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Pleistocene island connectivity did not enhance dispersal or impact population size change in Galápagos geckos.

Patterns of biodiversity on remote archipelagos are largely shaped by intra-archipelago colonization followed by in situ diversification. Pleistocene sea-level fluctuations purportedly enhanced gene flow among terrestrial organisms by increasing connectivity during periods of lower sea level. Furthermore, changes in sea-level are hypothesized to impact population sizes as a result of fluctuations in island sizes. Here, we used genomic data to test the role of Pleistocene island connectivity on the diversification and demographics of leaf-toed geckos (Phyllodactylus) endemic to the Galápagos. Consistent with previous studies, we found that present diversity of Galápagos Phyllodactylus stems from three independent dispersal events. Contrary to the hypothesis of Pleistocene-driven diversification, we found no correspondence between lineage divergence and island connectivity. Furthermore, we found no evidence of introgression; demographic modelling indicated that all species increased rapidly in effective population size in the period 20-150 ka, and these inferred demographic expansions were largely asynchronous and apparently unassociated with species or island age. Collectively, these results indicate that more complex abiotic and/or biotic factors may better explain the recent demographic history of Phyllodactylus and underscore the need for additional population genomic studies of terrestrial taxa to understand the impact of past climate cycles on Galápagos island communities.

Animals

De novo genome assemblies of threatened Asian hornbills (Bucerotidae) reveal declining population trajectories during the late Pleistocene.

BACKGROUND: Asian hornbills are flagship species of the wet tropics that face significant threats from hunting, habitat loss, and fragmentation. Despite being conservation flagships, whole genome information is available for only two of the 32 Asian hornbill species. In this study, we provide the first de novo genome assemblies for four hornbill species (Bucerotidae) in Asia. METHODS: We used a combination of long-read and short-read sequencing data to assemble and annotate de novo hybrid genomes of four species of hornbills. We also assembled and compared mitochondrial genomes of these species. Using a comparative genomics approach, we performed orthology assignment and gene evolution analyses to identify unique gene families in Asian hornbills, gene families that showed significant expansion, their functions and structural variation. Furthermore, using the Pairwise Sequentially Markov Coalescent (PSMC) method, we reconstructed demographic histories of hornbill species to examine changes in their population trajectories in the past. RESULTS: We present hybrid genome assemblies for Great Hornbill (B. bicornis - GH), Rufous-necked Hornbill (A. nipalensis- RNH), Malabar Pied Hornbill (A. coronatus- MPH) and Wreathed Hornbill (R. undulatus- WH). The genome sizes of these hornbills range from 1.1 Gb to 1.3 Gb, with over 95.9% completeness and gene prediction BUSCO. We reported 10,525 orthogroups shared among four Asian hornbill species and identified significant expansion in gene families associated with structural keratin development in Asian hornbills compared to their ancestors. We also provide annotated mitogenomes for each of these species. Furthermore, we found that the WH, a more abundant, widely distributed, and migratory species, showed a higher Ne than the other three hornbill species. However, an overall decline in Ne for all species was recorded during the Pleistocene climatic fluctuations. CONCLUSIONS: We present the first-ever, high-quality reference genomes for the threatened hornbill species from Asia. Hornbills have shown significant expansion in genes involved in structural keratin development. Our results indicate that Pleistocene climatic fluctuations have led to dramatic population declines in all four species. We believe that this study provides robust genomic resources to support future comparative and conservation genomics efforts for hornbills.

Animals

Hidden Diversity in the Sands: Genomic Footprints of Pleistocene Refugia and Fragile Futures of the Turkestan Ground-Jay (Podoces panderi) in Central Asia.

