Ectomycorrhizal (EcM)-mediated nitrogen (N) acquisition is one main strategy used by terrestrial plants to facilitate growth. Measurements of natural abundance nitrogen isotope ratios (denoted as δ(15) N relative to a standard) increasingly serve as integrative proxies for mycorrhiza-mediated N acquisition due to biological fractionation processes that alter (15) N:(14) N ratios. Current understanding of these processes is based on studies from high-latitude ecosystems where plant productivity is largely limited by N availability. Much less is known about the cause and utility of ecosystem δ(15) N patterns in the tropics. Using structural equation models, model selection and isotope mass balance we assessed relationships among co-occurring soil, mycorrhizal plants and fungal N pools measured from 40 high- and 9 low-latitude ecosystems. At low latitudes (15) N-enrichment caused ecosystem components to significantly deviate from those in higher latitudes. Collectively, δ(15) N patterns suggested reduced N-dependency and unique sources of EcM (15) N-enrichment under conditions of high N availability typical of the tropics. Understanding the role of mycorrhizae in global N cycles will require reevaluation of high-latitude perspectives on fractionation sources that structure ecosystem δ(15) N patterns, as well as better integration of EcM function with biogeochemical theories pertaining to climate-nutrient cycling relationships.