Transcriptome profiling reveals the strategy of thermal tolerance enhancement caused by heat-hardening in Mytilus coruscus

The thick-shell mussel Mytilus coruscus serves as a common sessile intertidal species and holds economic significance as an aquatic organism. M. coruscus often endure higher temperatures than their ideal range during consecutive low tides in the spring. This exposure to elevated temperatures provides them with a thermal tolerance boost, enabling them to adapt to high-temperature events caused by extreme low tides and adverse weather conditions. This phenomenon is referred to as heat-hardening. Some related studies showed the phenomenon of heat-hardening in sessile intertidal species but not reported at the mechanism level based on transcriptome so far. In this study, physiological experiments, gene family identification and transcriptome sequencing were performed to confirm the thermotolerance enhancement based on heat-hardening and explore the mechanism in M. coruscus. A total of 2935 DEGs were identified and the results of the KEGG enrichment showed that seven heat-hardening relative pathways were enriched, including Toll-like receptor signal pathway, Arachidonic acid metabolism, and others. Then, 24 HSP70 members and 36 CYP2 members, were identified, and the up-regulated members are correlated with increasing thermotolerance. Finally, we concluded that the heat-hardening M. coruscus have a better thermotolerance because of the capability of maintaining the integrity and the phenomenon of vasodilation of the gill under thermal stress. Further, the physiological experiments yielded the same conclusions. Overall, this study confirms the thermotolerance enhancement caused by heat-hardening and reveals the survival strategy in M. coruscus. In addition, the conclusion provides a new reference for studying the intertidal species' heat resistance mechanisms to combat extreme heat events and the strategies for dealing with extreme weather in aquaculture under the global warming trend.