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A comparison of ancestral state reconstruction methods for quantitative characters

Choosing an ancestral state reconstruction method among the alternatives available for quantitative characters may be puzzling. We present here a comparison of five of them, namely the maximum likelihood, restricted maximum likelihood, generalized least squares, phylogenetic independent contrasts and squared parsimony methods.\n\nA review of the relations between these methods shows that the first three ones infer the same ancestral states and can only be distinguished by the distributions accounting for the reconstruction uncertainty which they provide.\n\nThe respective accuracy of the methods is assessed over character evolution simulated under a Brownian motion with (and without) drift. We start by giving the general form of ancestral state distributions conditioned on leaf states under the simulation model.\n\nAncestral distributions are used first, to give a theoretical lower bound of the expected reconstruction error, and second, to develop an original evaluation scheme which is more efficient than comparing the reconstructed and the simulated states.\n\nOur simulations show that: (i) the methods do not perform well as the evolution drift increases; (ii) the maximum likelihood method is generally the most accurate and (iii) not all the distributions of the reconstruction uncertainty provided by the methods are equally relevant.

Evolutionary Biology

Evolutionary stability of jointly evolving traits in subdivided populations

The evolutionary stability of quantitative traits depends on whether a population can resist invasion by any mutant. While uninvadability is well understood in well-mixed populations, it is much less so in subdivided populations when multiple traits evolve jointly. Here, we investigate whether a spatially subdivided population at a monomorphic equilibrium for multiple traits can withstand invasion by any mutant, or is subject to diversifying selection. Our model also explores the among traits correlations arising from diversifying selection and how they depend on relatedness due to limited dispersal. We find that selection favours a positive (negative) correlation between two traits, when the selective effects of one trait on relatedness is positively (negatively) correlated to the indirect fitness effects of the other trait. We study the evolution of traits for which this matters: dispersal that decreases relatedness, and helping that has positive indirect fitness effects. We find that when dispersal cost is low and the benefits of helping accelerate faster than its costs, selection leads to the coexistence of mobile defectors and sessile helpers. Otherwise, the population evolves to a monomorphic state with intermediate helping and dispersal. Overall, our results highlight the importance of population subdivision for evolutionary stability and correlations among traits.

Evolutionary Biology

Genetic evidence for natural selection in humans in the contemporary United States

Recent findings from molecular genetics now make it possible to test directly for natural selection by analyzing whether genetic variants associated with various phenotypes have been under selection. I leverage these findings to construct polygenic scores that use individuals' genotypes to predict their body mass index, educational attainment (EA), glucose concentration, height, schizophrenia, total cholesterol, and (in females) age at menarche. I then examine associations between these scores and fitness to test whether natural selection has been occurring. My study sample includes individuals of European ancestry born between 1931 and 1953 in the Health and Retirement Study, a representative study of the US population. My results imply that natural selection has been slowly favoring lower EA in both females and males, and are suggestive that natural selection may have favored a higher age at menarche in females. For EA, my estimates imply a rate of selection of about -1.5 months of education per generation (which pales in comparison with the increases in EA observed in contemporary times). Though they cannot be projected over more than one generation, my results provide additional evidence that humans are still evolving--albeit slowly, especially when compared to the rapid secular changes that have occurred over the past few generations due to cultural and environmental factors.

Evolutionary Biology

Evolutionary inference across eukaryotes identifies specific pressures favoring mitochondrial gene retention

Since their endosymbiotic origin, mitochondria have lost most of their genes. Despite the central cellular importance of mtDNA, selective mechanisms underlying the evolution of its content across eukaryotes remain contentious, with no data-driven exploration of different hypotheses. We developed HyperTraPS, a powerful methodology coupling stochastic modelling with Bayesian inference to identify the ordering and causes of evolutionary events. Using a large dataset from over 2000 complete mitochondrial genomes, we inferred evolutionary trajectories of mtDNA gene loss across the eukaryotic tree of life. We find that proteins fulfilling central energetic roles in the assembly of mitochondrial complexes are prefentially retained in mitochondria across eukary-otes, biophysically supporting colocalization for redox regulation. We also provide the first quantitative evidence that a combination of GC content and protein hydrophobicity is required to explain mtDNA gene retention, and predicts the success of artificial gene transfer experiments. Our results demonstrate that a combination of these three characteristics explains most mtDNA gene variability, addressing the long-debated question of why particular genes are retained in mitochondrial genomes.

