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Please use this identifier to cite or link to this item: http://hdl.handle.net/10400.5/1523

Title: Agrobacterium-mediated transformation and insertional mutagenesis in Colletotrichum acutatum for investigating varied pathogenicity lifestyles
Authors: Oliveira, Helena
Talhinhas, Pedro
Martins, João Neves
Muthumeenakshi, S.
Sreenivasaprasad, S.
Keywords: colletotrichum acutatum
agrobacterium tumefaciens-mediated transformation
hygromycin B resistance
green fluorescent protein expression
insertional mutagenesis
pathogenicity mutants
Issue Date: 2008
Publisher: Humana Press Inc.
Citation: "Molecular Biotechnology". ISSN 1073-6085. 39:1 (2008) 57-67
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 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.
Description: Research
URI: http://hdl.handle.net/10400.5/1523
ISSN: 1073-6085
Appears in Collections:SSPV - Artigos de Revistas

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