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Research agrobacterium-mediated transformation and insertional mutagenesis in colletotrichum acutatum for investigating varied pathogenicity lifestyles.
| Content Provider | CiteSeerX |
|---|---|
| Author | Sreenivasaprasad, S. |
| Abstract | 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 labori-ous and inefficient. We have developed a protocol for efficient Agrobacterium tumefaciens-mediated transforma-tion (ATMT) of C. acutatum. Using this protocol we were able to transform C. acutatum isolates belonging to dif-ferent 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 identifi-cation of the genes associated with the varied lifestyles. |
| File Format | |
| Access Restriction | Open |
| Subject Keyword | Colletotrichum Acutatum Varied Pathogenicity Lifestyle Research Agrobacterium-mediated Transformation Insertional Mutagenesis Single Location Single Copy Integration Transformation Method Varied Lifestyle Wide Range Molecular Mechanism Transformation Efficiency Rapid Generation Acutatum Strain Green Fluorescence Protein Tagging Cosmopolitan Pathogen Dif-ferent Genetic Group Different Host Atmt Protocol Putative Pathogenicity Mutant Binary Vector Pbht2-gfp Important Disease Efficient Agrobacterium Tumefaciens-mediated Transforma-tion Pathogenicity Lifestyle Functional Analysis Involve Protoplast Transformation Histopathology Study Useful Tool Gene Discovery Large Panel Fungus Exhibit Abstract Colletotrichum Acutatum Insertional Mutant |
| Content Type | Text |
