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Isotopic signals in an agricultural watershed suggest denitrification is locally intensive in riparian areas but extensive in upland soilsAbstract Nitrogen loss from cultivated soils threatens the economic and environmental sustainability of agriculture. Nitrate (NO 3 − ) derived from nitrification of nitrogen fertilizer and ammonified soil organic nitrogen may be lost from soils via denitrification, producing dinitrogen gas (N 2 ) or the greenhouse gas nitrous oxide (N 2 O). Nitrate that accumulates in soils is also subject to leaching loss, which can degrade water quality and make NO 3 − available for downstream denitrification. Here we use patterns in the isotopic composition of NO 3 − observed from 2012 to 2017 to characterize N loss to denitrification within soils, groundwater, and stream riparian corridors of a non-irrigated agroecosystem in the northern Great Plains (Judith River Watershed, Montana, USA). We find evidence for denitrification across these domains, expressed as a positive linear relationship between δ 15 N and δ 18 O values of NO 3 − , as well as increasing δ 15 N values with decreasing NO 3 − concentration. In soils, isotopic evidence of denitrification was present during fallow periods (no crop growing), despite net accumulation of NO 3 − from the nitrification of ammonified soil organic nitrogen. We combine previous results for soil NO 3more »Free, publicly-accessible full text available March 1, 2023