Detection and identification of TMV infecting tomato under protected cultivation in Paraná State
subject: P30 protein, TMV, plasticulture
description: During an inspection in plastic houses in Sapopema, Paraná, 90% of tomato plants showed leaf abnormalities, probably associated with herbicide toxity. However, virus like symptoms developed in selected hosts after mechanical inoculatation. RT-PCR reactions using primers for an internal region within the movement protein gene of TMV and ToMV resulted in the amplification of a 409 bp cDNA fragment only by TMV primers. Deduced amino acids showed 100% identity when compared to TMV movement protein and 94% with ToMV. The RT-PCR protocol was efficient for quick and conclusive determination of virus species. The virus was purified and a polyclonal antiserum was raised for future surveys in tomato crops of Paraná. The partial genomic sequence obtained for TMV-Sapopema has been deposited under the accession number DQ173945, which is the first partial genomic sequence of an isolate of TMV from Brazil in the GenBank, and the first tomato virus isolate from Paraná to have some of its biological and molecular properties determined.Durante uma inspeção em cultivos protegidos de tomate em Sapopema, Paraná, foram observadas anormalidades foliares em 90% das plantas, indicando possivelmente a existência de um problema de fitotoxidade causada por herbicidas. Todavia, os sintomas manifestados nas hospedeiras após os ensaios de inoculação mecânica revelaram que os sintomas estariam relacionados a uma infecção por Tobamovirus. As reações de RT-PCR com oligonucleotídeos específicos para uma região interna da proteína de movimento de dois vírus comuns em tomate, TMV e ToMV, resultaram na amplificação de um fragmento de 409 pares de bases, apenas com os oligonucleotídeos específicos para o TMV. Após o sequenciamento, os aminoácidos deduzidos apresentaram identidade de 100% quando comparados com as seqüências das proteínas de movimento de outros isolados do TMV, e 94% de identidade com seqüências do ToMV. A RT-PCR demonstrou ser um método eficiente para a rápida e conclusiva determinação da…
creator: Rodrigo Martins da Silva, Eliezer Rodrigues de Souto, Júlio Cézar Pedroso, Ricardo Arakava, Álvaro Manuel Rodrigues Almeida, Antônio Augusto Lazarini Barboza, João Batista Vida
source: Brazilian Archives of Biology and Technology
publisher: Instituto de Tecnologia do Paraná (Tecpar)
date: 2008
relation: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000500005
coverage: Biotechnology