TY - CONF

T1 - Microbial biosynthesis of secondary metabolites involved in biocontrol

AU - Hennessy, Rosanna Catherine

AU - Michelsen, Charlotte Frydenlund

AU - Olsson, Stefan

AU - Stougaard, Peter

PY - 2015

Y1 - 2015

N2 - The impact of microbial diseases on crop production coupled with increasing antibioticresistance emphasizes the need for alternative agricultural methods that can reduceenvironmental impact and do not rely on chemical pesticides. Microbial biocontrol agents(mBCAs) could potentially provide effective and safe strategies to overcome currentconstraints. The Greenlandic soil-bacterium Pseudomonas fluorescens In5 is a promisingbiocontrol agent that has previously been shown to produce bioactive compounds againstfungal pathogens1,2. Genome sequencing and analysis of In5 identified large secondarymetabolite biosynthesis gene clusters. A combination of random and targeted mutagenesis,together with MALDI-TOF imaging mass spectrometry, linked two non-ribosomal peptides(NRPs) designated nunapeptin and nunamycin respectively, to antifungal activity againstRhizoctonia solani, Pythium aphanidermatum and Fusarium graminearum1, 2. In order tounravel the complex genetic regulation of these large NRP synthetase gene clusters,antisense RNAs (asRNAs) and CRISPR/Cas9 based systems are being tested anddeveloped as tools to target transcripts of interest and elucidate gene function3, 4. Toinvestigate the effect of purified nunamycin and nunapeptin at the omics level againstpathogenic fungi, an NRP production platform is being developed which, could additionallyprovide a source of antifungal compounds for industrial applications (e.g. food production,pharmaceutical, personal care). Methods for direct cloning based on either linear-linearhomologous recombination (LLHR) or direct assembly methods (e.g. Gibson assembly) willbe used to capture, clone and refactor In5 secondary metabolite gene clusters5. The overallaim of this project is to understand how mBCAs communicate in complex communities, theconditions under which these antifungal peptides are synthesised and the compound effectson fungal pathogens. The ultimate goal is to develop a novel and sustainable biocontroltechnology.

AB - The impact of microbial diseases on crop production coupled with increasing antibioticresistance emphasizes the need for alternative agricultural methods that can reduceenvironmental impact and do not rely on chemical pesticides. Microbial biocontrol agents(mBCAs) could potentially provide effective and safe strategies to overcome currentconstraints. The Greenlandic soil-bacterium Pseudomonas fluorescens In5 is a promisingbiocontrol agent that has previously been shown to produce bioactive compounds againstfungal pathogens1,2. Genome sequencing and analysis of In5 identified large secondarymetabolite biosynthesis gene clusters. A combination of random and targeted mutagenesis,together with MALDI-TOF imaging mass spectrometry, linked two non-ribosomal peptides(NRPs) designated nunapeptin and nunamycin respectively, to antifungal activity againstRhizoctonia solani, Pythium aphanidermatum and Fusarium graminearum1, 2. In order tounravel the complex genetic regulation of these large NRP synthetase gene clusters,antisense RNAs (asRNAs) and CRISPR/Cas9 based systems are being tested anddeveloped as tools to target transcripts of interest and elucidate gene function3, 4. Toinvestigate the effect of purified nunamycin and nunapeptin at the omics level againstpathogenic fungi, an NRP production platform is being developed which, could additionallyprovide a source of antifungal compounds for industrial applications (e.g. food production,pharmaceutical, personal care). Methods for direct cloning based on either linear-linearhomologous recombination (LLHR) or direct assembly methods (e.g. Gibson assembly) willbe used to capture, clone and refactor In5 secondary metabolite gene clusters5. The overallaim of this project is to understand how mBCAs communicate in complex communities, theconditions under which these antifungal peptides are synthesised and the compound effectson fungal pathogens. The ultimate goal is to develop a novel and sustainable biocontroltechnology.

M3 - Poster

T2 - Annual Plant Biotech Denmark meeting 2015

Y2 - 28 January 2015 through 29 January 2015

ER -