||Artificial light at night (ALAN) has been recently recognized as a threat for aquatic systems, but a comprehensive knowledge of its effects is still lacking. A fundamental question is whether and how ALAN might affect temporal variability of communities, thus undermining the stability of mature assemblages or influencing the colonization process. Here we investigated the role of ALAN on temporal variability of total biomass and maximum photosynthetic efficiency of marine autotrophic biofilms colonizing Mediterranean high-shore rock surfaces while controlling for density of their main grazers. Results showed stability in total biomass, but an increase in maximum photosynthetic efficiency from unlit to lit conditions, which suggested a temporal change in composition and/or abundance of different taxa within mature assemblages. The effect was weaker during the colonization process; in this case, density of grazers acted in the opposite direction of ALAN. We suggest that the addition of light at times when it would not be naturally present may affect the temporal variability of a variety of functioning in aquatic systems, depending on species-specific sensitivities to ALAN within microbial assemblages and/or indirect effects mediated by their consumers. We highlight to further investigate the role of this emergent topic in aquatic ecology.