Agrobacterium-mediated transformation and insertional mutagenesis in colletotrichum acutatum for investigating varied pathogenicity lifestyles
Authors
Talhinhas, PedroMuthumeenakshi, S.
Neves-Martins, João
Oliveira, Helena
Sreenivasaprasad, Surapareddy
Issue Date
2008Subjects
pathogenicity mutantscolletotrichum acutatum
agrobacterium tumefaciens-mediated transformation
green fluorescent protein expression
insertional mutagenesis
hygromycin B resistance
Metadata
Show full item recordAbstract
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.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.57-67Publisher
Humana PressJournal
Molecular BiotechnologyAdditional Links
http://link.springer.com/10.1007/s12033-007-9028-1Type
ArticleLanguage
enISSN
1073-60851559-0305
ae974a485f413a2113503eed53cd6c53
10.1007/s12033-007-9028-1