Ex-situ restoration of the Mediterranean forest-forming macroalga Ericaria amentacea: Optimizing growth in culture may not be the key to growth in the field

diverse marine forest communities on exposed Mediterranean coasts, have spurred restoration efforts focused on sustainable ex-situ techniques. The need to balance the costs of culture maintenance and the susceptibility of early life stages to stressors in the native habitat, including rapid, often extreme shifts in temperature, hydrodynamics and nutrient availability, have driven current efforts to create a culture environment that primes seedlings for outplant, increasing their resilience rather than maximizing growth. We tested the effects of 1) higher culture temperature (25 ◦C) combined with wave simulation and 2) reduced nutrient loads (10% of standard protocol) with wave simulation on post-culture and post-outplant outcomes relative to optimal growth conditions in established protocols (20 ◦C, no waves, high-nutrient culture medium). While increased temperature and water motion negatively affected seedling growth in culture, and higher nutrients caused oxidative stress likely associated with enhanced epiphyte overgrowth, these effects were not clearly translated into patterns of long-term growth in the field. Instead, survival in the initial days post-outplant appeared to be the bottleneck for restoration potential, where substrates with persisting seedlings at one month were generally found with flourishing juveniles at four months. Larger clumps of seedlings, in turn, were strongly associated with both initial survival and future growth. These results underscore the importance of the zygote settlement phase to establish high seedling densities, which may be optimized by phenological monitoring of the donor population. They also suggest that less-controlled, more environmentally-realistic culture conditions involving the introduction of mild stress may enhance the survival of early life stages of E. amentacea during the transition to the native environment, providing a means to simultaneously reduce human resource costs in culture and move toward scaling up.