Coalescent Simulations Research Articles

Discontinuous deformation analysis (DDA) simulations of crack propagation, branching and coalescence in brittle materials under dynamic loading

Cracking in brittle materials under dynamic loading is widely encountered in engineering. The discontinuous deformation analysis (DDA) is a numerical method initially proposed to calculate the static and dynamic behaviors of discrete deformable-block systems, which can also be used to simulate the cracking failure of materials via the sub-block element approach. In the present work, a sub-block element DDA method is applied to simulate the crack evolution problems in brittle materials under dynamic loading. The tensile and shear cracking along the artificial joints between glued sub-block elements is judged based on the stress levels of adjacent sub-blocks by the maximum tensile strength criterion and the Mohr-Coulomb criterion, respectively. Three numerical examples including the Kalthoff-Winkler experiment, the cracking in a tensile loaded plate and the double-hole blasting of a rectangular plate are simulated. The effect of the mesh size for the Kalthoff-Winkler experiment, the loading intensity for the Kalthoff-Winkler experiment and the tensile loaded plate, and the influence of the guiding hole for the double-hole blasting plate are specially investigated. The DDA simulation results of cracking routes and velocity are reasonably consistent with the corresponding experimental results or numerical results by other methods, which manifests the ability of the applied sub-block element DDA method in the simulations crack propagation, branching and coalescence of brittle materials under dynamic loading.

Numerical simulation of crack propagation and coalescence in marine cast iron materials using ordinary state-based peridynamics

Fracturing is one of the major failure modes in marine structures. It is important to investigate the crack propagation and fracture behavior of cast iron material to prevent brittle failure and protect the structural safety of ship equipment. Classical continuum mechanics (CCM) has intrinsic shortcomings in simulating the fracture of solid materials. Peridynamics (PD) is a new nonlocal form of CCM that is highly suitable for the investigations of material failure. In the present study, a damage model, fracture criterion, and nonlinear characteristic are incorporated into the extended ordinary state-based peridynamics (OSPD) to simulate the initiation, propagation and coalescence of the preexisting flaws in marine cast iron materials subjected to tensile loads. First, the linear and nonlinear force functions of the failure process of the extended OSPD were constructed, and a new damage model and corresponding failure criterion were adopted. Then, the initiation, propagation and coalescence of the cracks in cast iron specimens with a single notch, double notch and multiple flows were investigated. Finally, the effects of arrays of cracks, Poisson's ratio and plate thickness on the crack propagation behaviors and peak loads were analyzed. The numerical results are in good agreement with the previous numerical and experimental observations.

Numerical simulation of crack propagation and coalescence in rock materials by the peridynamic method based on strain energy density theory

Numerical simulation of crack propagation and coalescence in rock materials by the peridynamic method based on strain energy density theory

Coalescent simulations indicate that the São Francisco River is a biogeographic barrier for six vertebrates in a seasonally dry South American forest

The riverine barrier hypothesis has been extensively explored in Neotropical rainforests, while its importance in drier regions such as the Caatinga, a seasonally dry tropical forest in northeastern Brazil, has only recently received more attention. The Caatinga is bisected by the São Francisco River (SFR), which has long been suggested to be an important biogeographic feature in the region. However, recent studies have found mixed support for the role of the SFR as a hard barrier, most of them relying on the presence or absence of genetic breaks congruent with its course. Here, we used published multilocus and next-generation data from six vertebrate species to test the SFR’s strength as a barrier. Using model-based approaches (approximate Bayesian computation and supervised machine learning), we tested demographic models incorporating full, intermediate, and zero migration across the SFR, estimating divergence times and migration rates for each species. We found support for the SFR’s role as a barrier, allowing gene flow for some species. Estimated divergence times varied among species but are limited to the late Pleistocene, coherent with one of several proposed paleocourse changes in the river’s geological history. Contrary to the mixed results of previous studies, our study supports the SFR as an important phylogeographic barrier across different taxonomic groups, driving diversification in the Caatinga.