Evolutionary Biology

Signatures of positive selection and local adaptation to urbanization in white-footed mice (Peromyscus leucopus)

Urbanization significantly alters natural ecosystems and has accelerated globally as humans move into dense urban centers. Urban wildlife populations are often highly fragmented by an inhospitable matrix of human infrastructure. Isolated populations may adapt in response to novel urban pressures, but few studies have found evidence of selection in urban environments. We used multiple approaches to examine signatures of selection in transcriptomes from white-footed mice (Peromyscus leucopus) in New York City. We analyzed transcriptomes from 48 P. leucopus individuals from three urban and three rural populations for evidence of rapid local adaption in isolated urban habitats. We generated a dataset of 154,770 SNPs and analyzed patterns of genetic differentiation between urban and rural sites. We also used genome scans and genotype-by-environment (GEA) analyses to identify signatures of selection in a large subset of genes. Neutral demographic processes may create allele frequency patterns that are indistinguishable from positive selection. Thus, we accounted for demography by simulating a neutral SNP dataset under the inferred demographic history for the sampled P. leucopus populations to serve as a null model for outlier analysis. We then annotated outlier genes and further validated them by associating allele frequency differences with two urbanization variables: percent impervious surface and human population density. Many candidate genes were involved in metabolic functions, especially dietary specialization. A subset of these genes have well-established roles in metabolizing lipids and carbohydrates, including transport of cholesterol and desaturation of fatty acids. Our results reveal clear genetic differentiation between rural and urban sites that resulted from rapid local adaptation and drift in urbanizing habitats. The specific outlier loci that we identified suggest that populations of P. leucopus are using novel food resources in urban habitats and selection pressures are acting to change metabolic pathways. Our findings support the idea that cities represent novel ecosystems with a unique set of selective pressures.

Evolutionary Biology

The genetic architecture of local adaptation II: The QTL landscape of water-use efficiency for foxtail pine (Pinus balfouriana Grev. & Balf.)

Water availability is an important driver of the geographic distribution of many plant species, although its importance relative to other climatic variables varies across climate regimes and species. A common indirect measure of water-use efficiency (WUE) is the ratio of carbon isotopes ({delta}13C) fixed during photosynthesis, especially when analyzed in conjunction with a measure of leaf-level resource utilization ({delta}15N). Here, we test two hypotheses about the genetic architecture of WUE for foxtail pine (Pinus balfouriana Grev. & Balf.) using a novel mixture of double digest restriction site associated DNA sequencing, species distribution modeling, and quantitative genetics. First, we test the hypothesis that water availability is an important determinant of the geographical range of foxtail pine. Second, we test the hypothesis that variation in {delta}13C and {delta}15N is genetically based, differentiated between regional populations, and has genetic architectures that include loci of large effect. We show that precipitation-related variables structured the geographical range of foxtail pine, climate-based niches differed between regional populations, and {delta}13C and {delta}15N were heritable with moderate signals of differentiation between regional populations. A set of large-effect QTLs (n = 11 for {delta}13C; n = 10 for {delta}15N) underlying {delta}13C and {delta}15N variation, with little to no evidence of pleiotropy, was discovered using multiple-marker, half-sibling regression models. Our results represent a first approximation to the genetic architecture of these phenotypic traits, including documentation of several patterns consistent with {delta}13C being a fitness-related trait affected by natural selection.

