Dual isotope and isotopomer measurements for the understanding of N2O production and consumption during denitrification in an arable soil

The aim of our research was to obtain information on the isotopic fingerprint of nitrous oxide (N₂O) associated with its production and consumption during denitrification. An arable soil was preincubated at high moisture content and subsequently amended with glucose (400 kg C ha^?1) and KNO₃ (80 kg N ha^?1) and kept at 85% water‐filled pore space. Twelve replicate samples of the soil were incubated for 13 days under a helium‐oxygen atmosphere, simultaneously measuring gas fluxes (N₂O, N₂ and CO₂) and isotope signatures (δ¹⁸O‐N₂O, δ¹⁵N^bulk‐N₂O, δ¹⁵N^α, δ¹⁵N^β and ¹⁵N site preference) of emitted N₂O. The maximum N₂O flux (6.9 ± 1.8 kg N ha^?1 day^?1) occurred 3 days after amendment application, followed by the maximum N₂ flux on day 4 (6.6 ± 3.0 kg N ha^?1 day^?1). The δ¹⁵N^bulk was initially ?34.4‰ and increased to +4.5‰ during the periods of maximum N₂ flux, demonstrating fractionation during N₂O reduction, and then decreased. The δ¹⁸O‐N₂O also increased, peaking with the maximum N₂ flux and remaining stable afterwards. The site preference (SP) decreased from the initial +7.5 to ?2.1‰ when the N₂O flux peaked, and then simultaneously increased with the appearance of the N₂ peak to +8.6‰ and remained stable thereafter, even when the O₂ supply was removed. We suggest that this results from a non‐homogenous distribution of NO in the soil, possibly linked to the KNO₃ amendments to the soil, causing the creation of several NO pools, which affected differently the isotopic signature of N₂O and the N₂O and N₂ fluxes during the various stages of the process. The N₂O isotopologue values reflected the temporal patterns observed in N₂O and N₂ fluxes. A concurrent increase in ¹⁵N site preference and δ¹⁸O of N₂O was found to be indicative of N₂O reduction to N₂.