Use este identificador para citar ou linkar para este item: http://www.alice.cnptia.embrapa.br/alice/handle/doc/978312
Unidade da Embrapa/Coleção:: Embrapa Agrossilvipastoril - Artigo em periódico indexado (ALICE)
Data do documento: 3-Fev-2014
Tipo do Material: Artigo em periódico indexado (ALICE)
Autoria: RODRIGUES, M. B. C.
FAVARO, L. C. de L.
PALLU, A. P. de S.
FERREIRA, A.
SEBASTIANES, F. de S.
RODRIGUES, M. J. C.
SPÓSITO, M. B.
ARAÚJO, W. L. de
PIZZIRANI-KLEINER, A. A.
Informaçães Adicionais: Maria Beatriz Calderan Rodrigues, USP-ESALQ; LEIA CECILIA DE LIMA FAVARO, CNPAE; Ana Paula de Souza Pallu, USP-ESALQ; ANDERSON FERREIRA, CPAMT; Fernanda de Souza Sebastianes, USP-ESALQ; Maria Juliana Calderan Rodrigues, USP-ESALQ; Marcel Bellato Spósito, USP-ESALQ; Welington Luiz DE Araújo, UMC; Aline Aparecida Pizzirani-Kleiner, USP-ESALQ.
Título: Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis.
Fonte/Imprenta: Fungal Biology, Oxford, v. 117, n. 7/8, p. 556-568, jul./aug., 2013.
Idioma: en
Palavras-chave: Agrobacterium-Mediated
Citrus Black Spot
Genetic Transformation
Guignardia Citricarpa
Plant-Pathogen Interaction
Conteúdo: Guignardia citricarpa is the causal agent of Citrus Black Spot (CBS), an important disease in Citriculture. Due to the expressive value of this activity worldwide, especially in Brazil, understanding more about the functioning of this fungus is of utmost relevance, making possible the elucidation of its infection mechanisms, and providing tools to control CBS. This work describes for the first time an efficient and successful methodology for genetic transformation of G. citricarpa mycelia, which generated transformants expressing the gene encoding for the gfp (green fluorescent protein) and also their interaction with citrus plant. Mycelia of G. citricarpa were transformed via Agrobacterium tumefaciens, which carried the plasmid pFAT-gfp, contains the genes for hygromycin resistance (hph) as well as gfp. The optimization of the agrotransformation protocol was performed testing different conditions (type of membrane; inductor agent concentration [acetosyringonee - AS] and cocultivation time). Results demonstrated that the best condition occurred with the utilization of cellulose?s ester membrane; 200µM of AS and 96 h as cocultivation time. High mitotic stability (82 %) was displayed by transformants using Polymerase Chain Reaction (PCR) technique to confirm the hph gene insertion. In addition, the presence of gfp was observed inside mycelia by epifluorescence optical microscopy. This technique easy visualization of the behaviour of the pathogen interacting with the plant for the first time, allowing future studies on the pathogenesis of this fungus. The establishment of a transformation method for G. citricarpa opens a range of possibilities and facilitates the study of insertional mutagenesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plant-pathogen interaction.
Ano de Publicação: 2013
ISSN: 1878-6146
Aparece nas coleções:Artigo em periódico indexado (CPAMT)

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