Ultrahigh foraging rates of Baikal seals make tiny endemic amphipods profitable in Lake Baikal

Understanding what, how, and how often apex predators hunt is important due to their disproportionately large effects on ecosystems. In Lake Baikal with rich endemic fauna, Baikal seals appear to eat, in addition to fishes, a tiny (<0.1 g) endemic amphipod Macrohectopus branickii (the world’s only freshwater planktonic species). Yet, its importance as prey to seals is unclear. Globally, amphipods are rarely targeted by single-prey feeding (i.e., nonfilter-feeding) mammals, presumably due to their small size. If M. branickii is energetically important prey, Baikal seals would exhibit exceptionally high foraging rates, potentially with behavioral and morphological specializations. Here, we used animal-borne accelerometers and video cameras to record Baikal seal foraging behavior. Unlike the prevailing view that they predominantly eat fishes, they also hunted M. branickii at the highest rates (mean, 57 individuals per dive) ever recorded for single-prey feeding aquatic mammals, leading to thousands of catches per day. These rates were achieved by gradual changes in dive depth following the diel vertical migration of M. branickii swarms. Examining museum specimens revealed that Baikal seals have the most specialized comb-like postcanine teeth in the subfamily Phocinae, allowing them to expel water while retaining prey during high-speed foraging. Our findings show unique mammal–amphipod interactions in an ancient lake, demonstrating that organisms even smaller than krill can be important prey for single-prey feeding aquatic mammals if the environment and predators’ adaptations allow high foraging rates. Further, our finding that Baikal seals directly eat macroplankton may explain why they are so abundant in this ultraoligotrophic lake.

The world’s deepest ancient lake, Lake Baikal in Russia, has a diverse endemic fauna, including Baikal seals Pusa sibirica, the apex predator of the lake and the only pinniped species inhabiting exclusively freshwater systems. The prevailing view is that Baikal seals predominantly eat fishes (pelagic sculpins, Comephorus spp. and Cottocomephorus spp.) (1–3). An endemic amphipod Macrohectopus branickii, the main diet of pelagic sculpins (4, 5), can also be found in the stomach contents of seals (6). Yet, its small size (<0.1 g) and the lack of indigestible body parts (e.g., otolith in fishes) make quantitative assessments of its contribution difficult. Amphipods in Lake Baikal have rapidly evolved from a few immigrant groups to >340 endemic species, representing a textbook example of adaptive radiation (7, 8). Among them, M. branickii exhibits an extreme adaptation with a distinct slender body and fully pelagic lifestyle (Fig. 1A), unlike many other benthic species with more stout bodies. This unusual amphipod is the dominant macroplankton in the lake. It forms dense aggregations and exhibits diel vertical migration (i.e., staying deep during the day and migrating to shallow depths at night) (9), similar to other macroplanktons in different systems. Therefore, M. branickii could potentially be important prey readily accessible at night for Baikal seals, much like Antarctic krill for some Antarctic seals (10). However, to what extent Baikal seals eat M. branickii in addition to fishes, let alone how they catch them, are currently unclear.Open in a separate windowFig. 1.Baikal seals hunt endemic amphipods in Lake Baikal. (A) M. branickii, the world’s only freshwater amphipod with a fully pelagic lifestyle (Photo credit: S. Didorenko). (B) Image from animal-borne video footage, showing the seal about to hunt a M. branickii (yellow arrow) by stretching its neck. M. branickii is in an upside-down position with its paired antennae visible. (C) A foraging dive on M. branickii by a seal, showing depth, swim speed, pitch angle (with positive and negative values indicating upward and downward attitude, respectively), and body acceleration (i.e., the vectorial sum of triaxial accelerations). The simultaneously recorded video footage showed that the seal encountered and hunted M. branickii nearly continuously (total, 154 individuals) during the bottom phase (denoted by many red markers that appear as a line).Globally, amphipods are rarely targeted by aquatic mammals, except for a few filter-feeding baleen whales (11–13) and local populations of Arctic seals (14, 15), despite their high diversity and wide distribution. Amphipods are typically much smaller than Antarctic krill, a preferred crustacean prey of many aquatic mammals, including pinnipeds (10). Gaining an energy surplus by hunting small amphipods is thus likely difficult for aquatic mammals, especially those that catch prey individually (i.e., pinnipeds and toothed whales). If Baikal seals eat M. branickii as part of their main diet, they would exhibit exceptionally high foraging rates with specialized foraging strategies, so that this small crustacean becomes energetically profitable. Moreover, if M. branickii is part of their diet, Baikal seals might also exhibit morphological adaptations on their feeding apparatus (i.e., teeth). Because water inevitably enters the mouth while animals forage during dives, high foraging rates could lead to excessive water intake unless they can expel water effectively. Crabeater and leopard seals, the two krill-feeding phocid seals in the Southern Ocean, have specialized postcanine teeth with developed cusps, allowing them to expel water through the cusp spacing while retaining krill (16, 17). Baikal seals also have cusped postcanine teeth, which are “like a comb” (1); however, the possibility that their teeth represent adaptations for feeding amphipods has never been explored.Here, we used modern electronic tags (i.e., accelerometers and video cameras) to show that Baikal seals individually hunt thousands of tiny (<0.1 g) M. branickii per day, primarily by tracking the diel vertical migrations of its swarms. We also show that Baikal seals have the most developed cusped teeth in the subfamily Phocinae (“northern true seals”), suggesting the function of their teeth as a filter for expelling water while retaining prey during high-speed foraging. Our findings reveal the unique predator–prey interaction driven by the adaptive radiation of amphipods along with the behavioral and morphological specializations of seals in an ancient lake. Our results also have important implications for the function of the Lake Baikal ecosystem.     

Early Holocene greening of the Sahara requires Mediterranean winter rainfall

The greening of the Sahara, associated with the African Humid Period (AHP) between ca. 14,500 and 5,000 y ago, is arguably the largest climate-induced environmental change in the Holocene; it is usually explained by the strengthening and northward expansion of the African monsoon in response to orbital forcing. However, the strengthened monsoon in Early to Middle Holocene climate model simulations cannot sustain vegetation in the Sahara or account for the increased humidity in the Mediterranean region. Here, we present an 18,500-y pollen and leaf-wax δD record from Lake Tislit (32° N) in Morocco, which provides quantitative reconstruction of winter and summer precipitation in northern Africa. The record from Lake Tislit shows that the northern Sahara and the Mediterranean region were wetter in the AHP because of increased winter precipitation and were not influenced by the monsoon. The increased seasonal contrast of insolation led to an intensification and southward shift of the Mediterranean winter precipitation system in addition to the intensified summer monsoon. Therefore, a winter rainfall zone must have met and possibly overlapped the monsoonal zone in the Sahara. Using a mechanistic vegetation model in Early Holocene conditions, we show that this seasonal distribution of rainfall is more efficient than the increased monsoon alone in generating a green Sahara vegetation cover, in agreement with observed vegetation. This conceptual framework should be taken into consideration in Earth system paleoclimate simulations used to explore the mechanisms of African climatic and environmental sensitivity.

Moisture availability in northern Africa, from the Sahel to the Mediterranean coast, is a critical issue for both ecosystems and human societies yet represents one of the largest uncertainties in future climate simulations (1, 2). The humid time span in the African Sahel and Sahara, known as the African humid period (AHP) (3–13), occurred in northern Africa after the last glacial period (4, 10, 11, 14–16) and lasted from ca. 14.5 to 5 ky ago (ka), with an optimum between 11 and 6 ka (11, 16). This prominent climatic event allowed semiarid, subtropical, and tropical plant species to spread outside their modern ranges (14) into the Sahara and human populations to inhabit what is known as the green Sahara (5, 17).The green Sahara is an example of extreme environmental change, which highlights the region’s extraordinary sensitivity and the need to better understand its hydroclimatic variability. Current explanations for the greening of the Sahara point to the Earth’s orbital changes during the Early Holocene, leading to increased boreal summer (JJA) insolation, which drove the intensification and northward expansion of the JJA monsoon over northern Africa (15, 18), aided by strong positive feedbacks from the land surface (19–22). Reproducing the green Sahara has posed a lasting challenge for climate modelers. The influence of the African monsoon extends only to ∼24° N (with or without interactive vegetation) in most Middle Holocene simulations, which is insufficient to sustain a vegetated Sahara. Models that integrate vegetation, dust, and soil feedbacks push the monsoon influence further north but still have discrepancies with proxy data (18, 23, 24).When all surface feedbacks are prescribed, simulated precipitation in the northern Sahara is still too low compared to paleoclimatic evidence for substantially increased moisture at 31° N (11, 13) or too high in the 15 to 20° N range (20), creating incompatibility with prescribed vegetation (22). Additional sources of moisture (25, 26) may have contributed to an AHP that extended toward the Mediterranean borderlands through different mechanisms. However, identifying the moisture sources over North Africa during the AHP requires paleoclimate records of both winter (DJF) and JJA precipitation.In the High Atlas Mountains, we collected an 8.5-m sediment core from Lake Tislit (ca. 32° N). The lake traps pollen grains from the surrounding landscape and, as a closed lake, is highly sensitive to hydroclimate fluctuations. It is ideally located for capturing the climatic variability of the Mediterranean and northwestern Sahara (Fig. 1). The Tislit sequence yielded unique hydrological data from leaf-wax stable isotopes and ostracod stable oxygen isotopes (δ¹⁸O), as well as a quantified time series of seasonal rainfall from the fossil pollen assemblages. Based on the findings from the Tislit record, we propose a precipitation regime for the AHP, including both Mediterranean DJF precipitation and monsoon JJA precipitation increases. Using a dynamic vegetation model for a conceptual experiment with 9 ka boundary conditions, we evaluate how a change in the seasonal distribution of precipitation over the Sahara can affect its revegetation.Open in a separate windowFig. 1.Maps showing the location of Lake Tislit, core GC27 (11), and Lake Yoa (6), along with the schematic position of the Inter Tropical Convergence Zone, with modern mean JJA (A) and DJF (B) rainfall (56). Map C shows the correlation coefficients (r) between Tislit and northern Morocco for DJF precipitation variability over the 1901 to 2010 time period (using the 20th century reanalysis of National Oceanic and Atmospheric Administration; https://psl.noaa.gov/data/20thC_Rean/). The limit of statistical significance (0.05 level) is shown by the dashed black line. Gray contours indicate annual precipitation isohyets (millimeter/year).     

Stable metal anodes enabled by a labile organic molecule bonded to a reduced graphene oxide aerogel

Metallic anodes (lithium, sodium, and zinc) are attractive for rechargeable battery technologies but are plagued by an unfavorable metal–electrolyte interface that leads to nonuniform metal deposition and an unstable solid–electrolyte interphase (SEI). Here we report the use of electrochemically labile molecules to regulate the electrochemical interface and guide even lithium deposition and a stable SEI. The molecule, benzenesulfonyl fluoride, was bonded to the surface of a reduced graphene oxide aerogel. During metal deposition, this labile molecule not only generates a metal-coordinating benzenesulfonate anion that guides homogeneous metal deposition but also contributes lithium fluoride to the SEI to improve Li surface passivation. Consequently, high-efficiency lithium deposition with a low nucleation overpotential was achieved at a high current density of 6.0 mA cm⁻². A Li|LiCoO₂ cell had a capacity retention of 85.3% after 400 cycles, and the cell also tolerated low-temperature (−10 °C) operation without additional capacity fading. This strategy was applied to sodium and zinc anodes as well.

