Breaking the constraint on the number of cervical vertebrae in mammals: On homeotic transformations in lorises and pottos.

Mammals almost always have seven cervical vertebrae. The strong evolutionary constraint on changes in this number has been broken in sloths and manatees. We have proposed that the extremely low activity and metabolic rates of these species relax the stabilizing selection against changes in the cervical count. Our hypothesis is that strong stabilizing selection in other mammals is largely indirect and due to associated pleiotropic effects, including juvenile cancers. Additional direct selection can occur due to biomechanical problems (thoracic outlet syndrome). Low metabolic and activity rates are thought to diminish these direct and indirect effects. To test this hypothesis within the primates, we have compared the number of cervical vertebrae of three lorisid species with particularly low activity and metabolic rates with those of more active primate species, including with their phylogenetically closest active relatives, the galagids (bushbabies). In support of our hypothesis, we found that 37.6% of the lorisid specimens had an abnormal cervical count, which is a higher percentage than in the other nine primate families, in which the incidence varied from zero to 2.2%. We conclude that our data support the importance of internal selection in constraining evolvability and of a relaxed stabilizing selection for increasing evolvability. Additionally, we discuss that there is no support for a role of the muscularized diaphragm in the evolutionary constraint.

Mild drought in the vegetative stage induces phenotypic, gene expression, and DNA methylation plasticity in Arabidopsis but no transgenerational effects.

There is renewed interest in whether environmentally induced changes in phenotypes can be heritable. In plants, heritable trait variation can occur without DNA sequence mutations through epigenetic mechanisms involving DNA methylation. However, it remains unknown whether this alternative system of inheritance responds to environmental changes and if it can provide a rapid way for plants to generate adaptive heritable phenotypic variation. To assess potential transgenerational effects induced by the environment, we subjected four natural accessions of Arabidopsis thaliana together with the reference accession Col-0 to mild drought in a multi-generational experiment. As expected, plastic responses to drought were observed in each accession, as well as a number of intergenerational effects of the parental environments. However, after an intervening generation without stress, except for a very few trait-based parental effects, descendants of stressed and non-stressed plants were phenotypically indistinguishable irrespective of whether they were grown in control conditions or under water deficit. In addition, genome-wide analysis of DNA methylation and gene expression in Col-0 demonstrated that, while mild drought induced changes in the DNA methylome of exposed plants, these variants were not inherited. We conclude that mild drought stress does not induce transgenerational epigenetic effects.

Egg size does not universally predict embryonic resources and hatchling size across annual killifish species.

Egg size has a crucial impact on the reproductive success of a mother and the performance of her offspring. It is therefore reasonable to employ egg size as a proxy for egg content when studying variation in offspring performance. Here, we tested species differences in allometries of several egg content parameters with egg area. We measured individual eggs in five species of annual killifish (Cyprinodontiformes), a group of fish where egg banks permit population survival over dry season. Apart from comparing allometric scaling exponents, amounts and compositions of egg components across the different species, we assessed the explanatory power of egg area for egg wet and dry weight and for hatchling size. We found notable species-specific allometries between egg area and the other egg parameters (egg dry weight and water content, elemental composition and triglyceride content). Across species, egg area predicted egg wet weight with highest power. Within species, coefficients of determination were largest in A. elongatus, a large piscivorous species with large eggs. Our study shows that systematically using egg area as a proxy of egg content between different species can ignore relevant species-specific differences and mask within-species variability in egg content.

Fast running restricts evolutionary change of the vertebral column in mammals.

The mammalian vertebral column is highly variable, reflecting adaptations to a wide range of lifestyles, from burrowing in moles to flying in bats. However, in many taxa, the number of trunk vertebrae is surprisingly constant. We argue that this constancy results from strong selection against initial changes of these numbers in fast running and agile mammals, whereas such selection is weak in slower-running, sturdier mammals. The rationale is that changes of the number of trunk vertebrae require homeotic transformations from trunk into sacral vertebrae, or vice versa, and mutations toward such transformations generally produce transitional lumbosacral vertebrae that are incompletely fused to the sacrum. We hypothesize that such incomplete homeotic transformations impair flexibility of the lumbosacral joint and thereby threaten survival in species that depend on axial mobility for speed and agility. Such transformations will only marginally affect performance in slow, sturdy species, so that sufficient individuals with transitional vertebrae survive to allow eventual evolutionary changes of trunk vertebral numbers. We present data on fast and slow carnivores and artiodactyls and on slow afrotherians and monotremes that strongly support this hypothesis. The conclusion is that the selective constraints on the count of trunk vertebrae stem from a combination of developmental and biomechanical constraints.

