Transformação de Fusarium oxysporum f. sp. vasinfectum com os genes marcadores GFP e DsRed

• Ano de defesa: 2009
• Autor(a) principal: Pedrozo, Rodrigo
• Orientador(a): Machado, José da Cruz
• Banca de defesa: Guimarães, Renato Mendes
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Lavras; Escola de Ciências Agrárias de Lavras (ESAL)
• Programa de Pós-Graduação: Programa de Pós-Graduação em Agronomia/Fitopatologia
• Departamento: Departamento de Fitopatologia
• País: BRASIL
• Palavras-chave em Português: Algodão; Fusarium oxysporum f. sp. vasinfectum; Transformação genética; Gene repórter; Green Fluorescent Protein (GFP); Red Fluorescente Protein (DsRed); Cotton; Genetic transformation; Report gene
• Área do conhecimento CNPq: Ciências Agrárias
• Link de acesso: https://repositorio.ufla.br/handle/1/3105

Fusarium oxysporum f sp. vasinfectum, the casual agent of fusarium wilt disease of cotton, is frequent in most cotton-producing regions in Brazil. The fungus infects roots, colonizes the rhizomes and eventually blocks the vascular system of the pseudostems, leading to the plant collapse. The Green and Red Fluorescent Proteins (GFP and DsRed) emit green and red fluorescence respectively when excited by blue and green light, making them useful tools to study early pathogen infection and other aspects of interest in plant pathology. In this context the aim of this study was to transform Fusarium oxysporum f sp. vasinfectum isolates with those markers and to compare the transformants with the wild-types isolates. Two isolates, with different geographic origins in Brazil, were transformed with the GFP and DsRed derivative markers using hygromicin B as an indicative marker. The results showed that the transformed isolates did not differ from the wild-type isolates in relation to growth and morphological characteristics in vitro. Analyses in fluorescence microscope showed expression of the green and red fluorescent proteins in the fungal structures. Also, the presence of GFP and DsRed in the fungal isolates was confirmed by PCR using specific primers. In the pathogenicity tests it was observed that the virulence of the trnasformant isolates was maintained. From infected tissues of artificially inoculated plants of cotton the transformed isolates were re-isolated and they expressed both proteins markers in the fluorescence microscope.