Molecular Ecology

Vole and lemming population cycles are an enigma in ecology. Decades of field observations and experimental manipulations have revealed that cycles cannot always be explained by extrinsic factors in the environment, including food availability or predator numbers. Thus, it has been proposed that intrinsic mechanisms, such as adaptive alterations in phenotype during different phases of the cycle, drive population dynamics. However, the mechanisms underlying such phenotypic changes have not been elucidated. We test the hypothesis that epigenetic changes occur over population cycles by comparing whole epigenome DNA methylation changes in brain tissue collected from northern red-backed voles (Clethrionomys rutilus) in a wild, naturally cycling population during the peak, decline, and low years. Overall, the greatest number of differentially methylated CG sites (DMCs) and differentially methylated regions (DMRs) were detected in comparisons between voles from the peak phase and low phase of the cycle. We highlight methylation differences in the promoter region of ATP synthase subunit c (Atp5g3) and an intron of insulin-like growth factor 1 receptor (Igf1R), which may be associated with growth, development, and bioenergetics. There were additional changes in the promoters of members of the cytochrome P450 enzyme family, including Cyp1a1, associated with estrogen metabolism, as well as the promoter of macrophage migration inhibitory factor (Mif), and in an exon of serum/glucocorticoid regulated kinase (Sgk1), which may link changes in stressors to direct brain changes. Our study is the first interrogation into broad epigenetic changes associated with natural population cycle phase in a wild mammal.

Understanding the drivers of genomic health and their consequences for population viability is often overlooked but potentially important to effective conservation amidst the biodiversity crisis of the Anthropocene. Leatherback turtle (Dermochelys coriacea) populations have declined globally due to anthropogenic factors, with some populations losing over 90% of their abundance over the past 30-50 years. While conservation efforts have been successful in stabilizing some populations, others continue to decline, and the reasons for these differential trajectories remain unclear. To assess how recent demographic factors, such as population size and decline, influence population genomic health, we combined population monitoring information with medium depth whole-genome and reduced representation resequencing data from globally representative populations. We found that small-stable populations have lower genomic diversity and higher inbreeding than large declining populations, reflecting prolonged small population sizes and limited gene flow. Yet, small-stable populations also show evidence of deleterious allele purging, suggesting genetic resilience. This, combined with lack of detectable genomic erosion over the study period, provides hope for potential recovery of healthy leatherback populations provided that anthropogenic threats are effectively mitigated. However, potential time lags and possible recent increases in inbreeding among close relatives in recently declined populations warrant continued monitoring and assessment. Genomic and abundance-based metrics were less aligned following rapid population declines, emphasizing the different timescales of the evolutionary and demographic processes they reflect, respectively, and the strength in their complementary, integrative use for extinction risk assessments. This also supports that it is not too late to turn the tide for recently declined leatherback populations and that continued investment in conservation efforts and threat reductions are warranted. Collectively, our results highlight how recent and historical demography shapes current genomic health and recovery potential in leatherback turtles, aids understanding of current risks and informs future conservation and management strategies.

Genomic forecasting approaches based on genotype-environment associations (GEAs) are increasingly used to estimate genomic offsets (GOs), which predict population maladaptation and extinction risk under current or future climatic conditions. Despite their widespread use, only a subset of studies have evaluated how accurately GOs predict (mal)adaptation, limiting their interpretation and application in policy and management. Here, we used GEA analyses to estimate GOs for past, present, and future climates in Lycaeides butterflies, focusing on the causes of variation in GOs among populations and their relationships with demographic parameters inferred from population genomic data. Using multivariate linear regression and genotyping-by-sequencing data from 42 Lycaeides populations (922 butterflies), we found that mean annual temperature, cumulative annual precipitation, and hybridization history together explained 47.6% of variation in genome-wide allele frequencies. Genomic offsets differed substantially among populations and across past, present, and future climates, with evidence for increasing maladaptation under more distant future climate scenarios. We found no relationship between GOs for present climates and contemporary effective population size. In contrast, genetic diversity, which reflects long-term effective population size, and local rates of gene flow together explained 27.3% of variation in contemporary GOs. Populations with higher genetic diversity and more gene flow exhibited lower GOs, consistent with the hypothesis that genetic diversity enhances adaptive capacity and that gene flow may introduce adaptive alleles. Overall, our results support the utility of GO predictions, particularly when validated with independent measures of adaptation, while cautioning against simplistic interpretations of GO as a direct measure of maladaptation in conservation and management contexts.