Climate and atmosphere simulator for experiments on ecological systems in changing environments.

Grand challenges in global change research and environmental science raise the need for replicated experiments on ecosystems subjected to controlled changes in multiple environmental factors. We designed and developed the Ecolab as a variable climate and atmosphere simulator for multifactor experimentation on natural or artificial ecosystems. The Ecolab integrates atmosphere conditioning technology optimized for accuracy and reliability. The centerpiece is a highly contained, 13-m(3) chamber to host communities of aquatic and terrestrial species and control climate (temperature, humidity, rainfall, irradiance) and atmosphere conditions (O2 and CO2 concentrations). Temperature in the atmosphere and in the water or soil column can be controlled independently of each other. All climatic and atmospheric variables can be programmed to follow dynamical trajectories and simulate gradual as well as step changes. We demonstrate the Ecolab's capacity to simulate a broad range of atmospheric and climatic conditions, their diurnal and seasonal variations, and to support the growth of a model terrestrial plant in two contrasting climate scenarios. The adaptability of the Ecolab design makes it possible to study interactions between variable climate-atmosphere factors and biotic disturbances. Developed as an open-access, multichamber platform, this equipment is available to the international scientific community for exploring interactions and feedbacks between ecological and climate systems.

What life cycle graphs can tell about the evolution of life histories.

We analyze long-term evolutionary dynamics in a large class of life history models. The model family is characterized by discrete-time population dynamics and a finite number of individual states such that the life cycle can be described in terms of a population projection matrix. We allow an arbitrary number of demographic parameters to be subject to density-dependent population regulation and two or more demographic parameters to be subject to evolutionary change. Our aim is to identify structural features of life cycles and modes of population regulation that correspond to specific evolutionary dynamics. Our derivations are based on a fitness proxy that is an algebraically simple function of loops within the life cycle. This allows us to phrase the results in terms of properties of such loops which are readily interpreted biologically. The following results could be obtained. First, we give sufficient conditions for the existence of optimisation principles in models with an arbitrary number of evolving traits. These models are then classified with respect to their appropriate optimisation principle. Second, under the assumption of just two evolving traits we identify structural features of the life cycle that determine whether equilibria of the monomorphic adaptive dynamics (evolutionarily singular points) correspond to fitness minima or maxima. Third, for one class of frequency-dependent models, where optimisation is not possible, we present sufficient conditions that allow classifying singular points in terms of the curvature of the trade-off curve. Throughout the article we illustrate the utility of our framework with a variety of examples.

Field sampling of fig pollinator wasps across host species and host developmental phase: Implications for host recognition and specificity.

Previous genetic studies of pollinator wasps associated with a community of strangler figs (Ficus subgenus Urostigma, section Americana) in Central Panama suggest that the wasp species exhibit a range in host specificity across their host figs. To better understand factors that might contribute to this observed range of specificity, we used sticky traps to capture fig-pollinating wasp individuals at 13 Ficus species, sampling at different phases of the reproductive cycle of the host figs (e.g., trees with receptive inflorescences, or vegetative trees, bearing only leaves). We also sampled at other tree species, using them as non-Ficus controls. DNA barcoding allowed us to identify the wasps to species and therefore assign their presence and abundance to host fig species and the developmental phase of that individual tree. We found: (1) wasps were only very rarely captured at non-Ficus trees; (2) nonetheless, pollinators were captured often at vegetative individuals of some host species; (3) overwhelmingly, wasp individuals were captured at receptive host fig trees representing the fig species from which they usually emerge. Our results indicate that wasp occurrence is not random either spatially or temporally within the forest and across these hosts, and that wasp specificity is generally high, both at receptive and vegetative host trees. Therefore, in addition to studies that show chemicals produced by receptive fig inflorescences attract pollinator wasps, we suggest that other cues (e.g., chemicals produced by the leaves) can also play a role in host recognition. We discuss our results in the context of recent findings on the role of host shifts in diversification processes in the Ficus genus.