Rechargeable batteries based on metal anodes including lithium (Li), sodium (Na), and zinc (Zn) show great promise in achieving high energy density (1–3). Unfortunately, the electrochemical interface of the metal anodes is not favorable for metal deposition. Metal nucleation is inhomogeneous at the surface, leading to the growth of metal dendrites (4–7) and the formation of an unstable solid–electrolyte interphase (SEI) that is incapable of protecting metals from the side reactions with the electrolyte (8–12).Substantial efforts have been devoted to stabilizing the interface of metal anodes, especially for Li metal. These include the design of artificial protective layers (13–17), alternative electrolytes (18–24), and sacrificial additives (25–30) to stabilize the metal–electrolyte interface, the development of mechanically robust coatings (31–34) to block Li dendrite growth, and the use of structured scaffolds to host dendrite-free Li deposition by reducing local current densities (35–43). However, the performance of metal anodes remains poor under high-current or low-temperature conditions. This is because the inhomogeneous Li nucleation and unstable SEI problems have not been well addressed, and these problems at the interface are even exacerbated under critical operating conditions, especially high-current densities and low temperatures (5, 6, 44).Toward this end, we report a simple molecular approach for regulating the electrochemical interface of metal anodes, which enables even Li deposition and stable SEI formation in a conventional electrolyte. This was realized by bonding a labile organic molecule, benzenesulfonyl fluoride (BSF), to a reduced graphene oxide (rGO) aerogel surface as the Li anode host (Fig. 1A). During Li deposition, BSF molecules electrochemically decompose at the interface and generate benzenesulfonate anions bonded to the rGO aerogel (Fig. 1B). The conjugated anions have a strong binding affinity for Li, serving as lithiophilic sites on the rGO surface to synergistically induce homogeneous Li nucleation of Li on the rGO surface. At the same time, BSF molecules contribute LiF to the SEI layer, which facilitates Li surface passivation (Fig. 1C). As a result, high-efficiency (99.2%) Li deposition was achieved at a Li deposition amount of 6.0 mAh cm⁻² and a current density of 6.0 mA cm⁻²; the barrier to Li nucleation was markedly reduced, as evidenced by the low nucleation overpotentials at high-current density (6.0 mA cm⁻²) or at a low temperature (−10 °C). A 400-cycle life with a capacity retention of 83.6% was achieved for a Li|LiCoO₂ (LCO) cell in a conventional carbonate electrolyte. Moreover, with the organic molecule-tuned interface, the Li|LCO cell can be stably cycled at a low operating temperature (−10 °C). This approach was applied to Na and Zn metal anodes as well.Open in a separate windowFig. 1.Illustration of a stable interface for Li deposition using a labile organic molecule, benzenesulfonyl fluoride (BSF). (A) Covalently bonded BSF on the rGO aerogel surface. (B) In situ generation of a lithiophilic conjugated anion (benzenesulfonate) and LiF on the surface during Li deposition. (C) Li nucleation preferentially occurs at the conjugated anion sites owing to the strong Li binding affinity, which leads to uniform Li deposition. In addition, the LiF that is formed is in the SEI layer and passivates the Li surface.     

A solution to a sex ratio puzzle in Melittobia wasps

The puzzling sex ratio behavior of Melittobia wasps has long posed one of the greatest questions in the field of sex allocation. Laboratory experiments have found that, in contrast to the predictions of theory and the behavior of numerous other organisms, Melittobia females do not produce fewer female-biased offspring sex ratios when more females lay eggs on a patch. We solve this puzzle by showing that, in nature, females of Melittobia australica have a sophisticated sex ratio behavior, in which their strategy also depends on whether they have dispersed from the patch where they emerged. When females have not dispersed, they lay eggs with close relatives, which keeps local mate competition high even with multiple females, and therefore, they are selected to produce consistently female-biased sex ratios. Laboratory experiments mimic these conditions. In contrast, when females disperse, they interact with nonrelatives, and thus adjust their sex ratio depending on the number of females laying eggs. Consequently, females appear to use dispersal status as an indirect cue of relatedness and whether they should adjust their sex ratio in response to the number of females laying eggs on the patch.

Sex allocation has produced many of the greatest success stories in the study of social behaviors (1–4). Time and time again, relatively simple theory has explained variation in how individuals allocate resources to male and female reproduction. Hamilton’s local mate competition (LMC) theory predicts that when n diploid females lay eggs on a patch and the offspring mate before the females disperse, the evolutionary stable proportion of male offspring (sex ratio) is (n − 1)/2n (Fig. 1) (5). A female-biased sex ratio is favored to reduce competition between sons (brothers) for mates and to provide more mates (daughters) for those sons (6–8). Consistent with this prediction, females of >40 species produce female-biased sex ratios and reduce this female bias when multiple females lay eggs on the same patch (higher n; Fig. 1) (9). The fit of data to theory is so good that the sex ratio under LMC has been exploited as a “model trait” to study the factors that can constrain “perfect adaptation” (4, 10–13).Open in a separate windowFig. 1.LMC. The sex ratio (proportion of sons) is plotted versus the number of females laying eggs on a patch. The bright green dashed line shows the LMC theory prediction for the haplodiploid species (5, 39). A more female-biased sex ratio is favored in haplodiploids because inbreeding increases the relative relatedness of mothers to their daughters (7, 32). Females of many species adjust their offspring sex ratio as predicted by theory, such as the parasitoid Nasonia vitripennis (green diamonds) (82). In contrast, the females of several Melittobia species, such as M. australica, continue to produce extremely female-biased sex ratios, irrespective of the number of females laying eggs on a patch (blue squares) (15).In stark contrast, the sex ratio behavior of Melittobia wasps has long been seen as one of the greatest problems for the field of sex allocation (3, 4, 14–21). The life cycle of Melittobia wasps matches the assumptions of Hamilton’s LMC theory (5, 15, 19, 21). Females lay eggs in the larvae or pupae of solitary wasps and bees, and then after emergence, female offspring mate with the short-winged males, who do not disperse. However, laboratory experiments on four Melittobia species have found that females lay extremely female-biased sex ratios (1 to 5% males) and that these extremely female-biased sex ratios change little with increasing number of females laying eggs on a patch (higher n; Fig. 1) (15, 17–20, 22). A number of hypotheses to explain this lack of sex ratio adjustment have been investigated and rejected, including sex ratio distorters, sex differential mortality, asymmetrical male competition, and reciprocal cooperation (15–18, 20, 22–26).We tested whether Melittobia’s unusual sex ratio behavior can be explained by females being related to the other females laying eggs on the same patch. After mating, some females disperse to find new patches, while some may stay at the natal patch to lay eggs on previously unexploited hosts (Fig. 2). If females do not disperse, they can be related to the other females laying eggs on the same host (27–31). If females laying eggs on a host are related, this increases the extent to which relatives are competing for mates and so can favor an even more female-biased sex ratio (28, 32–35). Although most parasitoid species appear unable to directly assess relatedness, dispersal behavior could provide an indirect cue of whether females are with close relatives (36–38). Consequently, we predict that when females do not disperse and so are more likely to be with closer relatives, they should maintain extremely female-biased sex ratios, even when multiple females lay eggs on a patch (28, 35).Open in a separate windowFig. 2.Host nest and dispersal manners of Melittobia. (A) Photograph of the prepupae of the leaf-cutter bee C. sculpturalis nested in a bamboo cane and (B) a diagram showing two ways that Melittobia females find new hosts. The mothers of C. sculpturalis build nursing nests with pine resin consisting of individual cells in which their offspring develop. If Melittobia wasps parasitize a host in a cell, female offspring that mate with males inside the cell find a different host on the same patch (bamboo cane) or disperse by flying to other patches.We tested whether the sex ratio of Melittobia australica can be explained by dispersal status in a natural population. We examined how the sex ratio produced by females varies with the number of females laying eggs on a patch and whether or not they have dispersed before laying eggs. To match our data to the predictions of theory, we developed a mathematical model tailored to the unique population structure of Melittobia, where dispersal can be a cue of relatedness. We then conducted a laboratory experiment to test whether Melittobia females are able to directly access the relatedness to other females and adjust their sex ratio behavior accordingly. Our results suggest that females are adjusting their sex ratio in response to both the number of females laying eggs on a patch and their relatedness to the other females. However, relatedness is assessed indirectly by whether or not they have dispersed. Consequently, the solution to the puzzling behavior reflects a more-refined sex ratio strategy.     

Observation of non-Hermitian topology and its bulk–edge correspondence in an active mechanical metamaterial

Topological edge modes are excitations that are localized at the materials’ edges and yet are characterized by a topological invariant defined in the bulk. Such bulk–edge correspondence has enabled the creation of robust electronic, electromagnetic, and mechanical transport properties across a wide range of systems, from cold atoms to metamaterials, active matter, and geophysical flows. Recently, the advent of non-Hermitian topological systems—wherein energy is not conserved—has sparked considerable theoretical advances. In particular, novel topological phases that can only exist in non-Hermitian systems have been introduced. However, whether such phases can be experimentally observed, and what their properties are, have remained open questions. Here, we identify and observe a form of bulk–edge correspondence for a particular non-Hermitian topological phase. We find that a change in the bulk non-Hermitian topological invariant leads to a change of topological edge-mode localization together with peculiar purely non-Hermitian properties. Using a quantum-to-classical analogy, we create a mechanical metamaterial with nonreciprocal interactions, in which we observe experimentally the predicted bulk–edge correspondence, demonstrating its robustness. Our results open avenues for the field of non-Hermitian topology and for manipulating waves in unprecedented fashions.

The inclusion of non-Hermitian features in topological insulators has recently seen an explosion of activity. Exciting developments include tunable wave guides that are robust to disorder (1–3), structure-free systems (4, 5), and topological lasers and pumping (6–10). In these systems, active components are introduced to typically 1) break time-reversal symmetry to create topological insulators with unidirectional edge modes (1–5) and 2) pump topologically protected edge modes, thus harnessing Hermitian topology in non-Hermitian settings (6–9, 11). In parallel, extensive theoretical efforts have generalized the concept of a topological insulator to truly non-Hermitian phases that cannot be realized in Hermitian materials (12–14). However, such non-Hermitian topology and its bulk–edge correspondence remain a matter of intense debate. On the one hand, it has been argued that the usual bulk–edge correspondence breaks down in non-Hermitian settings, while on the other hand, new topological invariants specific to non-Hermitian systems have been proposed to capture particular properties of their edge modes (15–20).Here, focusing on a non-Hermitian version of the Su–Schrieffer–Heeger (SSH) model (15–17, 21) with an odd number of sites (Fig. 1A), we find that a change in the bulk non-Hermitian topological invariant is accompanied by a localization change in the zero-energy edge modes. This finding suggests the existence of a bulk–edge correspondence for this type of truly non-Hermitian topology. We further construct a mechanical analogue of the non-Hermitian quantum model (Fig. 1B) and create a mechanical metamaterial (Fig. 1C) in which we observe the predicted correspondence between the non-Hermitian topological invariant and the topological edge mode. In particular, we report that the edge mode in the non-Hermitian topological phase has a peculiar nature, as it is localized on the rigid rather than the floppy side of the mechanical metamaterial.Open in a separate windowFig. 1.Quantum-to-classical mapping of a chain with non-Hermitian topology. (A) An SSH chain with two sublattices, A (in red) and B (in blue), augmented with nonreciprocal variations in the hopping amplitudes (indicated by ±ε). (B) The nonreciprocal classical analog of the augmented SSH chain, in which the classical masses (in red) correspond to the A sites in the quantum model, while the nonreciprocal springs (in blue) are analogous to the B sites. (C) Picture of the mechanical metamaterial realizing the nonreciprocal classical analogue of the augmented SSH model.     

