Deep Timescales Research Articles

The Divergent Responses of Salinity Generalists to Hyposaline Stress Provide Insights Into the Colonisation of Freshwaters by Diatoms.

Environmental transitions, such as the salinity divide separating marine and fresh waters, shape biodiversity over both shallow and deep timescales, opening up new niches and creating opportunities for accelerated speciation and adaptive radiation. Understanding the genetics of environmental adaptation is central to understanding how organisms colonise and subsequently diversify in new habitats. We used time-resolved transcriptomics to contrast the hyposalinity stress responses of two diatoms. Skeletonema marinoi has deep marine ancestry but has recently invaded brackish waters. Cyclotella cryptica has deep freshwater ancestry and can withstand a much broader salinity range. Skeletonema marinoi is less adept at mitigating even mild salinity stress compared to Cyclotella cryptica, which has distinct mechanisms for rapid mitigation of hyposaline stress and long-term growth in low salinity. We show that the cellular mechanisms underlying low salinity tolerance, which has allowed diversification across freshwater habitats worldwide, includes elements that are both conserved and variable across the diatom lineage. The balance between ancestral and lineage-specific environmental responses in phytoplankton have shaped marine-freshwater transitions on evolutionary timescales and, on contemporary timescales, will affect which lineages survive and adapt to changing ocean conditions.

Climate Change Responses of High-Elevation Polylepis Forests

The high mountain forests on Earth are characterized by sharp environmental heterogeneity, high species endemism, and unique phenotypic adaptations. Yet, global warming is jeopardizing this ecosystem as no other as some forests may have nowhere to go beyond the mountain’s summits. One of the most human-endangered high-elevation ecosystems is the Polylepis forest. Despite its vulnerability, forecasting climate reactions in this distinctive high mountain forest type remains a formidable challenge. Therefore, in this study, we modeled climate change responses of high-elevation allopatric Polylepis sericea and P. quadrijuga (Rosaceae) in the northern Andes. The analysis took into account VIF-prioritized bioclimatic variables for near-to-present and future (2081–2100 MPI-ESM1-2-HR) conditions. Model selection was carried out following the AUC scores of 12 MaxEnt and six machine learning algorithms. Predictive power reached 0.97 for MaxEnt’s model fc.H_rm.1 (H-1). Results indicate a moderate vulnerability of P. sericea, with a 29% loss of area in the trailing edge, due to climate change for the period 2081–2100. On the other hand, P. quadrijuga is likely to experience even larger distribution losses, up to 99%, for the same period. The findings of this study suggest that P. quadrijuga, as compared to P. sericea, exhibits a restricted ability to maintain the corresponding habitat requirements in the face of climatic change. Higher niche specialization of P. quadrijuga in the environmentally heterogeneous Eastern Cordillera contrasts with the more generalist nature of P. sericea in the topographically less complex Central Cordillera. In addition to climate change, this trend may be exacerbated by the detrimental effects of agriculture, mining, and an expanding rural population, which represent significant human-driven pressures yet to be modeled in the northern Andean highlands. Yet, based on previous studies, the historical population dynamics during the past glacial cycles suggests that range shifts could play a more significant role at deeper time scales than previously forecasted in the species’ reaction to climate change. Additionally, Polylepis forests may be able to endure at the current locations via local adaptation and plasticity, major drivers of the phenotypic variation in long-lived trees, counteracting the vulnerability scenario under a niche conservatism hypothesis. Ultimately, the modeling procedure employed in this study provides insights into the potential effects of climate change on Polylepis forests 70 years from now. Oncoming studies may consider alternative responses inherent to the gene pool of the species and the interaction with edaphic and biotic agents. We advocate for the application of comparable estimations in other high-elevation tree communities found at the tree line.

An efficient and robust ABC approach to infer the rate and strength of adaptation.