Evolutionary Biology

Hybrid asexuality as a primary reproductive barrier: on the interconnection between asexuality and speciation

Speciation usually proceeds in a continuum from intensively hybridizing populations until the formation of irreversibly isolated species. Restriction of interspecific gene flow may often be achieved by gradual accumulation of intrinsic postzygotic incompatibilities with hybrid infertility typically evolving more rapidly than inviability. A reconstructed history of speciation in European loaches (Cobitis) reveals that accumulation of postzygotic reproductive incompatibilities may take an alternative, in the literature largely neglected, pathway through initiation of hybrids asexuality rather than through a decrease in hybrids fitness. Combined evidence shows that contemporary Cobitis species readily hybridize in hybrid zones, but their gene pools are isolated as hybridization produces infertile males and fertile but clonally reproducing females that cannot mediate introgressions. Nevertheless, coalescent analyses indicated intensive historical gene flow during earlier stages of Cobitis diversification, suggesting that non-clonal hybrids must have existed in the past. The revealed patterns imply that during the initial stages of speciation, hybridization between little diverged species produced recombinant hybrids mediating gene flow, but growing divergence among species caused disrupted meiosis in hybrids resulting in their clonality, which acts as a barrier to gene flow. Comparative analysis of published data on other fish hybrids corroborated the generality of our findings; the species pairs producing asexual hybrids were more genetically diverged than those pairs producing fertile sexual hybrids but less diverged than species pairs producing infertile hybrids. Hybrid asexuality therefore appears to evolve at lower divergence than other types of postzygotic barriers and might thus represent a primary reproductive barrier in many taxa.

Evolutionary Biology

How transposons drive evolution of virulence in a fungal pathogen

Genomic plasticity enables adaptation to changing environments, which is especially relevant for pathogens that engage in arms races with their hosts. In many pathogens, genes mediating aggressiveness cluster in highly variable, transposon-rich, physically distinct genomic compartments. However, understanding of the evolution of these compartments, and the role of transposons therein, remains limited. We now show that transposons are the major driving force for adaptive genome evolution in the fungal plant pathogen Verticillium dahliae. Highly variable genomic regions evolved by frequent segmental duplications mediated by erroneous homologous recombination, often utilizing transposons, leading to genetic material that is free to diverge. Intriguingly, the duplicated regions are enriched in active transposons that further contribute to local genome plasticity. Thus, we provide evidence for genome shaping by transposons, both in an active and passive manner, which impacts the evolution of pathogen aggressiveness.

Evolutionary Biology

Sequence amplification via cell passaging creates spurious signals of positive adaptation in influenza H3N2 hemagglutinin

Clinical influenza A isolates are frequently not sequenced directly. Instead, a majority of these isolates (~70% in 2015) are first subjected to passaging for amplification, most commonly in non-human cell culture. Here, we find that this passaging leaves distinct signals of adaptation in the viral sequences, which can confound evolutionary analyses of the viral sequences. We find distinct patterns of adaptation to Madin-Darby (MDCK) and monkey cell culture absent from unpassaged hemagglutinin sequences. These patterns also dominate pooled datasets not separated by passaging type, and they increase in proportion to the number of passages performed. By contrast, MDCK-SIAT1 passaged sequences seem mostly (but not entirely) free of passaging adaptations. Contrary to previous studies, we find that using only internal branches of the influenza phylogenetic trees is insufficient to correct for passaging artifacts. These artifacts can only be safely avoided by excluding passaged sequences entirely from subsequent analysis. We conclude that future influenza evolutionary analyses should appropriately control for potentially confounding effects of passaging adaptations.

