Sources and migration similarly determine nitrate concentrations: Integrating isotopic, landscape, and biological approaches

Rapid land use change has significantly increased nitrate (NO3−) loading to rivers, leading to eutrophication, and posing water security problems. Determining the sources of NO3− to waters and the underlying influential factors is critical for effectively reducing pollution and better managing water resources. Here, we identified the sources and influencing mechanisms of NO3− in a mixed land-use watershed by integrating stable isotopes (δ15N-NO3− and δ18O-NO3−), molecular biology, water chemistry, and landscape metrics measurements. Weak transformation processes of NO3− were identified in the river, as evinced by water chemistry, isotopes, species compositions, and predicted microbial genes related to nitrogen metabolism. NO3− concentrations were primarily influenced by exogenous inputs (i.e., from soil nitrogen (NS), nitrogen fertilizer (NF), and manure & sewage (MS)). The proportions of NO3− sources seasonally varied. In the wet season, the source contributions followed the order of NS (38.6 %) > NF (31.4 %) > atmospheric deposition (ND, 16.2 %) > MS (13.8 %). In the dry season, the contributions were in the order of MS (39.2 %) > NS (29.2 %) > NF (29 %) > ND (2.6 %). Farmland and construction land were the original factors influencing the spatial distribution of NO3− in the wet and dry seasons, respectively, while slope, basin relief (HD), hypsometric integral (HI), and COHESION, HD were the primary indicators associated with NO3− transport in the wet and dry seasons, respectively. Additionally, spatial scale differences were observed for the effects of landscape structure on NO3− concentrations, with the greatest effect at the 1000-m buffer zone scale in the wet season and at the sub-basin scale in the dry season. This study overcomes the limitation of isotopes in identifying nitrate sources by combining multiple approaches and provides new research perspectives for the determination of nitrate sources and migration in other watersheds. [Display omitted] •A “source-flow-sink” perspective was used to study the sources, transport, and transformation processes of NO3−.•Integrating stable isotope, molecular biology techniques, and water chemistry to determine NO3− sources and influencing mechanisms in a mixed land use watershed.•Nitrogen transformation processes showed a weak effect on NO3−.•Soil nitrogen and manure & sewage were the largest contributors of NO3− in the wet and dry seasons, respectively.•Seasonal and spatial scale differences in the effects