Demographic history is expected to play a central role in shaping population vulnerability to climate change through its lasting effects on effective population sizes and genetic connectivity. However, existing studies report contrasting outcomes, and the consequences of alternative demographic histories have seldom been assessed concurrently across multiple taxa. Here, we analysed population genomic data from six of the major European forest tree species to address this gap. We found that, across all species, more genetically isolated populations showed reduced adaptive potential, as measured by standing levels of genetic diversity. Furthermore, populations with reduced historical gene flow and higher genetic differentiation exhibited higher genetic load and suboptimal climate adaptation, particularly in small, fragmented populations, potentially increasing their sensitivity to climate change. Finally, the beneficial effects of gene flow were evidenced by the absence of greater suboptimal climate adaptation in highly connected populations across the six species. Altogether, our results provide valuable insights into how genetic differentiation, reflecting the combined effects of genetic drift and limited historical gene flow, influences current vulnerability of forest tree populations to climate change.

Trait-selective harvesting by fisheries can impose strong selective pressures on fish populations, driving changes in life history traits affecting fisheries productivity and ecosystem functioning. While the genetic consequences of harvesting have been extensively studied, the extent to which phenotypic variation reflects genomic evolution versus environmentally-induced plasticity remains unclear. Epigenetic mechanisms, such as DNA methylation, may mediate between these processes, serving as a rapid and reversible response to the selective pressures imposed by harvesting. In this study, we implemented an improved laboratory and bioinformatics protocol, epiGBS3, to examine genomic variation and DNA methylation patterns in the marine fish Xyrichtys novacula. The study spanned three replicated geographical areas each comprising two adjacent locations: an intensively exploited fishery and a no-take Marine Protected Area (ntMPA). A nested analysis design across the three areas revealed strong gene flow and no evidence of genetic structure. Nevertheless, nucleotide diversity was significantly reduced in fisheries relative to ntMPAs. We also found that DNA methylation levels differed between protected and exploited sites after controlling for age, suggesting that fishing may influence epigenetic changes independently of fisheries-induced age-truncation effects. This represents one of the first lines of evidence that fisheries can potentially shape epigenetic variation, supporting DNA methylation as contributor to local adaptation under high gene flow and strong anthropogenic selection.

Laboratory and field populations of insects can experience a decline in fitness and loss of genetic diversity due to inbreeding depression and genetic drift, respectively. Matings among related individuals and small population size may also influence insect host microbiomes with consequences for fitness. In the dengue vector mosquito, Aedes aegypti, the bacterial microbiome is largely environmentally determined but recent studies have also revealed host genetic components. We generated a panel of 55 inbred lines from either of two founding outbred populations of Ae. aegypti to test for associations between life history traits, inbreeding, allelic diversity and microbiome composition using ddRADseq and bacterial 16S rRNA gene sequencing on pools of mosquitoes. Effects of inbreeding were diverse with severe composite fitness costs in many lines but minimal costs in others despite similar low levels of genetic diversity. We found no strong relationship between major life history traits across inbred lines, suggesting that any costs due to inbreeding were trait specific. Bacterial microbiome analysis of pooled samples from a subset of populations revealed common microbes across populations, with Elizabethkingia, Aeromonas and Ralstonia being the most abundant. Despite bacterial composition varying widely, there was no clear relationship between microbiome composition and fitness or population origin. However, there were several significant positive correlations between the relative abundance of different microbial taxa across lines. Our results demonstrate diverse impacts of inbreeding on fitness of mosquito populations but with limited impacts on the microbiome.