Inferring the effects of positive selection on genomes remains a critical step in characterizing the ultimate and proximate causes of adaptation across species, and quantifying positive selection remains a challenge due to the confounding effects of many other evolutionary processes. Robust and efficient approaches for adaptation inference could help characterize the rate and strength of adaptation in nonmodel species for which demographic history, mutational processes, and recombination patterns are not currently well-described. Here, we introduce an efficient and user-friendly extension of the McDonald-Kreitman test (ABC-MK) for quantifying long-term protein adaptation in specific lineages of interest. We characterize the performance of our approach with forward simulations and find that it is robust to many demographic perturbations and positive selection configurations, demonstrating its suitability for applications to nonmodel genomes. We apply ABC-MK to the human proteome and a set of known virus interacting proteins (VIPs) to test the long-term adaptation in genes interacting with viruses. We find substantially stronger signatures of positive selection on RNA-VIPs than DNA-VIPs, suggesting that RNA viruses may be an important driver of human adaptation over deep evolutionary time scales.

Bone collagen from subtropical Australia is preserved for more than 50,000 years

Ancient protein studies have demonstrated their utility for looking at a wide range of evolutionary and historical questions. The majority of palaeoproteomics studies to date have been restricted to high latitudes with relatively temperate environments. A better understanding of protein preservation at lower latitudes is critical for disentangling the mechanisms involved in the deep-time survival of ancient proteins, and for broadening the geographical applicability of palaeoproteomics. In this study, we aim to assess the level of collagen preservation in the Australian fossil record. Collagen preservation was systematically examined using a combination of thermal age estimates, Fourier Transform Infrared Spectroscopy, Zooarchaeology by Mass Spectrometry, and protein deamidation calculations. We reveal unexpected subtropical survival of collagen in bones more than 50 thousand years old, showing that protein preservation can exceed chemical predictions of collagen survival in bone. These findings challenge preconceptions concerning the suitability of palaeoproteomics in subtropical Pleistocene environments. We explore potential causes of this unexpected result to identify the underlying mechanisms leading to this exceptional preservation. This study serves as a starting point for the analysis of ancient proteins in other (sub)tropical contexts, and at deeper timescales.

Inaccurate fossil placement does not compromise tip‐dated divergence times

AbstractTime‐scaled phylogenies underpin the interrogation of evolutionary processes across deep timescales, as well as attempts to link these to Earth's history. By inferring the placement of fossils and using their ages as temporal constraints, tip dating under the fossilized birth–death (FBD) process provides a coherent prior on divergence times. At the same time, it also links topological and temporal accuracy, as incorrectly placed fossil terminals should misinform divergence times. This could pose serious issues for obtaining accurate node ages, yet the interaction between topological and temporal error has not been thoroughly explored. We simulate phylogenies and associated morphological datasets using methodologies that incorporate evolution under selection, and are benchmarked against empirical datasets. We find that datasets of 300 characters and realistic levels of missing data generally succeed in inferring the correct placement of fossils on a constrained extant backbone topology, and that true node ages are usually contained within Bayesian posterior distributions. While increased fossil sampling improves the accuracy of inferred ages, topological and temporal errors do not seem to be linked: analyses in which fossils resolve less accurately do not exhibit elevated errors in node age estimates. At the same time, inferred divergence times are biased, probably due to a mismatch between the FBD prior and the shape of our simulated trees. While these results are encouraging, suggesting that even fossils with uncertain affinities can provide useful temporal information, they also emphasize that palaeontological information cannot overturn discrepancies between model priors and the true diversification history.

