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

Molecular Biotechnology
Pedro TalhinhasSurapareddy Sreenivasaprasad

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 o...Continue Reading

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Citations

Jan 10, 2009·Molecular Plant-microbe Interactions : MPMI·Aurélie HuserRichard O'Connell
Dec 14, 2011·AIDS Research and Treatment·Laura SaganicAnn Duerr
Nov 5, 2011·Journal of Basic Microbiology·Adelene Shu Mei AuyongPaul William James Taylor
Jun 30, 2015·Environmental Microbiology·Jian ZhangIrina S Druzhinina
Sep 3, 2013·Fungal Genetics and Biology : FG & B·Paulo de BoerRemko Offringa
Aug 13, 2013·Fungal Biology·Maria Beatriz Calderan RodriguesAline Aparecida Pizzirani-Kleiner
Jun 3, 2016·Journal of Basic Microbiology·Meenakshi SharmaSaurabh Kulshrestha

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