Landscape transcriptomic analysis detects thermal stress responses and potential adaptive variation in wild brook trout (Salvelinus fontinalis) during successive heatwaves

Extreme weather events, such as heatwaves, are becoming more frequent and intense as a result of climate change. Critically, such extreme weather events can be more important drivers of extirpation and selection than changes in annual or seasonal averages and they pose a particularly large threat to poikilothermic organisms. In this study, we evaluated the transcriptomic response of a coldwater adapted fish species, the eastern brook trout (Salvelinus fontinalis), to two successive heatwaves during July and August 2022. We sampled brook trout at eight time points from four streams (N = 116 fish), sequenced mRNA from gill samples using TagSeq, and quantified expression levels of 32,670 unique transcripts. Multivariate analyses found that overall expression patterns in response to water temperature change were similar among streams. These analyses further detected groups of genes involved in immune response and oxygen carrier activity that were upregulated and downregulated respectively at higher water temperatures. We also detected 43 genes that were differentially expressed at different time points and followed the same expression pattern during the two heatwaves. Of these genes, 42 covaried with water temperature and most (27, 62.8 %) exhibited responses that varied by stream. Some of the differentially expressed genes, including heat shock proteins and cold-inducible RNA binding proteins, have been widely linked to temperature responses in experimental studies, whereas other genes we identified have functions that have not been well-studied in relationship to temperature or have unknown functions. This study shows the utility of landscape transcriptomic approaches to identify important biological processes governing wild organismal responses to short-term stressors. The results of this study can guide future investigations to identify phenotypic and genetic diversity that contribute to adaptive responses to heatwaves and improve predictions of how populations will respond to future climate change.