Niche conservatism over deep timescales and diverse niche axes in land vertebrates

AbstractAimThere has been considerable interest in niche conservatism, the idea that ecological variables are similar among related species. Much research has focused on climatic niches of recently diverged species, rather than deeper timescales or non‐climatic niche axes. Furthermore, it has been suggested that conservatism disappears over deeper timescales, and is greater in alpha niche traits (like diet and microhabitat) than beta niche variables (like climate). Here, we test these latter two ideas by comparing patterns of phylogenetic conservatism among 10 niche variables across major clades of land vertebrates.LocationGlobal.Time PeriodPresent to 350 million years ago.Major Taxa StudiedTetrapods, including amphibians, mammals, lepidosaurs (including lizards and snakes), turtles, crocodilians and birds.MethodsThe 10 niche variables included four alpha niche components (diet, diel activity, habitat, body temperature) and six beta niche components (related to climatic temperature and precipitation). We analysed these variables on time‐calibrated phylogenies with similar taxon sampling (~1700 species), using phylogenetic signal (lambda) to estimate conservatism, along with the D statistic and estimates of evolutionary rates.ResultsPhylogenetic signal was generally strong across all variables, with lambda generally >0.80 (with 1.0 representing maximum signal). Nevertheless, mean phylogenetic signal was lower in beta niche traits than alpha niche traits (based on lambda and especially the D statistic), and alpha niche traits showed significantly slower rates of evolution.Main ConclusionsWe address two long‐held views in the literature on niche conservatism, rejecting one but supporting the other. We show that phylogenetic signal does not disappear over deep timescales for many important niche variables, even over 350 million years. We also generally support greater conservatism in alpha niche traits than beta niche traits over hundreds of millions of years, a pattern that was previously suggested (but not explicitly tested) based on closely related species.

Estimating transcriptome complexities across eukaryotes

BackgroundGenomic complexity is a growing field of evolution, with case studies for comparative evolutionary analyses in model and emerging non-model systems. Understanding complexity and the functional components of the genome is an untapped wealth of knowledge ripe for exploration. With the “remarkable lack of correspondence” between genome size and complexity, there needs to be a way to quantify complexity across organisms. In this study, we use a set of complexity metrics that allow for evaluating changes in complexity using TranD.ResultsWe ascertain if complexity is increasing or decreasing across transcriptomes and at what structural level, as complexity varies. In this study, we define three metrics – TpG, EpT, and EpG- to quantify the transcriptome's complexity that encapsulates the dynamics of alternative splicing. Here we compare complexity metrics across 1) whole genome annotations, 2) a filtered subset of orthologs, and 3) novel genes to elucidate the impacts of orthologs and novel genes in transcript model analysis. Effective Exon Number (EEN) issued to compare the distribution of exon sizes within transcripts against random expectations of uniform exon placement. EEN accounts for differences in exon size, which is important because novel gene differences in complexity for orthologs and whole-transcriptome analyses are biased towards low-complexity genes with few exons and few alternative transcripts.ConclusionsWith our metric analyses, we are able to quantify changes in complexity across diverse lineages with greater precision and accuracy than previous cross-species comparisons under ortholog conditioning. These analyses represent a step toward whole-transcriptome analysis in the emerging field of non-model evolutionary genomics, with key insights for evolutionary inference of complexity changes on deep timescales across the tree of life. We suggest a means to quantify biases generated in ortholog calling and correct complexity analysis for lineage-specific effects. With these metrics, we directly assay the quantitative properties of newly formed lineage-specific genes as they lower complexity.

Direct observation of the evolution of cell-type-specific microRNA expression signatures supports the hematopoietic origin model of endothelial cells.

