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Hunting down fungal secretomes using liquid-phase IEF prior to high resolution 2-DE

hal.structure.identifierBiodiversité, Gènes & Communautés [BioGeCo]
dc.contributor.authorVINCENT, Delphine
hal.structure.identifierBIOlogie et GEstion des Risques en agriculture [BIOGER]
dc.contributor.authorBALESDENT, Marie-Helene
hal.structure.identifierInteractions Arbres-Microorganismes [IAM]
dc.contributor.authorGIBON, Julien
hal.structure.identifierUniversité de Bordeaux Ségalen [Bordeaux 2]
dc.contributor.authorCLAVEROL, Stéphane
hal.structure.identifierUniversité de Bordeaux Ségalen [Bordeaux 2]
dc.contributor.authorLAPAILLERIE, Delphine
hal.structure.identifierUniversité de Bordeaux Ségalen [Bordeaux 2]
dc.contributor.authorLOMENECH, Anne-Marie
hal.structure.identifierBIOlogie et GEstion des Risques en agriculture [BIOGER]
dc.contributor.authorBLAISE, Francoise
hal.structure.identifierBIOlogie et GEstion des Risques en agriculture [BIOGER]
dc.contributor.authorROUXEL, Thierry
hal.structure.identifierInteractions Arbres-Microorganismes [IAM]
dc.contributor.authorMARTIN, Francis
hal.structure.identifierUniversité de Bordeaux Ségalen [Bordeaux 2]
dc.contributor.authorBONNEU, Marc
hal.structure.identifierUnité de Recherche Génomique Info [URGI]
dc.contributor.authorAMSELEM, Joelle
hal.structure.identifierUnité de Recherche Génomique Info [URGI]
dc.contributor.authorDOMINGUEZ, Victoria
hal.structure.identifierschool of botany
dc.contributor.authorHOWLETT, Barbara J.
hal.structure.identifier*
dc.contributor.authorWINCKER, Patrick
hal.structure.identifierGénétique Quantitative et Evolution - Le Moulon (Génétique Végétale) [GQE-Le Moulon]
hal.structure.identifierCentre National de la Recherche Scientifique [CNRS]
hal.structure.identifierAgroParisTech
dc.contributor.authorJOETS, Johann
hal.structure.identifierCentre National de la Recherche Scientifique [CNRS]
dc.contributor.authorLEBRUN, Marc-Henri
hal.structure.identifierBiodiversité, Gènes & Communautés [BioGeCo]
dc.contributor.authorPLOMION, Christophe
dc.date.issued2009
dc.identifier.issn0173-0835
dc.description.abstractEnThe secreted proteins (secretome) of fungi play a key role in interactions of pathogenic and symbiotic fungi with plants. Using the plant pathogenic fungus Leptosphaeria maculans and symbiont Laccaria bicolor grown in culture, we have established a proteomic protocol for extraction, concentration and resolution of the fungal secretome. As no proteomic data were available on mycelium tissues from both L. maculans and L. bicolor, mycelial proteins were studied; they also helped verifying the purity of secretome samples. The quality of protein extracts was initially assessed by both 1-DE and 2-DE using first a broad pH range for IEF, and then narrower acidic and basic pH ranges, prior to 2-DE. Compared with the previously published protocols for which only dozens of 2-D spots were recovered from fungal secretome samples, up to approximately 2000 2-D spots were resolved by our method. MS identification of proteins along several pH gradients confirmed this high resolution, as well as the presence of major secretome markers such as endopolygalacturonases, -glucanosyltransferases, pectate lyases and endoglucanases. Shotgun proteomic experiments evidenced the enrichment of secreted protein within the liquid medium. This is the first description of the proteome of L. maculans and L. bicolor, and the first application of liquid-phase IEF to any fungal extracts.
dc.language.isoen
dc.publisherWiley-VCH Verlag
dc.subjectLEPTOSPHAERIA MACULAN
dc.subject2-DE
dc.subjectSECRETOME FONGIQUE
dc.subject.enLACCARIA BICOLOR
dc.subject.enLIQUID-PHASE IEF
dc.subject.enPROTEOMIC
dc.title.enHunting down fungal secretomes using liquid-phase IEF prior to high resolution 2-DE
dc.typeArticle de revue
dc.identifier.doi10.1002/elps.200900415
dc.subject.halSciences du Vivant [q-bio]
bordeaux.journalElectrophoresis
bordeaux.page4118-4136
bordeaux.volume30
bordeaux.issue23
bordeaux.peerReviewedoui
hal.identifierhal-02656973
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-02656973v1
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