Birdsong mediates territory acquisition and mate choice. In agonistic interactions, local songs generally elicit stronger responses than songs from more distant populations. However, the molecular mechanisms associated with differential responses to local vs. foreign songs are poorly understood. We addressed this knowledge gap by combining behavioral assays in the field with blood transcriptomic analysis, using a within-subjects design to ask whether male song sparrows (Melospiza melodia) show differential gene expression when exposed to playback of local and foreign songs. Transcriptomic profiles reflected the difference in behavioral response to local vs. foreign songs, with individuals exposed to local songs showing greater expression of genes associated with song perception and production, anti-inflammatory responses and energy metabolism. Our study suggests that changes in expression of key molecular pathways correlate with behavioral responses to geographic song variation, providing insight into the potential mechanisms regulating signal recognition and response to social challenges. HighlightsO_LIGene expression in sparrow blood was measured after simulated territorial intrusion. C_LIO_LIStronger response to local songs was associated with differential gene expression. C_LIO_LISong-associated genes (FOXP2, NRXN1) had higher expression when birds heard local songs. C_LIO_LIGene expression in the blood contains potential biomarkers of song recognition. C_LI

Introgressive hybridization between wild and domestic animals is a widespread phenomenon with important implications for genetic diversity, local adaptation, and conservation management. The causes and consequences of this process are poorly understood. In Australia, hybridization between dingoes and domestic dogs presents a dual conservation challenge, threatening the genetic integrity of dingoes while allowing potential adaptive introgression. To investigate the environmental drivers of this process, we analyzed high-density SNP array data in 390 dingoes and 396 domestic dogs. Dingo populations showed regional genetic structure and were clearly differentiated from domestic dogs. Using local ancestry inference and genome-environment association analyses, we found low levels of dog introgression in dingoes from remote areas in Central and Western Australia, and moderate levels in Eastern and Southern populations. Climatic variables (maximum temperature of the warmest month, mean temperature of the driest quarter) and the Human Footprint Index (reflecting density of human populations and environmental modifications) were significant predictors of introgression. We identified four genomic regions with overrepresented dog ancestry, including a large introgressed block on chromosome 27, which contained an olfactory receptor gene showing signatures of positive selection, suggesting adaptive introgression. In addition, a chromosomal inversion previously described in dogs and absent in dingoes was initially identified as an introgressed block. We also detected eight genomic regions nearly free of dog ancestry, suggesting purifying selection against maladaptive variants. Together, these results highlight the complex interplay between introgression, human influence, and local adaptation in dingoes, offering valuable insights for conserving the evolutionary potential of this apex predator in increasingly modified landscapes.

Cryptic species diversity, overlooked due to extreme morphological similarity, is a common phenomenon among ants. The "honeypot ant" genus Myrmecocystus (Wesmael, 1838; Formicinae: Lasiini) likely features multiple cryptic species, as previously suggested by phylogenetic studies based on ultraconserved elements (UCEs). Here, this work is expanded upon by examining 140 specimens and 2,508 UCE loci, with a particular focus on the M. mendax species complex from the southwestern USA and northern Mexico. Phylogenomic and population genomic analyses revealed five distinct M. mendax-like lineages and identified two potential cases of cryptic species diversity, one within samples matching the morphology of M. mendax and another within samples conforming to M. placodops. Most specimens morphologically identified as M. mendax formed a well-supported monophyletic group sister to M. melliger assigned individuals, with evidence for ongoing hybridization between both species in the Madrean Sky Islands along the USA-Mexico border. Patterns in the main M. mendax clade also suggest adaptive divergence across ecological gradients, warranting further investigation. Overall, these findings highlight the power of UCE-based genomic data in phylogenetic reconstructions and population genetic analyses to better resolve cryptic species diversity, and clarify complex evolutionary histories shaped by introgression and incomplete lineage sorting.

Strong population contractions can leave a persistent genomic legacy that can influence populations long after their demographic recovery. While bottlenecks facilitate the removal of strongly deleterious mutations, the effectiveness of purging may be limited in historically small populations. The Kirtlands warbler (Setophaga kirtlandii) is a rare North American songbird with an ancestrally small population. After narrowly evading extinction, they are one of few species that have been delisted from federal protections in the USA. Despite their recovery, a previous study showed evidence for recent inbreeding and a high burden of deleterious mutations that may have not been purged despite strong bottlenecks. Historical DNA offers a unique opportunity to understand the consequences of recent demographic declines on genetic diversity. Here, we use DNA from over 100-year-old museum specimens to estimate changes in genetic load in the Kirtlands warblers pre- and post-bottleneck. We validate our results with forward-in-time genetic simulations and explore how sample size and missing data can affect estimates. Both empirical data and simulations suggest a reduced ability to purge deleterious mutations in this historically small population. Our simulations also highlight that limited sampling design and data quality can constrain the ability to detect changes.

