Aim To test the role of mountains as barriers to gene flow in co‐distributed taxa with different life history traits. Location Sierra de Guadarrama, Central Spain. Methods We used larval genotypes of four amphibian species (Epidalea calamita, Hyla molleri, Pelophylax perezi and Pelobates cultripes) sampled on northern and southern slopes of Sierra de Guadarrama to describe genetic structure with FST, migration rates per generation, clustering algorithms and resistance by elevation surfaces. We also recorded individual displacement events as a proxy of dispersal potential during a seven‐year monitoring project based on capture–mark–recapture (CMR). Results All species travelled longer cumulative distances than those reported in the study area for P. cultripes (0.71 km). Individuals of E. calamita travelled up to 3.55 km, followed by H. molleri (2.84 km) and P. perezi (1.51 km). Pairwise FST estimates showed lower overall connectivity in P. cultripes. Average migration rates per generation were low in all species, with exceptions in same‐slope populations of H. molleri and P. cultripes. Clustering algorithms consistently recovered well‐differentiated population groups of P. cultripes in northern versus southern slopes, but widely admixed areas were observed in the other species, especially near mountain passes. Resistance by elevation surfaces showed a strong barrier effect of Sierra de Guadarrama in P. cultripes and suggested a potential role of topography in the genetic structure of E. calamita and H. molleri. Main conclusions Sierra de Guadarrama currently acts as a strong barrier to gene flow for P. cultripes and, to a lesser extent, for E. calamita, H. molleri and P. perezi. This differential effect can be partly explained by differences in life history traits, including dispersal potential. Our findings support the general role of the Central System as a key feature shaping population connectivity and genetic variation in amphibian communities.

Accurate characterization of genetic diversity is essential for understanding population demography, predicting future trends and implementing efficient conservation policies. For that purpose, molecular markers are routinely developed for nonmodel species, but key questions regarding sampling design, such as calculation of minimum sample sizes or the effect of relatives in the sample, are often neglected. We used accumulation curves and sibship analyses to explore how these 2 factors affect marker performance in the characterization of genetic diversity. We illustrate this approach with the analysis of an empirical dataset including newly optimized microsatellite sets for 3 Iberian amphibian species: Hyla molleri, Epidalea calamita, and Pelophylax perezi. We studied 17-21 populations per species (total n = 547, 652, and 516 individuals, respectively), including a reference locality in which the effect of sample size was explored using larger samples (77-96 individuals). As expected, FIS and tests for Hardy-Weinberg equilibrium and linkage disequilibrium were affected by the presence of full sibs, and most initially inferred disequilibria were no longer statistically significant when full siblings were removed from the sample. We estimated that to obtain reliable estimates, the minimum sample size (potentially including full sibs) was close to 20 for expected heterozygosity, and between 50 and 80 for allelic richness. Our pilot study based on a reference population provided a rigorous assessment of marker properties and the effects of sample size and presence of full sibs in the sample. These examples illustrate the advantages of this approach to produce robust and reliable results for downstream analyses.

The ratio of the effective number of breeders (N b) to the adult census size (N a), N b/N a, approximates the departure from the standard capacity of a population to maintain genetic diversity in one reproductive season. This information is relevant for assessing population status, understanding evolutionary processes operating at local scales, and unraveling how life‐history traits affect these processes. However, our knowledge on N b/N a ratios in nature is limited because estimation of both parameters is challenging. The sibship frequency (SF) method is adequate for reliable N b estimation because it is based on sibship and parentage reconstruction from genetic marker data, thereby providing demographic inferences that can be compared with field‐based information. In addition, capture–mark–recapture (CMR) robust design methods are well suited for N a estimation in seasonal‐breeding species. We used tadpole genotypes of three pond‐breeding amphibian species (Epidalea calamita, Hyla molleri, and Pelophylax perezi, n = 73–96 single‐cohort tadpoles/species genotyped at 15–17 microsatellite loci) and candidate parental genotypes (n = 94–300 adults/species) to estimate N b by the SF method. To assess the reliability of N b estimates, we compared sibship and parentage inferences with field‐based information and checked for the convergence of results in replicated subsampled analyses. Finally, we used CMR data from a 6‐year monitoring program to estimate annual N a in the three species and calculate the N b/N a ratio. Reliable ratios were obtained for E. calamita (N b/N a = 0.18–0.28) and P. perezi (0.5), but in H. molleri, N a could not be estimated and genetic information proved insufficient for reliable N b estimation. Integrative demographic studies taking full advantage of SF and CMR methods can provide accurate estimates of the N b/N a ratio in seasonal‐breeding species. Importantly, the SF method provides results that can be readily evaluated for reliability. This represents a good opportunity for obtaining robust demographic inferences with wide applications for evolutionary and conservation research.