Numerical study on the coalescence of multiple argon bubbles at a horizontal plane with different orifice arrangements in liquid steel under laminar flow

ABSTRACT In the current study, the coalescence of multiple argon bubbles at a horizontal plane in liquid steel with different orifice arrangements was numerically investigated using the volume of the fluid model combined with the continuum surface force model. The numerical simulation of pairwise bubbles coalescence was verified with the experimental observation in the air–water system. The arrangements of equilateral triangle, square and rhombic orifices were considered. The effects of bubble size and orifice structure arrangement on the lateral coalescence of multiple argon bubbles were analysed. Argon bubbles coalesce into a bigger one when the initial bubble distance is less than the critical distance of bubble coalescence. The critical distance for bubble coalescence decreases with the increasing bubble size. When the orifice arrangements were arranged in square, rhombic and equilateral triangles, the coalescence of multiple bubbles become easier. In addition, the mechanisms of bubble coalescence and bounce were discussed.

Theoretical analysis of principal components in an umbrella model of intraspecific evolution

Principal component analysis (PCA) is one of the most frequently-used approach to describe population structure from multilocus genotype data. Regarding geographic range expansions of modern humans, interpretations of PCA have, however, been questioned, as there is uncertainty about the wave-like patterns that have been observed in principal components. It has indeed been argued that wave-like patterns are mathematical artifacts that arise generally when PCA is applied to data in which genetic differentiation increases with geographic distance. Here, we present an alternative theory for the observation of wave-like patterns in PCA. We study a coalescent model – the umbrella model – for the diffusion of genetic variants. The model is based on genetic drift without any particular geographical structure. In the umbrella model, splits from an ancestral population occur almost continuously in time, giving birth to small daughter populations at a regular pace. Our results provide detailed mathematical descriptions of eigenvalues and eigenvectors for the PCA of sampled genomic sequences under the model. When variants uniquely represented in the sample are removed, the PCA eigenvectors are defined as cosine functions of increasing periodicity, reproducing wave-like patterns observed in equilibrium isolation-by-distance models. Including singleton variants in the analysis, the eigenvectors corresponding to the largest eigenvalues exhibit complex wave shapes. The accuracy of our predictions is further investigated with coalescent simulations. Our analysis supports the hypothesis that highly structured wave-like patterns could arise from genetic drift only, and may not always be artificial outcomes of spatially structured data. Genomic data related to the peopling of the Americas are reanalyzed in the light of our new theory.

Simulation-Based Evaluation of Methods, Data Types, and Temporal Sampling Schemes for Detecting Recent Population Declines.

Understanding recent population trends is critical to quantifying species vulnerability and implementing effective management strategies. To evaluate the accuracy of genomic methods for quantifying recent declines (beginning <120 generations ago), we simulated genomic data using forward-time methods (SLiM) coupled with coalescent simulations (msprime) under a number of demographic scenarios. We evaluated both site frequency spectrum (SFS)-based methods (momi2, Stairway Plot) and methods that employ linkage disequilibrium information (NeEstimator, GONE) with a range of sampling schemes (contemporary-only samples, sampling two time points, and serial sampling) and data types (RAD-like data and whole-genome sequencing). GONE and momi2 performed best overall, with >80% power to detect severe declines with large sample sizes. Two-sample and serial sampling schemes could accurately reconstruct changes in population size, and serial sampling was particularly valuable for making accurate inferences when genotyping errors or minor allele frequency cutoffs distort the SFS or under model mis-specification. However, sampling only contemporary individuals provided reliable inferences about contemporary size and size change using either site frequency or linkage-based methods, especially when large sample sizes or whole genomes from contemporary populations were available. These findings provide a guide for researchers designing genomics studies to evaluate recent demographic declines.

A cold-water fish striving in a warming ocean: Insights from whole-genome sequencing of the Greenland halibut in the Northwest Atlantic

Characterizing the extent of genetic differentiation among individuals and its distribution across the genome is increasingly important to inform both conservation and management of exploited species. The Greenland Halibut is one of the main demersal fish species to be commercially exploited in Eastern Canada, and accurate information on geographic population structure and local adaptation is required to ensure the long-term presence of this species. We generated high-quality whole-genome sequencing data for 1,297 Greenland Halibut sampled across 32 locations throughout the Northwest Atlantic (from Arctic Canadian and Greenlandic coasts to the Gulf of St Lawrence). Population genetic structure was analyzed, revealing an absence of population differentiation between Canada and west Greenland but significant genetic differentiation between the Gulf of Saint Lawrence and the remainder of the Northwest Atlantic. Except for Gulf of Saint Lawrence, Greenland Halibut thus appear to be panmictic throughout the Northwest Atlantic. Environmental Association Analyses revealed that the environment explained up to 51 % might be replaced by 51% of the differentiation observed between the two stocks, with both ocean-bottom and surface variables (e.g., temperature and oxygen) involved in the observed genomic differentiation. Altogether, these results indicate that phenotypic differences previously observed between the Gulf of Saint Lawrence and the Northwest Atlantic likely resulted from functional adaptive divergence to their respective environmental conditions. Using coalescent simulations, we also assessed how high levels of migration between the two stocks would allow Greenland Halibut to potentially escape unfavorable environmental conditions in the Gulf of Saint Lawrence. In addition to supporting the management of this important exploited species, this work highlights the utility of using comprehensive genomic datasets to characterize the effects of climate change across a wider range of species.

