While plant litter is known to regulate soil moisture, little is known about the extent to which litter impacts moisture over and above the physical environment (i.e., ecosite) throughout the growing season, particularly in cool-temperate grasslands where moisture is considered less limiting for plant growth. In this study, we examined the relative impact of litter and ecosite on growing season soil moisture in a northern rough fescue (Festuca hallii) grassland. We also examined the relationship between litter and plant biomass throughout the growing season, including linkages between litter, plant growth, and the effects of litter on microclimate. During May, only ecosite was found to be associated with soil moisture, with a similar finding for plant biomass. Litter became important in maintaining greater soil moisture in June and July, however, likely through its corresponding negative impact on soil temperature and associated evaporation. In general, litter had a stronger and more consistent influence on soil moisture than ecosite. Finally, litter had a positive relationship with above-ground biomass, but only during June and July, the same months when litter exhibited the strongest relationship with soil moisture. Litter therefore appears to promote mid-season plant growth in these temperate grasslands, presumably through its ability to reduce evaporation and maintain greater soil water during seasonal moisture limitations.