in

Extracellular vesicle-mediated metabolic exchange shapes the seasonal assembly of aquatic bacterial communities


Abstract

Microorganisms secrete extracellular vesicles (EVs) that transport bioactive molecules, including proteins and metabolites. While their functions are well studied in model microbes, their ecological contributions to natural ecosystems remain largely unexplored. Here we performed an integrative study investigating the role of environmental EVs in shaping microbial community assembly in the Xinglinwan Reservoir. By combining genome-scale metabolic models and multi-omics of field EVs, we found that EVs mediated metabolite exchanges mainly through carrying amino acids, disaccharides, carbohydrate-active enzymes (CAZymes) and signals. EVs can facilitate the growth of amino acid auxotrophic strains. Moreover, EVs act as an external reservoir of functional traits, potentially reinforcing stochastic assembly processes and conferring functional redundancy to the ecosystem. Collectively, our integrative data demonstrate that EV-mediated metabolic exchange is an auxiliary mechanism supplementing classical nutrient transport in aquatic environments. EVs emerge here as a significant, distinct vector in biogeochemical cycling, offering a critical layer for resolving complex natural microbial interactions.

Access through your institution

Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Composition and diversity of bacterial communities vary among different microbial groups in the XLR across three seasons.
Fig. 2: The metabolic exchanges in different EV contributors.
Fig. 3: The metabolite profiling of field EVs from XLR across three seasons and EVs from FBEVs isolated from XLR.
Fig. 4: The metaproteomic profiling of EV-associated proteins.
Fig. 5: The deterministic and stochastic processes in the bacterial community assembly of different EV contributors and their environmental drivers.

Similar content being viewed by others

Spatial transcriptome uncovers rich coordination of metabolism in E. coli K12 biofilm

Extracellular vesicles as structured vectors of quorum sensing signals influence aquatic microbial communities

Evolutionary shift of a tipping point can precipitate, or forestall, collapse in a microbial community

Data availability

All the metagenomic sequencing data has been uploaded and is publicly available in the BIG Submission at https://ngdc.cncb.ac.cn/gsa/browse/CRA027247. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the iProX partner repository (accession: PXD065377). Full mass spectrometry metabolomic data were deposited to MetaboLights (MTBLS12874). Source data are provided with this paper.

References

  1. Chase, J. M. Community assembly: when should history matter?. Oecologia 136, 489–498 (2003).

    Article 
    PubMed 

    Google Scholar 

  2. Zhou, J. et al. Stochasticity, succession, and environmental perturbations in a fluidic ecosystem. Proc. Natl Acad. Sci. USA 111, E836–E845 (2014).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  3. Ke-Chang, N., Yi-Ning, L., Ze-Hao, S., Fang-Liang, H. & Jing-Yun, F. Community assembly: the relative importance of neutral theory and niche theory. Biodivers. Sci. 17, 579–593 (2009).

    Article 

    Google Scholar 

  4. Mo, Y. et al. Biogeographic patterns of abundant and rare bacterioplankton in three subtropical bays resulting from selective and neutral processes. ISME J. 12, 2198–2210 (2018).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  5. Liao, J. et al. The importance of neutral and niche processes for bacterial community assembly differs between habitat generalists and specialists. FEMS Microbiol. Ecol. 92, fiw174 (2016).

