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Metabarcoding is often presented as an alternative identification tool to compensate for coarse taxonomic resolution and misidentification encountered with traditional morphological approaches. However, metabarcoding comes with two major impediments which slow down its adoption. First, the picking and destruction of organisms for DNA extraction are time and cost consuming and do not allow organism conservation for further evaluations. Second, current metabarcoding protocols include a PCR enrichment step which induces errors in the estimation of species diversity and relative biomasses. In this study, we first evaluated the capacity of capture enrichment to replace PCR enrichment using controlled freshwater macrozoobenthos mock communities. Then, we tested if DNA extracted from the fixative ethanol (etDNA) of the same mock communities can be used as an alternative to DNA extracted from pools of whole organisms (bulk DNA). We show that capture enrichment provides more reliable and accurate representation of species occurrences and relative biomasses in comparison with PCR enrichment for bulk DNA. While etDNA does not permit to estimate relative biomasses, etDNA and bulk DNA provide equivalent species detection rates. Thanks to its robustness to mismatches, capture enrichment is already an efficient alternative to PCR enrichment for metabarcoding and, if coupled to etDNA, is a time‐saver option in studies where presence information only is sufficient. [ABSTRACT FROM AUTHOR]

Transition to novel environments, such as groundwater colonization by surface organisms, provides an excellent research ground to study phenotypic evolution. However, interspecific comparative studies on evolution to groundwater life are few because of the challenge in assembling large ecological and molecular resources for species‐rich taxa comprised of surface and subterranean species. Here, we make available to the scientific community an operational set of working tools and resources for the Asellidae, a family of freshwater isopods containing hundreds of surface and subterranean species. First, we release the World Asellidae database (WAD) and its web application, a sustainable and FAIR solution to producing and sharing data and biological material. WAD provides access to thousands of species occurrences, specimens, DNA extracts and DNA sequences with rich metadata ensuring full scientific traceability. Second, we perform a large‐scale dated phylogenetic reconstruction of Asellidae to support phylogenetic comparative analyses. Of 424 terminal branches, we identify 34 pairs of surface and subterranean species representing independent replicates of the transition from surface water to groundwater. Third, we exemplify the usefulness of WAD for documenting phenotypic shifts associated with colonization of subterranean habitats. We provide the first phylogenetically controlled evidence that body size of males decreases relative to that of females upon groundwater colonization, suggesting competition for rare receptive females selects for smaller, more agile males in groundwater. By making these tools and resources widely accessible, we open up new opportunities for exploring how phenotypic traits evolve in response to changes in selective pressures and trade‐offs during groundwater colonization. [ABSTRACT FROM AUTHOR]

All organisms on Earth are exposed to low doses of natural radioactivity but some habitats are more radioactive than others. Yet, documenting the influence of natural radioactivity on the evolution of biodiversity is challenging. Here, we addressed whether organisms living in naturally more radioactive habitats accumulate more mutations across generations using 14 species of waterlice living in subterranean habitats with contrasted levels of radioactivity. We found that the mitochondrial and nuclear mutation rates across a waterlouse species' genome increased on average by 60% and 30%, respectively, when radioactivity increased by a factor of three. We also found a positive correlation between the level of radioactivity and the probability of G to T (and complementary C to A) mutations, a hallmark of oxidative stress. We conclude that even low doses of natural bedrock radioactivity influence the mutation rate possibly through the accumulation of oxidative damage, in particular in the mitochondrial genome. [ABSTRACT FROM AUTHOR]