The evolution of specialized cell-types is a long-standing interest of biologists, but given the deep time-scales very difficult to reconstruct or observe. microRNAs have been linked to the evolution of cellular complexity and may inform on specialization. The endothelium is a vertebrate-specific specialization of the circulatory system that enabled a critical new level of vasoregulation. The evolutionary origin of these endothelial cells is unclear. We hypothesized that Mir-126, an endothelial cell-specific microRNA may be informative. We here reconstruct the evolutionary history of Mir-126. Mir-126 likely appeared in the last common ancestor of vertebrates and tunicates, which was a species without an endothelium, within an intron of the evolutionary much older EGF Like Domain Multiple (Egfl) locus. Mir-126 has a complex evolutionary history due to duplications and losses of both the host gene and the microRNA. Taking advantage of the strong evolutionary conservation of the microRNA among Olfactores, and using RNA in situ hybridization, we localized Mir-126 in the tunicate Ciona robusta. We found exclusive expression of the mature Mir-126 in granular amebocytes, supporting a long-proposed scenario that endothelial cells arose from hemoblasts, a type of proto-endothelial amoebocyte found throughout invertebrates. This observed change of expression of Mir-126 from proto-endothelial amoebocytes in the tunicate to endothelial cells in vertebrates is the first direct observation of the evolution of a cell-type in relation to microRNA expression indicating that microRNAs can be a prerequisite of cell-type evolution.

Widespread reticulate evolution in an adaptive radiation.

A fundamental assumption of evolutionary biology is that phylogeny follows a bifurcating process. However, hybrid speciation and introgression are becoming more widely documented in many groups. Hybrid inference studies have been historically limited to small sets of taxa, while exploration of the prevalence and trends of reticulation at deep time scales remains unexplored. We study the evolutionary history of an adaptive radiation of 109 gemsnakes in Madagascar (Pseudoxyrhophiinae) to identify potential instances of introgression. Using several network inference methods, we find 12 reticulation events within the 22-million-year evolutionary history of gemsnakes, producing 28% of the diversity for the group, including one reticulation that resulted in the diversification of an 18 species radiation. These reticulations are found at nodes with high gene tree discordance and occurred among parental lineages distributed along a north-south axis that share similar ecologies. Younger hybrids occupy intermediate contact zones between the parent lineages showing that post-speciation dispersal in this group has not eroded the spatial signatures of introgression. Reticulations accumulated consistently over time, despite drops in overall speciation rates during the Pleistocene. This suggests that while bifurcating speciation rates may decline as the result of species accumulation and environmental change, speciation by hybridization may be more robust to these processes.

Ruminant inner ear shape records 35 million years of neutral evolution

Extrinsic and intrinsic factors impact diversity. On deep-time scales, the extrinsic impact of climate and geology are crucial, but poorly understood. Here, we use the inner ear morphology of ruminant artiodactyls to test for a deep-time correlation between a low adaptive anatomical structure and both extrinsic and intrinsic variables. We apply geometric morphometric analyses in a phylogenetic frame to X-ray computed tomographic data from 191 ruminant species. Contrasting results across ruminant clades show that neutral evolutionary processes over time may strongly influence the evolution of inner ear morphology. Extant, ecologically diversified clades increase their evolutionary rate with decreasing Cenozoic global temperatures. Evolutionary rate peaks with the colonization of new continents. Simultaneously, ecologically restricted clades show declining or unchanged rates. These results suggest that both climate and paleogeography produced heterogeneous environments, which likely facilitated Cervidae and Bovidae diversification and exemplifies the effect of extrinsic and intrinsic factors on evolution in ruminants.

Dust against the Anthropocene: Yhonnie Scarce’s nuclear geo-fictions

Against the universalizing of the Anthropocene, radioactive dust affects specific communities more than others. At the same time, it carries particles from local sites to cosmic horizons. Uranium dust encodes deep timescales of planetary formation and extinction as they intersect with histories of violence and extraction, myth and current politics. This article analyses artwork by Yhonnie Scarce, descendant of the Kokatha and Nukunu peoples of South Australia, arguing for a particulate geo-fiction as method of engagement with colonial politics of deep time. By sampling and literally unearthing nuclear histories, Scarce’s work traces more-than-human toxic ecologies. Through a condensation of uranium-scale temporalities, the present moment of its exhibition is prised open. This becomes a speculative ethical encounter with responsibilities to deep histories and futures beyond itself, the lingering after effects of British colonial violence inscribed into the materiality of the work.