The Turkestan ground-jay (Podoces panderi), a corvid endemic to Central Asia's deserts and steppes, exemplifies how extreme environments drive speciation. Our study provides the first comprehensive high-resolution genomic analysis of this species, using complete mitochondrial genomes (49 individuals) to decode its population structure and demographic past. Our analyses revealed three highly divergent genetic clusters with strong geographic structure. The P. p. iliensis population (Cluster_3) showed particularly pronounced genetic distinctiveness, with significant differentiation from P. p. panderi (Cluster_2 and Cluster_1) populations. This clear genetic separation supports the taxonomic validity of P. p. iliensis as a distinct evolutionary lineage. Demographic reconstruction indicated that Cluster_2 likely represents the ancestral group, with subsequent southward expansion into the Karakum region. The isolated P. p. iliensis population exhibited signatures of long-term isolation, including reduced genetic diversity and absence of recent gene flow with other clusters. These results provide strong evidence that P. p. iliensis represents a distinct evolutionary unit. The genetic structuring into three clusters reflects historical isolation in desert refugia during Pleistocene climatic fluctuations. Notably, we detected asymmetric gene flow among three clusters. These findings redefine P. panderi as a model for desert adaptation, where climatic extremes forged genetic fragmentation amid limited dispersal. Beyond taxonomy, our work highlights how aridification sculpted biodiversity in Asia's interior, urging conservation attention for these evolutionarily distinct lineages.

Animals

Fore- and hindlimb proportions in Plio-Pleistocene hominids.

Associated fore- and hindlimb parts of five individuals are known from the hominid Plio-Pleistocene fossil collections in Africa. Four of these have been classified as Australopithecus and show definite evidence that in comparison with humans, forelimbs were relatively large and hindlimbs were relatively small. The fourth individual, placed in the genus Homo, has human proportions. These findings do not necessarily imply locomotor differences: the forelimbs may have been relatively long in Australopithecus simply because they were as yet not completely reduced from their generalized hominoid ancestral state.

Animals

Habitat Specialisation Impacts Clownfish Demographic Resilience to Pleistocene Sea-Level Fluctuations.

Habitat fragmentation and loss are key threats to biodiversity, yet their impacts on marine species remain poorly understood. Clownfishes, which rely on sea anemones for shelter and reproduction, provide an interesting model to explore how ecological specialisation mediates species responses to habitat perturbations. We used whole-genome data from 382 individuals across 10 species with varying host specialisations to reconstruct demographic histories and infer spatial genetic structure to assess the impact of Pleistocene sea-level fluctuations. Generalist species, associated with multiple hosts, maintained stable effective population sizes () and population connectivity during habitat fragmentation, reflecting resilience to environmental instability. In contrast, specialists experienced severedeclines and genetic structuring, driven by their dependence on specific hosts, without signs of population recovery following habitat reconnection. Spatial genomic analyses identified the Indonesian Through-Flow as a key dispersal corridor and the Coral Triangle as a critical hub of genetic diversity, while continental shelves and extensive open ocean regions appeared as barriers to gene flow. Our findings reveal how host specialisation shapes clownfish population dynamics, emphasising the importance of incorporating ecological dependencies into conservation assessments and deepening our understanding of species responses to ecological constraints and environmental changes over evolutionary timescales.

Animals

The proteome of the late Middle Pleistocene Harbin individual.

Denisovans are a hominin group primarily known through genomes or proteins, but their precise morphological features remain elusive because of the fragmentary nature of the discovered fossils. Here, we report 95 endogenous proteins retrieved from a nearly complete cranium from Harbin, China, dating to at least 146,000 years ago and previously assigned to a new species, Homo longi. This individual has three Denisovan-derived amino acid variants and clusters with Denisova 3, suggesting that the Harbin individual belongs to a Denisovan population. This study fills the gap between morphological and molecular evidence, enhancing our understanding of Denisovans' spatiotemporal dispersal and evolutionary history.

Animals

Climatic data sources and limitations of ecological niche models impact the estimations of historical ranges and niche overlaps in distantly related Korean salamanders.

