Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.09.30.678834

Aposematic color patterns are the dominant axis of phenotypic diversification in Nymphalid butterflies

Abstract

Butterfly wing patterns serve diverse roles in visual communication, from aposematic signaling and mimicry to mate attraction and camouflage. In brush-footed butterflies (Nymphalidae), this diversity can be traced to the wing pattern "ground plan" that generates phenotypes deterministic in origin yet highly multidimensional in form. Quantifying such complexity at scale has long been a challenge, limiting our understanding of how visual signals interact, constrain one another, and evolve. Here, we used computer vision to extract high-dimensional traits from standardized museum specimens and assembled the largest comparative dataset of wing color patterns to date, spanning over one third of all known Nymphalid species. We first tested whether chemically defended species occupy a distinct region in morphospace and then derived a quantitative score for aposematism from the principal color patterns associated with defense. Using this score, we examined whether aposematic signals are expressed consistently across wing surfaces and sexes, and whether their origins are linked to shifts in evolutionary rate. We found that the dominant axis of morphospace is defined by chromatic and achromatic contrast, along which defended and undefended species cluster. Validation with an expert-labeled moth dataset confirmed that this axis separates aposematic from non-aposematic phenotypes across Lepidoptera. Consistent with theory, strongly aposematic species showed greater visual similarity between dorsal and ventral surfaces, between sexes, and among individuals. Rate analyses further indicated that aposematic patterns evolved repeatedly and were associated with non-linear shifts in evolutionary tempo. Together, these results identify aposematism as the dominant organizing axis of wing color pattern evolution in Nymphalidae. Significance statementButterflies are renowned for their striking diversity of wing patterns, including the warning colors that signal chemical defense to predators. Yet whether such warning patterns share common features across lineages has remained unclear. Here, we applied a metric computer vision model to more than 16,000 museum specimens spanning one third of all Nymphalid species, encoding their wing patterns into a common morphospace. Within this space, high-contrast aposematic patterns emerged as the dominant axis of diversification, explaining up to 20% of phenotypic variance. These signals were expressed consistently across wing surfaces, sexes, and individuals, and evolved repeatedly across the family. Our approach demonstrates how computer vision enables meaningful comparative analyses of complex patterns, revealing general principles of butterfly diversification.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Luerig, M. D., Shirai, L. T., Mota, L., Willmott, K. R., Freitas, A. V. L., Porto, A.. 2025-10-01. Aposematic color patterns are the dominant axis of phenotypic diversification in Nymphalid butterflies. https://doi.org/10.1101/2025.09.30.678834

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Denisovan introgression left differential selection regimes in Humans and Neanderthals on the SLC30A9 gene

Signals of positive selection around the SLC30A9 gene have been reported in human populations outside Africa. Selection likely acted on a highly differentiated single-nucleotide polymorphism, rs1047626, leading to a non-synonymous substitution in the encoded zinc transporter. Because of the striking similarity between the putatively selected SLC30A9 haplotype observed in several current human populations and the Denisovan individual, previous work has proposed adaptive introgression. Yet alternative explanations, including ancient human variation, and the precise archaic source -Neanderthal or Denisovan- remained unresolved. Considering the potentially complex evolution of SLC30A9, we applied Approximate Bayesian Computation (ABC) algorithms coupled to machine learning to investigate the most plausible evolutionary origin of this substitution. After modelling different evolutionary scenarios with forward-in-time simulations, our results highlight that the most probable scenario is a Denisovan origin of the rs1047626 polymorphism. However, the allele likely introgressed into Neanderthals first and was then passed into non-African modern humans. Moreover, the derived allele frequency for rs1047626 across several African populations is consistent with back-to-Africa migrations. Finally, our ABC analyses indicate strong positive selection in East Asian populations and other out-of-Africa populations, whereas in Neanderthal populations, the selection coefficient was probably neutral or slightly deleterious.

evolutionary biology↗

RELAX does not reproduce its own estimates at default settings, and its output does not show it

Selection-intensity estimates from RELAX are reported as a point value of K with a likelihood-ratio P. We report that, at default settings and on data of ordinary size, the program does not reproduce its own fits. Of 27 enzyme entries refitted under two optimiser configurations, none reproduced its log-likelihood to within 0.01 units; the median change was 103 units, the largest over 3,400, and four verdicts reversed. Eighty null orthologues reproduced none. A byte-identical command returned a distinct likelihood on every repetition, single-threaded, across three releases, and on alignments simulated under the fitted model, where 3.3 per cent of replicates reproduced. The documented random-number seed never reaches the generator when assigned on the command line, yet reads back as the value supplied. PAML localises the cause: its two-ratio model, without site classes, reproduced its log-likelihood for all 288 genes; its site-class models agreed for 27 to 67 per cent. The instability follows the mixture over sites, not the program. The output does not show it: 46 of 410 fits ended with a negative likelihood-ratio statistic, impossible under convergence, and 123 of 410 report a K re-estimated under a domain restriction rather than the unconstrained maximum. Of 234 published studies using RELAX, none reported a seed. Seeding while holding the thread count at one reproduced sixty of sixty runs on twenty genes under two releases; the seed alone reproduced none of five, and no documentation states the second condition. We recommend that fits be repeated and their dispersion published.

evolutionary biology↗

Sequential accumulation of adaptive alleles forms an inversion supergene in deer mice

Supergenes are clusters of co-inherited loci that affect multiple or complex phenotypes. Despite the growing number of chromosomal inversions identified as supergenes in natural populations, their molecular basis and evolutionary history often remain obscure. Here, we identified two candidate genes, Slc45a2 and Npr3, within a 41-Mb inversion supergene in the deer mouse (Peromyscus maniculatus) that respectively drive darker coats and longer tails - two traits associated with forest adaptation. Mice homozygous for the inversion (inv/inv) exhibit elevated Slc45a2 expression in melanocytes relative to the congenic standard genotype (std/std), disrupting pheomelanin production. In parallel, downregulation of Npr3 in inv/inv mouse growth plates prolongs postnatal growth of caudal vertebrae, resulting in tail elongation. Population-level analyses further implicate that this supergene arose through the subsequent accumulation of the Npr3 allele within the inversion, rather than by capturing all beneficial mutations at its origin.

evolutionary biology↗