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On the Cover: A water hole in Australia. Aridification of Australia during the Pleistocene has influenced the divergence of a migratory freshwater obligate, the golden perch (Macquaria ambigua). Photo credit: Shutterstock/Tim Pryce. For more information see “Aridification-driven evolution of a migratory fish revealed by niche modelling and coalescence simulations” by Booth et al. (DOI:10.1111/jbi.14337), a contribution to the special section on Geogenomics.

Subgrain Coalescence Simulation by Means of an Advanced Statistical Model of Inelastic Deformation

The development of technological methods for processing and manufacturing of functional (with a priori targeted properties) polycrystalline materials and products made of these materials still remains an acute problem. A multilevel modeling approach offers researchers the opportunity to describe inelastic deformation by applying internal variables that give an effective characterization of the material structure at different structural scale levels. High temperature plastic deformation is accompanied by these processes, which leads to a significant rearrangement of the meso- and microstructure of the material. The most substantial contribution to changing the properties of polycrystals is made by the evolution of grain and defect structures at the expense of dynamic recrystallization, which significantly depends on dynamic recovery. In this paper, we consider the problem of the coalescence of subgrains undergoing rotation during inelastic hot deformation. This process is called subgrain coalescence, and it is one of the dynamic recovery mechanisms responsible for changes in the fine subgrain structure. Under applied thermomechanical loads, the coalescence process promotes the formation of recrystallization nuclei and their subsequent growth, which can greatly change the grain structure of a polycrystal. The problem was solved in terms of the advanced statistical model of inelastic deformation, modified to describe the subgrain coalescence process. The model takes into account the local interactions between contacting structural elements (subgrains). These have to be considered so that the grain coalescence caused by a decrease in subboundary energies during their progressive merging can be adequately analyzed. For this purpose, a subgrain structure quite similar to the real structure was modeled using Laguerre polyhedra. Subgrain rotations were investigated using the developed model, which relies on the consideration of the excess density edge component of the same sign dislocations on incidental subgrain boundaries. The results of modeling of a copper polycrystal are presented, and the effects of temperature and strain rate on the subgrain coalescence process is demonstrated.

Parallel local adaptation to an alpine environment in Arabidopsis arenosa

Abstract Parallel local adaptation, that is, when distinct genetic lineages independently adapt to the same selective environment, provides strong evidence for the action of natural selection. A few cases of parallel local adaptation were reported in plants but underlying mechanisms promoting or preventing the parallel response, such as the balance between migration and selection, were rarely quantified. Here, we conducted a transplant experiment to test whether distinct foothill–alpine population pairs of Arabidopsis arenosa exhibited similar adaptive responses to a contrasting alpine environment. We further investigated selection and migration patterns in these populations. Seedlings of 16 foothill and alpine populations of A. arenosa from four distinct mountain regions (one occupied by diploid and three by tetraploid populations) were transplanted into one low‐ and one high‐elevation site. We recorded fitness proxies over two growing seasons to test whether the elevation‐of‐origin advantage was manifested in the same way across the four regions of origin. Then, we quantified the strength of selection on the traits at each transplantation site and used coalescent simulations to estimate past gene flow intensity between each pair of foothill and alpine populations in each region. We demonstrated that the four pairs of populations exhibited similar adaptive responses to elevation difference in terms of survival, number of flowering plants, stem height and accumulation of above‐ground dry biomass; the other traits (rosette size, number of leaves, stems and flowers) exhibited rather regional‐specific patterns. In addition, we found minor effects of ploidy level on the fitness proxies recorded. Our selection and migration analysis revealed that parallel local adaptation was probably achieved by differential selective pressure at low versus high elevation in combination with lack or limited gene flow between foothill and alpine populations. Synthesis. We show that the previously documented strong morphological and genetic distinctness of alpine A. arenosa compared to their foothill counterparts, which has been hypothesized to be driven by natural selection, is indeed mirrored in fitness differences consistent with parallel local adaptation. Our results provide experimental support for the repeatability of adaptive evolution and highlight the prominent role of divergent selection.