BACKGROUND: Ecological niche models (ENMs) and analyses of niche overlap/divergence have become popular methods in ecology and evolutionary biology. These analyses rely on environmental data available from several databases. However, the influence of data sources on these analyses is rarely tested. Here, we test the impact of climatic data choice on the prediction of current and Plio-Pleistocene suitable habitats for two distantly related, but broadly sympatric, salamanders endemic to the Korean Peninsula. We ran MaxEnt separately on WorldClim and CHELSA climate data. We then hindcasted ENMs to five time periods of the Plio-Pleistocene, bracketing the estimated intraspecific divergence times for these species. We then quantified the differences in predictions between WorldClim- and CHELSA-based models. Also, given the sympatry and similar habitat requirements of the two species, we tested for niche overlaps using niche identity and background tests and tested the sensitivity of the results to climatic data choice. RESULTS: The ENMs successfully predicted contemporary suitable habitats for the two species. However, the predictions were highly sensitive to climatic data choice as well as variable combinations. The hindcasted ENMs produced contrasting predictions depending on the choice of climatic dataset and failed to predict suitable habitats for some Pleistocene time periods regardless of the climatic data choice. The niche analyses were also sensitive to climatic data choice, with results suggesting either niche overlaps or divergence depending on the climatic dataset used for the analyses. CONCLUSIONS: Our study highlights the influence of climatic data choice on the outcomes of ENMs and niche analyses. Our results also underscore the limitations of macroclimate-based ENMs, especially when the species is likely buffered from macroclimatic changes by microhabitat. We argue for the need for additional ecological, ecophysiological, and population genomic studies to better understand the range formation of these enigmatic species.

Animals

Florisbad and human population succession in Southern Africa.

The human cranium recovered at Florisbad in 1932 is compared with other Sub-Saharan African hominid remains from Broken Hill, the Omo and Klasies River Mouth. The Florisbad frontal is very broad, but despite this breadth and differences in zygomatic form, there is a definite resemblance to archaic Homo sapiens from Broken Hill. There is also some similarity to both Omo I and Omo II, while fragmentary remains from Klasies River are more lightly built and hence more modern in appearance. These impressions are strengthened by measurement and statistical analysis, which demonstrates that Florisbad and Broken Hill are distant from recent African populations. Even if Florisbad is less archaic than the earlier (Middle Pleistocene?) hominid, it is not noticeably Bushman-like. New dates suggestive of early Upper Pleistocene antiquity also place Florisbad securely in a lineage containing Broken Hill, and there is no evidence to support special ties with any one group of living Africans.

Biological Evolution

Fossil reptilian gelatins.

1. Studies have been made of products obtained by treating several reptilian fossil bones with dilute formic acid. 2. Their amounts and compositions have been compared with those of similarly treated fresh and Pleistocene bones. 3. Fewer and small peptides are obtained from the older fossils but in only one instance was the composition significantly different from that of the total protein. 4. For the collagenous proteins aspartic acid was the most readily freed by hydrolysis; for these older proteins it is alanine. 5. Unlike the Pleistocene proteins, the chromatograms from these older proteins have always had peaks in the position of beta-alanine, and usually others of unidentified origin.

Amino Acids

A Million Years of Mammoth Mitogenome Evolution.

