Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.09.09.675270

Genomic data confirms phenotypic predictions of hybridization between cryptic Hawaiian cricket species

Abstract

Hybridization is increasingly recognized as an important driver of species radiations, yet its detection in morphologically cryptic taxa remains challenging. The Hawaiian crickets of the genus Laupala represent a rapid non-adaptive radiation in which species are morphologically indistinguishable but differ in acoustic mating signals. Here we report the first discovery of natural hybridization in the Laupala radiation. During field surveys on Maui, we observed crickets with intermediate pulse rates and unusually high acoustic variation at sites where the ranges of Laupala makaio (slow singer) and Laupala orientalis (fast singer) converge. We hypothesized these represented natural hybrids and tested this prediction using genomic data from genotyping-by-sequencing (GBS). Maximum likelihood phylogenetic analysis showed that fast-singing putative hybrids clustered with L. orientalis and the slow-singing ones with L. makaio, yet all putative hybrids formed a cohesive geographic group. Using 9,345 ancestry-informative markers, we calculated hybrid indices and heterozygosity values for triangle plot analysis. All hybrids showed excess L. makaio ancestry (predicted from analysis of pulse rate patterns) and were classified as later-generation hybrids or backcrosses to L. makaio, with no F1 hybrids detected. Critically, we found a strong positive correlation (R = +0.74, p = 4.1x10-) between pulse rate and hybrid index at the site with highest pulse rate variation, showing that genomic ancestry predicts pulse rate phenotype. Morphological comparisons confirmed that hybrids were cryptic, reinforcing the difficulty of detecting hybridization without genomic data. Together, our results provide the first demonstration of natural hybridization in Laupala, showing that phenotypic predictions based on acoustic intermediacy can be confirmed with genomic evidence. The discovery of natural hybridization in Laupala opens new avenues for investigating how hybridization contributes to rapid diversification in non-adaptive radiations, paralleling patterns observed in some adaptive radiations.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Sen, R., Shaw, K. L.. 2025-09-15. Genomic data confirms phenotypic predictions of hybridization between cryptic Hawaiian cricket species. https://doi.org/10.1101/2025.09.09.675270

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

KEEP EXPLORING

Related preprints

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↗

Toxin structure shapes palatability in a chemically defended butterfly

The toxicity of chemical defences is well studied, but the potential contribution of compound structure to predator deterrence remains largely unexplored. Whether predation acts more strongly on toxicity or unpalatability remains largely untested, partly because few systems allow toxin structure to vary independently of quantity. Heliconius sara larvae provide such a system: those reared on Passiflora auriculata sequester cyclopentenyl cyanogenic glucosides (CGs), while those reared on P. biflora biosynthesise comparable quantities of aliphatic CGs. Using two invertebrate predators, Camponotus floridanus ants and Hierodula membranacea mantids, we tested whether this structural difference affects palatability independent of toxicity. Mantids rejected larvae with cyclopentenyl CGs more often than larvae with aliphatic CGs, despite no detectable difference in total CG content. This pattern was mirrored in extract-based assays with ants, independently of cyanide release: extracts with cyclopentenyl CGs remained deterrent, while extracts with aliphatic CGs did not differ in deterrence from water. Live larvae, by contrast, elicited similar responses from ants regardless of CG structure. These results show that variation in toxin structure can strongly affect palatability, with some compounds conferring greater protection than others. This demonstrates the importance of chemical structural diversity in the evolution of chemical defences.

evolutionary biology↗