1. Temperature is one of the primary environmental drivers of the distribution of species, and particularly high temperatures challenge physiological processes by disruption of cellular homoeostasis. This exerts selection on organisms to maintain cellular homoeostasis by adaptive physiological and/or behavioural responses. The social spider Stegodyphus dumicola occurs across several climate zones inSouthern Africa, and populations experience high and variable temperatures, suggesting a wide temperature niche, or alternatively that populations respond with plastic or locally adapted responses to temperature. Using a common garden design, we investigated complementary adaptive heat responses (behavioural thermoregulation and cuticle wax composition) in individuals from warmer and cooler locations.3. The spiders exhibited higher temperature tolerance than most ectotherms (CTmax almost 49℃), with the individuals from warmer locations showing the highest tolerance. Analyses of cuticle wax revealed chemical compositions consistent with a higher melting temperature (e.g. increased chain length and lower occurrence of branching) and therefore improved waterproofing in spiders originating from warmer locations and acclimated at a higher temperature, as expected if local temperature drives changes in the cuticle composition to improve waterproofing.4. The spiders exhibited a clear behavioural escape response from increasing temperature, with individuals from warmer locations and kept at higher acclimation temperature showing a lower threshold temperature at which this behaviour was triggered, suggesting that this threshold is under natural selection. Our study provides evidence of both local adaptation and phenotypic plasticity inphysiological and behavioural traits relating to temperature tolerance. Population differences in trait expression suggest local adaptation to different thermal environments, and individuals plastically adjust cuticle wax composition and cooling behaviour in response to temperature changes. 6. The Bogert effect predicts that behavioural thermoregulation may relax selection for physiological adaptations. Our study instead suggests that synergistic effects | 2729 Functional Ecology MALMOS et AL.

The environment fluctuates at a range of timescales and in space across species ranges. If environmental changes occur over periods that are many multiples of species generation times, or if there are restrictions on gene flow between locations, organisms can evolve naturally selected adaptations to this variation (Charlesworth et al., 2017). Additionally, and even in the absence of local

While metabarcoding of plant DNA from their environment is an exciting method that can supplement inventorying of live plant species, the accuracy and specificity has yet to be fully assessed over complex continuous landscapes. In this work, we evaluate plant community profiles produced via metabarcoding of soil by comparing them to a morphological survey. We assessed plant communities by metabarcoding of soil DNA in 130 sites along ecological gradients (nutrients, succession, moisture) in Denmark using chloroplast trnL region (10–143 bp) primer set and compared the resulting communities to communities produced with a longer nuclear ITS2 region (~216 bp) and a morphological survey. We found that the community variation observed within the morphological survey was well represented by molecular surveys, with significant correlation with both community composition and richness using both primer sets. While the majority of the ITS2 sequences could be assigned to species (over 80%), we had less success with the trnL sequences (70%), which was only possible after restricting the reference database to local species. We conclude that the community profiles produced by metabarcoding can be highly effective in performing large‐scale macroecological studies. However, the discovery rates and taxonomic assignments produced via metabarcoding remained inferior to morphological surveys, but manual curation of databases improves the specificity of assignments made by the trnL primers, and improves the accuracy of the assignments made with the ITS2 primers. Finally, we suggest that a greater percentage of named diversity would be recovered by increasing soil sampling with the use of additional universal primer sets.

Animals experience climatic variation in their natural habitats, which may lead to variation in phenotypic responses among populations through local adaptation or phenotypic plasticity. In ectotherm arthropods, the expression of thermoprotective metabolites such as free amino acids, sugars, and polyols, in response to temperature stress, may facilitate temperature tolerance by regulating cellular homeostasis. If populations experience differences in temperatures, individuals may exhibit population-specific metabolite profiles through differential accumulation of metabolites that facilitate thermal tolerance. Such thermoprotective metabolites may originate from the animals themselves or from their associated microbiome, and hence microbial symbionts may contribute to shape the thermal niche of their host. The social spider Stegodyphus dumicola has extremely low genetic diversity, yet it occupies a relatively broad temperature range occurring across multiple climate zones in Southern Africa. We investigated whether the metabolome, including thermoprotective metabolites, differs between populations, and whether population genetic structure or the spider microbiome may explain potential differences. To address these questions, we assessed metabolite profiles, phylogenetic relationships, and microbiomes in three natural populations along a temperature gradient. The spider microbiomes in three genetically distinct populations of S. dumicola showed no significant population-specific pattern, and none of its dominating genera (Borrelia, Diplorickettsia, and Mycoplasma) are known to facilitate thermal tolerance in hosts. These results do not support a role of the microbiome in shaping the thermal niche of S. dumicola. Metabolite profiles of the three spider populations were significantly different. The variation was driven by multiple metabolites that can be linked to temperature stress (e.g., lactate, succinate, or xanthine) and thermal tolerance (e.g., polyols, trehalose, or glycerol): these metabolites had higher relative abundance in spiders from the hottest geographic region. These distinct metabolite profiles are consistent with a potential role of the metabolome in temperature response.