    Article 
    PubMed 

    Google Scholar 

  6. Riddley, M. et al. Differential roles of deterministic and stochastic processes in structuring soil bacterial ecotypes across terrestrial ecosystems. Nat. Commun. 16, 2337 (2025).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  7. Zengler, K. & Zaramela, L. S. The social network of microorganisms–how auxotrophies shape complex communities. Nat. Rev. Microbiol. 16, 383–390 (2018).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  8. Wang, M., Chen, X., Tang, Y. Q., Nie, Y. & Wu, X. L. Substrate availability and toxicity shape the structure of microbial communities engaged in metabolic division of labor. mLife 1, 131–145 (2022).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  9. Zhu, L. T. et al. Diverse functional genes harboured in extracellular vesicles from environmental and human microbiota. J. Extracell. Vesicles 11, e12292 (2022).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  10. Biller, S. J. et al. Bacterial vesicles in marine ecosystems. Science 343, 183–186 (2014).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  11. Toyofuku, M., Schild, S., Kaparakis-Liaskos, M. & Eberl, L. Composition and functions of bacterial membrane vesicles. Nat. Rev. Microbiol. 21, 415–430 (2023).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  12. Kost, C., Patil, K. R., Friedman, J., Garcia, S. L. & Ralser, M. Metabolic exchanges are ubiquitous in natural microbial communities. Nat. Microbiol. 8, 2244–2252 (2023).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  13. Schatz, D. & Vardi, A. Extracellular vesicles–new players in cell–cell communication in aquatic environments. Curr. Opin. Microbiol. 43, 148–154 (2018).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  14. Toyofuku, M. et al. Membrane vesicle-mediated bacterial communication. ISME J. 11, 1504–1509 (2017).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  15. Bomberger, J. M. et al. Long-distance delivery of bacterial virulence factors by Pseudomonas aeruginosa outer membrane vesicles. PLoS Pathog. 5, e1000382 (2009).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  16. Weinberger, V. et al. Proteomic and metabolomic profiling of extracellular vesicles produced by human gut archaea. Nat. Commun. 16, 5094 (2025).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  17. Long, L. et al. Extracellular vesicles are prevalent and effective carriers of environmental allergens in indoor dust. Environ. Sci. Technol. 59, 1969–1983 (2025).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  18. Yan, X. et al. Community stability of free-living and particle-attached bacteria in a subtropical reservoir with salinity fluctuations over 3 years. Water Res. 254, 121344 (2024).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  19. Peng, X. et al. iNAP 2.0: harnessing metabolic complementarity in microbial network analysis. iMeta 3, e235 (2024).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  20. Hagemann, M. Molecular biology of cyanobacterial salt acclimation. FEMS Microbiol. Rev. 35, 87–123 (2011).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  21. Ruan, Y. L. Sucrose metabolism: gateway to diverse carbon use and sugar signaling. Annu. Rev. Plant Biol. 65, 33–67 (2014).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  22. Buchan, A., LeCleir, G. R., Gulvik, C. A. & González, J. M. Master recyclers: features and functions of bacteria associated with phytoplankton blooms. Nat. Rev. Microbiol. 12, 686–698 (2014).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  23. Qin, Y. L. et al. Heterotrophic nitrification by Alcaligenes faecalis links organic and inorganic nitrogen metabolism. ISME J. 18, wrae174 (2024).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  24. Starke, S. et al. Amino acid auxotrophies in human gut bacteria are linked to higher microbiome diversity and long-term stability. ISME J. 17, 2370–2380 (2023).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  25. Lima, S., Matinha-Cardoso, J., Tamagnini, P. & Oliveira, P. Extracellular vesicles: an overlooked secretion system in cyanobacteria. Life 10, 129 (2020).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  26. Stegen, J. C. et al. Quantifying community assembly processes and identifying features that impose them. ISME J. 7, 2069–2079 (2013).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  27. Yang, N. J. & Hinner, M. J. in Site-Specific Protein Labeling (Humana, 2015).

  28. Li, J., Azam, F. & Zhang, S. Outer membrane vesicles containing signalling molecules and active hydrolytic enzymes released by a coral pathogen Vibrio shilonii AK1. Environ. Microbiol. 18, 3850–3866 (2016).

