Navegando por Autor "Costa, Roberta de Paula Saturnino"

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    Validação de um sistema para análise funcional de genes em Phakopsora pachyrhizi via silenciamento gênico mediado por aplicação ectópica de dsRNA
    (Universidade Estadual do Norte do Paraná, 2018-07-31) Costa, Roberta de Paula Saturnino; Carvalho, Mayra Costa da Cruz Gallo de; https://orcid.org/0000-0002-1559-3828; http://lattes.cnpq.br/2844751503530944; Carvalho, Mayra Costa da Cruz Gallo de; https://orcid.org/0000-0002-1559-3828; http://lattes.cnpq.br/2844751503530944; Torres, João Pereira; https://orcid.org/0000-0002-7306-9872; http://lattes.cnpq.br/2582950118304206; Carvalho, Kenia de; http://lattes.cnpq.br/6278825285024494; Seiva, Fábio Rodrigues Ferreira; https://orcid.org/0000-0002-7461-8773; http://lattes.cnpq.br/2646377436835972; Carvalho, Sandremir de; http://lattes.cnpq.br/8684697240694261
    RNA interference (RNAi) induced by direct dsRNA application has emerged as a promising strategy both for functional gene analysis and for the management of pests and pathogens. Although the RNA-silencing machinery of Phakopsora pachyrhizi remains uncharacterized, the prospect of applying this approach to an obligate biotrophic pathogen that severely impacts soybean, one of the world’s most important commodities, is particularly attractive. We sprayed GFP-dsRNA directly onto detached soybean leaves and observed that dsRNA was taken up by the foliage, remained detectable throughout the fungal infection cycle, and was even transmitted to the first generation of spores. GFP-dsRNA fluorescence was observed predominantly in the spore germ tubes. We then targeted a predicted essential gene from P. pachyrhizi (de_novo_4692) and recorded a dose-dependent silencing efficiency of up to 86.6%, with the strongest effect occurring at the lowest dose tested (50 ng/μL). Disease-severity assays on silenced detached leaves revealed a significant reduction in symptom development. These results demonstrate that foliar dsRNA application effectively triggers target-gene silencing in P. pachyrhizi, indicating that this approach holds promise for functional genomic studies and represents an innovative alternative to conventional fungicides.