Nitrifier‐induced denitrification is an important source of soil nitrous oxide and can be inhibited by a nitrification inhibitor 3,4‐dimethylpyrazole phosphate

Soil ecosystem represents the largest contributor to global nitrous oxide (N₂ O) production, which is regulated by a wide variety of microbial communities in multiple biological pathways. A mechanistic understanding of these N₂ O production biological pathways in complex soil environment is essential for improving model performance and developing innovative mitigation strategies. Here, combined approaches of the ¹⁵ N-¹⁸ O labelling technique, transcriptome analysis, and Illumina MiSeq sequencing were used to identify the relative contributions of four N₂ O pathways including nitrification, nitrifier-induced denitrification (nitrifier denitrification and nitrification-coupled denitrification) and heterotrophic denitrification in six soils (alkaline vs. acid soils). In alkaline soils, nitrification and nitrifier-induced denitrification were the dominant pathways of N₂ O production, and application of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) significantly reduced the N₂ O production from these pathways; this is probably due to the observed reduction in the expression of the amoA gene in ammonia-oxidizing bacteria (AOB) in the DMPP-amended treatments. In acid soils, however, heterotrophic denitrification was the main source for N₂ O production, and was not impacted by the application of DMPP. Our results provide robust evidence that the nitrification inhibitor DMPP can inhibit the N₂ O production from nitrifier-induced denitrification, a potential significant source of N₂ O production in agricultural soils.