Evolutionary Biology

Punctuated evolution and transitional hybrid network in an ancestral cell cycle of fungi

Although cell cycle control is an ancient, conserved, and essential process, some core animal and fungal cell cycle regulators share no more sequence identity than non-homologous proteins. Here, we show that evolution along the fungal lineage was punctuated by the early acquisition and entrainment of the SBF transcription factor through horizontal gene transfer. Cell cycle evolution in the fungal ancestor then proceeded through a hybrid network containing both SBF and its ancestral animal counterpart E2F, which is still maintained in many basal fungi. We hypothesize that a virally-derived SBF may have initially hijacked cell cycle control by activating transcription via the cis-regulatory elements targeted by the ancestral cell cycle regulator E2F, much like extant viral oncogenes. Consistent with this hypothesis, we show that SBF can regulate promoters with E2F binding sites in budding yeast.

Evolutionary Biology

Skew in ovarian activation depends on domicile size in a facultatively social thrips

Costs and benefits of group living are a fundamental topic in behavioural ecology. Resource availability affects individuals breeding prospects alone and in groups, as well as how reproduction is distributed within groups (\"reproductive skew\"). Here, we provide correlational evidence in facultatively social thrips that breeding resources are associated with (1) whether solitary or social living is favoured, and (2) the degree of ovarian skew.\n\nDunatothrips aneurae (Thysanoptera, Phlaeothripidae) cooperatively build silk \"domiciles\" on Australian Acacias, feeding exclusively from internal phyllode surfaces. Per capita productivity scaled differently with group size depending on domicile volume - females in small domiciles did better alone than in groups, whereas in large domiciles single and group-nesting females did equally well. Ovarian dissections revealed that in small domiciles some females were nonreproductive, indicating ovarian (i.e. reproductive) skew. Skew increased as domicile size decreased and group size increased. Breeders had smaller oocyte volume in smaller domiciles, especially those containing nonreproductives.\n\nThese findings suggest group formation and reproductive skew in D. aneurae may be influenced by reproductive competition for breeding resources. Nonreproductive females in small domiciles may be reproductively suppressed, subfertile, or waiting to reproduce. We speculate they may avoid eviction by contributing as \"helpers\" to domicile maintenance.

Evolutionary Biology

Phenotypic plasticity promotes balanced polymorphism in periodic environments by a genomic storage effect

Phenotypic plasticity is known to evolve in perturbed habitats, where it alleviates the deleterious effects of selection. But the effects of plasticity on levels of genetic polymorphism, an important precursor to adaptation in temporally varying environments, are unclear. Here we develop a haploid, two-locus population-genetic model to describe the interplay between a plasticity modifier locus and a target locus subject to periodically varying selection. We find that the interplay between these two loci can produce a \"genomic storage effect\" that promotes balanced polymorphism over a large range of parameters, in the absence of all other conditions known to maintain genetic variation. The genomic storage effect arises as recombination allows alleles at the two loci to escape more harmful genetic backgrounds and associate in haplotypes that persist until environmental conditions change. Using both Monte Carlo simulations and analytical approximations we quantify the strength of the genomic storage effect across a range of selection pressures, recombination rates, plasticity modifier effect sizes, and environmental periods.