Heterogeneity of hyporheic fauna is associated with geomorphological features and related vertical water exchanges. Constrictions on river floodplain are known to induce groundwater inputs and increase stygobite fauna. Two floodplain constrictions were studied in a large braided river (the Drôme River): one linked to a natural process (valley narrowing), another to an artificial river regulation (early 20th embankment). Spatial distribution of hyporheic organisms were sampled upstream and downstream of the two constrained sections, at 9 stations, 3 positions (left and right sides, centre of the braided strip), 3 replication points and at a depth of 50 cm in the river sediment. The spatial heterogeneity in community composition was higher near the banks than at the centre of the braided strip, no matter the width of the strip. The artificial constriction induced a decrease in spatial heterogeneity of the benthic fraction of the hyporheic fauna, but no changes were detected for the stygofauna. The natural valley narrowing reduced width and thickness of the alluvium and induced an inflow of groundwater resulting in an increase in stygofauna abundance. Natural floodplain narrowing linked to geology thus control the distribution of stygobite species, while artificial constrictions only modify the spatial distribution of the benthic fraction of the hyporheic fauna. Unlabelled Image • Braided channel narrowing can be natural or artificial. • Artificial narrowing induces homogenization of the benthic fraction of the hyporheos. • Natural narrowing induces upwelling of groundwater and increase in stygobite fauna. [ABSTRACT FROM AUTHOR]

Ecologists increasingly rely on molecular delimitation methods (MMs) to identify species boundaries, thereby potentially increasing the number of putative species because of the presence of morphologically cryptic species. It has been argued that cryptic species could challenge our understanding of what determine large‐scale biodiversity patterns which have traditionally been documented from morphology alone. Here, we used morphology and three MMs to derive four different sets of putative species among the European groundwater crustaceans. Then, we used regression models to compare the relative importance of spatial heterogeneity, productivity and historical climates, in shaping species richness and range size patterns across sets of putative species. We tested three predictions. First, MMs would yield many more putative species than morphology because groundwater is a constraining environment allowing little morphological changes. Second, for species richness, MMs would increase the importance of spatial heterogeneity because cryptic species are more likely along physical barriers separating ecologically similar regions than along resource gradients promoting ecologically‐based divergent selection. Third, for range size, MMs would increase the importance of historical climates because of reduced and asymmetrical fragmentation of large morphological species ranges at northern latitudes. MMs yielded twice more putative species than morphology and decreased by 10‐fold the average species range size. Yet, MMs strengthened the mid‐latitude ridge of high species richness and the Rapoport effect of increasing range size at higher latitudes. Species richness predictors did not vary between morphology and MMs but the latter increased the proportion of variance in range size explained by historical climates. These findings demonstrate that our knowledge of groundwater biodiversity determinants is robust to overlooked cryptic species because the latter are homogeneously distributed along environmental gradients. Yet, our findings call for incorporating multiple species delimitation methods into the analysis of large‐scale biodiversity patterns across a range of taxa and ecosystems. [ABSTRACT FROM AUTHOR]

The hyporheic zone of rivers potentially acts as a dispersal corridor for groundwater organisms because it provides a spatially continuous interstitial habitat between isolated aquifers. Yet, the degree to which it can facilitate the movement of organisms has been hypothesized to vary in response to change in sediment regime, which determines channel morphology., In this study, we used microsatellite markers to test for a relationship between the genetic structure and diversity of the minute interstitial isopod Proasellus walteri and channel morphology along three nearby hyporheic corridors differing widely in their sediment regime. We predicted that genetic diversity would decrease and genetic structuring would increase as sediment supply-limited channels would become prominent features in the river corridor. The reason is that such channels have fewer and less suitable sedimentary habitats for migration because they lack large depositional bedforms such as gravel bars., Using genotypic data from seven microsatellite loci for a total of 713 individuals distributed among 25 demes, we found that demes had on average more alleles and were less differentiated in the river showing the most extensive alluvial deposits and shortest length of sediment supply-limited channels. Population clusters were also of greater size, reaching up to 30 km in length. The longitudinal pattern of genetic differentiation in this sediment-rich river was best explained by hydrologic distance and the longitudinal pattern of allelic richness was bell-shaped, as expected under a stepping-stone model with symmetrical migration., The length of sediment supply-channels was more important than hydrologic distance in explaining the longitudinal distribution of genetic differentiation in the two other corridors facing a sediment shortage. Allelic richness decreased monotonically upstream in the most sediment-poor river. This correlates with the expansion further downstream of sediment supply-limited channels in this river, which is likely to decrease animal movement and hence gene flow among demes., This study provides the first evidence that the degree to which the hyporheic zone facilitates the movement of groundwater organisms varies greatly among rivers of contrasted geomorphology. Extending the application of riverscape genetics across a range of interstitial taxa and geomorphic settings holds much promise for assessing the contribution of the hyporheic zone to the dispersal of groundwater organisms. [ABSTRACT FROM AUTHOR]

