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As principais espécies de vírus envolvidas na etiologia do enrolamento da folha da videira (Vitis spp.) são Grapevine leafroll-associated virus 1 e 3 (GLRaV-1 e -3). Neste estudo da variabilidade desses vírus, foram amplificados dois fragmentos de DNA (396 bp do GLRaV-1 e 602 bp do GLRaV-3) por RT-PCR, a partir de RNA total extraído de nervuras e pecíolos de videiras infetadas, utilizando-se dois pares de oligonucleotídeos. Os DNAs amplificados foram clonados e reamplificados, a partir dos clones recombinantes, e comparados quanto às diferenças conformacionais das fitas simples desnaturadas (SSCP). Foram observados dois padrões distintos de perfis eletroforéticos para cada vírus, tendo sido seqüenciado pelo menos um clone viral correspondente a cada padrão. As duas seqüências de nucleotídeos obtidas para o GLRaV-1 apresentaram maior homologia (79,8% e 87,4%) com um isolado australiano e as duas seqüências relativas ao GLRaV-3 exibiram maior homologia (75,1% e 81,8%) com um isolado norte-americano. Os resultados demonstraram a ocorrência de seqüências variantes destes vírus nas videiras analisadas. The main viral species associated with grapevine (Vitis spp.) leafroll etiology are Grapevine leafroll-associated virus 1 and 3 (GLRaV-1 and -3). To partially evaluate the variability of these viruses, two DNA fragments (396 bp of GLRaV-1 and 602 bp of GLRaV-3) were amplified by RT-PCR using two sets of primers after total RNA extraction from veins and petioles of infected grapevines. The amplified DNA fragments were cloned and the reamplified viral DNAs, obtained from the recombinant clones, were studied with regard to denatured single-strand DNA conformational differences. Two different electrophoretic profiles were observed for each virus species. At least one viral clone representing each mobility profile was sequenced. The two nucleotide sequences obtained for GLRaV-1 showed the highest homologies (79,8% and 87,4%) with an Australian isolate, whereas the two nucleotide sequences of GLRaV-3 exhibited the highest homologies (75,1% and 81,8%) with a North American isolate. These results demonstrated the occurrence of GLRaV-1 and -3 sequence variants infecting the analysed grapevines

fitopatologia brasileira

Avaliação da variabilidade do Grapevine leafroll-associated virus 1 e 3 por análise de seqüências de nucleotídeos e polimorfismo conformacional de fita simples