Structure,self-assembly,and properties of a truncated reflectin variant

Naturally occurring and recombinant protein-based materials are frequently employed for the study of fundamental biological processes and are often leveraged for applications in areas as diverse as electronics, optics, bioengineering, medicine, and even fashion. Within this context, unique structural proteins known as reflectins have recently attracted substantial attention due to their key roles in the fascinating color-changing capabilities of cephalopods and their technological potential as biophotonic and bioelectronic materials. However, progress toward understanding reflectins has been hindered by their atypical aromatic and charged residue-enriched sequences, extreme sensitivities to subtle changes in environmental conditions, and well-known propensities for aggregation. Herein, we elucidate the structure of a reflectin variant at the molecular level, demonstrate a straightforward mechanical agitation-based methodology for controlling this variant’s hierarchical assembly, and establish a direct correlation between the protein’s structural characteristics and intrinsic optical properties. Altogether, our findings address multiple challenges associated with the development of reflectins as materials, furnish molecular-level insight into the mechanistic underpinnings of cephalopod skin cells’ color-changing functionalities, and may inform new research directions across biochemistry, cellular biology, bioengineering, and optics.

Materials from naturally occurring and recombinant proteins are frequently employed for the study of fundamental biological processes and leveraged for applications in fields as diverse as electronics, optics, bioengineering, medicine, and fashion (1–13). Such broad utility is enabled by the numerous advantageous characteristics of protein-based materials, which include sequence modularity, controllable self-assembly, stimuli-responsiveness, straightforward processability, inherent biological compatibility, and customizable functionality (1–13). Within this context, unique structural proteins known as reflectins have recently attracted substantial attention because of their key roles in the fascinating color-changing capabilities of cephalopods, such as the squid shown in Fig. 1A, and have furthermore demonstrated their utility for unconventional biophotonic and bioelectronic technologies (11–40). For example, in vivo, Bragg stack-like ultrastructures from reflectin-based high refractive index lamellae (membrane-enclosed platelets) are responsible for the angle-dependent narrowband reflectance (iridescence) of squid iridophores, as shown in Fig. 1B (15–20). Analogously, folded membranes containing distributed reflectin-based particle arrangements within sheath cells lead to the mechanically actuated iridescence of squid chromatophore organs, as shown in Fig. 1C (15, 16, 21, 22). Moreover, in vitro, films processed from squid reflectins not only exhibit proton conductivities on par with some state-of-the-art artificial materials (23–27) but also support the growth of murine and human neural stem cells (28, 29). Additionally, morphologically variable coatings assembled from different reflectin isoforms can enable the functionality of chemically and electrically actuated color-changing devices, dynamic near-infrared camouflage platforms, and stimuli-responsive photonic architectures (27, 30–34). When considered together, these discoveries and demonstrations constitute compelling motivation for the continued exploration of reflectins as model biomaterials.Open in a separate windowFig. 1.(A) A camera image of a D. pealeii squid for which the skin contains light-reflecting cells called iridophores (bright spots) and pigmented organs called chromatophores (colored spots). Image credit: Roger T. Hanlon (photographer). (B) An illustration of an iridophore (Left), which shows internal Bragg stack-like ultrastructures from reflectin-based lamellae (i.e., membrane-enclosed platelets) (Inset). (C) An illustration of a chromatophore organ (Left), which shows arrangements of reflectin-based particles within the sheath cells (Inset). (D) The logo of the 28-residue-long N-terminal motif (RMN), which depicts the constituent amino acids (Upper) and their predicted secondary structures (Lower). (E) The logo of the 28-residue-long internal motif (RMI), which depicts the constituent amino acids (Upper) and their predicted secondary structures (Lower). (F) The logo of the 21-residue-long C-terminal motif (RMC), which depicts the constituent amino acids (Upper) and their predicted secondary structures (Lower). (G) The amino acid sequence of full-length D. pealeii reflectin A1, which contains a single RMN motif (gray oval) and five RMI motifs (orange ovals). (H) An illustration of the selection of the prototypical truncated reflectin variant (denoted as RfA1TV) from full-length D. pealeii reflectin A1.Given reflectins’ demonstrated significance from both fundamental biology and applications perspectives, some research effort has been devoted to resolving their three-dimensional (3D) structures (30, 31, 35–39). For example, fibers drawn from full-length Euprymna scolopes reflectin 1a and films processed from truncated E. scolopes reflectin 1a were shown to possess secondary structural elements (i.e., α-helices or β-sheets) (30, 31). In addition, precipitated nanoparticles and drop-cast films from full-length Doryteuthis pealeii reflectin A1 have exhibited β-character, which was seemingly associated with their conserved motifs (35, 36). Moreover, nanoparticles assembled from both full-length and truncated Sepia officinalis reflectin 2 variants have demonstrated signatures consistent with β-sheet or α-helical secondary structure, albeit in the presence of surfactants (38). However, such studies were made exceedingly challenging by reflectins’ atypical primary sequences enriched in aromatic and charged residues, documented extreme sensitivities to subtle changes in environmental conditions, and well-known propensities for poorly controlled aggregation (12, 14, 15, 30–32, 34–39). Consequently, the reported efforts have all suffered from multiple drawbacks, including the need for organic solvents or denaturants, the evaluation of only polydisperse or aggregated (rather than monomeric) proteins, a lack of consensus among different experimental techniques, inadequate resolution that precluded molecular-level insight, imperfect agreement between computational predictions and experimental observations, and/or the absence of conclusive correlations between structure and optical functionality. As such, there has emerged an exciting opportunity for investigating reflectins’ molecular structures, which remain poorly understood and the subject of some debate.Herein, we elucidate the structure of a reflectin variant at the molecular level, demonstrate a robust methodology for controlling this variant’s hierarchical assembly, and establish a direct correlation between its structural characteristics and optical properties. We first rationally select a prototypical reflectin variant expected to recapitulate the behavior of its parent protein by using a bioinformatics-guided approach. We next map the conformational and energetic landscape accessible to our selected protein by means of all-atom molecular dynamics (MD) simulations. We in turn produce our truncated reflectin variant with and without isotopic labeling, develop solution conditions that maintain the protein in a monomeric state, and characterize the variant’s size and shape with small-angle X-ray scattering (SAXS). We subsequently resolve our protein’s dynamic secondary and tertiary structures and evaluate its backbone conformational fluctuations with NMR spectroscopy. Finally, we demonstrate a straightforward mechanical agitation-based approach to controlling our truncated reflectin variant’s secondary structure, hierarchical self-assembly, and bulk refractive index distribution. Overall, our findings address multiple challenges associated with the development of reflectins as materials, furnish molecular-level insight into the mechanistic underpinnings of cephalopod skin cells’ color-changing functionalities, and appear poised to inform new directions across biochemistry, cellular biology, bioengineering, and optics.     

From the Cover: Evidence from South Africa for a protracted end-Permian extinction on land

Earth’s largest biotic crisis occurred during the Permo–Triassic Transition (PTT). On land, this event witnessed a turnover from synapsid- to archosauromorph-dominated assemblages and a restructuring of terrestrial ecosystems. However, understanding extinction patterns has been limited by a lack of high-precision fossil occurrence data to resolve events on submillion-year timescales. We analyzed a unique database of 588 fossil tetrapod specimens from South Africa’s Karoo Basin, spanning ∼4 My, and 13 stratigraphic bin intervals averaging 300,000 y each. Using sample-standardized methods, we characterized faunal assemblage dynamics during the PTT. High regional extinction rates occurred through a protracted interval of ∼1 Ma, initially co-occurring with low origination rates. This resulted in declining diversity up to the acme of extinction near the Daptocephalus–Lystrosaurus declivis Assemblage Zone boundary. Regional origination rates increased abruptly above this boundary, co-occurring with high extinction rates to drive rapid turnover and an assemblage of short-lived species symptomatic of ecosystem instability. The “disaster taxon” Lystrosaurus shows a long-term trend of increasing abundance initiated in the latest Permian. Lystrosaurus comprised 54% of all specimens by the onset of mass extinction and 70% in the extinction aftermath. This early Lystrosaurus abundance suggests its expansion was facilitated by environmental changes rather than by ecological opportunity following the extinctions of other species as commonly assumed for disaster taxa. Our findings conservatively place the Karoo extinction interval closer in time, but not coeval with, the more rapid marine event and reveal key differences between the PTT extinctions on land and in the oceans.

