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

2.50
Hdl Handle:
http://hdl.handle.net/10547/302152
Title:
Agrobacterium-mediated transformation and insertional mutagenesis in colletotrichum acutatum for investigating varied pathogenicity lifestyles
Authors:
Talhinhas, Pedro; Muthumeenakshi, S.; Neves-Martins, João; Oliveira, Helena; Sreenivasaprasad, Surapareddy
Abstract:
Colletotrichum 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 10(4) 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.
Affiliation:
Technical University of Lisbon; University of Warwick
Citation:
Talhinhas, P., Muthumeenakshi, S., Neves-Martins, J., Oliveira, H. and Sreenivasaprasad, S. (2008) 'Agrobacterium-Mediated Transformation and Insertional Mutagenesis in Colletotrichum acutatum for Investigating Varied Pathogenicity Lifestyles, Molecular Biotechnology, 39(1),pp.57-67
Publisher:
Humana Press
Journal:
Molecular Biotechnology
Issue Date:
2008
URI:
http://hdl.handle.net/10547/302152
DOI:
10.1007/s12033-007-9028-1
Additional Links:
http://link.springer.com/10.1007/s12033-007-9028-1
Type:
Article
Language:
en
ISSN:
1073-6085; 1559-0305
Appears in Collections:
Environmental Monitoring Research Group

Full metadata record

DC FieldValue Language
dc.contributor.authorTalhinhas, Pedroen_GB
dc.contributor.authorMuthumeenakshi, S.en_GB
dc.contributor.authorNeves-Martins, Joãoen_GB
dc.contributor.authorOliveira, Helenaen_GB
dc.contributor.authorSreenivasaprasad, Surapareddyen_GB
dc.date.accessioned2013-09-24T09:08:43Z-
dc.date.available2013-09-24T09:08:43Z-
dc.date.issued2008-
dc.identifier.citationTalhinhas, P., Muthumeenakshi, S., Neves-Martins, J., Oliveira, H. and Sreenivasaprasad, S. (2008) 'Agrobacterium-Mediated Transformation and Insertional Mutagenesis in Colletotrichum acutatum for Investigating Varied Pathogenicity Lifestyles, Molecular Biotechnology, 39(1),pp.57-67en_GB
dc.identifier.issn1073-6085-
dc.identifier.issn1559-0305-
dc.identifier.doi10.1007/s12033-007-9028-1-
dc.identifier.urihttp://hdl.handle.net/10547/302152-
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 10(4) 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.en_GB
dc.language.isoenen
dc.publisherHumana Pressen_GB
dc.relation.urlhttp://link.springer.com/10.1007/s12033-007-9028-1en_GB
dc.rightsArchived with thanks to Molecular Biotechnologyen_GB
dc.subjectpathogenicity mutantsen_GB
dc.subjectcolletotrichum acutatumen_GB
dc.subjectagrobacterium tumefaciens-mediated transformationen_GB
dc.subjectgreen fluorescent protein expressionen_GB
dc.subjectinsertional mutagenesisen_GB
dc.subjecthygromycin B resistanceen_GB
dc.titleAgrobacterium-mediated transformation and insertional mutagenesis in colletotrichum acutatum for investigating varied pathogenicity lifestylesen
dc.typeArticleen
dc.contributor.departmentTechnical University of Lisbonen_GB
dc.contributor.departmentUniversity of Warwicken_GB
dc.identifier.journalMolecular Biotechnologyen_GB
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