Thermal limits along tropical elevational gradients: Poison frog tadpoles show plasticity but maintain divergence across elevation.

Temperature is arguably one of the most critical environmental factors impacting organisms at molecular, organismal, and ecological levels. Temperature variation across elevation may cause divergent selection in physiological critical thermal limits (CTMAX and CTMIN). Generally, high elevation populations are predicted to withstand lower environmental temperatures than low elevation populations. Organisms can also exhibit phenotypic plasticity when temperature varies, although theory and empirical evidence suggest that tropical ectotherms have relatively limited ability to acclimate. To study the effect of temperature variation along elevational transects on thermal limits, we measured CTMAX and CTMIN of 934 tadpoles of a poison frog species, Epipedobates anthonyi, along two elevational gradients (200-1700 m asl) in southwestern Ecuador to investigate their thermal tolerance across elevation. We also tested if tadpoles could plastically shift their critical thermal limits in response to exposure to different temperatures representing the range of temperatures they experience in nature (20 °C, 24 °C, and 28 °C). Overall, we found that CTMAX did not change across elevation. In contrast, CTMIN was lower at higher elevations, suggesting that elevational variation in temperature influences this thermal trait. Moreover, all populations shifted their CTMAX and CTMIN according to treatment temperatures, demonstrating an acclimation response. Overall, trends in CTMIN among high, mid, and low elevation populations were maintained despite plastic responses to treatment temperature. These results demonstrate that, for tadpoles of E. anthonyi across tropical elevational gradients, temperature acts as a selective force for CTMIN, even when populations show acclimation abilities in both, CTMAX and CTMIN. Our findings advance our understanding on how environmental variation affects organisms' evolutionary trajectories and their abilities to persist in a changing climate in a tropical biodiversity hotspot.

Contrasting environmental drivers of genetic and phenotypic divergence in an Andean poison frog (Epipedobates anthonyi).

Phenotypic and genetic divergence are shaped by the homogenizing effects of gene flow and the differentiating processes of genetic drift and local adaptation. Herein, we examined the mechanisms that underlie phenotypic (size and color) and genetic divergence in 35 populations (535 individuals) of the poison frog Epipedobates anthonyi along four elevational gradients (0-1800 m asl) in the Ecuadorian Andes. We found phenotypic divergence in size and color despite relatively low genetic divergence at neutral microsatellite loci. Genetic and phenotypic divergence were both explained by landscape resistance between sites (isolation-by-resistance, IBR), likely due to a cold and dry mountain ridge between the northern and southern elevational transects that limits dispersal and separates two color morphs. Moreover, environmental differences among sites also explained genetic and phenotypic divergence, suggesting isolation-by-environment (IBE). When northern and southern transects were analyzed separately, genetic divergence was predicted either by distance (isolation-by-distance, IBD; northern) or environmental resistance between sites (IBR; southern). In contrast, phenotypic divergence was primarily explained by environmental differences among sites, supporting the IBE hypothesis. These results indicate that although distance and geographic barriers are important drivers of population divergence, environmental variation has a two-fold effect on population divergence. On the one hand, landscape resistance between sites reduces gene flow (IBR), while on the other hand, environmental differences among sites exert divergent selective pressures on phenotypic traits (IBE). Our work highlights the importance of studying both genetic and phenotypic divergence to better understand the processes of population divergence and speciation along ecological gradients.

Isolation and characterization of 12 microsatellite loci in Epipedobates anthonyi (Amphibia: Anura: Dendrobatidae) for population genetic analysis.

Tropical anurans are among the most diverse and vulnerable organisms on Earth, yet the evolutionary mechanisms behind their diversity remain relatively unexplored. Epipedobates anthonyi is a poison frog that inhabits southern Ecuador and northern Peru along a broad elevational range (0-1800 m). Throughout its range, this species exhibits variation in phenotypic traits, such as color, advertisement calls, and alkaloid composition. The aim of this study is to isolate and characterize microsatellite loci to investigate patterns of genetic variation within the species. Using a next-generation sequencing approach to screen an enriched genomic library, we report twelve polymorphic microsatellite loci. The number of alleles per locus ranged from 7 to 15 per population. For the two populations tested, mean observed heterozygosity was 0.69 and 0.79, and mean expected heterozigosity was 0.84 and 0.85 respectively. Only locus EAN002 showed significant departure of HWE in both populations. None of the loci showed consistent null alleles in both populations. Also, no evidence of linkage disequilibrium was found across loci. In this paper, we report for the first time 12 microsatellite loci for E. anthonyi. These markers will be used to further elucidate evolutionary mechanisms underlying genetic and phenotypic variation across the species' range.

Regional variation in drivers of connectivity for two frog species (Rana pretiosa and R. luteiventris) from the U.S. Pacific Northwest.

Comparative landscape genetics has uncovered high levels of variability in which landscape factors affect connectivity among species and regions. However, the relative importance of species traits versus environmental variation for predicting landscape patterns of connectivity is unresolved. We provide evidence from a landscape genetics study of two sister taxa of frogs, the Oregon spotted frog (Rana pretiosa) and the Columbia spotted frog (Rana luteiventris) in Oregon and Idaho, USA. Rana pretiosa is relatively more dependent on moisture for dispersal than R. luteiventris, so if species traits influence connectivity, we predicted that connectivity among R. pretiosa populations would be more positively associated with moisture than R. luteiventris. However, if environmental differences are important drivers of gene flow, we predicted that connectivity would be more positively related to moisture in arid regions. We tested these predictions using eight microsatellite loci and gravity models in two R. pretiosa regions and four R. luteiventris regions (n = 1,168 frogs). In R. pretiosa, but not R. luteiventris, connectivity was positively related to mean annual precipitation, supporting our first prediction. In contrast, connectivity was not more positively related to moisture in more arid regions. Various temperature metrics were important predictors for both species and in all regions, but the directionality of their effects varied. Therefore, the pattern of variation in drivers of connectivity was consistent with predictions based on species traits rather than on environmental variation.