Volatile organic compounds (VOCs) produced by soil bacteria have been shown to exert plant pathogen biocontrol potential owing to their strong antimicrobial activity. While the impact of VOCs on soil microbial ecology is well established, their effect on plant pathogen evolution is yet poorly understood. Here we experimentally investigated how plant-pathogenic Ralstonia solanacearum bacterium adapts to VOC-mixture produced by a biocontrol Bacillus amyloliquefaciens T-5 bacterium and how these adaptations might affect its virulence. We found that VOC selection led to a clear increase in VOC-tolerance, which was accompanied with cross-tolerance to several antibiotics commonly produced by soil bacteria. The increasing VOC-tolerance led to trade-offs with R. solanacearum virulence, resulting in almost complete loss of pathogenicity in planta. At the genetic level, these phenotypic changes were associated with parallel mutations in genes encoding lipopolysaccharide O-antigen (wecA) and type-4 pilus biosynthesis (pilM), which both have been linked with outer membrane permeability to antimicrobials and plant pathogen virulence. Reverse genetic engineering revealed that both mutations were important, with pilM having a relatively larger negative effect on the virulence, while wecA having a relatively larger effect on increased antimicrobial tolerance. Together, our results suggest that microbial VOCs are important drivers of bacterial evolution and could potentially be used in biocontrol to select for less virulent pathogens via evolutionary trade-offs.
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The genome sequences of ancestral, No-VOC and H-VOC clones of Ralstonia solanacearum QL-Rs1115 are available at NCBI Sequence Read Archive (SRA) under the accession number SAMN29421919. The rest of the data are available in the article and its online supplementary material.
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This work was supported by the National Natural Science Foundation of China (Nos. 42090064, 42090062, 42007038, 31972504, 32170180, 42277113, 41922053) to ZW, GJ and QS, and the National Key Research and Development Program of China (No. 2018YFD1000800), the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement (No. 838710-ReproDev to WR), the Royal Society (No. RSG\R1\180213 and No. CHL\R1\180031 to VPF) and jointly by a grant from UKRI, Defra, and the Scottish Government, under the Strategic Priorities Fund Plant Bacterial Diseases Programme (No. BB/T010606/1 to VPF), the Natural Science Foundation of Jiangsu Province (No. BK20190518), the Fundamental Research Funds for the Central Universities (grant no. XUEKEN2022025), and the Jiangxi Branch of China National Tobacco Corporation, China (grant no. 2021.01.010).
The authors declare no competing interests.
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Wang, J., Raza, W., Jiang, G. et al. Bacterial volatile organic compounds attenuate pathogen virulence via evolutionary trade-offs. ISME J 17, 443–452 (2023). https://doi.org/10.1038/s41396-023-01356-6