Whole genome sequencing and RNA-seq evaluation allowed to detect Cd adaptation footprint in Chironomus riparius

Evolutionary adaptation and phenotypic plasticity are important processes on how organisms respond to pollutant exposure. We dissected here the contribution of both processes to increased tolerance in Chironomus riparius to cadmium (Cd) exposure in a multi-generation experiment and inferred the underlying genomic basis. We simulated environmentally realistic conditions by continuously increasing contaminant concentration in six replicates initiated with 1000 larvae each, three pre-exposed to Cd and three not exposed to Cd (no-Cd) over eight generations. We measured life-cycle traits, transcriptomic responses and genome-wide allele frequency changes from this evolve and resequencing (E&R) experiment. Overall, life cycle tests revealed little phenotypic adaptation to Cd exposure, but a slightly increase in survival in the first larval stage was observed. Population genomic analyses showed a strong genome-wide selective response in all replicates, highlighting two main biological functions involved in development and growth of the chironomids. Emphasizing that laboratory conditions continually exert selective pressure. However, the integration of the transcriptomic to the genomic data allowed to distinguish pathways specifically selected by the Cd exposure related to microtubules and organelles and cellular movement. Those pathways could be functionally related to an excretion of metals. Thus, our results indicate that genetic adaptation to Cd in C. riparius can happen within few generations under an environmentally relevant exposure scenario, but substantial phenotypic tolerance might take more time to arise. With our approach, we introduce an experimental setup to fill the existing gap in evolutionary ecotoxicology to investigate these early signs of genetic adaptation.