Context Robust assessment of functional connectivity in amphibian population networks is essential to address their global decline. The potential of graph theory to characterize connectivity among amphibian populations has already been confirmed, but the movement data on which modelled graphs rely are often scarce and inaccurate. While probabilistic methods that account for intraspecific variability in dispersal better reflect the biological reality of functional connectivity, they must be informed by systematically recorded individual movement data, which are difficult to obtain for secretive taxa like amphibians. Objectives Our aim is to assess the applied potential of probabilistic graph theory to characterize overall connectivity across amphibian pondscapes using fine-scale capture-recapture data, and to inform conservation management based on the role of ponds on functional connectivity. Methods We monitored an amphibian community in a pondscape located in a Spanish “dehesa” for 2 years. Photoidentification was used to build capture histories for individuals of six species, from which dispersal kernels and population sizes were estimated to model probabilistic graphs. Results We obtained kernels of variable robustness for six species. Node importance for connectivity varied between species, but with common patterns such as shared road crossing areas and the presence of coincident interconnected pond clusters. Conclusions The combination of photoidentification, capture-recapture data and graph theory allowed us to characterize functional connectivity across the pondscape of study accounting for dispersal variability and identify areas where conservation actions could be most efficient. Our results highlight the need to account for interspecific differences in the study and management of amphibian pondscapes.

Aim: To reconstruct the historical biogeography of Hyla molleri, a tree frog endemic to the Eurosiberian and Mediterranean bioclimatic zones in the Iberian Peninsula.Location: Iberian Peninsula. Methods:We used molecular data (mtDNA and species-specific, polymorphic microsatellite loci) and species distribution modelling (SDM) from the Last Interglacial (LIG) to the present to characterize spatial patterns of genetic diversity in this species and assess their relationship with climatically favourable areas through time.Results: Genetic diversity is heterogeneously distributed across the range of H. molleri, with two main genetic reservoirs located in (a) central and southern Portugal and (b) a fragmented area encompassing mountainous areas in northern Spain.According to SDM, the Iberian Peninsula has experienced a progressive and continuous decrease in climatically favourable areas for H. molleri since the LIG, especially in southern and eastern Iberia, where the species is currently absent. However, we found no correlation between areas that have remained climatically favourable since the LIG and current genetic diversity.Main conclusions: Our results suggest that the demographic history of H. molleri since the Pleistocene has been characterized by relative stability, contrasting with the large-scale cycles of extinction-recolonization inferred for other more thermophilous, co-distributed amphibian species in Iberia. Accounting for discordant demographic responses to climatic changes across syntopic species provides new insights about the evolutionary history of amphibian communities in southern Europe.

Comparative studies of mortality in the wild are necessary to understand the evolution of aging; yet, ectothermic tetrapods are underrepresented in this comparative landscape, despite their suitability for testing evolutionary hypotheses. We present a study of aging rates and longevity across wild tetrapod ectotherms, using data from 107 populations (77 species) of nonavian reptiles and amphibians. We test hypotheses of how thermoregulatory mode, environmental temperature, protective phenotypes, and pace of life history contribute to demographic aging. Controlling for phylogeny and body size, ectotherms display a higher diversity of aging rates compared with endotherms and include phylogenetically widespread evidence of negligible aging. Protective phenotypes and life-history strategies further explain macroevolutionary patterns of aging. Analyzing ectothermic tetrapods in a comparative context enhances our understanding of the evolution of aging.

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Sex‐related differences in mortality are widespread in the animal kingdom. Although studies have shown that sex determination systems might drive lifespan evolution, sex chromosome influence on aging rates have not been investigated so far, likely due to an apparent lack of demographic data from clades including both XY (with heterogametic males) and ZW (heterogametic females) systems. Taking advantage of a unique collection of capture–recapture datasets in amphibians, a vertebrate group where XY and ZW systems have repeatedly evolved over the past 200 million years, we examined whether sex heterogamy can predict sex differences in aging rates and lifespans. We showed that the strength and direction of sex differences in aging rates (and not lifespan) differ between XY and ZW systems. Sex‐specific variation in aging rates was moderate within each system, but aging rates tended to be consistently higher in the heterogametic sex. This led to small but detectable effects of sex chromosome system on sex differences in aging rates in our models. Although preliminary, our results suggest that exposed recessive deleterious mutations on the X/Z chromosome (the “unguarded X/Z effect”) or repeat‐rich Y/W chromosome (the “toxic Y/W effect”) could accelerate aging in the heterogametic sex in some vertebrate clades.