Detection of Closterovirus in grapes and partial characterization of an isolate of Grapevine leafroll-associated virus 3

O enrolamento da folha da videira (Vitis spp.) é uma doença causada por até oito vírus, Grapevine leafroll-associated virus (GLRaV) 1 a 8, sorologicamente distintos e associados ao floema de videiras infetadas. Neste trabalho, foram detectados GLRaV-1 e -3 por DAS-ELISA em 6,9 e 14,7% das amostras analisadas, respectivamente, e provenientes de duas importantes regiões vitícolas do Brasil (Serra Gaúcha e Vale do São Francisco). Os GLRaV-2, -5 e -7 não foram detectados. O GLRaV-3 também foi detectado por dot-ELISA e western blot, observando-se a provável proteína capsidial com cerca de 36 kDa. Um fragmento de 340 pb, compreendendo o terminal 3' do gene da polimerase viral de GLRaV-3, foi amplificado por PCR e seqüenciado. As seqüências de nucleotídeos e aminoácidos deduzidos deste isolado apresentaram alta homologia, 95,0 e 97,1%, respectivamente, com outro isolado de GLRaV-3 (NY1).Grapevine (Vitis spp.) leafroll is a disease caused by up to eight viruses, denominated Grapevine leafroll-associated viruses (GLRaV- 1 to -8), which are serologically unrelated and associated with phloem tissue. In the present paper, GLRaV-1 and -3 were detected by DAS-ELISA in 6.9 and 14.7% of the samples, respectively, and GLRaV-2, -5 and -7 were not detected in grapes. The analysed samples came from two of the most important grape growing regions in Brazil (Serra Gaúcha and Vale do São Francisco). The GLRaV-3 was also identified by dot-ELISA and western blot, associated with a probable coat protein subunit of approximately 36 kDa. A fragment of 340 bp, comprising the 3' terminal of the viral polymerase gene, was PCR-amplified and sequenced. The nucleotide and deduced amino acid sequences of this isolate showed high homology, 95 and 97.1%, respectively, with another GLRaV-3 isolate (NY1)