| |
|
|
|
Registro Completo |
|
Biblioteca(s): |
Embrapa Agrossilvipastoril. |
|
Data corrente: |
03/02/2014 |
|
Data da última atualização: |
10/08/2017 |
|
Tipo da produção científica: |
Artigo em Periódico Indexado |
|
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. |
|
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. |
|
Título: |
Agrobacterium-mediated transformation of Guignardia citricarpa: an efficient tool to gene transfer and random mutagenesis. |
|
Ano de publicação: |
2013 |
|
Fonte/Imprenta: |
Fungal Biology, Oxford, v. 117, n. 7/8, p. 556-568, jul./aug., 2013. |
|
ISSN: |
1878-6146 |
|
DOI: |
http://dx.doi.org/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 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 |
|
Palavras-Chave: |
Agrobacterium-Mediated; Plant-Pathogen Interaction. |
|
Thesaurus Nal: |
citrus black spot; genetic transformation; Guignardia citricarpa. |
|
Categoria do assunto: |
-- |
|
URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/96268/1/cpamt-Ferreira-1878-6146-2013.pdf
|
|
Marc: |
LEADER 02874naa a2200301 a 4500
001 1978312
005 2017-08-10
008 2013 bl uuuu u00u1 u #d
022 $a1878-6146
024 7 $ahttp://dx.doi.org/10.1016/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 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.
Download
Esconder MarcMostrar Marc Completo |
|
Registro original: |
Embrapa Agrossilvipastoril (CPAMT) |
|
|
Biblioteca |
ID |
Origem |
Tipo/Formato |
Classificação |
Cutter |
Registro |
Volume |
Status |
URL |
Voltar
|
|
|
| Registros recuperados : 8 | |
| 4. |  | RIBEIRO, M. A. C.; CARVALHO, J. E. U. de; NASCIMENTO, W. M. O. do. Enxertia da gravioleira (Annona muricata L.) em porta-enxertos dos gêneros Annona e Rollinia. In: SEMINÁRIO DE INICIAÇÃO CIENTÍFICA DA FCAP, 10.; SEMINÁRIO DE INICIAÇÃO CIENTÍFICA DA EMBRAPA AMAZÔNIA ORIENTAL, 4., 2000, Belém, PA. Resumos. Belém, PA: FCAP, 2000. p. 137-139.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Amazônia Oriental. |
|    |
| 5. | | RIBEIRO, M. A. C.; CARVALHO, J. E. U. de; NASCIMENTO, W. M. O. do. Efeitos do dessecamento sobre a germinação de sementes de Araticum-do-brejo (Annona glabra). In: SEMINÁRIO DE INICIAÇÃO CIENTÍFICA DA FCAP, 9.; SEMINÁRIO DE INICIAÇÃO CIENTÍFICA DA EMBRAPA AMAZÔNIA ORIENTAL, 3., 1999, Belém, PA. Resumos. Belém, PA: FCAP: Embrapa Amazônia Oriental, 1999. p. 68-69.| Biblioteca(s): Embrapa Amazônia Oriental. |
| |
| 6. | | CARVALHO, C. J. R. de; CARVALHO, J. E. U. de; RIBEIRO, M. A. C. Relações hídricas de gravioleiras (Annona muricata L.) jovens em porta-enxertos dos gêneros Annona e Rollinia. In: CONGRESSO BRASILEIRO DE FRUTICULTURA, 17., 2002, Belém, PA. Anais. Belém: SBF, 2002. 4 f. Publicado também: FRAZÃO, D. A. C.; HOMMA, A. K. O; VIÉGAS, I. de J. M. (Ed.). Contribuição ao desenvolvimento da fruticultura na Amazônia. Belém, PA: Embrapa Amazônia Oriental, 2006. p. 543-548.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Amazônia Oriental. |
| |
| 8. | | RIBEIRO, M. A. C.; CARVALHO, J. E. U. de; MÜLLER, C. H.; NASCIMENTO, W. M. O. do. Enxertia da gravioleira (Annona muricata L.) em porta-enxertos dos generos Annona e Rollinia. In: CONGRESSO BRASILEIRO DE FRUTICULTURA, 17., 2002, Belém, PA. Anais. Belém: SBF, 2002. 3f. Publicado também: FRAZÃO, D. A. C.; HOMMA, A. K. O; VIÉGAS, I. de J. M. (Ed.). Contribuição ao desenvolvimento da fruticultura na Amazônia. Belém, PA: Embrapa Amazônia Oriental, 2006. p. 537-542.| Tipo: Artigo em Anais de Congresso |
| Biblioteca(s): Embrapa Amazônia Oriental. |
| |
| Registros recuperados : 8 | |
|
| Nenhum registro encontrado para a expressão de busca informada. |
|
|