Using gridCoal to assess whether standard population genetic theory holds in the presence of spatio-temporal heterogeneity in population size.

Spatially explicit population genetic models have long been developed, yet have rarely been used to test hypotheses about the spatial distribution of genetic diversity or the genetic divergence between populations. Here, we use spatially explicit coalescence simulations to explore the properties of the island and the two-dimensional stepping stone models under a wide range of scenarios with spatio-temporal variation in deme size. We avoid the simulation of genetic data, using the fact that under the studied models, summary statistics of genetic diversity and divergence can be approximated from coalescence times. We perform the simulations using gridCoal, a flexible spatial wrapper for the software msprime (Kelleher et al., 2016, Theoretical Population Biology, 95, 13) developed herein. In gridCoal, deme sizes can change arbitrarily across space and time, as well as migration rates between individual demes. We identify different factors that can cause a deviation from theoretical expectations, such as the simulation time in comparison to the effective deme size and the spatio-temporal autocorrelation across the grid. Our results highlight that FST , a measure of the strength of population structure, principally depends on recent demography, which makes it robust to temporal variation in deme size. In contrast, the amount of genetic diversity is dependent on the distant past when Ne is large, therefore longer run times are needed to estimate Ne than FST . Finally, we illustrate the use of gridCoal on a real-world example, the range expansion of silver fir (Abies alba Mill.) since the last glacial maximum, using different degrees of spatio-temporal variation in deme size.

Synthesis of geological data and comparative phylogeography of lowland tetrapods suggests recent dispersal through lowland portals crossing the Eastern Andean Cordillera.

Vicariance is the simplest explanation for divergence between sister lineages separated by a potential barrier, and the northern Andes would seem to provide an ideal example of a vicariant driver of divergence. We evaluated the potential role of the uplift of the Eastern Cordillera (EC) of the Colombian Andes and the Mérida Andes (MA) of Venezuela as drivers of vicariance between lowland populations co-distributed on both flanks. We synthesized published geological data and provided a new reconstruction showing that the EC-MA grew from north to south, reaching significant heights and separating drainages and changing sediment composition by 38–33 million years ago (Ma). A few lowland passes across the EC-MA may have reached their current heights (~1,900 m a.s.l.) at 3–5 Ma. We created a comparative phylogeographic data set for 37 lineages of lowland tetrapods. Based on molecular phylogenetic analyses, most divergences between sister populations or species across the EC-MA occurred during Pliocene and the Quaternary and a few during the latest Miocene, and coalescent simulations rejected synchronous divergence for most groups. Divergence times were on average slightly but significantly more recent in homeotherms relative to poikilotherms. Because divergence ages are mostly too recent relative to the geological history and too asynchronous relative to each other, divergence across the northern Andes may be better explained by organism-environment interactions concomitant with climate oscillations during the Pleistocene, and/or dispersal across portals through the Andes.

EggLib 3: A python package for population genetics and genomics.

Rapid and repeatable polymorphism analyses have become a necessity with the current amount of genomic data that can be collected in many organisms. Traditionally, such analyses are conducted using a variety of tools in combination, often requiring numerous format translation and manipulation. Here, we present a massively updated version of our previous software package egglib, intended to alleviate such costly and error-prone tinkering with the data. egglib has been streamlined into a python package and thoroughly updated and optimized to accommodate modern-day sized dataset. We show the main characteristics of the package making it a tool of choice to perform population genetics analyses. Once the data are imported (whatever their encoding), they can be filtered, edited, analysed and compared to coalescent simulations very easily and efficiently. Furthermore, the list of diversity and polymorphism statistics that can now be calculated has been greatly expanded. The software and its full documentation are available at https://egglib.org/.