The genomic study of specimens dating to the Early and Middle Pleistocene (EP and MP), a period spanning from 2.6 million years ago (Ma) to 126 thousand years ago (ka), has the potential to elucidate the evolutionary processes that shaped present-day biodiversity. Obtaining genomic data from this period is challenging, but mitochondrial DNA, given its higher abundance compared to nuclear DNA, could play an important role to understand evolutionary processes at this time scale. In this study, we report 34 new mitogenomes, including two EP and nine MP mammoth (Mammuthus spp.) specimens from Siberia and North America and analyze them jointly with >200 publicly available mitogenomes to reconstruct a transect of mammoth mitogenome diversity throughout the last million years. We find that our EP mitogenomes fall outside the diversity of all Late Pleistocene (LP) mammoths, while those derived from MP mammoths are basal to LP mammoth Clades 2 and 3, supporting an ancient Siberian origin of these lineages. In contrast, the geographical origin of Clade 1 remains unresolved. With these new deep-time mitogenomes, we observe diversification events across all clades that appear consistent with previously hypothesized MP and LP demographic changes. Furthermore, we improve upon an existing methodology for molecular clock dating of specimens >50 ka, demonstrating that specimens need to be individually dated to avoid biases in their age estimates. Both the molecular and analytical improvements presented here highlight the importance of deep-time genomic data to discover long-lost genetic diversity, enabling better assessments of evolutionary histories.

Animals

Suid evolution and correlation of African hominid localities.

Recently discovered Plio-Pleistocene vertebrate assemblages have allowed complete systematic revision of the sub-Saharan African Suidae. New phylogenies are proposed for the 7 genera and 16 species of fossil and extant representatives. Suids are common elements of African Plio-Pleistocene faunas, and their evolutionary trends, particularly in the species Mesochoerus limnetes and Metridiochoerus andrewsi, are of great correlative value. Suid data are employed in a refinement of stratigraphic correlations at Omo Shungura, Olduvai, and east of Lake Turkana (formerly East Rudolf) and in a correlation of East African and South African sites, with important implications for interpretation of hominid evolution. The suid record also bears significantly on questions of theoretical evolutionary biology.

Africa

Paleogenomics and habitat modeling reveal temperate Eurasian origins of woolly rhinoceroses.

The woolly rhinoceros was a prominent Ice Age megafaunal species, and there is limited knowledge regarding its origin and responses to past glacial cycles. We sequenced 29 mitochondrial and 14 nuclear genomes from Pleistocene specimens across Eurasia and modeled the species' habitats over the past 500,000 years. Our results suggest that its maternal genetic diversity mainly evolved in temperate Eurasia around 460 thousand to 420 thousand years ago during a prolonged glacial-interglacial transition. We found that a ~170-thousand-year-old East Asian individual was ancestral to all later populations, indicating East Asia as one possible origin of Late Pleistocene ancestry. We also identified the Altai region as a major climatic refugium. These findings highlight the crucial role of temperate Eurasia in the evolution of woolly rhinoceroses and the diversification of cold-adapted megafauna.

Animals

Evolutionary history of Aotearoa New Zealand's extinct mātuhituhi | bush wren.

The reconstruction of ecosystem responses to past climate change has historically focused on large vertebrates. In contrast, small vertebrates with potentially stricter habitat preferences have been neglected in ancient DNA studies despite their potential utility as proxies for inferring geographic and temporal changes in habitat. Aotearoa New Zealand's acanthisittid wrens are a speciose group of tiny perching birds, including the mātuhituhi | bush wren (Xenicus longipes ssp.). Despite its relatively recent extinction in the 1970s, very little is known about this enigmatic bird. Here we sequence mitochondrial genomes and nuclear ultra conserved genomic elements from 32 historical bush wren specimens to reconstruct their evolutionary history. We also genetically sex specimens and reanalyse their plumage to reconstruct aspects of bush wren plumage variation. Our analyses suggest North and South Island bush wren populations diverged 2.6 million years ago when narrowing and closure of Plio-Pleistocene seaways allowed colonisation of new habitats, followed by rapid glaciation-driven diversification of South Island populations 470,000-94,000 years ago. Genetic sexing allowed an accurate reconstruction of ontogenetic, sexual, and geographic variation in plumage. Our multidisciplinary data supports recognition of North and South Island populations as separate species, and the description of a new subspecies X. longipes perditus subsp. nov. This research shows how ecosystems can buffer against the impacts of climate change up to an ecological tipping point, which has important lessons for conservation management in a fast-changing world.

Acanthisittidae

Homoploid Hybrid Speciation in a Marine Pelagic Fish.