Mass extinctions are major perturbations of the biosphere resulting from a wide range of different causes including glaciations and sea level fall (1), large igneous provinces (2), and bolide impacts (3, 4). These events caused permanent changes to Earth’s ecosystems, altering the evolutionary trajectory of life (5). However, links between the broad causal factors of mass extinctions and the biological and ecological disturbances that lead to species extinctions have been difficult to characterize. This is because ecological disturbances unfold on timescales much shorter than the typical resolution of paleontological studies (6), particularly in the terrestrial record (6–8). Coarse-resolution studies have demonstrated key mass extinction phenomena including high extinction rates and lineage turnover (7, 9), changes in species richness (10), ecosystem instability (11), and the occurrence of disaster taxa (12). However, finer time resolutions are central to determining the association and relative timings of these effects, their potential causal factors, and their interrelationships. Achieving these goals represents a key advance in understanding the ecological mechanisms of mass extinctions.The end-Permian mass extinction (ca. 251.9 Ma) was Earth’s largest biotic crisis as measured by taxon last occurrences (13–15). Large outpourings from Siberian Trap volcanism (2) are the likely trigger of calamitous climatic changes, including a runaway greenhouse effect and ocean acidification, which had profound consequences for life on land and in the oceans (16–18). An estimated 81% of marine species (19) and 89% of tetrapod genera became extinct as established Permian ecosystems gave way to those of the Triassic. In the ocean, this included the complete extinction of reef-forming tabulate and rugose corals (20, 21) and significant losses in previously diverse ammonoid, brachiopod, and crinoid families (22). On land, many nonmammalian synapsids became extinct (16), and the glossopterid-dominated floras of Gondwana also disappeared (23). Stratigraphic sequences document a global “coral gap” and “coal gap” (24, 25), suggesting reef and forest ecosystems were rare or absent for up to 5 My after the event (26). Continuous fossil-bearing deposits documenting patterns of turnover across the Permian–Triassic transition (PTT) on land (27) and in the oceans (28) are geographically widespread (29, 30), including marine and continental successions that are known from China (31, 32) and India (33). Continental successions are known from Russia (34), Australia (35), Antarctica (36), and South Africa’s Karoo Basin (Fig. 1 and 37–40), the latter providing arguably the most densely sampled and taxonomically scrutinized (41–43) continental record of the PTT. The main extinction has been proposed to occur at the boundary between two biostratigraphic zones with distinctive faunal assemblages, the Daptocephalus and Lystrosaurus declivis assemblage zones (Fig. 1), which marks the traditional placement of the Permian–Triassic geologic boundary [(37) but see ref. 44]. Considerable research has attempted to understand the anatomy of the PTT in South Africa (38, 39, 45–52) and to place it in the context of biodiversity changes across southern Gondwana (53, 54) and globally (29, 31, 32, 44, 47, 55).Open in a separate windowFig. 1.Map of South Africa depicting the distribution of the four tetrapod fossil assemblage zones (Cistecephalus, Daptocephalus, Lystrosaurus declivis, Cynognathus) and our two study sites where fossils were collected in this study (sites A and B). Regional lithostratigraphy and biostratigraphy within the study interval are shown alongside isotope dilution–thermal ionization mass spectrometry dates retrieved by Rubidge et al., Botha et al., and Gastaldo et al. (37, 44, 80). The traditional (dashed red line) and associated PTB hypotheses for the Karoo Basin (37, 44) are also shown. Although traditionally associated with the PTB, the Daptocephalus–Lystrosaurus declivis Assemblage Zone boundary is defined by first appearances of co-occurring tetrapod assemblages, so its position relative to the three PTB hypotheses is unchanged. The Ripplemead member (*) has yet to be formalized by the South African Committee for Stratigraphy.Decades of research have demonstrated the richness of South Africa’s Karoo Basin fossil record, resulting in hundreds of stratigraphically well-documented tetrapod fossils across the PTT (37, 39, 56). This wealth of data has been used qualitatively to identify three extinction phases and an apparent early postextinction recovery phase (39, 45, 51). Furthermore, studies of Karoo community structure and function have elucidated the potential role of the extinction and subsequent recovery in breaking the incumbency of previously dominant clades, including synapsids (11, 57). Nevertheless, understanding patterns of faunal turnover and recovery during the PTT has been limited by the scarcity of quantitative investigations. Previous quantitative studies used coarsely sampled data (i.e., assemblage zone scale, 2 to 3 Ma time intervals) to identify low species richness immediately after the main extinction, potentially associated with multiple “boom and bust” cycles of primary productivity based on δ¹³C variation during the first 5 My of the Triassic (41, 58). However, many details of faunal dynamics in this interval remain unknown. Here, we investigate the dynamics of this major tetrapod extinction at an unprecedented time resolution (on the order of hundreds of thousands of years), using sample-standardized methods to quantify multiple aspects of regional change across the Cistecephalus, Daptocephalus, and Lystrosaurus declivis assemblage zones.     

Seasonal drought events in tropical East Asia over the last 60,000 y

The cause of seasonal hydrologic changes in tropical East Asia during interstadial/stadial oscillations of the last glaciation remains controversial. Here, we show seven seasonal drought events that occurred during the relatively warm interstadials by phytolith and pollen records. These events are significantly manifested as high percentages of bilobate phytoliths and are consistent with the large zonal sea-surface temperature (SST) gradient from the western to eastern tropical Pacific, suggesting that the reduction in seasonal precipitation could be interpreted by westward shifts of the western Pacific subtropical high triggered by changes of zonal SST gradient over the tropical Pacific and Hadley circulation in the Northern Hemisphere. Our findings highlight that both zonal and meridional ocean–atmosphere circulations, rather than solely the Intertropical Convergence Zone or El Niño-Southern Oscillation, controlled the hydrologic changes in tropical East Asia during the last glaciation.

The tropics are home to regions of central importance to global hydrologic cycles. As an important hydrological parameter, seasonal precipitation underwent profound changes in the tropical oceans and their adjacent continents on different timescales during the last glaciation (1). However, the features and mechanisms that caused seasonal precipitation changes during interstadial/stadial oscillations of the last glacial–interglacial cycle in tropical regions remain controversial (2). Two contrasting hypotheses were proposed to explain these changes. Both of these hypotheses involve interactions between the meridional Hadley and zonal Walker circulations as well as changes in sea-surface temperature (SST) (3).The first hypothesis invokes tropical SST changes in response to a slowdown of the Atlantic Meridional Overturning Circulation (AMOC) triggered by large iceberg discharges in the North Atlantic during the stadials (4, 5) and the resulting shift of the intertropical convergence zone (ITCZ). This AMOC-forced southward movement of the ITCZ caused antiphase changes in seasonal rainfall between north and south of the equator in the tropical Pacific and its adjacent continents on the millennium scale (6–9). The second hypothesis attributes the changes in the tropical atmosphere–ocean dynamics stimulated by tropical SST changes (e.g., El Niño-Southern Oscillation, ENSO) (10, 11) as the cause of seasonal heavy/light rainfall in the eastern/western tropical Pacific during the Heinrich stadials (12).However, most of the evidence supporting these two hypotheses comes from the Atlantic (13) and the tropical Pacific (1, 12, 14). Limited evidence is from tropical continents, and particularly little evidence is available from terrestrial tropical East Asia, where the mechanisms responsible for seasonal precipitation changes are still under debate. Although the oxygen isotopes of stalagmites on land have been viewed as one of the most robust East Asian summer monsoon records, the interpretation of speleothem δ¹⁸O in China remains controversial (15–17). Therefore, to demonstrate these hypotheses, both unique research sites, which could simultaneously receive continental and tropical ocean signals, and unambiguous proxies, which could reflect annual and seasonal precipitation, are urgently needed.The core studied in this paper is located at Huguangyan Maar Lake (HML) in Guangdong, southern coast of China (110°17′ E, 21°9′ N, 23 m above sea level) (Fig. 1). This region is closely influenced by Hadley circulation (HC) over the continent and Walker circulation (WC) over the tropical Pacific (18, 19), as well as the related changes in the western Pacific subtropical high (WPSH) (19, 20). Here, we present two sets of proxy datasets. One proxy is the phytolith record, a reliable indicator for the evaluation of seasonal–annual Poaceae in ecosystems (21, 22); and Poaceae taxa are more sensitive than arboreal taxa in revealing seasonal hydrological changes (21, 22). The other proxy is the pollen record, which is an indicator of annual precipitation and reflects the general paleoclimate features at low latitudes in East Asia (23–29). Therefore, based on the two contrasting proxies, the successive 60,000-y phytolith and pollen sequences in this study could substantially reveal the mechanism of regional seasonal precipitation changes during stadials/interstadials since the last glacial period in northern tropical East Asia.Open in a separate windowFig. 1.Geographical and climatological settings of study site and region. (A) Topographic map from ENVI 5.1 showing the location of study site (red star, Huguangyan Maar Lake, HML) in south China and seasonal positions of ITCZ (30). (B) Schematic map showing the settings of atmospheric circulations and SSTs in the tropical Pacific. Black solid circles indicate cores mentioned in this paper: 1 = MD98-2181, 2 = ODP806B, 3 = MD02-2529, 4 = ME0005A-24JC, 5 = ODP846, 6 = TR163-22, and 7 = TR163-19. Colors are annual mean SSTs from the period 1981–2010 obtained from https://psl.noaa.gov.     

Intolerance of uncertainty modulates brain-to-brain synchrony during politically polarized perception

Political partisans see the world through an ideologically biased lens. What drives political polarization? Although it has been posited that polarization arises because of an inability to tolerate uncertainty and a need to hold predictable beliefs about the world, evidence for this hypothesis remains elusive. We examined the relationship between uncertainty tolerance and political polarization using a combination of brain-to-brain synchrony and intersubject representational similarity analysis, which measured committed liberals’ and conservatives’ (n = 44) subjective interpretation of naturalistic political video material. Shared ideology between participants increased neural synchrony throughout the brain during a polarizing political debate filled with provocative language but not during a neutrally worded news clip on polarized topics or a nonpolitical documentary. During the political debate, neural synchrony in mentalizing and valuation networks was modulated by one’s aversion to uncertainty: Uncertainty-intolerant individuals experienced greater brain-to-brain synchrony with politically like-minded peers and lower synchrony with political opponents—an effect observed for liberals and conservatives alike. Moreover, the greater the neural synchrony between committed partisans, the more likely that two individuals formed similar, polarized attitudes about the debate. These results suggest that uncertainty attitudes gate the shared neural processing of political narratives, thereby fueling polarized attitude formation about hot-button issues.

Countries around the world are experiencing the strain of growing political polarization (1–5). Opposing partisans come to see the world through different eyes. Where one sees the freedom to choose, another sees murder; where one sees the right to protest, another sees violent conduct (6–8). Such a polarized perception of reality hampers bipartisan cooperation and can even undermine the basic principles of democracy (8, 9).How does polarization arise? One popular theory posits that a need to have certain, structured, and stable beliefs about the world drives people toward political extremes (10–13). Rather than seeing the world in nuanced shades of gray, cognitively rigid individuals perceive information in black and white, painting the world in categorical and predictable terms (14)—a view that dovetails with the immutable taxonomy of political ideologues (15–19). The rigid mind is characterized by a trait-like tendency to find unpredictable and uncertain events aversive and threatening (14, 20, 21) and has long been theorized to play an outsized role in shaping polarized perceptions (22–26). Although recent work suggests that uncertainty can impact the evaluation of political candidates (27) and policy positions (28, 29) and is a major factor contributing to political conservatism (30, 31), the link between uncertainty and political polarization remains unclear. Here, we examine whether individual differences in intolerance of uncertainty (IUS) (20, 21) shape how naturalistic political information is processed in the brain at the time of perception. We test the hypothesis that uncertainty-intolerant individuals interpret polarizing political information through an ideologically biased, subjective “lens” that produces clear-cut judgments of the issue at hand (20, 32). We further examine whether the neural fingerprint of these uncertainty-driven polarized perceptions—that is, increased brain-to-brain synchrony between like-minded partisans—predicts the formation of polarized attitudes.We combine two techniques to measure polarized perceptions of political information. First, intersubject correlation [ISC (33)] provides a direct measure of the similarity in subjective interpretations of naturalistic social stimuli (e.g., video narratives) among participants (34, 35). This technique capitalizes on the neural processes triggered by incoming auditory and visual information. If two individuals exhibit similar neural profiles when processing the same incoming information (e.g., synchronized blood oxygen level-dependent [BOLD] responses in functional MRI [fMRI]), they likely have a shared perception and understanding of that information (36–40). Given that ISC offers an established metric to gauge whether individuals are processing information in a similar way, we can use it to test whether two individuals who share the same political ideology also have similar subjective perceptions of political information, which circumvents issues with demand characteristics and explicit self-report (41). Second, to make neural synchrony analyses sensitive to more subtle differences along the ideological continuum than simple left–right groupings and to test for interactive effects between ideology and intolerance to uncertainty, we combine ISC with intersubject representational similarity analysis [IS-RSA (42–44)]. This versatile approach enables us to leverage continuous individual differences and test whether uncertainty attitudes exacerbate the processing of political information in the brain to fuel polarized political attitude formation.Using a combination of targeted online and field recruiting (n = 360), we invited 22 liberals and 22 conservatives to participate in a study on political cognition (Fig. 1A). While undergoing fMRI, participants viewed three types of videos: a neutrally worded news segment on a politically charged topic (abortion; taken from Public Broadcasting Service [PBS] News), an inflammatory debate segment (police brutality and immigration; taken from the 2016 Cable News Network [CNN] Vice-Presidential debate), and a nonpolitical nature video (taken from British Broadcasting Corporation [BBC] Earth; Fig. 1B). Neural data analysis consisted of time locking the fMRI BOLD signal to the onset of the videos and computing voxel-wise time course correlations between each possible pairing of subjects across the entire participant pool, resulting in a “neural synchrony” measurement that indexes shared subjective interpretations of dynamic, naturalistic stimuli (35, 45, 46). We first analyzed behavioral responses to the videos to test whether ideology, IUS, or both predicted similarities in attitude formation about the presented political videos. Next, we analyzed variation in neural synchrony across participant dyads using IS-RSA (Fig. 1D) to test three interrelated hypotheses: 1) shared ideology between subjects will predict brain-to-brain synchrony during the perception of political stimuli, 2) IUS will modulate this neural synchrony in committed partisans, and 3) increasing neural synchrony will predict the subsequent expression of shared polarized attitudes about the political stimuli.Open in a separate windowFig. 1.(A) Participants underwent fMRI and behavioral testing as part of a larger study on political cognition. (B) Participants viewed three videos in a fixed order while undergoing fMRI. (C) Participants were clearly divided on political ideology. (D) Analytical approach. We tested for variation in neural synchrony as a function of ideology and IUS. The statistical map slice is taken from Fig. 2C.     

