First Report of Grapevine Virus A in Spain

Grapevine trichovirus A (GVA), a flexuous, filamentous, phloem-limited virus with an approximately 7.3-kbp RNA genome, is widespread in grapevines showing symptoms of leafroll and/or rugose wood. The virus can be mechanically inoculated to Nicotiana benthamiana and N. clevelandii. A field survey of diseased Vitis vinifera white and red cultivars was carried out in Pontevedra (northwest Spain) during the autumn of 1993. We detected the presence of GVA in vines showing leafroll symptoms by an immunocapture-reverse transcription-polymerase chain reaction (PCR) method (2) with GVA-specific primers (1). Bands of the expected size of 430 bp were obtained with extracts from petioles and stem bark as reaction substrates. To verify these results, Northern (RNA) blots with double-stranded (ds) RNAs isolated from grapevines were prepared. Hybridization was positive in two out of 10 samples analyzed. The probe was a ³²P-labeled 430-bp PCR product amplified from extracts of N. benthamiana plants infected with GVA strain Is151 (gift of A. Mina-fra). The specificity of this probe was confirmed in dot blot hybridization, as a positive signal was obtained with extracts from GVA-inoculated N. benthamiana, but not with extracts of phosphate buffer-inoculated N. benthamiana, turnip mosaic potyvirus-inoculated Arabidopsis thaliana, or potato potyvirus Y-inoculated N. xanthi plants. The probe did not hybridize to dsRNAs extracted from enzyme-linked immunosorbent assay-positive GLRaV-III-infected grapevines. GVA has been identified in other Mediterranean countries, but to our knowledge this is the first report of the detection of GVA in Spain. References: (1) A. Minafra and A. Hadidi. J. Virol. Methods 47:175, 1994. (2) G. Nolasco et al. J. Virol. Methods 45:201, 1993.