| |
|
|
 | Acesso ao texto completo restrito à biblioteca da Embrapa Agroenergia. Para informações adicionais entre em contato com biblioteca@embrapa.br. |
|
Registro Completo |
|
Biblioteca(s): |
Embrapa Agroenergia. |
|
Data corrente: |
12/11/2013 |
|
Data da última atualização: |
26/12/2018 |
|
Tipo da produção científica: |
Artigo em Periódico Indexado |
|
Autoria: |
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. |
|
Afiliação: |
Maria Beatriz Calderan Rodrigues, Universidade de São Paulo; LEIA CECILIA DE LIMA FAVARO, CNPAE; Ana Paula de Souza Pallu, Universidade de São Paulo; ANDERSON FERREIRA, CPAMT; Fernanda de Souza Sebastianes, Universidade de São Paulo; Maria Juliana Calderan Rodrigues, Universidade de São Paulo; Marcel Bellato Sposito, Universidade de São Paulo; Welington Luiz de Araujo, University of Mogi das Cruzes; Aline Aparecida Pizzirani-Kleiner, Universidade de São Paulo. |
|
Título: |
Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis. |
|
Ano de publicação: |
2013 |
|
Fonte/Imprenta: |
Fungal Biology, v. 117, n. 7-8, p. 463-576, 2013. |
|
DOI: |
10.1016/j.funbio.2013.06.005 |
|
Idioma: |
Inglês |
|
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 con-ditions (type of membrane; inductor agent concentration [acetosyringone e AS] and cocul-tivation time). Results demonstrated that the best condition occurred with the utilization of cellulose?s ester membrane; 200 m M 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 muta-genesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plante 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 con-ditions (type of membrane; inductor agent concentration [acetosyringone e AS] and cocul-tivation time). Results demonstrated that the best condition occurred with the utilization of cellulose?s ester membrane; 200 m M 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 f... Mostrar Tudo |
|
Palavras-Chave: |
Agrobacterium-mediated; Plant e pathogen interaction; Transformation. |
|
Thesaurus Nal: |
citrus black spot; genetic transformation; Guignardia citricarpa. |
|
Categoria do assunto: |
-- |
|
Marc: |
LEADER 02846naa a2200301 a 4500
001 1970991
005 2018-12-26
008 2013 bl uuuu u00u1 u #d
024 7 $a10.1016/j.funbio.2013.06.005$2DOI
100 1 $aRODRIGUES, M. B. C.
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 con-ditions (type of membrane; inductor agent concentration [acetosyringone e AS] and cocul-tivation time). Results demonstrated that the best condition occurred with the utilization of cellulose?s ester membrane; 200 m M 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 muta-genesis and genetic knockouts, in order to identify the most important genes involved in the pathogenesis mechanisms and plante pathogen interaction.
650 $acitrus black spot
650 $agenetic transformation
650 $aGuignardia citricarpa
653 $aAgrobacterium-mediated
653 $aPlant e pathogen interaction
653 $aTransformation
700 1 $aFAVARO, L. C. de L.
700 1 $aPALUU, A. P. de S.
700 1 $aFERREIRA, A.
700 1 $aSEBASTIANES, F. de S.
700 1 $aRODRIGUES, M. J. C.
700 1 $aSPOSITO, M. B.
700 1 $aARAUJO, W. L. de
700 1 $aPIZZIRANI-KLEINER, A. A.
773 $tFungal Biology$gv. 117, n. 7-8, p. 463-576, 2013.
Download
Esconder MarcMostrar Marc Completo |
|
Registro original: |
Embrapa Agroenergia (CNPAE) |
|
|
Biblioteca |
ID |
Origem |
Tipo/Formato |
Classificação |
Cutter |
Registro |
Volume |
Status |
URL |
Voltar
|
|
|
| Registros recuperados : 2 | |
| Registros recuperados : 2 | |
|
| Nenhum registro encontrado para a expressão de busca informada. |
|
|
