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Significance Nitrogen (N) losses from terrestrial ecosystems can occur as inert forms or heat-trapping greenhouse gases, and via nitrate (NO3 − ) leaching to drainage waters, which can contribute to eutrophication and anoxia in downstream ecosystems. Here, we use natural isotopes to demonstrate that microbial gaseous N production via denitrification is the dominant pathway of NO3 − removal from forest ecosystems, with gaseous N losses that are up to ∼60-fold higher than those based on traditional techniques. Denitrification becomes less efficient compared with NO3 − leaching in more N-polluted ecosystems, which has important implications for assessing the connections between terrestrial soils and downstream ecosystems under rising anthropogenic N deposition.

Microbial denitrification dominates nitrate losses from forest ecosystems