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bioRxiv · 10.1101/2020.08.05.238063

Genomic and chemical diversity of Bacillus subtilis secondary metabolites against plant pathogenic fungi

Kiesewalter, H. T.·Lozano Andrade, C. N.·Wibowo, M.·Lenz Strube, M.·Maroti, G.·Snyder, D.·Jorgensen, T. S.·Larsen, T. O.·Cooper, V. S.·Weber, T.·Kovacs, A. T.
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Abstract

Bacillus subtilis produces a wide range of secondary metabolites providing diverse plant-growth-promoting and biocontrol abilities. These secondary metabolites include non-ribosomal peptides (NRPs) with strong antimicrobial properties, causing either cell lysis, pore formation in fungal membranes, inhibition of certain enzymes, or bacterial protein synthesis. However, the natural products of B. subtilis are mostly studied either in laboratory strains or in individual isolates and therefore, a comparative overview of B. subtilis secondary metabolites is missing. In this study, we have isolated 23 B. subtilis strains from eleven sampling sites, compared the fungal inhibition profiles of wild types and their NRPs mutants, followed the production of targeted lipopeptides, and determined the complete genomes of 13 soil isolates. We discovered that non-ribosomal peptide production varied among B. subtilis strains co-isolated from the same soil samples. In vitro antagonism assays revealed that biocontrol properties depend on the targeted plant pathogenic fungus and the tested B. subtilis isolate. While plipastatin alone is sufficient to inhibit Fusarium sp., a combination of plipastatin and surfactin is required to hinder the growth of Botrytis cinerea. Detailed genomic analysis revealed that altered NRP production profiles in certain isolates is due to missing core genes, nonsense mutation, or potentially altered gene regulation. Our study combines microbiological antagonism assays with chemical NRPs detection and biosynthetic gene cluster predictions in diverse B. subtilis soil isolates to provide a broader overview of the secondary metabolite chemodiversity of B. subtilis. IMPORTANCESecondary or specialized metabolites with antimicrobial activities define the biocontrol properties of microorganisms. Members of the Bacillus genus produce a plethora of secondary metabolites, of which non-ribosomally produced lipopeptides in particular display strong antifungal activity. To facilitate prediction of the biocontrol potential of new Bacillus subtilis isolates, we have explored the in vitro antifungal inhibitory profiles of recent B. subtilis isolates, combined with analytical natural product chemistry, mutational analysis, and detailed genome analysis of biosynthetic gene clusters. Such a comparative analysis helped to explain why selected B. subtilis isolates lack production of certain secondary metabolites.

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Kiesewalter, H. T., Lozano Andrade, C. N., Wibowo, M., Lenz Strube, M., Maroti, G., Snyder, D., Jorgensen, T. S., Larsen, T. O., Cooper, V. S., Weber, T., Kovacs, A. T.. 2020-08-05. Genomic and chemical diversity of Bacillus subtilis secondary metabolites against plant pathogenic fungi. https://doi.org/10.1101/2020.08.05.238063

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