    Article 
    PubMed 

    Google Scholar 

  29. Biller, S. J. et al. Prochlorococcus extracellular vesicles: molecular composition and adsorption to diverse microbes. Environ. Microbiol. 24, 420–435 (2022).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  30. Valguarnera, E., Scott, N. E., Azimzadeh, P. & Feldman, M. F. Surface exposure and packing of lipoproteins into outer membrane vesicles are coupled processes in bacteroides. mSphere. 3, e00559–18 (2018).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  31. Biller, S. J. et al. Environmental and taxonomic drivers of bacterial extracellular vesicle production in marine ecosystems. Appl. Environ. Microbiol. 89, e0059423 (2023).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  32. Zhang, H. & Yang, C. Arginine and nitrogen mobilization in cyanobacteria. Mol. Microbiol. 111, 863–867 (2019).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  33. Braakman, R., Follows, M. J. & Chisholm, S. W. Metabolic evolution and the self-organization of ecosystems. Proc. Natl Acad. Sci. USA 114, E3091–E3100 (2017).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  34. Ramoneda, J., Jensen, T. B. N., Price, M. N., Casamayor, E. O. & Fierer, N. Taxonomic and environmental distribution of bacterial amino acid auxotrophies. Nat. Commun. 14, 7608 (2023).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  35. Cackovic, A., Pjevac, P., Orlic, S. & Reintjes, G. Selective heterotopic bacteria can selfishly process polysaccharides in freshwater lakes. Cell Rep. 44, 115415 (2025).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  36. Allison, S. D. Cheaters, diffusion and nutrients constrain decomposition by microbial enzymes in spatially structured environments. Ecol. Lett. 8, 626–635 (2005).

    Article 

    Google Scholar 

  37. Sartorio, M. G., Pardue, E. J., Scott, N. E. & Feldman, M. F. Human gut bacteria tailor extracellular vesicle cargo for the breakdown of diet- and host-derived glycans. Proc. Natl Acad. Sci. USA 120, e2306314120 (2023).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  38. Ramond, P., Galand, P. E. & Logares, R. Microbial functional diversity and redundancy: moving forward. FEMS Microbiol. Rev. 49, fuae031 (2025).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  39. Burke, C., Steinberg, P., Rusch, D., Kjelleberg, S. & Thomas, T. Bacterial community assembly based on functional genes rather than species. Proc. Natl Acad. Sci. USA 108, 14288–14293 (2011).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  40. Rice, E. W., Baird, R. B. & Eaton, A. D. Standard Methods for the Examination of Water and Wastewater 23rd edn (American Water Works Association, 2017).

  41. Mo, Y. et al. Low shifts in salinity determined assembly processes and network stability of microeukaryotic plankton communities in a subtropical urban reservoir. Microbiome 9, 128 (2021).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  42. Bolyen, E. et al. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2. Nat. Biotechnol. 37, 852–857 (2019).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  43. Callahan, B. J. et al. DADA2: high-resolution sample inference from Illumina amplicon data. Nat. Methods 13, 581–583 (2016).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  44. McDonald, D. et al. Greengenes2 unifies microbial data in a single reference tree. Nat. Biotechnol. 42, 715–718 (2024).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  45. Chen, S., Zhou, Y., Chen, Y. & Gu, J. fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 34, i884–i890 (2018).

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  46. Li, D., Liu, C. M., Luo, R., Sadakane, K. & Lam, T. W. MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph. Bioinformatics 31, 1674–1676 (2015).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  47. Fu, L., Niu, B., Zhu, Z., Wu, S. & Li, W. CD-HIT: accelerated for clustering the next-generation sequencing data. Bioinformatics 28, 3150–3152 (2012).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  48. Li, R., Li, Y., Kristiansen, K. & Wang, J. SOAP: short oligonucleotide alignment program. Bioinformatics 24, 713–714 (2008).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  49. Buchfink, B., Xie, C. & Huson, D. H. Fast and sensitive protein alignment using DIAMOND. Nat. Methods 12, 59–60 (2015).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  50. Zhou, Z. et al. Metabolite annotation from knowns to unknowns through knowledge-guided multi-layer metabolic networking. Nat. Commun. 13, 6656 (2022).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  51. Breiman, L. Random forests. Mach. Learn. 45, 5–32 (2001).