Phylogenetic reconstruction and shell evolution of the Diplommatinidae (Gastropoda: Caenogastropoda).

The fascinating and often unlikely shell shapes in the terrestrial micromollusc family Diplommatinidae (Gastropoda: Caenogastropoda) provide a particularly attractive set of multiple morphological traits to investigate evolutionary patterns of shape variation. Here, a molecular phylogenetic reconstruction, based on five genes and 2700 bp, was undertaken for this family, integrated with ancestral state reconstruction and phylogenetic PCA of discrete and quantitative traits, respectively. We found strong support for the Diplommatininae as a monophyletic group, separating the Cochlostomatidae into a separate family. Five main clades appear within the Diplommatininae, corresponding with both coiling direction and biogeographic patterns. A Belau clade (A) with highly diverse (but always sinistral) morphology comprised Hungerfordia, Palaina, and some Diplommatina. Arinia (dextral) and Opisthostoma (sinistroid) are sister groups in clade B. Clade C and D solely contain sinistral Diplommatina that are robust and little ornamented (clade C) or slender and sculptured (clade D). Clade E is dextral but biogeographically diverse with species from all sampled regions save the Caroline Islands. Adelopoma, Diplommatina, Palaina, and Hungerfordia require revision to allow taxonomy to reflect phylogeny, whereas Opisthostoma is clearly monophyletic. Ancestral state reconstruction suggests a sinistral origin for the Diplommatinidae, with three reversals to dextrality.

Lifespan prolonging mechanisms and insulin upregulation without fat accumulation in long-lived reproductives of a higher termite.

Kings and queens of eusocial termites can live for decades, while queens sustain a nearly maximal fertility. To investigate the molecular mechanisms underlying their long lifespan, we carried out transcriptomics, lipidomics and metabolomics in Macrotermes natalensis on sterile short-lived workers, long-lived kings and five stages spanning twenty years of adult queen maturation. Reproductives share gene expression differences from workers in agreement with a reduction of several aging-related processes, involving upregulation of DNA damage repair and mitochondrial functions. Anti-oxidant gene expression is downregulated, while peroxidability of membranes in queens decreases. Against expectations, we observed an upregulated gene expression in fat bodies of reproductives of several components of the IIS pathway, including an insulin-like peptide, Ilp9. This pattern does not lead to deleterious fat storage in physogastric queens, while simple sugars dominate in their hemolymph and large amounts of resources are allocated towards oogenesis. Our findings support the notion that all processes causing aging need to be addressed simultaneously in order to prevent it.

Pollination and protection against herbivory of Nepalese Coelogyninae (Orchidaceae).

PREMISE OF THE STUDY: Although many species in the orchid genus Coelogyne are horticulturally popular, hardly anything is known about their pollination. Pollinators of three species were observed in the field in Nepal. This information is urgently needed because many orchid species in Nepal are endangered. Whether the exudates produced by extrafloral nectaries played a role in protection against herbivory was also investigated. METHODS: Pollinators of C. flaccida, C. nitida, and Otochilus albus were filmed, captured, and identified. Ant surveys and exclusion experiments were carried out. To investigate whether pollinators are needed for fruit set, plants were wrapped in mesh wire bags. Inflorescence stems were examined with microscopy. Fehling's reagent was used to detect sugars in extrafloral exudates. KEY RESULTS: Coelogyne flaccida and C. nitida need pollinators to set fruit and are pollinated by wild bees identified as Apis cerana. Otochilus albus was found to be pollinated by Bombus kashmirensis. Extrafloral nectar was found to be exuded by nectary-modified stomata and contained high amounts of sugars. Different species of ants were observed collecting these exudates. A significant difference was found in damage inflicted by flower and leaf-eating beetles between C. nitida plants living in trees with ant nests and those in ant-free trees. CONCLUSIONS: Floral syndromes include scented and colored trap flowers without reward to their pollinators. All orchids investigated exude extrafloral nectar by nectary-modified stomata. This nectar was found to flow from the phloem to the stomata through intercellular spaces in the outer parenchymatous layer of the inflorescence.

