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

Summary Soil ecosystem represents the largest contributor to global nitrous oxide (N 2 O) production, which is regulated by a wide variety of microbial communities in multiple biological pathways. A mechanistic understanding of these N 2 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 N 2 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 N 2 O production, and application of the nitrification inhibitor 3,4‐dimethylpyrazole phosphate (DMPP) significantly reduced the N 2 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 N 2 O production, and was not impacted by the application of DMPP. Our results provide robust evidence that the nitrification inhibitor DMPP can inhibit the N 2 O production from nitrifier‐induced denitrification, a potential significant source of N 2 O production in agricultural soils.