    Article 

    Google Scholar 

  52. Pedregosa, F. et al. Scikit-learn: machine learning in Python. J. Mach. Learn. Res. 12, 2825–2830 (2011).

    Google Scholar 

  53. Stegen, J. C., Lin, X., Konopka, A. E. & Fredrickson, J. K. Stochastic and deterministic assembly processes in subsurface microbial communities. ISME J. 6, 1653–1664 (2012).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  54. Kembel, S. W. et al. Picante: R tools for integrating phylogenies and ecology. Bioinformatics 26, 1463–1464 (2010).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  55. Machado, D., Andrejev, S., Tramontano, M. & Patil, K. R. Fast automated reconstruction of genome-scale metabolic models for microbial species and communities. Nucleic Acids Res. 46, 7542–7553 (2018).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  56. Bowers, R. M. et al. Minimum information about a single amplified genome (MISAG) and a metagenome-assembled genome (MIMAG) of bacteria and archaea. Nat. Biotechnol. 35, 725–731 (2017).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  57. Hyatt, D. et al. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinf. 11, 119 (2010).

    Article 

    Google Scholar 

  58. Yin, Q. et al. Ecological dynamics of Enterobacteriaceae in the human gut microbiome across global populations. Nat. Microbiol. 10, 541–553 (2025).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  59. Peng, X. et al. Metabolic interdependencies in thermophilic communities are revealed using co-occurrence and complementarity networks. Nat. Commun. 15, 8166 (2024).

    Article 
    CAS 
    PubMed 
    PubMed Central 

    Google Scholar 

  60. Yuan, M. M. et al. Climate warming enhances microbial network complexity and stability. Nat. Clim. Change 11, 343–348 (2021).

    Article 

    Google Scholar 

  61. Ofek-Lalzar, M. et al. Niche and host-associated functional signatures of the root surface microbiome. Nat. Commun. 5, 4950 (2014).

    Article 
    CAS 
    PubMed 

    Google Scholar 

  62. Kenny, D. A., Kaniskan, B. & McCoach, D. B. The performance of RMSEA in models with small degrees of freedom. Sociol. Methods Res. 44, 486–507 (2014).

    Article 

    Google Scholar 

Download references

Acknowledgements

This work was supported by National Natural Science Foundation of China (42177362) and National Basic Science Data Center ‘Environment Health DataBase’ (number NBSDC-DB-21). We thank Y. Tao and N. Xue from Institute of Microbiology, Chinese Academy of Sciences, for providing the auxotrophic E. coli strains. We thank Z. Guo and Y. Duan for their help in sampling and data analysis.

Author information

Authors and Affiliations

Authors

Contributions

X.X. conducted the experiments, generated and analysed the data and wrote the paper. M.W. wrote the paper. A.U.O. and L.-T.Z. helped with the data analysis. F.L. and Y.G. provided the suggestions for computational adjustments. X.L., Z.S. and M.Z. contributed to extract EV experiments. Q.H. wrote the paper and got the funding.

Corresponding authors

Correspondence to
Miaoxiao Wang or Qiansheng Huang.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Nature Water thanks the anonymous reviewers for their contribution to the peer review of this work. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information (download PDF )

Supplementary Figs. 1–26.

Reporting Summary (download PDF )

Peer Review File (download PDF )

Supplementary Tables 1–10 (download XLSX )

Supplementary tables as separate files.

Supplementary Data (download ZIP )

Source data for Supplementary Figs. 1, 2, 4, 5, 8–10, 12, 16–18, 20–24 and 26.

Source data

Source Data Fig. 1 (download XLSX )

Statistical source data for Fig. 1.

Source Data Fig. 2 (download XLSX )

Statistical source data for Fig. 2.

Source Data Fig. 3 (download XLSX )

Statistical source data for Fig. 3.

Source Data Fig. 4 (download XLSX )

Statistical source data for Fig. 4.

Source Data Fig. 5 (download XLSX )

Statistical source data for Fig. 5.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Cite this article

Xu, X., Obeten, A.U., Zhu, LT. et al. Extracellular vesicle-mediated metabolic exchange shapes the seasonal assembly of aquatic bacterial communities.
Nat Water (2026). https://doi.org/10.1038/s44221-026-00605-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Version of record:

  • DOI: https://doi.org/10.1038/s44221-026-00605-0


Source: Ecology - nature.com

River interlinking and biodiversity risks in Indian freshwater ecosystems

Inferring sperm whale (Physeter macrocephalus) sex and developmental stage using aerial photogrammetry