Numerical assessment of ultrasound supported coalescence of water droplets in crude oil

In this study, a numerical assessment of the coalescence of binary water droplets in water-in-oil emulsion was conducted. The investigation addressed the effect of various parameters on the acoustic pressure and coalescence time of water droplets in oil phase. These include transducer material, initial droplet diameter (0.05–0.2 in), interfacial tension (0.012–0.082 N/m), dynamic viscosity (10.6–530 mPas), temperature (20–100 °C), US (ultra sound) frequency (26.04–43.53 kHz) and transducer power (2.5–40 W). The materials assessed are lead zirconate titanate (PZT), lithium niobate (LiNbO3), zinc oxide (ZnO), aluminum nitride (AlN), polyvinylidene fluoride (PVDF), and barium titanate (BaTiO3). The numerical simulation of the binary droplet coalescence showed good agreement with experimental data in the literature. The US implementation at a fixed frequency produced enhanced coalescence (t = 5.9–8.5 ms) as compared to gravitational settling (t = 9.8 ms). At different ultrasound (US) frequencies and transducer materials, variation in the acoustic pressure distribution was observed. Possible attenuation of the US waves, and the subsequent inhibitive coalescence effect under various US frequencies and viscosities, were discussed. Moreover, the results showed that the coalescence time reduced across the range of interfacial tensions which was considered. This reduction can be attributed to the fact that lower interfacial tension produces emulsions which are relatively more stable. Hence, at lower interface tension between the water and crude oil, there was more resistance to the coalescence of the water droplets due to their improved emulsion stability. The increment of the Weber number at higher droplet sizes leads to a delay in the recovery of the droplet to spherical forms after their starting deformation. These findings provide significant insights that could aid further developments in demulsification of crude oil emulsions under varying US and emulsion properties.

A phylotranscriptome study using silica gel-dried leaf tissues produces an updated robust phylogeny of Ranunculaceae

The utility of transcriptome data in plant phylogenetics has gained popularity in recent years. However, because RNA degrades much more easily than DNA, the logistics of obtaining fresh tissues has become a major limiting factor for widely applying this method. Here, we used Ranunculaceae to test whether silica-dried plant tissues could be used for RNA extraction and subsequent phylogenomic studies. We sequenced 27 transcriptomes, 21 from silica gel-dried (SD-samples) and six from liquid nitrogen-preserved (LN-samples) leaf tissues, and downloaded 27 additional transcriptomes from GenBank. Our results showed that although the LN-samples produced slightly better reads than the SD-samples, there were no significant differences in RNA quality and quantity, assembled contig lengths and numbers, and BUSCO comparisons between two treatments. Using these data, we conducted phylogenomic analyses, including concatenated- and coalescent-based phylogenetic reconstruction, molecular dating, coalescent simulation, phylogenetic network estimation, and whole genome duplication (WGD) inference. The resulting phylogeny was consistent with previous studies with higher resolution and statistical support. The 11 core Ranunculaceae tribes grouped into two chromosome type clades (T- and R-types), with high support. Discordance among gene trees is likely due to hybridization and introgression, ancient genetic polymorphism and incomplete lineage sorting. Our results strongly support one ancient hybridization event within the R-type clade and three WGD events in Ranunculales. Evolution of the three Ranunculaceae chromosome types is likely not directly related to WGD events. By clearly resolving the Ranunculaceae phylogeny, we demonstrated that SD-samples can be used for RNA-seq and phylotranscriptomic studies of angiosperms.

Ancient mitochondrial and modern whole genomes unravel massive genetic diversity loss during near extinction of Alpine ibex.

Population bottlenecks can have dramatic consequences for the health and long‐term survival of a species. Understanding of historic population size and standing genetic variation prior to a contraction allows estimating the impact of a bottleneck on the species' genetic diversity. Although historic population sizes can be modelled based on extant genomics, uncertainty is high for the last 10–20 millenia. Hence, integrating ancient genomes provides a powerful complement to retrace the evolution of genetic diversity through population fluctuations. Here, we recover 15 high‐quality mitogenomes of the once nearly extinct Alpine ibex spanning 8601 BP to 1919 CE and combine these with 60 published modern whole genomes. Coalescent demography simulations based on modern whole genomes indicate population fluctuations coinciding with the last major glaciation period. Using our ancient and historic mitogenomes, we investigate the more recent demographic history of the species and show that mitochondrial haplotype diversity was reduced to a fifth of the prebottleneck diversity with several highly differentiated mitochondrial lineages having coexisted historically. The main collapse of mitochondrial diversity coincides with elevated human population growth during the last 1–2 kya. After recovery, one lineage was spread and nearly fixed across the Alps due to recolonization efforts. Our study highlights that a combined approach integrating genomic data of ancient, historic and extant populations unravels major long‐term population fluctuations from the emergence of a species through its near extinction up to the recent past.

