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Registro Completo |
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Biblioteca(s): |
Embrapa Agrossilvipastoril. |
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Data corrente: |
03/02/2014 |
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Data da última atualização: |
10/08/2017 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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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. |
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Afiliação: |
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. |
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Título: |
Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis. |
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Ano de publicação: |
2013 |
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Fonte/Imprenta: |
Fungal Biology, Oxford, v. 117, n. 7/8, p. 556-568, jul./aug., 2013. |
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ISSN: |
1878-6146 |
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DOI: |
http://dx.doi.org/10.1016/j.funbio.2013.06.005 |
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Idioma: |
Inglês |
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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. MenosGuignardia 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 ... Mostrar Tudo |
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Palavras-Chave: |
Agrobacterium-Mediated; Plant-Pathogen Interaction. |
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Thesaurus Nal: |
citrus black spot; genetic transformation; Guignardia citricarpa. |
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Categoria do assunto: |
-- |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/96268/1/cpamt-Ferreira-1878-6146-2013.pdf
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Marc: |
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024 7 $ahttp://dx.doi.org/10.1016/j.funbio.2013.06.005$2DOI
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245 $aAgrobacterium-mediated transformation of Guignardia citricarpa$ban efficient tool to gene transfer and random mutagenesis.$h[electronic resource]
260 $c2013
520 $aGuignardia 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.
650 $acitrus black spot
650 $agenetic transformation
650 $aGuignardia citricarpa
653 $aAgrobacterium-Mediated
653 $aPlant-Pathogen Interaction
700 1 $aFAVARO, L. C. de L.
700 1 $aPALLU, A. P. de S.
700 1 $aFERREIRA, A.
700 1 $aSEBASTIANES, F. de S.
700 1 $aRODRIGUES, M. J. C.
700 1 $aSPÓSITO, M. B.
700 1 $aARAÚJO, W. L. de
700 1 $aPIZZIRANI-KLEINER, A. A.
773 $tFungal Biology, Oxford$gv. 117, n. 7/8, p. 556-568, jul./aug., 2013.
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Registro original: |
Embrapa Agrossilvipastoril (CPAMT) |
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| Registros recuperados : 6 | |
| 2. |  | MARTINS, P. C.; CAMPOS, M. M.; JAYME, C. G.; MACHADO, F. S.; TOMICH, T. R.; PEREIRA, L. G. R.; SILVA, R. B. e; ARAÚJO, W. L. de P. Levantamento das técnicas zootécnicas utilizadas na criação de bezerras leiteiras no município de Rio Pomba, MG. In: WORKSHOP DE INICIAÇÃO CIENTÍFICA DA EMBRAPA GADO DE LEITE, 15., 2015, Juiz de Fora, MG. Anais... Juiz de Fora: Embrapa Gado de Leite, 2015. (Embrapa Gado de Leite. Documentos, 176.). 1 p.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Gado de Leite. |
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| 3. | | 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. Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis. Fungal Biology, Oxford, v. 117, n. 7/8, p. 556-568, jul./aug., 2013.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Agrossilvipastoril. |
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| 4. | | RODRIGUES, M. B. C.; FAVARO, L. C. de L.; PALUU, A. P. de S.; FERREIRA, A.; SEBASTIANES, F. de S.; RODRIGUES, M. J. C.; SPOSITO, M. B.; ARAUJO, W. L. de; PIZZIRANI-KLEINER, A. A. Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis. Fungal Biology, v. 117, n. 7-8, p. 463-576, 2013.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Agroenergia. |
|  |
| 5. | | ZWAR, I. P.; TROTTA, C. do V.; ZIOTTI, A. B. S.; LIMA NETO, M.; ARAÚJO, W. L. de; MELO, I. S. de; OTTONI, C. A.; SOUZA, A. O. de. Biosynthesis of silver nanoparticles using actinomycetes, phytotoxicity on rice seeds, and potential application in the biocontrol of phytopathogens. Journal of Basic Microbiology, v. 63, n. 1, p. 64-74, 2022.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
| Biblioteca(s): Embrapa Meio Ambiente. |
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| 6. | | MENDES, T. D.; MORAIS, K. dos S.; MENEZES, B. dos S.; QUIRINO, B. F.; RODRIGUES, D. de S.; FAVARO, L. C. de L.; ARAUJO, W. L. de; MELO, I. S. de; SALUM, T. F. C. Produção de celulases por novas cepas fúngicas isoladas de biomas brasileiros. In: SEMINÁRIO BRASILEIRO DE TECNOLOGIA ENZIMÁTICA, 12., 2014, Rio de Janeiro. Anais... São Paulo: Croda; São Paulo: Analítica, 2014.| Tipo: Resumo em Anais de Congresso |
| Biblioteca(s): Embrapa Agroenergia. |
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| Registros recuperados : 6 | |
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| Nenhum registro encontrado para a expressão de busca informada. |
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