Dimensions of invasiveness: Links between local abundance,geographic range size,and habitat breadth in Europe’s alien and native floras

Understanding drivers of success for alien species can inform on potential future invasions. Recent conceptual advances highlight that species may achieve invasiveness via performance along at least three distinct dimensions: 1) local abundance, 2) geographic range size, and 3) habitat breadth in naturalized distributions. Associations among these dimensions and the factors that determine success in each have yet to be assessed at large geographic scales. Here, we combine data from over one million vegetation plots covering the extent of Europe and its habitat diversity with databases on species’ distributions, traits, and historical origins to provide a comprehensive assessment of invasiveness dimensions for the European alien seed plant flora. Invasiveness dimensions are linked in alien distributions, leading to a continuum from overall poor invaders to super invaders—abundant, widespread aliens that invade diverse habitats. This pattern echoes relationships among analogous dimensions measured for native European species. Success along invasiveness dimensions was associated with details of alien species’ introduction histories: earlier introduction dates were positively associated with all three dimensions, and consistent with theory-based expectations, species originating from other continents, particularly acquisitive growth strategists, were among the most successful invaders in Europe. Despite general correlations among invasiveness dimensions, we identified habitats and traits associated with atypical patterns of success in only one or two dimensions—for example, the role of disturbed habitats in facilitating widespread specialists. We conclude that considering invasiveness within a multidimensional framework can provide insights into invasion processes while also informing general understanding of the dynamics of species distributions.

Human socioeconomic activities are altering species’ global distributions, bridging natural dispersal barriers through the accidental and intentional relocation of organisms, and opening opportunities for them to expand into new regions beyond their historic native ranges (1). The outcome of any given introduction event, however, is dependent on ecological and stochastic processes, and many introduced alien species fail to establish and persist (2, 3). Even species that do achieve persistent, self-sustaining populations (i.e., become naturalized sensu ref. 4) show varying degrees of success (i.e., invasiveness) in newly occupied regions. This has been true for natural colonization events throughout Earth’s history [e.g., on islands (5, 6) and during continental biotic interchanges (7–9)] and is certainly the case for the ongoing surge of human-mediated introductions (10–12). Disentangling the factors that lead to invasion success provides an opportunity not only for anticipating and mediating future anthropogenic invasions but also for better understanding the dynamics underlying natural range expansions (13).Quantifying a species’ success in invading the alien range is complex, a fact reflected in the diverse criteria applied by different authorities when deciding whether or not to classify naturalized species as invasive (14). Recent efforts have therefore recognized that invasiveness cannot be captured by a single metric but rather encompasses multiple aspects of ecological success and impact (15, 16). Some proposed metrics, such as spread rate and socioeconomic impacts, are difficult to quantify for large numbers of species (4, 17). However, Rabinowitz’s three-dimensional scheme for characterizing the rarity or commonness of species in their native distributions (18, 19) has been successfully co-opted as a valuable perspective for better understanding the success of alien species (16, 20, 21). Applied in the context of introduced species, this framework recognizes the potential for established aliens to vary along at least three demographic dimensions of invasiveness: 1) in local abundance within the naturalized range, 2) in geographic range size or extent of the naturalized range, and 3) in habitat breadth in the naturalized range (16). We subsequently distinguish these metrics as dimensions of invasiveness when measured in the naturalized distributions of alien species and dimensions of commonness when measured in species native distributions.Considering invasiveness within a multidimensional framework is particularly important if species vary independently among different dimensions (16, 21). Such a scenario opens the possibility for aliens to achieve invasion success in many different ways (Fig. 1). In other words, there could exist different forms of invasiveness, similar to the different forms of rarity or commonness originally proposed by Rabinowitz (19). On the other hand, theoretical concepts and empirical examples suggest correlations between Rabinowitz’s dimensions of commonness among species in their native distributions (6, 22, 23). For example, a positive relationship between local abundance and extent of geographic occurrence or range size has been documented at various scales for numerous taxa (24–26), including plants (24, 27–31), with niche breadth proposed as a linking mechanism (24, 26, 32). If the processes that generate these patterns in native distributions act similarly in species alien distributions, some of the forms of invasiveness outlined in Fig. 1 should be less likely to occur than others. More specifically, if the invasiveness dimensions are correlated, species should vary from excelling (abundant, widespread, generalists; form AWG in Fig. 1) to performing poorly (scarce, restricted, specialists; form 0 in Fig. 1) in all three invasiveness dimensions (33). On the other hand, these macroecological patterns are not without exception, and a recent assessment found little support for correlations among commonness dimensions in Europe’s native flora (34). Alien distributions may further differ because aliens vary in their residence time, and particularly recently introduced species may be in disequilibrium and still increasing along one or more of the invasiveness dimensions (21, 35–37). In line with these alternatives, a continuum from overall poor invaders to species succeeding in all three dimensions has been documented for the regional alien flora of French grassland communities (20), while associations among dimensions were found to be low for the herbaceous alien flora of Southeast Australia (16). The correspondence among different invasiveness dimensions at broader geographic scales has yet to be assessed.Open in a separate windowFig. 1.Conceptual diagram outlining the eight different forms of invasiveness depending on success in zero, one, two, or three dimensions of invasiveness (based on refs. 16, 18, and 20). Forms of invasiveness within the cyan polygon are associated with high naturalized abundance, within the magenta polygon with widespread naturalized geographic extent, and within the yellow polygon with high naturalized habitat breadth. The overlap between magenta and cyan is blue, between cyan and yellow is green, between magenta and yellow is red, and between all three is black. The forms of invasiveness are comparable to analogous forms of commonness used to describe species in their native distributions, and we refer to the same abbreviations in both cases.Functional traits play a role in mediating invasion processes, but efforts to identify characteristics of successful invaders have generally resulted in few or inconsistent associations (38, 39). However, distinguishing between different components of invasiveness may provide additional clarity if each is influenced by different traits or if the same trait has contrasting effects on different dimensions (15, 16, 21, 40, 41). For example, many plant traits are associated with general trade-offs between rapid growth (i.e., acquisitive growth strategies) versus stress tolerance and survival (i.e., conservative growth strategies) (42–44), and one can hypothesize scenarios where these divergent strategies are associated with success in different dimensions of invasiveness (40, 41). Another example are specialized adaptations for long-distance dispersal that may promote rapid range expansion, both in extent and into new habitats, but likely do not provide any advantages that would influence local abundances (45, 46). For habitat specialists, their specific habitat associations may additionally be important for determining whether or not they become widespread (31).A number of hypotheses for invasion success additionally emphasizes the importance of unique ecological dynamics that emerge when species are decoupled from constraints experienced in their native environments (47). For example, because species are able to occupy unfilled niches where introduced [i.e., Darwin’s naturalization hypothesis (48, 49)] or because they leave behind important herbivores, competitors, or pathogens that limit populations in the native distribution [i.e., enemy release (50, 51)]. These mechanisms may be less likely when species expand into areas near the native range, for example, during natural range expansions or intracontinental introductions, as the alien individuals are more likely to encounter conditions similar to those that limited their native distribution compared to species introduced from further abroad (e.g., those with extracontinental origins) (52–54).Here, we combine vegetation plot data covering Europe (55) with databases of alien and native distributions (56, 57), plant traits (58, 59), and historical dates of introduction (60) to provide a comprehensive assessment of multidimensional invasion success for the European alien seed plant flora. First, we test for correlations among local abundance, geographic extent, and habitat breadth of alien species in their naturalized distributions and classify species into one of the eight forms of invasiveness (Fig. 1). We ask whether some forms of invasiveness rarely occur and specifically whether species tend to fit along a continuum ranging from generally poor invaders to super invaders—species excelling in all three dimensions. In addition, we compare relationships among dimensions of invasiveness to those among dimensions of commonness measured for Europe’s native flora, assessing similarities and differences in patterns of distribution between contexts. Next, we explore likely drivers of each invasiveness dimension, testing whether the year of first alien occurrence in Europe, functional traits related to ecological strategies, specialized adaptations for long-distance dispersal, habitat associations, and region of origin explain different forms of invasion success.     

High-throughput developability assays enable library-scale identification of producible protein scaffold variants

Proteins require high developability—quantified by expression, solubility, and stability—for robust utility as therapeutics, diagnostics, and in other biotechnological applications. Measuring traditional developability metrics is low throughput in nature, often slowing the developmental pipeline. We evaluated the ability of 10 variations of three high-throughput developability assays to predict the bacterial recombinant expression of paratope variants of the protein scaffold Gp2. Enabled by a phenotype/genotype linkage, assay performance for 10⁵ variants was calculated via deep sequencing of populations sorted by proxied developability. We identified the most informative assay combination via cross-validation accuracy and correlation feature selection and demonstrated the ability of machine learning models to exploit nonlinear mutual information to increase the assays’ predictive utility. We trained a random forest model that predicts expression from assay performance that is 35% closer to the experimental variance and trains 80% more efficiently than a model predicting from sequence information alone. Utilizing the predicted expression, we performed a site-wise analysis and predicted mutations consistent with enhanced developability. The validated assays offer the ability to identify developable proteins at unprecedented scales, reducing the bottleneck of protein commercialization.