Evolutionary Biology

Evolutionary games of multiplayer cooperation on graphs

There has been much interest in studying evolutionary games in structured populations, often modelled as graphs. However, most analytical results so far have only been obtained for two-player or linear games, while the study of more complex multiplayer games has been usually tackled by computer simulations. Here we investigate evolutionary multiplayer games on graphs updated with a Moran death-Birth process. For cycles, we obtain an exact analytical condition for cooperation to be favored by natural selection, given in terms of the payoffs of the game and a set of structure coefficients. For regular graphs of degree three and larger, we estimate this condition using a combination of pair approximation and diffusion approximation. For a large class of cooperation games, our approximations suggest that graph-structured populations are stronger promoters of cooperation than populations lacking spatial structure. Computer simulations validate our analytical approximations for random regular graphs and cycles, but show systematic differences for graphs with many loops such as lattices. In particular, our simulation results show that these kinds of graphs can even lead to more stringent conditions for the evolution of cooperation than well-mixed populations. Overall, we provide evidence suggesting that the complexity arising from many-player interactions and spatial structure can be captured by pair approximation in the case of random graphs, but that it need to be handled with care for graphs with high clustering.\n\nAuthor SummaryCooperation can be defined as the act of providing fitness benefits to other individuals, often at a personal cost. When interactions occur mainly with neighbors, assortment of strategies can favor cooperation but local competition can undermine it. Previous research has shown that a single coefficient can capture this trade-off when cooperative interactions take place between two players. More complicated, but also more realistic models of cooperative interactions involving multiple players instead require several such coefficients, making it difficult to assess the effects of population structure. Here, we obtain analytical approximations for the coefficients of multiplayer games in graph-structured populations. Computer simulations show that, for particular instances of multiplayer games, these approximate coefficients predict the condition for cooperation to be promoted in random graphs well, but fail to do so in graphs with more structure, such as lattices. Our work extends and generalizes established results on the evolution of cooperation on graphs, but also highlights the importance of explicitly taking into account higher-order statistical associations in order to assess the evolutionary dynamics of cooperation in spatially structured populations.

Evolutionary Biology

The stasis that wasn’t: Adaptive evolution goes against phenotypic selection in a wild rodent population

In natural populations, quantitative trait dynamics often do not appear to follow evolutionary predictions: Despite abundant examples of natural selection acting on heritable traits, conclusive evidence for contemporary adaptive evolution remains rare for wild vertebrate populations, and phenotypic stasis seems to be the norm. This so-called stasis paradox highlights our inability to predict evolutionary change, which is especially concerning within the context of rapid anthropogenic environmental change. While the causes underlying the stasis paradox are hotly debated, comprehensive attempts aiming at a resolution are lacking. Here we apply a quantitative genetic framework to individual-based long-term data for a wild rodent population and show that despite a positive association between body mass and fitness, there has been a genetic change towards lower body mass. The latter represents an adaptive response to viability selection favouring juveniles growing up to become relatively small adults, i.e. with a low potential adult mass, which presumably complete their development earlier. This selection is particularly strong towards the end of the snow-free season, and it has intensified in recent years, coinciding which a change in snowfall patterns. Importantly, neither the negative evolutionary change, nor the selective pressures that drive it, are apparent on the phenotypic level, where they are masked by phenotypic plasticity and a non-causal (i.e. non-genetic) positive association between body mass and fitness, respectively. Estimating selection at the genetic level thereby enabled us to uncover adaptive evolution in action, and to identify the corresponding phenotypic selective pressure. We thereby demonstrate that natural populations can show a rapid and adaptive evolutionary response to a novel selective pressure, and that explicitly (quantitative) genetic models are able to provide us with an understanding of the causes and consequences of selection that is superior to purely phenotypic estimates of selection and evolutionary change.

Evolutionary Biology

Combining Shigella Tn-seq data with Gold-standard E. coli Gene Deletion Data Suggests Rare Transitions between Essential and Non-essential Gene Functionality

Gene essentiality - whether or not a gene is necessary for cell growth - is a fundamental component of gene function. It is not well established how quickly gene essentiality can change, as few studies have compared empirical measures of essentiality between closely related organisms. Here we present the results of a Tn-seq experiment designed to detect essential protein coding genes in the bacterial pathogen Shigella flexneri 2a 2457T on a genome-wide scale. Superficial analysis of this data suggested that 451 protein-coding genes in this Shigella strain are critical for robust cellular growth on rich media. Comparison of this set of genes with a gold-standard data set of essential genes in the closely related Escherichia coli K12 BW25113 suggested that an excessive number of genes appeared essential in Shigella but non-essential in E. coli. Importantly, and in converse to this comparison, we found no genes that were essential in E. coli and non-essential in Shigella, suggesting that many genes were artefactually inferred as essential in Shigella. Controlling for such artefacts resulted in a much smaller set of discrepant genes. Among these, we identified three sets of functionally related genes, two of which have previously been implicated as critical for Shigella growth, but which are dispensable for E. coli growth. The data presented here highlight the small number of protein coding genes for which we have strong evidence that their essentiality status differs between the closely related bacterial taxa E. coli and Shigella. A set of genes involved in acetate utilization provides a canonical example. These results leave open the possibility of developing strain-specific antibiotic treatments targeting such differentially essential genes, but suggest that such opportunities may be rare in closely related bacteria.

