Affiliation:
1. Instituto Potosino de Investigación Científica y Tecnológica, A.C., San Luis Potosí, S.L.P., México 78216
2. Facultad de Agronomía, Universidad Autónoma de San Luis Potosí, P.O. Box 32, San Luis Potosí, S. L. P. México 78321
Abstract
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.
Subject
Plant Science,Agronomy and Crop Science