A common constraint across diagnostic, therapeutic, and industrial proteins is the ability to manufacture, store, and use intact and active molecules. These protein properties, collectively termed developability, are often associated to quantitative metrics such as recombinant yield, stability (chemical, thermal, and proteolytic), and solubility (1–5). Despite this universal importance, developability studies are performed late in the commercialization pipeline (2, 4) and limited by traditional experimental capacity (6). This is problematic because 1) proteins with poor developability limit practical assay capacity for measuring primary function, 2) optimal developability is often not observed with proteins originally found in alternative formats [such as display or two-hybrid technologies (7)], and 3) engineering efforts are limited by the large gap between observation size (∼10²) and theoretical mutational diversity (∼10²⁰). Thus, efficient methods to measure developability would alleviate a significant bottleneck in the lead selection process and accelerate protein discovery and engineering.Prior advances to determine developability have focused on calculating hypothesized proxy metrics from existing sequence and structural data or developing material- and time-efficient experiments. Computational sequence-developability models based on experimental antibody data have predicted posttranslational modifications (8, 9), solubility (10, 11), viscosity (12), and overall developability (13). Structural approaches have informed stability (14) and solubility (10, 15). However, many in silico models require an experimentally solved structure or suffer from computational structure prediction inaccuracies (16). Additionally, limited developability information allows for limited predictive model accuracy (17). In vitro methods have identified several experimental protocols to mimic practical developability requirements [e.g., affinity-capture self-interaction nanoparticle spectroscopy (18) and chemical precipitation (19) as metrics for solubility]. However, traditional developability quantification requires significant amounts of purified protein. Noted in both fronts are numerous in silico and/or in vitro metrics to fully quantify developability (1, 5).We sought a protein variant library that would benefit from isolation of proteins with increased developability and demonstrate the broad applicability of the process. Antibodies and other binding scaffolds, comprising a conserved framework and diversified paratope residues, are effective molecular targeting agents (20–24). While significant progress has been achieved with regards to identifying paratopes for optimal binding strength and specificity (25, 26), isolating highly developable variants remains plagued. One particular protein scaffold, Gp2, has been evolved into specific binding variants toward multiple targets (27–29). Continued study improved charge distribution (30), hydrophobicity (31), and stability (28). While these studies have suggested improvements for future framework and paratope residues (including a disulfide-stabilized loop), a poor developability distribution is still observed (32) (Fig. 1 A and B). Assuming the randomized paratope library will lack similar primary functionality, the Gp2 library will simulate the universal applicability of the proposed high-throughput (HT) developability assays.Open in a separate windowFig. 1.HT assays were evaluated for the ability to identify protein scaffold variants with increased developability. (A and B) Gp2 variant expression, commonly measured via low-throughput techniques such as the dot blot shown, highlights the rarity of ideal developability. (C and D) The HT on-yeast protease assay measures the stability of the POI by proteolytic extent. (E and F) The HT split-GFP assay measures POI expression via recombination of a genetically fused GFP fragment. (G and H) The HT split β-lactamase assay measures the POI stability by observing the change in cell-growth rates when grown at various antibiotic concentrations. (I and J) Assay scores, assigned to each unique sequence via deep sequencing, were evaluated by predicting expression (Fig. 3). (K and L) HT assay capacity enables large-scale developability evaluation and can be used to identify beneficial mutations (Fig. 4).We sought HT assays that allow protein developability differentiation via cellular properties to improve throughput. Variations of three primary assays were examined: 1) on-yeast stability (Fig. 1 C and D)—previously validated to improve the stability of de novo proteins (33), antimicrobial lysins (34), and immune proteins (35)—measures proteolytic cleavage of the protein of interest (POI) on the yeast cell surface via fluorescence-activated cell sorting (FACS). We extend the assay by performing the proteolysis at various denaturing combinations to determine if different stability attributes (thermal, chemical, and protease specificity) can be resolved; 2) Split green fluorescent protein (GFP, Fig. 1 E and F)—previously used to determine soluble protein concentrations (36)—measures the assembled GFP fluorescence emerging from a 16–amino acid fragment (GFP₁₁) fused to the POI after recombining with the separably expressed GFP_1-10. We extend the assay by utilizing FACS to separate cells with differential POI expression to increase throughput over the plate-based assay; and 3) Split β-lactamase (Fig. 1 G and H)—previously used to improve thermodynamic stability (37) and solubility (38)—measures cell growth inhibition via ampicillin to determine functional lactamase activity achieved from reconstitution of two enzyme fragments flanking the POI. We expand assay capacity by deep sequencing populations grown at various antibiotic concentrations to relate change in cell frequency to functional enzyme concentration.In this paper, we determined the HT assays’ abilities to predict Gp2 variant developability. We deep sequenced the stratified populations and calculated assay scores (correlating to hypothesized developability) for ∼10⁵ Gp2 variants (Fig. 1I). We then converted the assay scores into a traditional developability metric by building a model that predicts recombinant yield (Fig. 1J). The assays’ capacity enabled yield evaluations for >100-fold traditional assay capacity (Fig. 1K, compared to Fig. 1B) and provide an introductory analysis of factors driving protein developability by observing beneficial mutations via predicted developable proteins (Fig. 1L).     

Ancient plant DNA reveals High Arctic greening during the Last Interglacial

Summer warming is driving a greening trend across the Arctic, with the potential for large-scale amplification of climate change due to vegetation-related feedbacks [Pearson et al., Nat. Clim. Chang. (3), 673–677 (2013)]. Because observational records are sparse and temporally limited, past episodes of Arctic warming can help elucidate the magnitude of vegetation response to temperature change. The Last Interglacial ([LIG], 129,000 to 116,000 y ago) was the most recent episode of Arctic warming on par with predicted 21st century temperature change [Otto-Bliesner et al., Philos. Trans. A Math. Phys. Eng. Sci. (371), 20130097 (2013) and Post et al., Sci. Adv. (5), eaaw9883 (2019)]. However, high-latitude terrestrial records from this period are rare, so LIG vegetation distributions are incompletely known. Pollen-based vegetation reconstructions can be biased by long-distance pollen transport, further obscuring the paleoenvironmental record. Here, we present a LIG vegetation record based on ancient DNA in lake sediment and compare it with fossil pollen. Comprehensive plant community reconstructions through the last and current interglacial (the Holocene) on Baffin Island, Arctic Canada, reveal coherent climate-driven community shifts across both interglacials. Peak LIG warmth featured a ∼400-km northward range shift of dwarf birch, a key woody shrub that is again expanding northward. Greening of the High Arctic—documented here by multiple proxies—likely represented a strong positive feedback on high-latitude LIG warming. Authenticated ancient DNA from this lake sediment also extends the useful preservation window for the technique and highlights the utility of combining traditional and molecular approaches for gleaning paleoenvironmental insights to better anticipate a warmer future.

The Arctic is greening as shrub biomass increases and vegetation ranges shift north in response to summer warming (1, 2). This process—one of the clearest terrestrial manifestations of climate change thus far—has major implications both for local ecosystems and for global energy balance and biogeochemical systems (3–5). In particular, taller shrubs darken otherwise snow-covered surfaces, contributing to the albedo feedback (6, 7), and enhanced evapotranspiration is expected to result in a positive greenhouse feedback (8). Shrub cover also impacts soil thermal regime, which may impact permafrost vulnerability (9–11). Because feedbacks related to Arctic greening are complex and potentially large in magnitude, estimating the extent and rate of northward shrub migration is a vital component of predicting future warming.Past warm periods serve as valuable analogs for understanding the extent of Arctic greening under well-constrained climate conditions. The Last Interglacial (LIG; Marine Isotope Stage [MIS] 5e, 129 to 116 ka [thousands of years before present]) was ∼1 °C warmer than the preindustrial period globally, but the Arctic experienced amplified warming due to higher summer insolation anomalies and positive feedbacks at high latitudes (12, 13). The Eastern Canadian Arctic and Greenland, in particular, were likely ∼4 to 8 °C warmer in summer than present (Fig. 1) (14–18). LIG sediment records from this region thus provide an archive of the vegetation response to Arctic warming at levels comparable to predicted 21^st-century climate change (19).Open in a separate windowFig. 1.Map of Baffin Island and Lake CF8 study area. The symbols represent maximum LIG temperature anomalies based on terrestrial proxy records (shape indicates proxy type) from Baffin Island and Greenland (see SI Appendix, Table S1 for metadata). The shaded regions indicate Arctic bioclimate subzones delineations (29), including modern Betula range in subzones D and E. We note that a small outlier population of Betula occurs east of the D/E boundary on Baffin Island (not captured by vegetation map resolution) (38).While most High Arctic lake basins were scraped clean by ice sheet erosion during the last glaciation and thus only contain postglacial sediments, lakes with small, low-relief catchments within regions of cold-based, slow-flowing ice were protected from erosion. Several such sites have been discovered on eastern Baffin Island, Arctic Canada and contain stratified records of multiple interglacials (20–22). Previous work from Lake CF8 on northeastern Baffin Island (Fig. 1 and SI Appendix, Fig. S1) demonstrates that its sediment record spans at least three interglacials (∼200 ka), including a substantially warmer-than-present LIG as indicated by chironomids, diatoms, and geochemical proxies (15, 23).We targeted the multi-interglacial record from Lake CF8 to assess the vegetation response to pronounced warmth during the LIG and moderate warmth during the Holocene. Pollen produced by some key shrubs and trees, including Betula (birch), is efficiently wind-transported and thus present in lake sediments far north of their ranges (24, 25). We therefore analyzed both sedimentary ancient DNA (sedaDNA), which is sourced locally from within the lake catchment and does not include pollen-derived DNA (26), and fossil pollen to generate a robust vegetation record spanning the last ∼130 ka. Taken together, DNA-inferred plant communities and pollen-inferred July air temperatures provide insight into Arctic plant range shifts under strong summer warming.     

Developmental influence on evolutionary rates and the origin of placental mammal tooth complexity

Development has often been viewed as a constraining force on morphological adaptation, but its precise influence, especially on evolutionary rates, is poorly understood. Placental mammals provide a classic example of adaptive radiation, but the debate around rate and drivers of early placental evolution remains contentious. A hallmark of early dental evolution in many placental lineages was a transition from a triangular upper molar to a more complex upper molar with a rectangular cusp pattern better specialized for crushing. To examine how development influenced this transition, we simulated dental evolution on “landscapes” built from different parameters of a computational model of tooth morphogenesis. Among the parameters examined, we find that increases in the number of enamel knots, the developmental precursors of the tooth cusps, were primarily influenced by increased self-regulation of the molecular activator (activation), whereas the pattern of knots resulted from changes in both activation and biases in tooth bud growth. In simulations, increased activation facilitated accelerated evolutionary increases in knot number, creating a lateral knot arrangement that evolved at least ten times on placental upper molars. Relatively small increases in activation, superimposed on an ancestral tritubercular molar growth pattern, could recreate key changes leading to a rectangular upper molar cusp pattern. Tinkering with tooth bud geometry varied the way cusps initiated along the posterolingual molar margin, suggesting that small spatial variations in ancestral molar growth may have influenced how placental lineages acquired a hypocone cusp. We suggest that development could have enabled relatively fast higher-level divergence of the placental molar dentition.

