTAAGGTCAAGCTCACC3' and NR5'TTATGCAACACCTGAAATTTTGGC3'), respectively. These fragments were sequenced (KF420087 and KF420088) and comparisons revealed 99, 83, and 80% identities with N gene sequences of TCSV, GRSV, and TSWV, respectively. Portions of the L, M, and S RNAs were amplified from symptomatic leaves by RT-PCR with degenerate L (TOSPO L For: CWGARGATRTDATWATAAATAAYAATGC and TOSPO L Rev: GCATCNACAGAWATYTTCCA), M (TOSPO M For: AGAGCAATCAGTGCATC and TOSPO M Rev: CTTRCAGGCTTCAATRAAKGC), and S (3) primers. The expected L, M, and S RNA fragments of 450, 849, and 871 bp, respectively, were amplified and sequenced (KF420089, KF420090, and KF420091). Sequence comparisons revealed 98, 83, and 78%; 99, 94, and 82%; and 99, 83, and 77% identities with TCSV-, GRSV-, and TSWV-L, M, and S RNA sequences, respectively. Weed surveys around tomato fields revealed tospovirus symptoms (chlorosis, mosaic/mottle, and necrosis) in leaves of two common species, Boerhavia erecta and Cleome viscosa. Symptomatic leaves were positive with TSWV immunostrips, whereas RT-PCR and sequence analyses of these leaves from C. viscosa (one each from the North and South) and B. erecta (one from the South) revealed infection with TCSV (99% identities for L, M, and S RNA fragments). In contrast, leaves from pepper plants with tospovirus symptoms (chlorosis, ringspots, and necrosis) in a commercial greenhouse in the North (Villa Gonzales) were positive for TSWV based on immunostrips and RT-PCR and sequence analyses. Dot blot hybridization tests with the cloned TCSV L RNA fragment confirmed TCSV infection in PCR-positive tomato plants and weeds, whereas no hybridization signal was detected for TSWV-infected peppers or uninfected tomatoes. Identification of thrips collected from symptomatic tomato plants at Navarrete and Hatillo Palma revealed that tomato thrips (Frankliniella schultzei) was predominant (90%) along with Western flower thrips (F. occidentalis) (10%), whereas only F. schultzei was identified from weeds in the South. Thus, TCSV is causing the tospovirus disease of processing tomato, and this is the first report of this virus in the Dominican Republic. This is also consistent with F. schultzei being an efficient vector of TCSV. An IPM program for TCSV based on planting thrips- and virus-free transplants and resistant varieties, roguing symptomatic plants, thrips monitoring and management, and area-wide sanitation is being implemented. References: (1) H. R. Pappu et al. Tobacco Sci. 40:74, 1996. (2) C. G. Webster et al. Virol. 413:216, 2011. (3) R. J. Weeks et al. Acta Hort. 431:159, 1996.

摘要

加工番茄(番茄)是多米尼加共和国的一项重要产业。2012年11月,北部众多加工番茄田(部分田块发病率超过50%)以及南部少数田块(发病率低于1%)出现了番茄斑萎病毒感染的典型症状(叶片青铜化、黄化和坏死)。受影响田块的植株叶片和花朵上有大量蓟马。用AgDia免疫试纸检测时,北部四个田块(瓜尤宾、胡安·戈麦斯、哈蒂略·帕尔马和纳瓦雷特)以及南部一个田块(阿苏阿)的有症状叶片对番茄斑萎病毒(TSWV)呈阳性感染。然而,用TSWV N基因引物对(1)对这些样品进行RT-PCR检测结果为阴性,而用花生环斑病毒(GRSV,2)和番茄褪绿斑点病毒(TCSV;NF5'ATGTCTAAGGTCAAGCTCACC3' 和NR5'TTATGCAACACCTGAAATTTTGGC3')的N基因引物分别扩增出了预期大小为590和777 bp的片段。对这些片段进行测序(KF420087和KF420088)并比较后发现,它们与TCSV、GRSV和TSWV的N基因序列的同源性分别为99%、83%和80%。用简并引物L(TOSPO L For: CWGARGATRTDATWATAAATAAYAATGC和TOSPO L Rev: GCATCNACAGAWATYTTCCA)、M(TOSPO M For: AGAGCAATCAGTGCATC和TOSPO M Rev: CTTRCAGGCTTCAATRAAKGC)和S(3)从有症状叶片中通过RT-PCR扩增出L、M和S RNA的部分片段。分别扩增出了预期大小为450、849和871 bp的L、M和S RNA片段并进行测序(KF420089、KF420090和KF420091)。序列比较显示,它们与TCSV、GRSV和TSWV的L、M和S RNA序列的同源性分别为98%、83%和78%;99%、94%和82%;以及99%、83%和77%。对番茄田周围的杂草调查发现,两种常见杂草直立博落回和粘毛醉蝶花的叶片出现了番茄斑萎病毒症状(黄化、花叶/斑驳和坏死)。有症状叶片用TSWV免疫试纸检测呈阳性,而对来自南部和北部的各一株粘毛醉蝶花以及来自南部的一株直立博落回的这些叶片进行RT-PCR和序列分析后发现感染了TCSV(L、M和S RNA片段的同源性为99%)。相比之下,北部(比利亚·冈萨雷斯)一个商业温室中出现番茄斑萎病毒症状(黄化、环斑和坏死)的辣椒植株叶片,基于免疫试纸检测以及RT-PCR和序列分析,对TSWV呈阳性。用克隆的TCSV L RNA片段进行斑点杂交试验证实了PCR阳性的番茄植株和杂草感染了TCSV,而未检测到TSWV感染的辣椒或未感染的番茄有杂交信号。对从纳瓦雷特和哈蒂略·帕尔马有症状番茄植株上采集的蓟马进行鉴定发现,番茄蓟马(西花蓟马)占主导(90%),西部花蓟马(西花蓟马)占10%,而从南部杂草中仅鉴定出了番茄蓟马。因此,TCSV正在引发加工番茄的番茄斑萎病毒病,这是该病毒在多米尼加共和国的首次报道。这也与番茄蓟马是TCSV的有效传播媒介相一致。正在实施一项针对TCSV的综合虫害管理计划,该计划包括种植无蓟马和无病毒的移栽苗及抗性品种、拔除有症状植株、监测和管理蓟马以及进行区域卫生清理。参考文献:(1)H. R. Pappu等人,《烟草科学》40:74,1996年。(2)C. G. Webster等人,《病毒学》413:216,2011年。(3)R. J. Weeks等人,《园艺学报》431:159,1996年。

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