Mauricio-Castillo J A, Argüello-Astorga G R, Ambriz-Granados S, Alpuche-Solís A G, Monreal-Vargas C T
Instituto Potosino de Investigación Científica y Tecnológica, A.C., San Luis Potosí, S.L.P., México 78216.
Facultad de Agronomía, Universidad Autónoma de San Luis Potosí, P.O. Box 32, San Luis Potosí, S. L. P. México 78321.
Plant Dis. 2007 Nov;91(11):1513. doi: 10.1094/PDIS-91-11-1513B.
The Rioverde Valley is an important farming area of the San Luis Potosi State in the north-central region of Mexico, where a variety of horticultural crops (i.e., tomato, pepper, cucumber, and watermelon) are annually cultivated. In the summer of 2005, a number of plants exhibiting a variety of symptoms, including leaf yellowing, curling, and stunted growth, were observed in several tomato (Lycopersicon esculentum L.) fields. The presence of whiteflies (Bemisia tabaci Genn.) and symptoms seemed to suggest a begomoviral etiology. Leaves of 12 symptomatic tomato plants and seven plants of the weed Solanum rostratum (Dunal) growing into the same area were collected in July and September from several fields throughout the Rioverde area and assessed for the presence of begomoviruses (genus Begomovirus, family Geminiviridae) by PCR using the degenerate primers prRepDGR (CCTCCTCTAGCASWTCTNCCGTC), SL2050 (2), and prC889 (3). Amplicons of 1.4 kb were derived from viral DNA-A present in all examined S. rostratum and tomato samples, which were cloned into pGEM-T Easy Vector (Promega, Madison, WI) and subsequently analyzed by restriction fragment length polymorphism (RFLP) using MspI and HinfI. Several restriction fragment patterns were observed among the cloned PCR products, hence indicating the occurrence of different begomoviruses in the sampled fields. Sequencing of amplicons derived from one S. rostratum plant revealed the concurrent presence of Tomato severe leaf curl virus (ToSLCV; GenBank Accession No. DQ347946; [2]) and a distinct virus (GenBank Accession No. EF501978) displaying a high sequence identity with Tomato golden mottle virus from Guatemala (ToGMoV-GT94-R2; GenBank Accession No. AF32852). Restriction fragment patterns identical to that of the ToGMoV-like isolate were found in PCR clones from three additional S. rostratum plants and five tomato samples. A set of partially overlapping PCR products of 1.8 and 1.4 kb encompassing the complete DNA-A component of ToGMoV were obtained from one tomato sample by using two pairs of degenerate primers, prRepQGR-rev and prCP70 (1) and prRepDGR and prC889. Amplicons were cloned, sequenced, and compared with viral sequences available in the GenBank database using BlastN and Clustal V alignments (MegAlign, DNASTAR, Madison, WI). The 2,614-bp DNA-A sequence of the Rioverde isolate (GenBank Accession No. DQ520943) displays 93% sequence identity with the Guatemalan isolate of ToGMoV. In addition, a number of B. tabaci specimens of unidentified biotype were collected in one tomato field and total DNA was isolated from them by a modified Dellaporta method. Amplification of viral DNA present in the whiteflies was carried out and the PCR products were cloned and sequenced. One of the begomoviral DNA-A genomes isolated from the whiteflies (GenBank Accession No. EF501976) displayed 99% sequence identity with the virus isolated from plants. Previously, ToGMoV had been found only in Central America ( http://gemini.biosci.arizona.edu/viruses ), but this report considerably expands its known geographical distribution. References: (1) R. De La Torre-Almaraz et al. Plant Dis. 90:378, 2006. (2) J. A. Mauricio-Castillo et al. Plant Dis. 90:1116, 2006. (3) S. D. Wyatt and J. K. Brown. Phytopathology 86:1288, 1996.
里奥韦尔代河谷是墨西哥中北部圣路易斯波托西州的一个重要农业区,这里每年都会种植多种园艺作物(如番茄、辣椒、黄瓜和西瓜)。2005年夏天,在几个番茄(Lycopersicon esculentum L.)田块中观察到许多出现各种症状的植株,包括叶片发黄、卷曲和生长发育迟缓。烟粉虱(Bemisia tabaci Genn.)的存在以及这些症状似乎表明病因是双生病毒。7月和9月,从里奥韦尔代地区的几个田块中采集了12株有症状的番茄植株以及生长在同一区域的7株杂草刺萼龙葵(Solanum rostratum (Dunal))的叶片,使用简并引物prRepDGR(CCTCCTCTAGCASWTCTNCCGTC)、SL2050 (2) 和prC889 (3) 通过聚合酶链反应(PCR)评估双生病毒(双生病毒属,双生病毒科)的存在情况。所有检测的刺萼龙葵和番茄样本中均存在大小为1.4 kb的扩增子,其来源于病毒DNA-A,将其克隆到pGEM-T Easy载体(普洛麦格公司,威斯康星州麦迪逊市)中,随后使用MspI和HinfI通过限制性片段长度多态性(RFLP)进行分析。在克隆的PCR产物中观察到几种限制性片段模式,因此表明在采样田块中存在不同的双生病毒。对来自一株刺萼龙葵植株的扩增子进行测序,结果显示同时存在番茄严重卷叶病毒(ToSLCV;GenBank登录号:DQ347946;[2])和一种与来自危地马拉的番茄金色斑驳病毒(ToGMoV-GT94-R2;GenBank登录号:AF32852)具有高度序列同一性的独特病毒(GenBank登录号:EF501978)。在另外三株刺萼龙葵植株和五个番茄样本的PCR克隆中发现了与ToGMoV样分离株相同的限制性片段模式。通过使用两对简并引物prRepQGR-rev和prCP70 (1) 以及prRepDGR和prC889,从一个番茄样本中获得了一组大小分别为1.8 kb和1.4 kb的部分重叠PCR产物,其包含ToGMoV完整的DNA-A组分。对扩增子进行克隆、测序,并使用BlastN和Clustal V比对(MegAlign,DNASTAR公司,威斯康星州麦迪逊市)与GenBank数据库中可用的病毒序列进行比较。里奥韦尔代分离株的2614 bp DNA-A序列(GenBank登录号:DQ520943)与危地马拉ToGMoV分离株具有93%的序列同一性。此外,在一个番茄田块中采集了一些未鉴定生物型的烟粉虱样本,并通过改良的Dellaporta方法从它们中分离出总DNA。对烟粉虱中存在的病毒DNA进行扩增,将PCR产物进行克隆和测序。从烟粉虱中分离出的一种双生病毒DNA-A基因组(GenBank登录号:EF501976)与从植物中分离出的病毒具有99%的序列同一性。此前,ToGMoV仅在中美洲被发现(http://gemini.biosci.arizona.edu/viruses),但本报告极大地扩展了其已知的地理分布范围。参考文献:(1) R. De La Torre-Almaraz等人,《植物病害》90:378,2006年。(2) J. A. Mauricio-Castillo等人,《植物病害》90:1116,2006年。(3) S. D. Wyatt和J. K. Brown,《植物病理学》86:1288,1996年。