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Agrobacterium-mediated transformation and insertional mutagenesis in Colletotrichum acutatum for investigating varied pathogenicity lifestyles

dc.contributor.authorOliveira, Helena
dc.contributor.authorTalhinhas, Pedro
dc.contributor.authorMartins, João Neves
dc.contributor.authorMuthumeenakshi, S.
dc.contributor.authorSreenivasaprasad, S.
dc.date.accessioned2009-11-24T15:56:48Z
dc.date.available2009-11-24T15:56:48Z
dc.date.issued2008
dc.descriptionResearchpt
dc.description.abstractColletotrichum acutatum is a cosmopolitan pathogen causing economically important diseases known as anthracnose on a wide range of hosts. This fungus exhibits varied pathogenicity lifestyles and the tools essential to understand the molecular mechanisms are still being developed. The transformation methods currently available for this species for gene discovery and functional analysis involve protoplast transformation and are laborious and inefficient. We have developed a protocol for efficient Agrobacterium tumefaciens-mediated transformation (ATMT) of C. acutatum. Using this protocol we were able to transform C. acutatum isolates belonging to different genetic groups and originating from different hosts. The transformation efficiency was up to 156 transformants per 104 conidia, with[70% transformants showing single location/single copy integration of T-DNA. Binary vector pBHt2-GFP was constructed, enabling green fluorescence protein tagging of C. acutatum strains, which will be a useful tool for epidemiology and histopathology studies. The ATMT protocol developed was used to identify putative pathogenicity mutants, suggesting the applicability of this technique for rapid generation of a large panel of insertional mutants of C. acutatum leading to the identification of the genes associated with the varied lifestyles.pt
dc.identifier.citation"Molecular Biotechnology". ISSN 1073-6085. 39:1 (2008) 57-67pt
dc.identifier.issn1073-6085
dc.identifier.urihttp://hdl.handle.net/10400.5/1523
dc.language.isoengpt
dc.publisherHumana Press Inc.pt
dc.subjectcolletotrichum acutatumpt
dc.subjectagrobacterium tumefaciens-mediated transformationpt
dc.subjecthygromycin B resistancept
dc.subjectgreen fluorescent protein expressionpt
dc.subjectinsertional mutagenesispt
dc.subjectpathogenicity mutantspt
dc.titleAgrobacterium-mediated transformation and insertional mutagenesis in Colletotrichum acutatum for investigating varied pathogenicity lifestylespt
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage67pt
oaire.citation.startPage57pt
oaire.citation.titleMolecular Biotechnologypt
person.familyNameOliveira
person.familyNameTalhinhas
person.givenNameHelena
person.givenNamePedro
person.identifier.ciencia-id001D-2C12-36BE
person.identifier.ciencia-id5A1E-9983-ADD5
person.identifier.orcid0000-0002-8274-9757
person.identifier.orcid0000-0001-6694-7502
person.identifier.ridJ-4784-2013
person.identifier.scopus-author-id8551918500
person.identifier.scopus-author-id6506057702
rcaap.rightsopenAccesspt
rcaap.typearticlept
relation.isAuthorOfPublication99632a6b-4e76-4dee-bc6c-65b8dd136dc4
relation.isAuthorOfPublicationeba8a4d6-04bf-4aa6-8a7d-e0bcef764ae7
relation.isAuthorOfPublication.latestForDiscovery99632a6b-4e76-4dee-bc6c-65b8dd136dc4

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