Over the past decade an increasing amount of research has sought to understand how the diversity of species in an ecosystem can influence fluxes of biologically important materials, such as the decomposition of organic matter and recycling of nutrients. Generalities among studies have remained elusive, perhaps because experimental manipulations have been performed at relatively small spatial scales where site-specific variation generates patterns that appear idiosyncratic. One approach for seeking generality is to perform parallel experiments at different sites using an identical species pool. Here we report results from a study where we manipulated the diversity of leaf litter from the same six dominant tree species in the litter layer of three forested ecosystems. These ecosystems spanned a 300 km latitudinal transect in Wisconsin, USA, and were characterized by a large gradient in temperature and moisture, and thus, rates of decomposition. After allowing combinations of one, two, four, and six species of leaf litter to decompose for 1 year, we found that increasing leaf litter richness led to slower rates of decomposition and higher fractions of nitrogen lost from litter. Across all sites, climate and initial litter chemistry explained more of the variation in decomposition rates than did litter richness. Effects of leaf litter diversity were non-additive, meaning they were greater than expected from the impacts of individual species, and appeared to be strongly influenced by the presence/absence of just 1-2 species (Tilia americana and Acer saccharum). The rate of decomposition of these two species was highly site-specific, which led to strong negative effects of litter richness only being observed at the southernmost sites where T. americana and A. saccharum decomposed more quickly. In contrast, litter diversity increased nitrogen loss at the northernmost sites where decomposition of T. americana was notably slowed. Our study shows that species diversity affected at least one of the two litter processes at each site along this 300-km gradient, but the exact nature of these effects were spatially variable because the performance of individual species changed across the heterogeneous landscape.