Contribution of Ammonium-Induced Nitrifier Denitrification to N2O in Paddy Fields

Paddy fields are one of the most important sources of nitrous oxide (N₂O), but biogeochemical N₂O production mechanisms in the soil profile remain unclear. Our study used incubation, dual-isotope (¹⁵N-¹⁸O) labeling methods, and molecular techniques to elucidate N₂O production characteristics and mechanisms in the soil profile (0-60 cm) during summer fallow, rice cropping, and winter fallow periods. The results pointed out that biotic processes dominated N₂O production (72.2-100%) and N₂O from the tillage layer accounted for 91.0-98.5% of total N₂O in the soil profile. Heterotrophic denitrification (HD) was the main process generating N₂O, contributing between 53.4 and 96.6%, the remainder being due to ammonia oxidation pathways, which was further confirmed by metagenomics and quantitative polymerase chain reaction (qPCR) assays. Nitrifier denitrification (ND) was an important N₂O production source, contributing 0-46.6% of total N₂O production, which showed similar trends with N₂O emissions. Among physicochemical and biological factors, ammonium content and the ratio of total organic matter to nitrate were the main driving factors affecting the contribution ratios of the ammonia oxidation pathways and HD pathway, respectively. Moisture content and pH affect norC-carrying Spirochetes and thus the N₂O production rate. These findings confirm the importance of ND to N₂O production and help to elucidate the impact of anthropogenic activities, including tillage, fertilization, and irrigation, on N₂O production.