Evolutionary Biology

No evidence for the positive relationship between genetic correlations and heritabilities

Quantitative genetics predicts, that traits subjected to strong selection should show low heritability and may yield biased estimates of genetic correlations (rg). Similar pattern may also appear if genetic sources of variation are confounded with non-genetic sources. Thus, a positive relationship between genetic correlations and heritabilities (h2) of underlying traits might be observed. Here we test this prediction using a large dataset of published estimates of genetic correlations and employing a powerful meta-analytical approach. We considered both between-traits and cross-sex genetic correlations. We failed to find support for the prediction about a positive rg - h2 relationship: our analysis based on nearly 1000 published estimates of genetic parameters indicates that the predicted relationship is weak and statistically non-significant. Thus, low heritability does not preclude the possibility of detecting substantial genetic correlations. Our meta-analysis indicates that published estimates of genetic parameters coming from various experimental designs and obtained using different statistical techniques are not significantly biased in case of weakly-heritable traits.

Evolutionary Biology

Inferring the frequency spectrum of derived variants to quantify adaptive molecular evolution in protein-coding genes of Drosophila melanogaster

Many approaches for inferring adaptive molecular evolution analyze the unfolded site frequency spectrum (SFS), a vector of counts of sites with different numbers of copies of derived alleles in a sample of alleles from a population. Accurate inference of the high copy number elements of the SFS is difficult, however, because of misassignment of alleles as derived versus ancestral. This is a known problem with parsimony using outgroup species. Here, we show that the problem is particularly serious if there is variation in the substitution rate among sites brought about by variation in selective constraint levels. We present a new method for inferring the SFS using one or two outgroups, which attempts to overcome the problem of misassignment. We show that two outgroups are required for accurate estimation of the SFS if there is substantial variation in selective constraints, which is expected to be the case for nonsynonymous sites of protein-coding genes. We apply the method to estimate unfolded SFSs for synonymous and nonsynonymous sites from Phase 2 of the Drosophila Population Genomics Project. We use the unfolded spectra to estimate the frequency and strength of advantageous and deleterious mutations, and estimate that {small tilde}50% of amino acid substitutions are positively selected, but that less than 0.5% of new amino acid mutations are beneficial, with a scaled selection strength of Nes {approx} 12.

Evolutionary Biology

Social selection maintains honesty of a dynamic visual signal in cichlid fish

How honest signals evolve is a question that has been hotly debated by animal communication theoreticians and for which empirical evidence has been difficult to obtain. Theory predicts that, due to strong conflicts of interest, communication in aggressive contexts should be under strong selection for clear and reliable signaling. On the other hand, context-dependent signaling increases cheating opportunities, depending on how senders and receivers use, acquire and process signal information. Using animal signaling theory, theoretical visual models and behavioral experimentation, we characterize and determine proximate honesty mechanisms of the facial coloration in the Princess of Burundi cichlid, Neolamprologus brichardi, a species with complex social interactions. We show that this facial color pattern evolved stable chromatic conspicuousness for efficient transmission in the aquatic environment, while context-dependent plasticity in luminance of the horizontal black stripe element is used to signal switches in aggressive intent. Importantly, using experimental signal manipulation we demonstrate that social selection by receiver retaliation is the mechanism responsible for maintaining signal honesty. We suggest that by affecting the evolution of pigmentation patterns in sexually monochromatic cichlid species, social selection can have potential impacts on diversification dynamics.

Evolutionary Biology