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Abstract

Accurate inferences of diversification and evolutionary processes depend on knowing how many independently evolving lineages exist within nominally widespread taxa. Uncertainty in lineage number and composition also limits our ability to meaningfully prioritise conservation efforts. Addressing these knowledge gaps requires frameworks that sequentially identify distinct lineages, assess mechanisms that contribute to their divergence, and quantify spatial and environmental determinants of their genomic diversity. With these goals in mind, we apply whole-genome, ecological, and morphological analyses to the American rubyspot damselfly (Hetaerina americana), a widespread and well-studied North American insect that has been long regarded as a single species of least conservation concern, despite some evidence of declines across the southwestern US. Whole-genome sequencing of 136 individuals revealed three deeply divergent lineages representing distinct species. Two of these lineages are putatively endemic to the California floristic province and a third clusters with H. americana sensu stricto across the central-eastern US. Comparative species distribution models and morphological analyses revealed significant niche and trait divergence among lineages, suggesting adaptive variation exists among species pairs—a relatively unique result for damselfly congeners. Genome-wide heterozygosity was extremely low within both newly discovered lineages, and landscape genomic analyses suggested that isolation by resistance—reflecting connectivity among rapidly disappearing lotic habitats—structures gene flow within each. Our field resurveys corroborated accelerating population losses, particularly in southern California, reshaping conservation priorities within this species complex. Together, our analyses comprise an integrative framework that links genomic, ecological, and morphological divergence to landscape and environmental variation, enabling reliable lineage delimitation, evolutionary inference, and conservation recommendations.

Document Type

Article

Publication Date

6-1-2026

Notes/Citation Information

Publisher Copyright: © 2026 The Author(s). Molecular Ecology published by John Wiley & Sons Ltd.

Digital Object Identifier (DOI)

10.1111/mec.70385

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