Aim Disentangling the roles of the interacting processes that shape species' ranges requires independent measurements of dispersal, physiological traits and habitat use. Multifaceted approaches of range determinants are, however, still rare, despite the widespread recognition that correlative modelling approaches alone are not sufficient to understand and predict species' distributions. Here, we combined genetic, distributional and physiological data to reveal the processes that cause the disjunct distribution of the groundwater isopod Proasellus valdensis in isolated Alpine mountains previously covered by Pleistocene glaciers. Location The Alps and Jura Mountains, France. Methods Phylogenetic/phylogeographical methods based on mitochondrial and nuclear genes were used to test for recent dispersal between mountains. A logistic regression on presence-absence data was performed to quantify variation in the probability of occurrence with temperature. Variation in survival and respiration over a range of temperatures was measured within four populations to test for a causal effect of temperature on species distribution. Results Despite the disjunct distribution, genetic analyses supported recent dispersal between mountains, as indicated by weak divergence among sequences of cytochrome c oxidase subunit I ( COI), a single haplotype network showing no spatial structuring, and a small proportion of molecular variance distributed between mountains. The probability of occurrence of P. valdensis decreased significantly with increasing temperature, although physiological experiments indicated that occurrence in warmer habitats was probably restricted by thermally dependent biotic interactions rather than by temperature itself. All populations maintained a high survival rate over a wide range of temperatures (3-15 °C), with a weak but detectable tendency for local adaptation. Main conclusions Combining phylogeographical, physiological and habitat modelling methods reveals the interacting processes that drive range dynamics. A broad thermal tolerance helps P. valdensis to colonize vacant habitats during the onset of glacial melting, but range fragmentation and local adaptation, leading to thermal niche narrowing, proceed during warmer interglacials as biotic interactions progressively intensify. [ABSTRACT FROM AUTHOR]

A key challenge for biologists is to document and explain global patterns of diversification in a wide range of environments. Here, we explore patterns of continental-scale diversification in a groundwater species-rich clade, the superfamily Aselloidea (Pancrustacea: Isopoda). Our analyses supported a constant diversification rate during most of the course of Aselloidea evolution, until 4-15 Ma when diversification rates started to decrease. This constant accumulation of lineages challenges the view that groundwater species diversification in temperate regions might have been primarily driven by major changes in physical environment leading to the extinction of surface populations and subsequent synchronous isolation of multiple groundwater populations. Rather than acting synchronously over broad geographic regions, factors causing extinction of surface populations and subsequent reproductive isolation of groundwater populations may act in a local and asynchronous manner, thereby resulting in a constant speciation rate over time. Our phylogeny also revealed several cases of parapatric distributions among closely related surface-water and groundwater species suggesting that species diversification could also arise from a process of disruptive selection along the surface-subterranean environmental gradient. Our results call for re-evaluating the spatial scale and timing of factors causing diversification events in groundwater. [ABSTRACT FROM AUTHOR]

Effective population size (Ne) is one of the most important parameters in, ecology, evolutionary and conservation biology; however, few studies of Ne in surface freshwater organisms have been published to date. Even fewer studies have been carried out in groundwater organisms, although their evolution has long been considered to be particularly constrained by small Ne. In this study, we estimated the contemporary effective population size of the obligate groundwater isopod: Proasellus walteri (Chappuis, 1948). To this end, a genomic library was enriched for microsatellite motifs and sequenced using 454 GS-FLX technology. A total of 54,593 reads were assembled in 10,346 contigs or singlets, of which 245 contained candidate microsatellite sequences with suitable priming sites. Ninety-six loci were tested for amplification, polymorphism and multiplexing properties, of which seven were finally selected for Ne estimation. Linkage disequilibrium and approximate Bayesian computation methods revealed that Ne in this small interstitial groundwater isopod could reach large sizes (> 585 individuals). Our results suggest that environmental conditions in groundwater, while often referred to as extreme, are not necessarily associated with small Ne. [ABSTRACT FROM AUTHOR]