Is Poaceae pollen size a useful proxy in palaeoecological studies? New insights from a Poaceae pollen morphological study in the Amazon

Poaceae (the grass family) is highly diverse, geographically widespread, and an important component of many terrestrial ecosystems. Poaceae pollen size has previously been suggested as a proxy to reconstruct the past vegetation and climates in the Amazon area, but it is still unclear if this variable can be used at broader spatial and deep-time scales. Here we set out to perform a comprehensive assessment and to test the robustness of this proxy. We generated pollen size data, as well as associated measurements of the pollen wall and pore, using 127 plant specimens from 58 species across the Poaceae phylogeny from the Amazon drainage basin (ADB). We tested the relationship between pollen size and a range of abiotic and biotic variables (vegetation type, soil composition, climate conditions, photosynthetic pathway and genome size), and fitted models using phylogenetic generalized least squares (P-GLS) and linear mixed models (LMM). Our measurement data shows that Poaceae pollen size presents a very wide range (18.77–71.62 μm), and varies not only among genera and species but also within species. There is no obvious relationship between pollen size and the explanatory variables considered here. We suggest that pollen size cannot be used as generally applicable proxy to reconstruct past vegetation and climate.

A common mechanism drives the alignment between the micro- and macroevolution of primate molars.

A central challenge for biology is to reveal how different levels of biological variation interact and shape diversity. However, recent experimental studies have indicated that prevailing models of evolution cannot readily explain the link between micro- and macroevolution at deep timescales. Here, we suggest that this paradox could be the result of a common mechanism driving a correlated pattern of evolution. We examine the proportionality between genetic variance and patterns of trait evolution in a system whose developmental processes are well understood to gain insight into how such alignment between morphological divergence and genetic variation might be maintained over macroevolutionary time. Primate molars present a model system by which to link developmental processes to evolutionary dynamics because of the biased pattern of variation that results from the developmental architecture regulating their formation. We consider how this biased variation is expressed at the population level, and how it manifests through evolution across primates. There is a strong correspondence between the macroevolutionary rates of primate molar divergence and their genetic variation. This suggests a model of evolution in which selection is closely aligned with the direction of genetic variance, phenotypic variance, and the underlying developmental architecture of anatomical traits.

Assessing Modern Calluna Heathland Fire Temperatures Using Raman Spectroscopy: Implications for Past Regimes and Geothermometry

Charcoal geothermometry continues to offer considerable potential in the study of palaeowildfires over decadal, centennial, millennial, and deep time scales—with substantial implications for the understanding of modern wildfire intensification. Recent developments in the application of Raman spectroscopy to carbonaceous organic material have indicated its capability to potentially reconstruct the palaeocharcoal formation temperature, and equivalent palaeowildfire pyrolysis intensity. Charcoal reflectance geothermometry (which also relies upon microstructural change with thermal maturation) has also been the subject of extensive modern evaluation, with multiple studies highlighting the key influence of energy flux on the resultant charcoal microstructure. The ability to accurately quantify modern wildfire temperatures based upon novel Raman-charcoal analyses has not yet been attempted. Using Raman band width-ratios (i.e., FWHMRa) and accompanying geothermometric trends to natural wildfire charcoals, our results identify differences between microstructurally-derived fire temperatures compared to those recorded during the fire event itself. Subsequent assessments of wildfire energy flux over time indicate no dominant influence for the observed differences, due to the inherent complexity of natural fire systems. Further analysis within this study, regarding the influence of reference pyrolysis methodology on microstructural change, also highlights the difficulty of creating accurate post-fire temperature reconstructions. The application of Raman spectroscopy, however, to the quantification of relative changes in fire temperature continues to prove effective and insightful.

Uncovering Cryptic Coevolution.

