A genotype-phenotype association approach to reveal thermal adaptation in Daphnia galeata.

Altering thermal environments impose strong selection pressures on organisms, whose local persistence depends on adaptive phenotypic plastic and genetic responses. Thus far, adaptive change is monitored using phenotypic shifts or molecular markers, although inevitable obstacles are inherent in both methods. In order to circumvent these, it is necessary to find a causal link between adaptive alleles and fitness. Combining both approaches by linking genetic analyses and life-history measurements, a potential genotype-phenotype relationship can be assessed and adaptation at the molecular level demonstrated. For our study, clonal lineages of the freshwater keystone species D. galeata from seven different populations distributed along a latitudinal gradient across Europe were tested for local thermal adaptation in common garden experiments. Fitness-related life-history responses were quantified under different thermal regimes and experimental clones were genotyped at three candidate gene marker loci to investigate a potential genotype-phenotype association. The analyses of the life-history data showed a significant temperature effect on several fitness-related life-history traits recorded in our experiments. However, we could not detect evidence for a direct association at neither candidate gene locus between genotypes and life-history traits. The observed phenotypic shifts might therefore not be based on the tested marker loci EA, M and TF, or in general not coding sequence-based and thus rather reveal phenotypic plasticity in response to thermal variation. Nonetheless, we revealed significant genotype by environment (GxE) interactions at all tested loci, potentially reflecting a contribution of marker loci to certain life-history trait values and contribution of multiple genetic loci to phenotypic traits.

Phenotypic plasticity in life-history traits of Daphnia galeata in response to temperature - a comparison across clonal lineages separated in time.

Climatic changes are projected to result in rapid adaptive events with considerable phenotypic shifts. In order to reconstruct the impact of increased mean water temperatures during past decades and to reveal possible thermal micro-evolution, we applied a resurrection ecology approach using dormant eggs of the freshwater keystone species Daphnia galeata. To this end, we compared the adaptive response of D. galeata clones from Lake Constance of two different time periods, 1965-1974 ("historical") versus 2000-2009 ("recent"), to experimentally increased temperature regimes. In order to distinguish between genetic versus environmentally induced effects, we performed a common garden experiment in a flow-through system and measured variation in life-history traits. Experimental thermal regimes were chosen according to natural temperature conditions during the reproductive period of D. galeata in Central European lakes, with one additional temperature regime exceeding the currently observable maximum (+2°C). Increased water temperatures were shown to significantly affect measured life-history traits, and significant "temperature × clonal age" interactions were revealed. Compared to historical clones, recent clonal lineages exhibited a shorter time to first reproduction and a higher survival rate, which may suggest temperature-driven micro-evolution over time but does not allow an explicit conclusion on the adaptive nature of such responses.