Kelp forests are widely distributed along temperate and polar coastlines worldwide and are among the worlds most productive and diverse marine ecosystems. Yet, due in part to ocean warming, they are declining and even disappearing in many parts of the world. While genomic tools can identify local adaptation and predict species responses to global change, these predictions have rarely been validated in the field, hampering their widespread use in conservation practice. Here, we applied a seascape genomics approach to investigate environmental adaptation in the two main canopy-forming species of the Northeast Pacific, Macrocystis tenuifolia and Nereocystis luetkeana. We leveraged whole-genome sequences of 598 individuals across 94 sites along the British Columbia and Washington coasts, together with 37 environmental variables. Both species showed genomic signatures of local adaptation, with distinct environmental drivers shaping adaptation in each species despite their co-occurrence across much of the studied area. Using gradient forests, we modelled the genetic turnover across environmental gradients and predicted populations vulnerability (genomic offset) under projected environmental conditions. Genomic offsets differed greatly among regions and were positively correlated with kelp declines observed to date, especially in Macrocystis, validating the link between genomic models and outcomes in the field and allowing us to translate genomic predictions into an ecologically meaningful metric: the risk of extirpation under global change. Our models predict that assisted migration could significantly attenuate kelps vulnerability to global change. Across environmentally heterogenous coastlines, short-distance migration can often substantially reduce future genomic-environmental mismatches, but in many cases, long-distance migration would be most beneficial. Our results highlight the potential of seascape genomics to predict vulnerability of populations to global change. Importantly, the validated link between our genomic models and ecological outcomes allows quantification of climate-driven extirpation risk and can inform conservation strategies to improve the resilience and sustainable management of these vulnerable ecosystems.

Hybridisation between wild and domestic taxa can favour the spread of domestic alleles into wild populations through backcrossing. The complex interplay of random genetic drift, recombination, and selection can shape the fate of introgressed alleles. Maladaptive domestic variants are likely to be purged by natural selection, but others may persist across generations. It has long been known that the Apennine Italian wolf population, exposed to large numbers of free-ranging dogs, has experienced extensive introgression. The unusually high frequency of black wolves observed in Italy, compared to other European populations, may parallel patterns documented in North American wolves, where the melanistic KB allele at the CBD103 gene, of domestic origin, has spread over thousands of years of introgression. We tested whether the KB mutation entered the peninsular Italian wolf population via hybridisation and spread through adaptive introgression. Genome-wide analyses of black and wild-type (grey-coated) Apennine wolves showed no clear signatures of recent dog ancestry in most melanistic animals. Our ancestry reconstruction approaches identified two distinct KB haplogroups of domestic origin, suggesting multiple introgression events. Notably, we found molecular evidence consistent with balancing selection on the KB haplotypes, whose functional role, nonetheless, warrants further research. Therefore, the microevolutionary genomic and ecological consequences of wolf-dog hybridisation in Italy should be carefully investigated to inform appropriate science-based conservation management strategies.