Homoploid hybrid speciation (HHS) is an enigmatic evolutionary process where new species arise through hybridisation of divergent lineages without changes in chromosome number. Although increasingly documented in various taxa and ecosystems, convincing cases of HHS in marine fishes have been lacking. This study presents a possible case of HHS in a pelagic marine fish based on comprehensive genomic, morphological, and ecological analyses. Population genomics, species tree estimation, and tests of introgression and admixture identified three sympatric clusters in Megalaspis cordyla in the western Pacific and the admixed nature of one cluster between the others. Moreover, model-based demographic inference favoured a hybrid speciation scenario over introgression for the origin of the admixed cluster. While contemporary gene flow suggested partial reproductive isolation, examination of occurrence data and ecologically relevant morphological characters suggested ecological differences between the clusters, potentially contributing to the reproductive isolation and niche partitioning in sympatry. The clusters are also morphologically distinguishable and thus can be taxonomically recognised as separate species. The hybrid cluster is restricted to the coasts of Taiwan and Japan, where all three clusters coexist. The parental clusters are additionally found in lower latitudes, where they display non-overlapping distributions. Given the geographical distributions, estimated times of species formation, and patterns of historical demographic changes, we propose that the Pleistocene glacial cycles were the primary driver of HHS in this system. We also develop an ecogeographic model of HHS in marine coastal ecosystems, including a novel hypothesis to explain the initial stages of HHS.

Animals

Regional variation in tooth size and pathology in fossil hominids.

Tooth size and dental pathology in fossil hominids were studied to test for regional differences in these parameters. The results showed little regional variation in tooth size for the Middle and Upper Pleistocene sites compared (except for Krapina) but considerable differences in the severity of attrition and dental pathology. These differences were considered indicative of regional differences in the functional load borne by the teeth, and in view of the similar technological status of the groups studied, were attributed to environmental differences in the diet. Since, in all regions, reduction in tooth size appeared to continue at the same rate for the periods investigated, no association can be established between presumed selective pressures related to differences in functional demands made on the dentition, and tooth reduction.

Adult

Early hominid enamel hypoplasia.

Enamel hypoplasia in the South African Plio-Pleistocene fossil hominid sample is examined. The Swartkrans hominids are shown to have a higher incidence of hypoplasia than the Sterkfontein hominids. Within the Swartkrans sample, individuals with hypoplasia of the first upper molar have lower-than-expected ages of death. Possible taphonomic explanations for these observations are discussed.

Animals

Paleogenomic sex inference of mammoth remains sheds light on the anthropogenic nature of bone accumulations.

Whether large accumulations of woolly mammoth (Mammuthus primigenius) bones reflect natural mortality or deliberate human resource exploitation has long been debated, with major implications for understanding Late Pleistocene human-megafaunal interactions.1,2,3,4,5 Here, we use ancient DNA to investigate site-formation hypotheses by comparing genetic sex ratios from mammoth remains recovered in putative anthropogenic bone accumulations and from geographically dispersed, non-anthropogenic contexts. We studied genome-wide data from 521 woolly mammoths-including 100 mammoths from bone accumulation sites and 421 mammoths from natural depositional settings across Eurasia and North America-of which 448 are newly generated. Genetic sex determination reveals a striking contrast between contexts: mammoths from dispersed sites show a male bias (∼66.5%), which is consistent with the heightened vulnerability of solitary males to hazards such as natural traps, where bones are more likely to be preserved, whereas mammoths from anthropogenic bone accumulations are predominantly female (∼70%). This female bias is pervasive across multiple sites, indicating an anthropogenic origin for these accumulations as a result of Upper Paleolithic hunters preferentially exploiting female mammoths, possibly derived from herd contexts. Together, these results provide population-scale genetic evidence that highlights the central role of mammoths in the subsistence and material economies of some Paleolithic communities.

Animals