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.
Keywords: dual-isotope (15N−18O) labeling method; nitrous oxide production pathways; paddy soil; tillage layer.