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ABSTRACT Plants are colonized by phylogenetically diverse microorganisms that affect plant growth and health. Representative genome-sequenced culture collections of bacterial isolates from

model plants, including _Arabidopsis thaliana_, have recently been established. These resources provide opportunities for systematic interaction screens combined with genome mining to

discover uncharacterized natural products. Here, we report on the biosynthetic potential of 224 strains isolated from the _A. thaliana_ phyllosphere. Genome mining identified more than 1,000

predicted natural product biosynthetic gene clusters (BGCs), hundreds of which are unknown compared to the MIBiG database of characterized BGCs. For functional validation, we used a

high-throughput screening approach to monitor over 50,000 binary strain combinations. We observed 725 inhibitory interactions, with 26 strains contributing to the majority of these. A

combination of imaging mass spectrometry and bioactivity-guided fractionation of the most potent inhibitor, the BGC-rich _Brevibacillus_ sp. Leaf182, revealed three distinct natural product

scaffolds that contribute to the observed antibiotic activity. Moreover, a genome mining-based strategy led to the isolation of a _trans_-acyltransferase polyketide synthase-derived

antibiotic, macrobrevin, which displays an unprecedented natural product structure. Our findings demonstrate that the phyllosphere is a valuable environment for the identification of

antibiotics and natural products with unusual scaffolds. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS

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Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS EXPLORING THE SPECIALIZED METABOLOME OF THE PLANT PATHOGEN _STREPTOMYCES_ SP. 11-1-2 Article Open access 06

May 2024 GENOMIC AND METABOLOMIC INSIGHTS INTO THE ANTIMICROBIAL COMPOUNDS AND PLANT GROWTH-PROMOTING POTENTIAL OF _BACILLUS VELEZENSIS_ B115 Article Open access 28 March 2025 CORRELATIVE

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financially supported by SNF grant NRP72 to J.P. and J.A.V. and by European Research Council Advanced Grants (PhyMo to J.A.V. and SynPlex to J.P.). AUTHOR INFORMATION Author notes * These

authors contributed equally: Eric J. N. Helfrich, Christine M. Vogel. AUTHORS AND AFFILIATIONS * Institute of Microbiology, ETH Zurich, Zurich, Switzerland Eric J. N. Helfrich, Christine M.

Vogel, Reiko Ueoka, Martin Schäfer, Florian Ryffel, Daniel B. Müller, Silke Probst, Markus Kreuzer, Jörn Piel & Julia A. Vorholt Authors * Eric J. N. Helfrich View author publications

You can also search for this author inPubMed Google Scholar * Christine M. Vogel View author publications You can also search for this author inPubMed Google Scholar * Reiko Ueoka View

author publications You can also search for this author inPubMed Google Scholar * Martin Schäfer View author publications You can also search for this author inPubMed Google Scholar *

Florian Ryffel View author publications You can also search for this author inPubMed Google Scholar * Daniel B. Müller View author publications You can also search for this author inPubMed 

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author inPubMed Google Scholar * Jörn Piel View author publications You can also search for this author inPubMed Google Scholar * Julia A. Vorholt View author publications You can also

search for this author inPubMed Google Scholar CONTRIBUTIONS E.J.N.H., C.M.V., R.U., M.S., F.R., D.B.M., J.P. and J.A.V. designed the research. C.M.V., M.S., F.R., D.B.M. and M.K. performed

binary interaction screens. E.J.N.H., C.M.V., F.R. and S.P. performed genome mining studies. C.M.V. and D.B.M. conducted statistical analyses. E.J.N.H., C.M.V. and M.S. conducted MALDI

imaging experiments. E.J.N.H., C.M.V., M.S., F.R. and S.P. conducted bioassays. E.J.N.H., C.M.V., R.U., F.R. and S.P. isolated and structure-elucidated metabolites. M.S. generated

_Brevibacillus_ knockout mutants. E.J.N.H., C.M.V., D.B.M., J.P. and J.A.V. wrote the manuscript with contributions from all authors. CORRESPONDING AUTHORS Correspondence to Jörn Piel or

Julia A. Vorholt. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing interests. ADDITIONAL INFORMATION PUBLISHER’S NOTE: Springer Nature remains neutral with regard to

jurisdictional claims in published maps and institutional affiliations. SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary Figures 1–48, Supplementary Results 1, Supplementary

Methods, Supplementary References. REPORTING SUMMARY SUPPLEMENTARY TABLE Supplementary Tables 1–26. RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE

Helfrich, E.J.N., Vogel, C.M., Ueoka, R. _et al._ Bipartite interactions, antibiotic production and biosynthetic potential of the _Arabidopsis_ leaf microbiome. _Nat Microbiol_ 3, 909–919

(2018). https://doi.org/10.1038/s41564-018-0200-0 Download citation * Received: 22 September 2017 * Accepted: 18 June 2018 * Published: 23 July 2018 * Issue Date: August 2018 * DOI:

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