Exploring copy number variants in deceased fetuses and neonates with abnormal vertebral patterns and cervical ribs.

BACKGROUND: Cervical patterning abnormalities are rare in the general population, but one variant, cervical ribs, is particularly common in deceased fetuses and neonates. The discrepancy between the incidence in the general population and early mortality is likely due to indirect selection against cervical ribs. The cause for the co-occurrence of cervical ribs and adverse outcome remains unidentified. Copy number variations resulting in gain or loss of specific genes involved in development and patterning could play a causative role. METHODS: Radiographs of 374 deceased fetuses and infants, including terminations of pregnancies, stillbirths and neonatal deaths, were assessed. Copy number profiles of 265 patients were determined using single nucleotide polymorphism array. RESULTS: 274/374 patients (73.3%) had an abnormal vertebral pattern, which was associated with congenital abnormalities. Cervical ribs were present in 188/374 (50.3%) and were more common in stillbirths (69/128 [53.9%]) and terminations of pregnancies (101/188 [53.7%]), compared to live births (18/58, 31.0%). Large (likely) deleterious copy number variants and aneuploidies were prevalent in these patients. None of the rare copy number variants were recurrent or overlapped with candidate genes for vertebral patterning. CONCLUSIONS: The large variety of copy number variants in deceased fetuses and neonates with similar abnormalities of the vertebral pattern probably reflects the etiological heterogeneity of vertebral patterning abnormalities. This genetic heterogeneity corresponds with the hypothesis that cervical ribs can be regarded as a sign of disruption of critical, highly interactive stages of embryogenesis. The vertebral pattern can probably provide valuable information regarding fetal and neonatal outcome.

Assessing species richness trends: Declines of bees and bumblebees in the Netherlands since 1945.

Estimating and predicting temporal trends in species richness is of general importance, but notably difficult because detection probabilities of species are imperfect and many datasets were collected in an opportunistic manner. We need to improve our capabilities to assess richness trends using datasets collected in unstandardized procedures with potential collection bias. Two methods are proposed and applied to estimate richness change, which both incorporate models for sampling effects and detection probability: (a) nonlinear species accumulation curves with an error variance model and (b) Pradel capture-recapture models. The methods are used to assess nationwide temporal trends (1945-2018) in the species richness of wild bees in the Netherlands. Previously, a decelerating decline in wild bee species richness was inferred for part of this dataset. Among the species accumulation curves, those with nonconstant changes in species richness are preferred. However, when analyzing data subsets, constant changes became selected for non-Bombus bees (for samples in collections) and bumblebees (for spatial grid cells sampled in three periods). Smaller richness declines are predicted for non-Bombus bees than bumblebees. However, when relative losses are calculated from confidence intervals limits, they overlap and touch zero loss. Capture-recapture analysis applied to species encounter histories infers a constant colonization rate per year and constant local species survival for bumblebees and other bees. This approach predicts a 6% reduction in non-Bombus species richness from 1945 to 2018 and a significant 19% reduction for bumblebees. Statistical modeling to detect species richness time trends should be systematically complemented with model checking and simulations to interpret the results. Data inspection, assessing model selection bias, and comparisons of trends in data subsets were essential model checking strategies in this analysis. Opportunistic data will not satisfy the assumptions of most models and this should be kept in mind throughout.

Embryonal life histories: Desiccation plasticity and diapause in the Argentinean pearlfish Austrolebias bellottii.