AbstractStudies of coevolution in the wild have largely focused on reciprocally specialized species pairs with striking and exaggerated phenotypes. Textbook examples include interactions between toxic newts and their garter snake predators, long-tongued flies and the flowers they pollinate, and weevils with elongated rostra used to bore through the defensive pericarp of their host plants. Although these studies have laid a foundation for understanding coevolution in the wild, they have also contributed to the widespread impression that coevolution is a rare and quirky sideshow to the day-to-day grind of ecology and evolution. In this perspective, we argue that the focus of coevolution has been biased toward the obvious and ignored the cryptic. We have focused on the obvious-studies of reciprocally specialized species pairs with exaggerated phenotypes-mainly because we have lacked the statistical tools required to study coevolution in more generalized and phenotypically mundane systems. Building from well-established coevolutionary theory, we illustrate how model-based approaches can be used to remove this barrier and begin estimating the strength of coevolutionary selection indirectly using routinely collected data, thus uncovering cryptic coevolution in more typical communities. By allowing the distribution of coevolutionary selection to be estimated across genomes, phylogenies, and communities and over deep timescales, these novel approaches have the potential to revolutionize the way we study coevolution. As we develop a road map to these next-generation approaches, we highlight recent studies making notable progress in this direction.

Elemental Memory: The Solid Fluidity of the Elements in the Nuclear Era

The epistemological challenges of the Anthropocene trouble distinctions of solid and fluid. In this contribution, the author proposes, after Gabrielle Hecht, that the nuclearity of the Anthropocene contributes significantly to destabilising these categories. Nuclear materials and ideas of nuclearity force (re)consideration of deep timescales and imperceptible processes, problematising fixed material ontologies. The article engages with nuclear matters and queries the logic of solids and fluids by developing the notion of elemental memory. An attention to elemental memory – an element’s capacity to auto-affect over time – reveals the inadequacy of terms like solid and fluid, and highlights the expressiveness of solid fluid substances. Empirically, the author demonstrates, first, how elemental memory informs the solid-fluid processuality of radioactive glasses, especially trinitite. Second, engaging with the work of artists Mari Keto and Erich Berger, she addresses the slow auto-transformations of radioactive minerals.

Evolution of male genitalia in the Drosophila repleta species group (Diptera: Drosophilidae).

The Drosophila repleta group comprises more than one hundred species that inhabit several environments in the Neotropics and use different hosts as rearing and feeding resources. Rather homogeneous in their external morphology, they are generally distinguished by the male genitalia, seemingly their fastest evolving morphological trait, constituting an excellent model to study patterns of genital evolution in the context of a continental adaptive radiation. Although much is known about the evolution of animal genitalia at population level, surveys on macroevolutionary scale of this phenomenon are scarce. This study used a suite of phylogenetic comparative methods to elucidate the macroevolutionary patterns of genital evolution through deep time and large continental scales. Our results indicate that male genital size and some aspects of shape have been evolving by speciational evolution, probably due to the microevolutionary processes involved in species mate recognition. In contrast, several features of the aedeagus shape seemed to have evolved in a gradual fashion, with heterogeneous evolutionary phenotypic rates among clades. In general, the tempo of the evolution of aedeagus morphology was constant from the origin of the group until the Pliocene, when it accelerated in some clades that diversified mainly in this period. The incidence of novel ecological conditions in the tempo of aedeagus evolution and the relationship between species mate recognition and speciation in the Drosophila repleta group are discussed.

Fathoming postnatural oceans: Towards a low trophic theory in the practices of feminist posthumanities

