Saprotrophic fungi are the primary decomposers of plant litter in temperate forests, and their activity is critical for carbon (C) and nitrogen (N) cycling. Simulated atmospheric N deposition is associated with reduced fungal biomass, shifts in fungal community structure, slowed litter decay, and soil C accumulation. Although rarely studied, N deposition may also result in novel selective pressures on fungi, affecting evolutionary trajectories. To directly test if long-term N enrichment reshapes fungal behaviors, we isolated decomposer fungi from a long-term (28 year) N addition experiment and used a common garden approach to compare growth rates and decay abilities of isolates from control and N amended plots. Both growth and decay were significantly altered by long-term exposure to N enrichment. Changes in growth rates were idiosyncratic, but litter decay by N isolates was generally lower compared to control isolates of the same species, a response not readily reversed when N isolates were grown in control (low N) environments. Changes in fungal behaviors accompany and perhaps drive previously observed N-induced shifts in fungal diversity, community composition, and litter decay dynamics.