Embryos of annual killifish diapause in soil egg banks while ponds are dry. Their rates of development and survival in different developmental stages determine the numbers and stages of embryos at rewetting. In the Argentinean pearlfish Austrolebias bellottii, we investigated plasticity for desiccation in such embryonal life history components across phases of mild desiccation and rewetting and also effects of life history on hatching. In comparison with nonannuals, our data suggest that incidences of diapause have become relatively independent of the occurrence of desiccation, as if they have become genetically assimilated. We found limited survival effects of desiccation, limited developmental delays, and an acceleration of development into the prehatching stage. This response can be adaptive when desiccation informs that an opportunity to hatch approaches. Embryos arrest development in the prehatching stage (diapause DIII) or in the dispersed-cell phase (diapause DI). Parental pair variation in rates of development and survival in the earliest developmental stages affects the fraction of embryos that are in DI at rewetting and the number surviving. Given such effects on life history fitness components, rates during embryonal development seem "visible" to selection and the developmental system can thus adapt when pair variation contains a heritable component. In agreement with expectations for the presence of diversified bet-hedging, some embryos hatched and others not in over half of the clutches with several developed embryos at the moment of rewetting. Hatching probabilities increased for eggs produced later in the experiment, and they increased when embryos were rewetted a second time after two months. This response is opposite of what is expected when age-dependent hatching would be adapted to exploit opportunities for completing another generation before the dry season.

Conservation of the segmented germband stage: robustness or pleiotropy?

Gene expression patterns of the segment polarity genes in the extended and segmented germband stage are remarkably conserved among insects. To explain the conservation of these stages, two hypotheses have been proposed. One hypothesis states that the conservation reflects a high interactivity between modules, so that mutations would have several pleiotropic effects in other parts of the body, resulting in stabilizing selection against mutational variation. The other hypothesis states that the conservation is caused by robustness of the segment polarity network against mutational changes. When evaluating the empirical evidence for these hypotheses, we found strong support for pleiotropy and little evidence supporting robustness of the segment polarity network. This points to a key role for stabilizing selection in the conservation of these stages. Finally, we discuss the implications for robustness of organizers and long-term conservation in general.

The interplay between behavior and morphology in the evolutionary dynamics of resource specialization.

We analyze the consequences of diet choice behavior for the evolutionary dynamics of foraging traits by means of a mathematical model. The model is characterized by the following features. Consumers feed on two different substitutable resources that are distributed in a fine-grained manner. On encounter with a resource item, consumers decide whether to attack it so as to maximize their energy intake. Simultaneously, evolutionary change occurs in morphological traits involved in the foraging process. The assumption here is that evolution is constrained by a trade-off in the consumer's ability to forage on the alternative resources. The model predicts that flexible diet choice behavior can guide the direction of evolutionary change and mediate coexistence of different consumer types. Such polymorphisms can evolve from a monomorphic population at evolutionary branching points and also at points where a small genetic change in a trait can provoke a sharp instantaneous and nongenetic change in choice behavior. In the case of weak trade-offs, the evolutionary dynamics of a dimorphic consumer population can lead to alternative evolutionarily stable communities. The robustness of these predictions is checked with individual-based simulations and by relaxing the assumption of optimally foraging consumers.

Protected polymorphism and evolutionary stability in pleiotropic models with trait-specific dominance.

When alleles have pleiotropic effects on a number of quantitative traits, the degree of dominance between a pair of alleles can be different for each trait. Such trait-specific dominance has been studied previously in models for the maintenance of genetic variation by antagonistic effects of an allele on two fitness components. By generalizing these models to an arbitrary number of fitness components or other phenotypic traits with different degrees of dominance, I show that genetic polymorphism is generally impossible without antagonistic fitness effects of different traits and without trait-specific dominance. I also investigate dominance and pleiotropy from a more long-term evolutionary perspective, allowing for the study of general ecological scenarios, and I discuss the effects of trait-specific dominance on evolutionary stability criteria. When selection is mainly directional and only trait-specific dominance and antagonism cause the emergence of polymorphism, then these polymorphisms can be overtaken by single mutants again, such that they are probably short-lived on an evolutionary time scale. Near evolutionarily singular points where directional selection is absent, trait-specific dominance and overdominance facilitate the emergence of polymorphism and cause evolutionary divergence in some cases. An important outcome of these models is that trait-specific dominance allows for the emergence of genetic polymorphisms without a selective disadvantage for heterozygotes. This removes the scope for the evolution of assortative mate choice and affects dominance modification. Sympatric speciation by disruptive ecological selection requires this heterozygote disadvantage in order to evolve, and therefore it becomes less plausible if the emergence of genetic polymorphism usually occurs via trait-specific dominance and antagonistic effects.