Climate change is causing rapid changes to freshwater environments, driving selection for phenotypes that can cope with altered thermal conditions, while also changing developmental environments. This may promote local adaptation, migration to new habitats, and/or phenotypic plasticity. Climate change may also increase hybridization rates between locally adapted phenotypes, as populations migrate and spatially mix in new ways. Consequently, these conditions may facilitate the production of variation, including both adaptive and maladaptive outcomes. To examine this, we leveraged threespine stickleback (Gasterosteus aculeatus) that have locally adapted to either geothermally warmed or ambient environments in Iceland. We tested the effects of hybridization between thermal ecotypes on gene expression using a common garden experiment, where pure-strain ecotypes and their hybrids were reared under 12{degrees}C and 18{degrees}C. We performed RNA-seq on brain and liver to assess 1) ecotype divergence, 2) plasticity, and 3) the effects of hybridization and inheritance patterns. We identified a low degree of expression divergence between locally adapted ecotypes, despite a high degree of plasticity across rearing environments. Hybrid ecotypes were highly divergent from both geothermal and ambient ecotypes and exhibited transgressive expression under both rearing temperatures. Transgressive expression disrupted gene networks extensively, with broader effects at 18{degrees}C than at 12{degrees}C, primarily associated with metabolism and mitochondrial function. Hybridization between locally adapted thermal ecotypes appears to largely disrupt genes associated with energy balance and metabolic function. While demonstrating mechanisms underlying the rapid evolution of reproductive isolation, these findings also provide insights for how populations may cope or fail within a warming world.

Microbial communities are essential for host health and ecosystem stability. However, whether host identity or shared foraging resources shapes microbiome structure among co-occurring species remains poorly understood. We studied bacterial and fungal communities of four Indian honeybee species in a mustard monoculture resource condition, integrating behavioural observation-based pollinator data with microbial co-occurrence networks derived from metabarcoding. Microbiome composition was linked to host identity rather than foraging behaviour, bee abundance, or landscape use. While core bacterial taxa were shared, relationships among bacterial cobionts, unlike those among fungal genera, remained species-specific. Microbial diversity, along with community structure and function, influenced network stability, with a highly modular microbial network of Apis cerana exhibiting more resilience to simulated perturbations. In summary, host-specific filtering shaped the microbiome more than resource homogenisation, with closely related species facing unique risks of microbial collapse, with broader implications for vulnerability to microbiome imbalance, environmental stress, and emerging infections.

Populations of the Caribbean reef-building coral, Acropora palmata, have declined sharply since their population genetic structure was first characterized in the early 2000s. Previous analyses comprehensively sampled coral colonies across the Caribbean and western North Atlantic but genomic resolution was limited by the number of loci assayed. These analyses indicated extensive asexual reproduction via fragmentation, high outcrossing at the genet level, and a distinct east-west population split. To advance basic research and inform genetic management of this endangered species, we present an updated population genomic assessment using a species-specific microarray to analyze over 4,000 samples representing [~]1,500 genets from 12 geographic regions. Data were contributed by more than 30 research and restoration groups. Our analysis identifies nine spatially structured genetic clusters, with low average pairwise FST values of between 0.01 to 0.125. Interestingly, legacy genets from the Florida Reef Tract were admixed between two clusters, one dominant in the Mesoamerican Reef Tract on the western flank and the other cluster appearing in genets from Cuba to the south. Migration surface analyses highlight the influence of major current systems on gene flow. Isolation by distance was evident along the Greater Antilles but weak along the Florida Reef Tract. Kinship among wild genets was low across sites, suggesting limited local relatedness; however, assisted sexual reproduction in restoration efforts may disrupt natural kinship patterns. These findings refine our understanding of A. palmatas genetic architecture and underscore the importance of incorporating genomic data into conservation strategies.

Conservation interventions are increasingly required for species threatened by population declines and isolation due to anthropogenic pressures. Small, isolated populations are particularly vulnerable to the loss of genetic diversity, increased inbreeding, and the accumulation of deleterious mutations. Translocations or supplementation of allopatric individuals for genetic rescue may be the only way to increase genetic diversity to increase population persistence via increased adaptive potential. Here, we use an experimentally admixed population of sand lizards on a small island in Sweden as a valuable model of genetic rescue. This population was established approximately 20 years ago (5-6 generations) resulting in increased fecundity and hatchling viability. This population was founded from crossings between individuals from an inbred population from the nearby mainland and individuals sourced from populations in southern Sweden. Low-coverage whole-genome sequencing revealed elevated genetic diversity and reduced realized genetic load in this admixed population relative to the source populations. Ancestry analyses indicated a greater contribution of southern Swedish genetic variation, potentially reflecting contribution of beneficial adaptive variation from this region that may underlie the positive population effects. This system provides valuable empirical insights into the long-term genomic consequences of genetic rescue in this model vertebrate population.

