Constitutive up-regulation of Turandot genes rather than changes in acclimation ability is associated with the evolutionary adaptation to temperature fluctuations in Drosophila simulans.

Most research on thermal adaptation of ectotherms is based on experiments performed at constant temperatures. However, for short-lived insects daily fluctuations of temperature could be an important environmental parameter involved in evolutionary adaptation to thermal heterogeneity. In this study we investigated the mechanisms underlying evolutionary adaptation to daily fluctuating temperatures. We studied replicated selection lines of Drosophila simulans evolved in a constant or a daily fluctuating thermal regime. Previous studies of these lines have shown clear acclimation benefits to heat tolerance induced by the fluctuating regime. First, we tested the existence of an evolved circadian controlled adjustment of heat resistance in selected flies. This was done by investigating the daily variation in time to heat knockdown in flies from both selection regimes when exposed to either a constant or a daily fluctuating thermal regime for a single generation. While daily variation in heat resistance was found, the results suggest that there was neither an evolved adaptive circadian controlled adjustment of heat resistance nor a continuous acclimation response induced by fluctuating temperatures in these lines. Second, in order to reveal functional candidates for adaptation to the fluctuating thermal regime, we investigated the global transcriptomic response to a high temperature exposure in flies from both regimes. We found that flies selected both in constant and fluctuating thermal regimes responded similarly to increasing temperature. However, we found that evolutionary adaptation to the fluctuating thermal regime led to transcriptional enrichment of the GO terms eggshell chorion assembly and cellular response to heat. The latter category was constituted by a constitutive up-regulation of four Turandot genes and not heat shock protein genes, suggesting that Turandot genes could play a prominent role for adaptation to daily fluctuating thermal conditions.