Whether developmental processes bias or constrain morphological adaptation is a long-standing question in evolutionary biology (1–4). Many of the distinctive features of a species derive from pattern formation processes that establish the position and number of anatomical structures (5). If developmental processes like pattern formation are biased toward generating only particular kinds of variation, adaptive radiations may often be directed along developmental–genetic “lines of least resistance” (2, 4, 6, 7). Generally, the evolutionary consequences of this developmental bias have been considered largely in terms of how it might influence the pattern of character evolution (e.g., refs. 1, 2, 8–10). But development could also influence evolutionary rates by controlling how much variation is accessible to natural selection in a given generation (11).For mammals, the dentition is often the only morphological system linking living and extinct species (12). Correspondingly, tooth morphology plays a crucial role in elucidating evolutionary relationships, time calibrating phylogenetic trees, and reconstructing adaptive responses to past environmental change (e.g., refs. 13–15). One of the most pervasive features of dental evolution among mammals is an increase in the complexity of the tooth occlusal surface, primarily through the addition of new tooth cusps (16, 17). These increases in tooth complexity are functionally and ecologically significant because they enable more efficient mechanical breakdown of lower-quality foods like plant leaves (18).Placental mammals are the most diverse extant mammalian group, comprising more than 6,000 living species spread across 19 extant orders, and this taxonomic diversity is reflected in their range of tooth shapes and dietary ecologies (12). Many extant placental orders, especially those with omnivorous or herbivorous ecologies (e.g., artiodactyls, proboscideans, rodents, and primates), convergently evolved a rectangular upper molar cusp pattern from a placental ancestor with a more triangular cusp pattern (19–21). This resulted from separate additions in each lineage of a novel posterolingual cusp, the "hypocone'''' [sensu (19)], to the tritubercular upper molar (Fig. 1), either through modification of a posterolingual cingulum (“true” hypocone) or another posterolingual structure, like a metaconule (pseudohypocone) (19). The fossil record suggests that many of the basic steps in the origin of this rectangular cusp pattern occurred during an enigmatic early diversification window associated with the divergence and early radiation of several placental orders (20, 21; Fig. 1). However, there remains debate about the rate and pattern of early placental divergence (22–24). On the one hand, most molecular phylogenies suggest that higher-level placental divergence occurred largely during the Late Cretaceous (25, 26), whereas other molecular phylogenies and paleontological analyses suggest more rapid divergence near the Cretaceous–Paleogene (K–Pg) boundary (21, 24, 27–29). Most studies agree that ecological opportunity created in the aftermath of the K–Pg extinction probably played an important role in ecomorphological diversification within the placental orders (30, 31). But exactly how early placentals acquired the innovations needed to capitalize on ecological opportunity remains unclear. Dental innovations, especially those which facilitated increases in tooth complexity, may have been important because they would have promoted expansion into plant-based dietary ecologies left largely vacant after the K–Pg extinction event (32).Open in a separate windowFig. 1.Placental mammal lineages separately evolved complex upper molar teeth with a rectangular cusp pattern composed of two lateral pairs of cusps from a common ancestor with a simpler, triangular cusp pattern. Many early relatives of the extant placental orders, such as Eritherium, possessed a hypocone cusp and a more rectangular primary cusp pattern. Examples of complex upper molars are the following: Proboscidea, the gomphothere Anancus; Rodentia, the wood mouse Apodemus; and Artiodactyla, the suid Nyanzachoerus.Mammalian tooth cusps form primarily during the “cap” and “bell” stage of dental development, when signaling centers called enamel knots establish the future sites of cusp formation within the inner dental epithelium (33, 34). The enamel knots secrete molecules that promote proliferation and changes in cell–cell adhesion, which facilitates invagination of the dental epithelium into an underlying layer of mesenchymal cells (34, 35). Although a range of genes are involved in tooth cusp patterning (36–38), the basic dynamics can be effectively modeled using reaction–diffusion models with just three diffusible morphogens: an activator, an inhibitor, and a growth factor (39–41). Candidate activator genes in mammalian tooth development include Bmp4, Activin A, Fgf20, and Wnt genes, whereas potential inhibitors include Shh and Sostdc, and Fgf4 and Bmp2 have been hypothesized to act as growth factors (38, 40–43). In computer models of tooth development, activator molecules up-regulated in the underlying mesenchyme stimulate differentiation of overlying epithelium into nondividing enamel knot cells. These in turn secrete molecules that inhibit further differentiation of epithelium into knot cells, while also promoting cell proliferation that creates the topographic relief of the cusp (40). Although many molecular, cellular, and physical processes have the potential to influence cusp formation, and thereby tooth complexity (35, 37), parameters that control the strength and conductance of the activator and inhibitor signals, the core components of the reaction–diffusion cusp patterning mechanism (39, 40) are likely to be especially important.Here, we integrate a previous computer model of tooth morphogenesis called ToothMaker (41), with simulations of trait evolution and data from the fossil record (Fig. 2), to examine the developmental origins of tooth complexity in placental mammals. Specifically, we ask the following: 1) What developmental processes can influence how many cusps form? 2) How might these developmental processes influence the evolution of tooth cusp number, especially rates? And 3) what developmental changes may have been important in the origins of the fourth upper molar cusp, the hypocone, in placental mammal evolution?Open in a separate windowFig. 2.Workflow for simulations of tooth complexity evolution. (A) Tooth shape is varied for five signaling and growth parameters in ToothMaker. (B) From an ancestral state, each parameter is varied in 2.5% increments up to a maximum of ± 50% of the ancestral state. (C) Tooth complexity and enamel knot (EK) pattern were quantified for each parameter combination. Tooth complexity was measured using cusp number/EK number and OPC. ToothMaker and placental upper second molars were classified into categories based on EK/cusp pattern. (D) The parameter space was populated with pattern and tooth complexity datums to build a developmental landscape. (E) Tooth complexity evolution was simulated on each developmental landscape. (F) Resulting diversity and pattern of tooth complexity was compared with placental mammal molar diversity.     

Network motifs involving both competition and facilitation predict biodiversity in alpine plant communities

Biological diversity depends on multiple, cooccurring ecological interactions. However, most studies focus on one interaction type at a time, leaving community ecologists unsure of how positive and negative associations among species combine to influence biodiversity patterns. Using surveys of plant populations in alpine communities worldwide, we explore patterns of positive and negative associations among triads of species (modules) and their relationship to local biodiversity. Three modules, each incorporating both positive and negative associations, were overrepresented, thus acting as "network motifs." Furthermore, the overrepresentation of these network motifs is positively linked to species diversity globally. A theoretical model illustrates that these network motifs, based on competition between facilitated species or facilitation between inferior competitors, increase local persistence. Our findings suggest that the interplay of competition and facilitation is crucial for maintaining biodiversity.

Identifying the processes that maintain natural biodiversity is a longstanding goal of ecology (1–4). Theoretical and empirical explanations of coexistence in plant communities have largely proceeded along two parallel lines: one focused on competition (5–7) and the other on facilitation (8–10). Regarding the former, recent theoretical models highlight the role of intransitive competition in preventing individual plant species from excluding inferior competitors (7, 11). Regarding the latter, studies have shown that key plant species (ecosystem engineers) can support many other species through the amelioration of local environmental conditions, a process widely referred to as direct facilitation (12–15). However, focusing on either competition or facilitation independently may be inadequate for fully understanding species coexistence. In fact, the balance between positive and negative effects plays a crucial role in regulating nutrient flow and driving population and community response to environmental change (2, 13).There is substantial evidence indicating that both competition and facilitation occur simultaneously within the same communities (16, 17). For instance, facilitated species may compete against one another (18), or species can facilitate each other in ways that outcompete other species (19, 20). Yet, because facilitation and competition are rarely considered together at the community level, we have little understanding of the degree to which the interplay of competition and facilitation affects biodiversity. A recent empirical study found that the overall frequency of positive and negative associations between plant species was only weakly correlated to plant diversity in drylands (21). It is unknown, however, whether it is the combination of positive and negative associations at fine spatial scales (orders of centimeters) that matters for biodiversity maintenance. This expectation is consistent with theoretical evidence on the impact of network modules on community dynamics (22).Here, we explore the prevalence and importance of community-level positive and negative associations among species in a global set of alpine plant communities. We analyzed whole-community population data (abundance of individuals per species) for 166 alpine plant communities at 83 sites worldwide (10) (Fig. 1). The dataset includes 2,252 plant species and more than 13,000 observations. Within the dataset, microhabitats with and without ecosystem engineers (an average of 81 paired plots per site) are treated separately (10, 14). We inferred positive and negative associations among all plant species using a Bayesian model in which the expected population size of a species is the sum of proportional changes in the population size of all other species (ref. 23; see Materials and Methods).Open in a separate windowFig. 1.Global map of alpine plant networks studied here. Red dots on the map indicate the spatial location of the networks, with a few networks plotted for reference. In the networks, green dots represent plant species, and blue and red arrows represent negative- and positive species associations, respectively. Dot size is proportional to species abundance. The four network modules analyzed here are represented at the bottom of the figure, from left to right: intransitive competition, facilitation-driven competition, and competition-driven facilitation 1 and 2.According to Abrams (24) and Wootton (25), and following the plant ecology literature (12, 15), network interactions derived from species associations are defined according to long-term effects measured on population size. In general, it is hard to infer interactions from association data alone (20, 26). Still, we have confidence that inferred categorical associations are indicative of the long-term outcome of species interactions to a large extent because: 1) alpine plant communities are relatively simple and spatial patterns closely reflect the net effects of plant–plant interactions (8, 12, 15, 27); 2) population data (number of plants per species) were collected at a fine spatial scale on the order of centimeters, where direct interactions take place (4, 8, 12), minimizing the influence of environmental heterogeneity and maximizing the imprint of direct neighbor effects (17, 27); 3) each network was built for each microhabitat at each site, with species growing in the same homogeneous environmental conditions (temperature, soil, aridity, etc.) (10), thus excluding spatial gradients that might mask or confound the correct inference of interactions from plant–plant associations (26); 4) we used high-resolution community data (as opposed to presence/absence data) that allow us to derive robust estimates of plant associations (28), as demonstrated in the sensitivity analysis (SI Appendix, Data and Code); 5) in addition to having highly resolved local communities, the survey was also replicated on a global scale, which allows us to generalize our findings since patterns are observed over a broad range of environmental and biogeographic contexts, from tropical to arctic latitudes (Fig. 1); and 6) inference was carried out with a Bayesian model that describes association strengths among plants with a similar power to existing models of plant fecundity and growth (5). This includes joint-posterior distributions of parameters, which allows us to infer associations among many species at the community level (23, 28).     

Maternal death and offspring fitness in multiple wild primates

Primate offspring often depend on their mothers well beyond the age of weaning, and offspring that experience maternal death in early life can suffer substantial reductions in fitness across the life span. Here, we leverage data from eight wild primate populations (seven species) to examine two underappreciated pathways linking early maternal death and offspring fitness that are distinct from direct effects of orphaning on offspring survival. First, we show that, for five of the seven species, offspring face reduced survival during the years immediately preceding maternal death, while the mother is still alive. Second, we identify an intergenerational effect of early maternal loss in three species (muriquis, baboons, and blue monkeys), such that early maternal death experienced in one generation leads to reduced offspring survival in the next. Our results have important implications for the evolution of slow life histories in primates, as they suggest that maternal condition and survival are more important for offspring fitness than previously realized.