Allopatric isolation under contrasting environments can drive rapid phenotypic divergence, even over contemporary timescales. Rapid changes in morphology or physiology can allow organisms to adapt to biotic and abiotic characteristics of their habitats. While studying metabolism, growth and resources needs may allow to understand adaptation to several selective pressures, these traits are rarely jointly considered. We investigated morphological, growth, and metabolic divergence in two allopatric populations of Arctic charr (Salvelinus alpinus) sharing a common evolutionary origin but inhabiting contrasting environments. We combined field observations, common garden and quantitative genetic approaches to disentangle contributions of genetic divergence and plasticity to phenotypic variability. Wild adults differed in body shape and growth trajectories, potentially reflecting plasticity related to resource availability and temperature variations. Under common garden conditions, juveniles displayed inter-population differences in routine metabolic rate, its allometric scaling with body mass. These patterns suggest divergent selection on physiological traits. Despite low neutral genetic differentiation, phenotypic divergence unfolded in fewer than 100 years, suggesting that plasticity and selection can promote rapid multi-trait changes. These findings highlight that considering changes in physiological, growth and morphological traits can reveal the adaptive potential of small, isolated populations facing rapid environmental change.

Sex ratio distorters (SRDs) are heritable elements that bias offspring sex ratios to enhance their transmission. In the terrestrial isopod Armadillidium vulgare, feminization of genetic males can occur through vertical transmission of the sex ratio distorter known as the f-element, as well as through infection by Wolbachia, a maternally inherited bacterial endosymbiont that can alter host reproduction. Previous studies have focused on the distribution of SRDs and their associations with mitochondrial haplotypes in native European populations, but these patterns are poorly understood in the United States. In this study, we sampled A. vulgare in 12 states, screening individuals for Wolbachia infection, the presence of the f-element, and mitochondrial haplotypes. We found that Wolbachia shows a heterogeneous distribution across populations and haplotypes, in contrast with stronger associations in Europe. The f-element occurred in lower overall frequencies but showed a strong association with mitochondrial haplotype VI. These results indicate that patterns associated with SRD differ from those observed in Europe and suggest that multiple introductions and population mixing have shaped these distributions.

As purifying selection becomes less effective and inbreeding increases, small populations frequently develop an increased load of genome-wide deleterious mutations. Reflecting this pattern, deleterious mutations in genes associated with fertility and immunity have previously been identified in the cheetah (Acinonyx jubatus), which has had a low effective population size for at least the last 10,000 years. However, the distribution of deleterious mutations across cheetah populations is currently unknown. Here, we analysed novel whole genome resequencing data from 30 ex situ and 9 wild cheetahs. We investigated variation in genetic diversity, genomic measures of inbreeding, and the distribution of deleterious mutations across cheetah populations. South Sudanese and Tanzanian cheetahs showed higher inbreeding and realized load, while Namibian cheetahs had a higher proportion of population-specific deleterious mutations. Genes containing high- or moderate-impact deleterious mutations were significantly enriched for sperm-related functions, highlighting putative causative loci associated with poor sperm quality in cheetahs. Similar levels of genetic diversity and inbreeding were observed in ex situ cheetahs compared to their wild counterparts, providing empirical evidence of the efficacy of captive breeding programmes in maintaining genetic variation in ex situ populations.

Despite the coppery titi monkey (Plecturocebus cupreus) being a model system for the study of neurodevelopment and behavior, the evolutionary forces shaping observed levels and patterns of genetic variation in the species have remained poorly studied. In order to illuminate the pervasive eCects of purifying and background selection, we have fit a distribution of fitness eCects of newly arising exonic mutations, utilizing patterns of polymorphism and divergence based on a recently published high-quality genome assembly. To further characterize episodically acting selective processes, we additionally performed the first whole-genome scans for recent positive and balancing selection in this species, reducing false-positive rates by incorporating the demographic history of the population into an evolutionary null model. These scans identified a small number of biomedically-relevant genes with strong statistical support for having experienced recent selective sweeps or long-term balancing selection. In addition, we identified four genomic deletions bearing the signatures of balancing selection. Taken together, this study provides the first insights into patterns of persistent and episodic selective processes in this species.