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Effects of urbanization on local adaptation and eco-evolutionary feedbacks in white clover

Nature Ecology & Evolution volume 9, pages 2322–2332 (2025)Cite this article

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Abstract

Urbanization is causing unprecedented ecological and evolutionary change worldwide. However, empirical evidence of local adaptation to urban environments is limited. Here we conduct a reciprocal transplant experiment using white clover (Trifolium repens) originating from urban and rural populations and distributed equally into five urban common gardens and five rural common gardens. Half of the plants in each garden produced the antiherbivore chemical defence hydrogen cyanide (HCN), and the other half lacked the defence, as this trait is known to exhibit genetic clines along urbanization gradients. Based on measurements of multiple vegetative and sexual fitness traits as well as ecological interactions with herbivores, pollinators and mutualistic root bacteria, we detect divergent selection on HCN between urban and rural environments, where HCN improved fitness in rural environments and reduced fitness in urban environments. Urban and rural white clover populations show genetic divergence that drove a tradeoff in life history strategies: urban plants invested more in vegetative growth, whereas rural plants produced more flowers and seeds in both urban and rural gardens. Finally, we demonstrate eco-evolutionary feedbacks, with increased herbivory at rural sites, and increased pollinator visitation to acyanogenic plants at urban sites. This study contributes to understanding how urbanization affects evolution and feeds back to influence broader ecosystem processes.

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Fig. 1: Experimental design and hypotheses.
Fig. 2: Map of common garden sites across the Greater Toronto Area, Canada and photographs of an example urban site (top, outlined in grey) and rural site (bottom, outlined in green).
Fig. 3: Effects of HCN on various components of fitness, at rural and urban common garden sites.
Fig. 4: Effects of source population on multiple components of fitness, at rural and urban common garden sites.
Fig. 5: Trade-offs between mean seed mass (g) and mean biomass (g) for plants sourced from rural populations (green, n = 385) and urban populations (grey, n = 383) at each common garden site (n = 10).
Fig. 6: Effects of HCN, source population and common garden site on species interactions.

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Data availability

The data associated with this article are available via figshare at https://doi.org/10.6084/m9.figshare.28848680 (ref. 60).

Code availability

The reproducible code for this article is available via Code Ocean at https://doi.org/10.24433/CO.0924253.v1 (ref. 61).

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Acknowledgements

We thank B. Wong, V. Vuong, S. Ahmad, P. Libert, A. Alencar, K. Bhachu, H. Du and Z. Nassrullah for help with plant preparation and data collection on this project and V. Velasco and UTM Growth Facilities for the use of the growth chamber and greenhouse. We thank the members of the EvoEco lab past and present, especially J. Santangelo, D. Murray-Stoker and L. Albano for the helpful advice and feedback throughout this project. We thank H. Rodd, P. Atto, D. Hall, E. Agbuya, the Da Silva family, J. Levitt, D. Goodman, D. Gwynne, G. Redshaw, N. Collins, M. Pallett, the Rawle family and N. and M. Roberts for hosting the common gardens. This work was funded by an NSERC Discovery grant (grant no. RGPIN-2022-04913) and Canada Research Chair (grant no. CRC-2022-00111) to M.T.J.J., a University of Toronto FAST Doctoral Fellowship and NSERC CGS-D award to E.M. and the University of Toronto Mississauga Department of Biology.

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Authors and Affiliations

  1. Department of Ecology and Evolutionary Biology, University of Toronto, Toronto, Ontario, Canada

    Ella Martin

  2. Department of Biology, University of Toronto Mississauga, Mississauga, Ontario, Canada

    Ella Martin & Marc T. J. Johnson

  3. Centre for Urban Environments, University of Toronto Mississauga, Mississauga, Ontario, Canada

    Ella Martin & Marc T. J. Johnson

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  1. Ella Martin
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  2. Marc T. J. Johnson
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Contributions

This project was designed by E.M. and M.T.T.J. E.M. conducted the experiment and collected the data. Analyses and initial drafts of the paper were done by E.M., with guidance from M.T.T.J., and both authors contributed to the final draft of the paper.

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Correspondence to Ella Martin.

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Extended data

Extended Data Fig. 1 Map of source populations and common garden sites.

Map of source populations (small orange points) and common garden sites (larger points) across the Greater Toronto Area, Canada. Credit: Basemap from Esri, USGS | Esri, TomTom, FAO, NOAA, USGS | Earthstar Geographics62.

Extended Data Fig. 2 Comparison of rural and urban field sites.

Horizontal lines represent rural and urban means, and points represent individual sites. Site colours correspond to points in the map in Fig. 1. A) Percent impervious surface within a 250 m radius around each site using 30 m resolution data from Landsat imagery63. B) Mean Human influence index over a 250 m radius around each site64. C) Mean NDVI value from MODIS Terra satellite (MOD13Q1, MODIS/Terra Vegetation Indices (NDVI/EVI) 16-Day L3 Global 250 m SIN Grid) between beginning and end of field season (May 25, 2023 – September 14, 2023)65,66. D) Percent tree cover within a 250 m radius around each site67. E) Minimum temperature (°C) between December 2023-March 2024 recorded on iButtons (Thermochron, Sunnyvale, USA) at each site. F) Mean temperature (°C) between December 2023-March 2024 recorded on iButtons at each site. G) Frequency of cyanogenic white clover from 50 stolons sampled from a 250 m radius around each site. H) Distance(km) of each site from Yonge-Dundas Square, Toronto, Canada.

Extended Data Fig. 3 Effects of source population on sexual fitness at urban and rural sites.

Effects of source population (green points: rural, grey points: urban) on sexual fitness at urban and rural sites. A: mean number of inflorescences produced by plants (n = 802); B: proportion of plants that produced seeds (n = 801). Points are mean values error bars show standard errors. P-values < 0.1 for Source (that is, source population from which populations were collected) and Site (that is, common garden habitat sites where plants were grown) fixed effects and interactions from LMMs or GLMMs are shown in top left, with significant (P < 0.05) effects in bold. Effects where P > 0.1 are not shown and can be found in Table S1.

Extended Data Fig. 4 Effects of HCN, source population, and common garden site on the percentage of leaf area eaten by herbivores in July 2023.

Effects of HCN, source population, and common garden site on the percentage of leaf area eaten by herbivores in July (n = 752). Points show means and error bars show standard errors. Significant P-values (P < 0.05) for fixed effects and interactions from LMM shown in top left. Additional statistics are not shown and can be found in Table S1.

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Martin, E., Johnson, M.T.J. Effects of urbanization on local adaptation and eco-evolutionary feedbacks in white clover. Nat Ecol Evol 9, 2322–2332 (2025). https://doi.org/10.1038/s41559-025-02892-4

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