Mammalian life history is marked by a strong dependent relationship between offspring and their mothers (1). The quantity or quality of maternal allocation to offspring, particularly during the gestation and lactation periods, is often related to maternal physical condition, and a range of offspring fitness outcomes are compromised if gestating or lactating mothers are in poor condition (2–8). In addition, infants that experience maternal loss prior to weaning face an enormous, acute risk of death in both nonhuman mammals and in humans (9–13) (Fig. 1, blue arrow).Open in a separate windowFig. 1.Four ways in which the death of a female primate mother (M) may be linked to her offspring’s fitness (F₁), if the death of M occurs while F₁ is still dependent on M. First, F₁ should display reduced survival during the immature period, following the death of M (especially before weaning but also after), because F₁ will lack the critically important social, nutritional, and/or protective resources that M provided (blue arrow). Second, F₁ should display reduced survival in the period before M actually dies, because, on average, mothers are in worse condition shortly before their death compared to mothers that survive the same period. We therefore expect M to provide lower-quality maternal care to F₁ during the weeks to years immediately preceding M’s death (purple arrow). Third, if F₁ survives these first two challenges, she is likely to be in chronically worse condition during adulthood because of reductions in maternal allocation that she received during development (red arrow). F₁ should therefore face reduced survival in adulthood, years (or even decades) after the death of M occurred. Fourth, this chronic reduction in F₁’s condition may have an intergenerational effect, such that F₂ (F₁’s offspring) also experience reduced immature survival (gold arrow). The blue and red arrows have been previously tested in several species; the analyses presented here focus on the purple and gold arrows.In some species, including primates, hyenas, whales, and some ungulates, mothers and offspring continue to associate after weaning, and mothers may provide substantial social and energetic input as well as protection during some or all of the remainder of the predispersal, immature period (hereafter the “immature period”) (12, 14–21). Thus, loss of the mother can continue to heighten the risk of death even in weaned, immature offspring (17, 19, 21) (Fig. 1, blue arrow). However, because offspring are less dependent on mothers during this phase of life, the effects of maternal loss after weaning can be sublethal (9, 16, 22). If an offspring that is weaned (but still partially dependent on its mother) survives its mother’s death, the offspring may experience long-lasting negative effects, including adverse behavioral or social outcomes in adolescence or adulthood [humans (23, 24); nonhumans (19, 20, 25–31)]. In baboons, chimpanzees, and elephants, motherless offspring may experience reduced survival during adolescence and adulthood, well after the maternal loss occurs, presumably because maternal loss results in a chronic reduction in body condition (13, 21, 26, 32) (Fig. 1, red arrow).These observations (see citations in previous paragraphs) combine to provide a strong framework that describes the dependent relationship between mammalian offspring and their mothers and allows us to make predictions about the expected effects of maternal death on offspring fitness. This framework relies on the following assumptions, which are based on the observations described above: Mammalian offspring are critically dependent on their mothers for nutrition, protection, transport, and learning. In many species, the period of dependence is not restricted to infancy and may extend well past weaning. During this dependent period, poor maternal body condition (defined here as an unmeasured physical state that predicts an animals’ ability to perform functions necessary for reproduction and survival) can lead to reduced maternal allocation to offspring and hence poor offspring body condition. Poor offspring condition, in turn, may have both immediate and later-life consequences for offspring fitness outcomes, including survival. As a result, mothers in poor condition (even those that survive to wean their offspring) are likely to produce offspring in poor condition that experience compromised survival. Maternal death at any time during this dependent period can therefore result in both short-term and chronic reductions in offspring physical condition and survival.This framework yields four main predictions about how maternal death experienced in early life affects offspring fitness outcomes across the life span. First, immature offspring that lose their mother will face reduced survival throughout the remainder of their immature period (Fig. 1, blue arrow). Although the impact of maternal loss will be especially strong if the mother dies before the offspring is weaned, the immature offspring may continue to face reduced survival if its mother dies any time before the offspring matures. Second, because the loss of the mother in early life results in developmental constraints that persist throughout the offspring’s lifetime, offspring that experienced early maternal loss will continue to experience reduced survival in adulthood, leading to shortened adult life spans (Fig. 1, red arrow). These two predictions are important for offspring fitness outcomes; they have been previously tested in several species (see above summary) and are therefore not the focus of our study.We focus instead on two additional predictions, which have received little previous attention. First, we expect offspring to face reduced survival if their mothers are going to die in the near future, because, on average, a mother whose death is imminent is more likely to be in poor condition compared to those mothers that survive the same period. Thus, in this prediction, imminent maternal death serves as a proxy for poor maternal condition. We can test this prediction by measuring the association between offspring survival and impending maternal death, while the mother is still alive (Fig. 1, purple arrow). Second, we predict an intergenerational effect of early maternal loss on offspring survival (Fig. 1, gold arrow). That is, we predict that female offspring that experience maternal loss but still survive to adulthood (F₁ generation in Fig. 1) will produce offspring with compromised survival (F₂ generation). We expect the proximate mechanism leading to this intergenerational effect to be that F₁’s compromised condition causes her to be less able to allocate adequate resources to her offspring.These latter two predictions about survival patterns have been previously confirmed in wild baboons (33), but otherwise we have little knowledge of the generality of these two links between maternal survival and the survival of offspring (F₁ generation) and grand offspring (F₂ generation) in natural populations of primates or other mammals. Here, we leverage long-term longitudinal data from eight wild populations of seven primate species to assess 1) the extent to which offspring suffer reduced survival when their mothers will soon die (Fig. 1, purple arrow), and 2) the extent to which the effects of early maternal loss carry over from one generation to the next, resulting in reduced immature survival for offspring whose mothers experienced early maternal loss (Fig. 1, gold arrow).     

A source for awareness-dependent figure–ground segregation in human prefrontal cortex

Figure–ground modulation, i.e., the enhancement of neuronal responses evoked by the figure relative to the background, has three complementary components: edge modulation (boundary detection), center modulation (region filling), and background modulation (background suppression). However, the neuronal mechanisms mediating these three modulations and how they depend on awareness remain unclear. For each modulation, we compared both the cueing effect produced in a Posner paradigm and fMRI blood oxygen-level dependent (BOLD) signal in primary visual cortex (V1) evoked by visible relative to invisible orientation-defined figures. We found that edge modulation was independent of awareness, whereas both center and background modulations were strongly modulated by awareness, with greater modulations in the visible than the invisible condition. Effective-connectivity analysis further showed that the awareness-dependent region-filling and background-suppression processes in V1 were not derived through intracortical interactions within V1, but rather by feedback from the frontal eye field (FEF) and dorsolateral prefrontal cortex (DLPFC), respectively. These results indicate a source for an awareness-dependent figure–ground segregation in human prefrontal cortex.

Figure–ground segregation is a fundamental process by which the visual system segments images into figures and background (1, 2). Previous neurophysiological and brain imaging studies of figure–ground segregation have shown neuronal responses are enhanced in the region perceived to be the figure and suppressed in the region perceived to be the background, an effect known as figure–ground modulation (1, 3–9). Figure–ground modulation plays a key role in identifying and localizing visual objects (7) and capturing focused attention (10). Remarkably, evidence from numerous neurophysiological (5, 11–16), psychophysical (17, 18), and brain imaging (19, 20) studies, as well as computational models (2, 17, 21) have suggested that figure–ground modulation relies on three complementary processes: boundary detection (i.e., edge modulation), region filling (i.e., center modulation), and background suppression (i.e., background modulation). During figure–ground segregation, boundary detection is the process that detects feature discontinuities that signal boundaries between the figures and background, while region filling is the process that groups figural regions with the same (or similar) features together (17), and the background suppression is the process that inhibits homogeneous features in the background (5, 22, 23). However, little is known how these three processes depend on awareness.A number of previous studies have supported an early feedforward processing phase for the boundary-detection and later feedback-processing phases for both region filling and background suppression in figure–ground modulation (2, 5, 21). These findings thus suggest that boundary detection is independent of awareness, whereas both region filling and background suppression are strongly modulated by awareness (12). Specifically, boundary detection is thought to be achieved through iso-feature inhibition (2, 21, 24, 25) within early visual areas, as early as the primary visual cortex (V1) (9, 14, 26–30), in which neurons preferring the same or similar features are more likely to suppress each other via lateral connections (31). The region-filling process, however, requires iso-feature excitation in which neurons representing the similar features enhance each other’s activity. In contrast to several studies suggesting the existence of the region-filling process within V1 (1, 9), most previous studies indicate that the region-filling process arises from feedback projections to V1 from a higher cortical area(s) (11, 12, 14, 15, 20, 32). Similarly, several neurophysiological studies have suggested that the background-suppression process may also be derived by feedback to V1 from a higher cortical area(s); it is the later processing phase in which neural activity elicited by the background is suppressed by the preceding segregated figures (5, 22, 23). However, it remains unknown which and how the top-down feedback from a higher cortical area(s) drive the region-filling and background-suppression processes in V1 that enhances the response of neurons tuned to the same feature and suppresses the neural activity elicited by the background, respectively. Also, it is unknown whether and how these feedback processes interact with awareness.Furthermore, another unclear but related issue is how boundary detection and region filling in figure–ground segregation attract focused attention. In fact, it is well known that successful segregation of a figure, as defined by an orientation contrast (Fig. 1A) from the background, leads to pop-out, which automatically attracts bottom-up attention to this salient figure location (10, 21, 25). However, it is not known whether there are different neural mechanisms by which the boundary detection and region filling attract our focused attention and whether the attentional attraction triggered by these two processes interacts with awareness.Open in a separate windowFig. 1.Stimuli, psychophysical protocol, and data. (A) Two sample orientation-defined figures presented in the upper visual field (Left: large figure; Right: small figure). The orientation contrasts between the figure bars and the background bars was 60° (the yellow dot indicates the fixation point). (B) Large (Left) and small (Right) grating probes, with the same diameter as the large and small figures, respectively. (C) Low- (Left) and high- (Right) luminance mask stimuli used in the visible and invisible conditions, respectively. (D) Psychophysical protocol. A figure–ground stimulus was presented for 50 ms, followed by a 100-ms mask and another 50-ms fixation interval. Then a large or small grating probe, with the same diameter as the large figure and small figure, respectively, was randomly presented for 50 ms with equal probability and presented randomly at either the figure location (valid cue condition) or its contralateral counterpart (invalid cue condition) with equal probability. The grating probe was orientated at 45° or 135° away from the vertical. Subjects were asked to press one of two buttons as rapidly and correctly as possible to indicate the orientation of the grating probe (45° or 135°). The psychophysical cueing effect for the large (E) and small (F) figures and the large and small gratings in both visible and invisible conditions. Each cueing effect was quantified as the difference between the reaction time of the probe task performance in the invalid cue condition and that in the valid cue condition. Error bars denote 1 SEM calculated across subjects and colored dots denote the data from each subject.To address these questions, we used a modified version of the Posner paradigm (33, 34) to measure the spatial cueing effect induced by figure boundary (edge modulation) or figure center (center modulation) of the large figure (Fig. 1 A, Left), and by the whole small figure or its surround background (background modulation, Fig. 1 A, Right). Blood oxygen level-dependent (BOLD) signals evoked by the figure boundary and figure center of the large figure, as well as the whole small figure and its surround background were also measured. Using a backward masking paradigm (10) with low- or high-luminance masks to render the whole figure–ground stimulus visible or invisible (confirmed by a two-alternative forced choice [2AFC]) to subjects, respectively, we examined how figure–ground segregation interacts with awareness. We also performed interregional correlation and effective connectivity analyses to examine the neural mechanisms of this potential interaction.