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Complex range expansion and selective regime in the introduced Florida cane toad

Heredity (2026)Cite this article

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Abstract

Introduced species that successfully establish in new areas are a powerful system for investigating the genetic, ecological, and adaptive processes underlying range expansion. Rhinella marina is the focus of many studies of invasion dynamics, rapid evolution, and range limits. However, comparatively little is known about the nearly simultaneous establishment of closely related R. horribilis in Florida, USA. We sequenced 280 individuals using double-digest restriction-associated DNAseq (ddRAD) to investigate the role of introduction history, standing genetic diversity, and adaptation in R. horribilis’ establishment in Florida. We test the hypothesis of a single introduction event versus the alternative of several cryptic introductions. Second, we characterize population structure and genetic diversity to elucidate the roles of genetic bottlenecks and subsequent gene flow. Third, we use redundancy analyses to identify climate-associated genetic variants that may play a role in adaptation in Florida, which is colder than the cane toad’s native range. Lastly, we analyze a morphological trait, limb length, to investigate potential evolution of dispersal at the range edge. We find evidence for a single introduction of R. horribilis and complex range expansion characterized by range-wide gene flow, a lack of isolation by distance or environment, and no range edge dispersal phenotype. We also find evidence of selection related to range-wide gradients of precipitation, temperature, and urbanization. Together, our results indicate that range-wide gene flow maintains genetic diversity and adaptive capacity, likely supporting the neotropical species’ success in adapting to and establishing in this temperate environment.

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Fig. 1: Map of hypothesized Rhinella introduction sources and dates of introduction to Florida based on anecdotal historical accounts.
Fig. 2: Map of sample localities and environmental gradients with cooler colors representing lower values, warmer colors representing higher values.
Fig. 3: Ancestry assignment for K = 9–12.
Fig. 4: Greater gene flow from core to edge than edge to core.
Fig. 5: Outliers detected by Redundancy Analysis (RDA).
Fig. 6: Boxplot of relative limb length (mean femur length divided by Snout Vent Length) of each Florida population, ordered by nearest to farthest distance to a core population.

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Data availability

Sequence data are available on the NCBI Sequence Read Archive under BioProject ID PRJNA1413252. Florida specimens are archived at the Cornell University Museum of Vertebrates.

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Acknowledgements

The authors thank the Florida Natural History Museum and the Cornell University Museum of Vertebrates for access to specimens, as well as Sean McHugh and Dylan Tarbox for measuring specimens. The authors also thank three anonymous reviewers for feedback, which greatly improved the manuscript.

Funding

This work was supported by the National Science Foundation (grant number 81023/A001) to CSM; the Andrew W. Mellon Foundation to CSM; and Sigma Xi to CSM.

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Authors and Affiliations

  1. Department of Ecology and Evolutionary Biology, Cornell University, Ithaca, NY, USA

    Cinnamon S. Mittan-Moreau & Kelly R. Zamudio

  2. W.K. Kellogg Biological Station, Michigan State University, Hickory Corners, MI, USA

    Cinnamon S. Mittan-Moreau

  3. Department of Biology, Colorado State, University Pueblo, Pueblo, CO, USA

    Daryl Trumbo

  4. Department of Integrative Biology, The University of Texas at Austin, Austin, TX, USA

    Kelly R. Zamudio

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  1. Cinnamon S. Mittan-Moreau
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Contributions

CSM: Conceptualization; data curation; formal analysis; funding acquisition; investigation, methodology; project administration; validation; visualization; writing—original draft; writing—review and editing. DT: Methodology, writing—review and editing. KRZ: Conceptualization, funding acquisition, methodology, project administration, resources, supervision, writing—review and editing.

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Correspondence to Cinnamon S. Mittan-Moreau.

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Mittan-Moreau, C.S., Trumbo, D. & Zamudio, K.R. Complex range expansion and selective regime in the introduced Florida cane toad. Heredity (2026). https://doi.org/10.1038/s41437-026-00823-y

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