Incomplete lineage sorting and local extinction shaped the complex evolutionary history of the Paleogene relict conifer genus, Chamaecyparis (Cupressaceae)

Inferring accurate biogeographic history of plant taxa with an East Asia (EA)-North America (NA) is usually hindered by conflicting phylogenies and a poor fossil record. The current distribution of Chamaecyparis (false cypress; Cupressaceae) with four species in EA, and one each in western and eastern NA, and its relatively rich fossil record, make it an excellent model for studying the EA-NA disjunction. Here we reconstruct phylogenomic relationships within Chamaecyparis using > 1400 homologous nuclear and 61 plastid genes. Our phylogenomic analyses using concatenated and coalescent approaches revealed strong cytonuclear discordance and conflicting topologies between nuclear gene trees. Incomplete lineage sorting (ILS) and hybridization are possible explanations of conflict; however, our coalescent analyses and simulations suggest that ILS is the major contributor to the observed phylogenetic discrepancies. Based on a well-resolved species tree and four fossil calibrations, the crown lineage of Chamaecyparis is estimated to have originated in the upper Cretaceous, followed by diversification events in the early and middle Paleogene. Ancestral area reconstructions suggest that Chamaecyparis had an ancestral range spanning both EA and NA. Fossil records further indicate that this genus is a relict of the “boreotropical” flora, and that local extinctions of European species were caused by global cooling. Overall, our results unravel a complex evolutionary history of a Paleogene relict conifer genus, which may have involved ILS, hybridization and the extinction of local species.

Mathematical constraints on FST: multiallelic markers in arbitrarily many populations.

Interpretations of values of the FST measure of genetic differentiation rely on an understanding of its mathematical constraints. Previously, it has been shown that FST values computed from a biallelic locus in a set of multiple populations and FST values computed from a multiallelic locus in a pair of populations are mathematically constrained as a function of the frequency of the allele that is most frequent across populations. We generalize from these cases to report here the mathematical constraint on FST given the frequency M of the most frequent allele at a multiallelic locus in a set of multiple populations. Using coalescent simulations of an island model of migration with an infinitely-many-alleles mutation model, we argue that the joint distribution of FST and M helps in disentangling the separate influences of mutation and migration on FST. Finally, we show that our results explain a puzzling pattern of microsatellite differentiation: the lower FST in an interspecific comparison between humans and chimpanzees than in the comparison of chimpanzee populations. We discuss the implications of our results for the use of FST.This article is part of the theme issue ‘Celebrating 50 years since Lewontin's apportionment of human diversity’.

Population genomics for symbiotic anthozoans: can reduced representation approaches be used for taxa without reference genomes?

Population genetic studies of symbiotic anthozoans have been historically challenging because their endosymbioses with dinoflagellates have impeded marker development. Genomic approaches like reduced representation sequencing alleviate marker development issues but produce anonymous loci, and without a reference genome, it is unknown which organism is contributing to the observed patterns. Alternative methods such as bait-capture sequencing targeting Ultra-Conserved Elements are now possible but costly. Thus, RADseq remains attractive, but how useful are these methods for symbiotic anthozoan taxa without a reference genome to separate anthozoan from algal sequences? We explore this through a case-study using a double-digest RADseq dataset for the sea anemone Bartholomea annulata. We assembled a holobiont dataset (3854 loci) for 101 individuals, then used a reference genome to create an aposymbiotic dataset (1402 loci). For both datasets, we investigated population structure and used coalescent simulations to estimate demography and population parameters. We demonstrate complete overlap in the spatial patterns of genetic diversity, demographic histories, and population parameter estimates for holobiont and aposymbiotic datasets. We hypothesize that the unique combination of anthozoan biology, diversity of the endosymbionts, and the manner in which assembly programs identify orthologous loci alleviates the need for reference genomes in some circumstances. We explore this hypothesis by assembling an additional 21 datasets using the assembly programs pyRAD and Stacks. We conclude that RADseq methods are more tractable for symbiotic anthozoans without reference genomes than previously realized.