In the Mediterranean Sea, brown macroalgae represent the dominant species in intertidal and subtidal habitats. Despite conservation efforts, these canopy-forming species showed a dramatic decline, highlighting the urge for active intervention to regenerate self-sustaining populations. For this reason, the restoration of macroalgae forests through transplantation has been recognized as a promising approach. However, the potential stress caused by the handling of thalli has never been assessed. Here, we used a manipulative approach to assess the transplant-induced stress in the Mediterranean Ericaria amentacea, through the analysis of biochemical proxies, i.e., phenolic compounds, lipids, and fatty acids in both transplanted and natural macroalgae over time. The results showed that seasonal environmental variability had an important effect on the biochemical composition of macroalgae, suggesting the occurrence of acclimation responses to summer increased temperature and light irradiance. Transplant-induced stress appears to have only amplified the biochemical response, probably due to increased sensitivity of the macroalgae already subjected to mechanical and osmotic stress (e.g., handling, wounding, desiccation). The ability of E. amentacea to cope with both environmental and transplant-induced stress highlights the high plasticity of the species studied, as well as the suitability of transplantation of adult thalli to restore E. amentacea beds.