As the planet’s largest ecosystem, oceans stabilise climate, produce oxygen, store CO2 and host unfathomable biodiversity at a deep time-scale. In recent decades, scientific assessments have indicated that the oceans are seriously degraded to the detriment of most near-future societies. Human-induced impacts range from climate change, ocean acidification, loss of biodiversity, eutrophication and marine pollution to local degradation of marine and coastal environments. Such environmental violence takes form of both ‘spectacular’ events, like oil spills and ‘slow violence’, occurring gradually and out of sight. The purpose of this paper is to show four cases of coastal and marine forms of slow violence and to provide counter-accounts of how to reinvent our consumer imaginary at such locations, as well as to develop what is here referred to as ‘low-trophic theory,’ a situated ethical stance that attends to entanglements of consumption, food, violence, environmental adaptability and more-than-human care from the co-existential perspective of multispecies ethics. We combine field-philosophical case studies with insights from marine science, environmental art and cultural practices in the Baltic and North Sea region and feminist posthumanities. The paper shows that the oceanic imaginary is not a unified place, but rather, a set of forces, which requires renewed ethical approaches, conceptual inventiveness and practical creativity. Based on the case studies and examples presented, the authors conclude that the consideration of more-than-human ethical perspectives, provided by environmental arts and humanities is crucial for both research on nature and space, and for the flourishing of local multispecies communities. This paper thus inaugurates thinking and practice along the proposed here ethical stance of low-trophic theory, developed it along the methodological lines of feminist environmental posthumanities.

Phylogenetic history of vascular plant metabolism revealed using a macroevolutionary common garden.

While the fundamental biophysics of C3 photosynthesis is highly conserved across plants, substantial leaf structural and enzymatic variation translates into variability in rates of carbon assimilation. Although this variation is well documented, it remains poorly understood how photosynthetic rates evolve, and whether macroevolutionary changes are related to the evolution of leaf morphology and biochemistry. A substantial challenge in large-scale comparative studies is disentangling evolutionary adaptation from environmental acclimation. We overcome this by using a 'macroevolutionary common garden' approach in which we measured metabolic traits (Jmax and Vcmax) from 111 phylogenetically diverse species in a shared environment. We find substantial phylogenetic signal in these traits at moderate phylogenetic timescales, but this signal dissipates quickly at deeper scales. Morphological traits exhibit phylogenetic signal over much deeper timescales, suggesting that these are less evolutionarily constrained than metabolic traits. Furthermore, while morphological and biochemical traits (LMA, Narea and Carea) are weakly predictive of Jmax and Vcmax, evolutionary changes in these traits are mostly decoupled from changes in metabolic traits. This lack of tight evolutionary coupling implies that it may be incorrect to use changes in these functional traits in response to global change to infer that photosynthetic strategy is also evolving.

Testing the Utility of Alternative Metrics of Branch Support to Address the Ancient Evolutionary Radiation of Tunas, Stromateoids, and Allies (Teleostei: Pelagiaria)

The use of high-throughput sequencing technologies to produce genome-scale data sets was expected to settle some long-standing controversies across the Tree of Life, particularly in areas where short branches occur at deep timescales. Instead, these data sets have often yielded many well-supported but conflicting topologies, and highly variable gene-tree distributions. A variety of branch-support metrics beyond the nonparametric bootstrap are now available to assess how robust a phylogenetic hypothesis may be, as well as new methods to quantify gene-tree discordance. We applied multiple branch-support metrics to a study of an ancient group of marine fishes (Teleostei: Pelagiaria) whose interfamilial relationships have proven difficult to resolve due to a rapid accumulation of lineages very early in its history. We analyzed hundreds of loci including published ultraconserved elements and newly generated exonic data along with their flanking regions to represent all 16 extant families for more than 150 out of 284 valid species in the group. Branch support was typically lower at inter- than intra-familial relationships regardless of the type of marker used. Several nodes that were highly supported with bootstrap had a very low site and gene-tree concordance, revealing underlying conflict. Despite this conflict, we were able to identify four consistent interfamilial clades, each comprised of two or three families. Combining exons with their flanking regions also produced increased branch lengths at the deep branches of the pelagiarian tree. Our results demonstrate the limitations of employing current metrics of branch support and species-tree estimation when assessing the confidence of ancient evolutionary radiations and emphasize the necessity to embrace alternative measurements to explore phylogenetic uncertainty and discordance in phylogenomic data sets.[Concatenation; exons; introns; phylogenomics; species-tree methods; target capture.].