DNA methylation plasticity drives copepod resilience to coastal high pCO₂ and cadmium pollution under...

The vast majority of coastal organisms have been facing multigenerational scenarios of fluctuatingly high pCO₂ and Cd pollution in their natural habitats. However, the adaptive capacity of these organisms to such combined stressors and the underlying mechanisms remain poorly understood. In this study, we conducted a multigenerational experiment (F1-F4) to investigate the adaptive responses of the marine copepod Tigriopus japonicus to combined fluctuatingly high pCO₂ and Cd exposure, along with the associated mechanisms. Our findings revealed that steady high pCO₂ aggravated Cd multigenerational toxicity, and it was more under fluctuating acidification. Notably, by the F4 generation, copepods potentially adapted to the combined stressors. Through transcriptomic and DNA methylation analyses of copepods from the F1 and F4 generations, we found that under combined exposure, F1 copepods likely reallocated more energy to counteract Cd toxicity; however, DNA hypermethylation inhibited Cd exclusion and detoxification/stress response pathways, ultimately compromising development and reproduction. In contrast, in the F4 generation, DNA hypomethylation enhanced processes such as cuticle repair program, compensatory mechanism (e.g., detoxification and immune response), and reproduction, consequently increasing the copepod's fitness. These findings reveal an epigenetic basis for phenotypic acclimatization, offering marine copepods a supplementary mechanism to cope with combined stressors.