Extreme selection in humans against homeotic transformations of cervical vertebrae.

Why do all mammals, except for sloths and manatees, have exactly seven cervical vertebrae? In other vertebrates and other regions, the vertebral number varies considerably. We investigated whether natural selection constrains the number of cervical vertebrae in humans. To this end, we determined the incidence of cervical ribs and other homeotic vertebral changes in radiographs of deceased human fetuses and infants, and analyzed several existing datasets on the incidence in infants and adults. Our data show that homeotic transformations that change the number of cervical vertebrae are extremely common in humans, but are strongly selected against: almost all individuals die before reproduction. Selection is most probably indirect, caused by a strong coupling of such changes with major congenital abnormalities. Changes in the number of thoracic vertebrae appear to be subject to weaker selection, in good correspondence with the weaker evolutionary constraint on these numbers. Our analysis highlights the role of prenatal selection in the conservation of our common body plan.

A new perspective on developmental plasticity and the principles of adaptive morph determination.

Organisms can have divergent paths of development leading to alternative phenotypes, or morphs. The choice of developmental path may be set by environmental cues, the individual's genotype, or a combination of the two. Using individual-based simulation and analytical investigation, we explore the idea that from the viewpoint of a developmental switch, genetic morph determination can sometimes be regarded as adaptive developmental plasticity. We compare the possibilities for the evolution of environmental and genetic morph determination and combinations of the two in situations with spatial variation in conditions. We find that the accuracy of environmental cues in predicting coming selective conditions is important for environmental morph determination, in accordance with previous results, and that genetic morph determination is favored in a similar way by the accuracy of genetic cues, in the form of selectively maintained gene frequency differences between local populations. Restricted gene flow and strong selection acting on the phenotypic alternatives produce clearer gene frequency differences and lead to greater accuracy of genetic cues. For combined environmental and genetic morph determination, we show that the developmental machinery can evolve toward efficiently combining information in environmental and genetic cues for the purpose of predicting coming selective conditions.

The evolution of resource specialization through frequency-dependent and frequency-independent mechanisms.

Levins's fitness set approach has shaped the intuition of many evolutionary ecologists about resource specialization: if the set of possible phenotypes is convex, a generalist is favored, while either of the two specialists is predicted for concave phenotype sets. An important aspect of Levins's approach is that it explicitly excludes frequency-dependent selection. Frequency dependence emerged in a series of models that studied the degree of character displacement of two consumers coexisting on two resources. Surprisingly, the evolutionary dynamics of a single consumer type under frequency dependence has not been studied in detail. We analyze a model of one evolving consumer feeding on two resources and show that, depending on the trait considered to be subject to evolutionary change, selection is either frequency independent or frequency dependent. This difference is explained by the effects different foraging traits have on the consumer-resource interactions. If selection is frequency dependent, then the population can become dimorphic through evolutionary branching at the trait value of the generalist. Those traits with frequency-independent selection, however, do indeed follow the predictions based on Levins's fitness set approach. This dichotomy in the evolutionary dynamics of traits involved in the same foraging process was not previously recognized.

Viviparity stimulates diversification in an order of fish.

Species richness is distributed unevenly across the tree of life and this may be influenced by the evolution of novel phenotypes that promote diversification. Viviparity has originated ∼150 times in vertebrates and is considered to be an adaptation to highly variable environments. Likewise, possessing an annual life cycle is common in plants and insects, where it enables the colonization of seasonal environments, but rare in vertebrates. The extent to which these reproductive life-history traits have enhanced diversification and their relative importance in the process remains unknown. We show that convergent evolution of viviparity causes bursts of diversification in fish. We built a phylogenetic tree for Cyprinodontiformes, an order in which both annualism and viviparity have arisen, and reveal that while both traits have evolved multiple times, only viviparity played a major role in shaping the patterns of diversity. These results demonstrate that changes in reproductive life-history strategy can stimulate diversification.