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与墨西哥下加利福尼亚州番茄病害相关的番茄传染性褪绿病毒

Tomato infectious chlorosis virus Associated with Tomato Diseases in Baja California, Mexico.

作者信息

Méndez-Lozano J, Magallanes-Tapia M A, Romero-Romero J L, Camacho-Beltrán E, Vega W L Orduño, Leyva-López N E, Santos-Cervantes M E, Félix-Gastélum R

机构信息

Instituto Politécnico Nacional, CIIDIR, Unidad Sinaloa, Departamento de Biotecnología Agrícola, Maestría en Recursos Naturales y Medio Ambiente, Blvd. Juan de Dios Bátiz Paredes No. 250, Col. San Joachín, Guasave, Sinaloa, México C.P. 81101.

Universidad de Occidente, Unidad Los Mochis, Blvd. Macario Gaxiola y Carretera Internacional s/n, Los Mochis, Sinaloa, México C.P. 81223.

出版信息

Plant Dis. 2012 Aug;96(8):1229. doi: 10.1094/PDIS-02-12-0196-PDN.

Abstract

Tomato (Solanum lycopersicum L.) is an important vegetable crop in Mexico. The national production in 2009 was 2,043,814 metric tons with a value of $163,560,636 US. Since 2007, abnormal yellow and crispy leaves were observed in commercial tomato fields in Ensenada County, Baja California, Mexico. In affected fields from two localities (San Quintín Valley and Ensenada), symptomatic plants were randomly distributed and symptoms resembled previous descriptions of crinivirus infections in tomato (3). The symptoms and the presence of whiteflies (Bemisia tabaci and Trialeurodes vaporariorum) in the affected fields suggested a viral etiology. Leaf samples of 143 symptomatic tomato plants were collected in the 2007 and 2008 growing seasons. Total RNA was extracted and analyzed by reverse transcription (RT)-PCR assay for simultaneous detection of Tomato infectious chlorosis virus (TICV) and Tomato chlorosis virus (ToCV). Degenerate primers (HS-11/HS-12) were used in combination with specific primers (TIC-3/TIC-4 and ToC-5/ToC-6) for detection of these viruses by nested-PCR (2). A PCR fragment of the expected size for TICV (223 bp) was amplified in 26 of 143 samples. None of the samples tested positive for ToCV. In addition, considering that whiteflies are vectors of begomoviruses, samples were also tested for presence of viral DNA. Results showed 30 positive samples and one with mixed infection. It is therefore possible that the viral disease symptoms observed could be caused in part by viruses other than TICV. Three amplicons from RT-PCR of tomato samples were cloned into the pGEM-T easy vector system II (Promega Corporation, Madison, WI) and sequenced. The sequence of one amplicon (GenBank Accession No. FJ609651) was compared with the sequences of other criniviruses reported in the NCBI/GenBank database using the Clustal V alignment method of the sequence analysis software suite Lasergene (MegAling, DNASTAR Inc., Madison, WI). Sequence analysis of the 223-bp PCR fragment corresponding to TICV showed 99.1% identity with a TICV isolate from Japan (GenBank Accession No. AB085602) and 100% identity with TICV isolates from the United States (GenBank Accession No. TIU67449). Although the presence of another crinivirus, ToCV, was reported previously in Mexico associated with tomato crops and two native weeds, S. nigrescens and Datura stramonium (1), this virus was not detected in Baja California during the present work. To our knowledge, this is the first report of TICV associated with tomato diseases in Mexico. The emerging of a previously unreported virus disease in tomato production areas of Mexico complicates disease management efforts. References: (1) P. Álvarez-Ruíz et al. Plant Pathol. 56:1043, 2007. (2) C. I. Dovas et al. Plant Dis. 86:1345, 2002. (3) G. C. Wisler et al. Plant Dis. 82:270, 1998.

摘要

番茄(Solanum lycopersicum L.)是墨西哥一种重要的蔬菜作物。2009年全国产量为2,043,814公吨,价值163,560,636美元。自2007年以来,在墨西哥下加利福尼亚州恩塞纳达县的商业番茄田中观察到叶片异常发黄和发脆的现象。在来自两个地区(圣昆廷谷和恩塞纳达)的受影响田块中,有症状的植株随机分布,症状与先前描述的番茄褪绿病毒感染相似(3)。受影响田块中出现的症状以及粉虱(烟粉虱和温室白粉虱)表明该病由病毒引起。在2007年和2008年生长季收集了143株有症状番茄植株的叶片样本。提取总RNA并通过逆转录(RT)-PCR检测法同时检测番茄传染性褪绿病毒(TICV)和番茄褪绿病毒(ToCV)。使用简并引物(HS-11/HS-12)与特异性引物(TIC-3/TIC-4和ToC-5/ToC-6)通过巢式PCR检测这些病毒(2)。在143个样本中的26个样本中扩增出了预期大小(223 bp)的TICV PCR片段。没有样本检测出ToCV呈阳性。此外,考虑到粉虱是双生病毒的传播媒介,还对样本进行了病毒DNA检测。结果显示有30个阳性样本,其中1个为混合感染。因此,观察到的病毒病症状可能部分是由TICV以外的病毒引起的。将番茄样本RT-PCR的三个扩增子克隆到pGEM-T easy载体系统II(Promega公司,威斯康星州麦迪逊)中并进行测序。使用序列分析软件套件Lasergene(MegAling,DNASTAR公司,威斯康星州麦迪逊)的Clustal V比对方法,将一个扩增子的序列(GenBank登录号FJ609651)与NCBI/GenBank数据库中报道的其他褪绿病毒序列进行了比较。对与TICV对应的223 bp PCR片段的序列分析表明,其与来自日本的TICV分离株(GenBank登录号AB085602)的同一性为99.1%,与来自美国的TICV分离株(GenBank登录号TIU67449)的同一性为100%。尽管先前报道在墨西哥与番茄作物以及两种本地杂草(龙葵和曼陀罗)相关的另一种褪绿病毒ToCV存在(1),但在本次研究中在下加利福尼亚州未检测到该病毒。据我们所知,这是墨西哥首次报道TICV与番茄病害相关。墨西哥番茄产区出现一种先前未报道的病毒病使病害管理工作变得复杂。参考文献:(1)P. Álvarez-Ruíz等人,《植物病理学》56:1043,2007年。(2)C. I. Dovas等人,《植物病害》86:1345,2002年。(3)G. C. Wisler等人,《植物病害》82:270,1998年。

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