Litter decomposition is an ecological process of key importance for forest headwater stream functioning, with repercussions for the global carbon cycle. The process is directly and indirectly mediated by microbial decomposers, mostly aquatic hyphomycetes, and influenced by environmental and biological factors such as water temperature and litter quality. These two factors are forecasted to change globally within the next few decades, in ways that may have contrasting effects on microbial-induced litter decomposition: while warming is expected to enhance microbial performance, the reduction in litter quality due to increased atmospheric carbon dioxide and community composition alteration may have the opposite outcome. We explored this issue through a microcosm experiment focused on early microbial-mediated litter decomposition under stream oligotrophic conditions, by simultaneously manipulating water temperature (10 °C and 15 °C) and litter quality (12 broadleaf plant species classified into 4 categories based on initial concentrations of nitrogen and tannins). We assessed potential changes in microbial-mediated litter decomposition and the performance of fungal decomposers (i.e., microbial respiration, biomass accrual, and sporulation rate) and species richness. We found stronger effects of litter quality, which enhanced the performance of microbial decomposers and decomposition rates, than temperature, which barely influenced any of the studied variables. Our results suggest that poorer litter quality associated with global change will have a major repercussion on stream ecosystem functioning.