N₂O emission contributions by different pathways and associated microbial community dynamics in a typical calcareous...

Nitrous oxide, one of the powerful long-lived greenhouse gases, is emitted mainly through biological processes, especially from fertilized soil. It is critical to partition the contribution of different pathways to N₂O emissions and the relevant characteristics of microbial communities to identify the key N₂O processes. An microcosm was conducted to partition the N₂O emissions from different pathways, and the changes in soil mineral nitrogen and various nitrifiers (amoA bacteria and amoA archaea) and denitrifiers (nirS, nirK, and nosZ) were also determined using qPCR and high-throughput sequencing methods. Different gas inhibitor combinations (i.e., 0.06% acetylene, pure oxygen, 0.06% acetylene in pure oxygen, and pure helium) were used to partition the N₂O pathways. A 5% oxygen treatment, with and without acetylene, was also included so that the N₂O emissions could be measured under lower oxygen partial pressure. Results showed that ammonia-oxidation (AO) and successive nitrifier denitrification (NiD) were the main pathways contributing to N₂O emissions at the earlier period after ammonium sulfate application with the cumulative N₂O emissions accounting for 30.9% and 59.2% of the total N₂O emissions, respectively. The higher NiD N₂O contributions occurred when the soil nitrite concentration appeared higher, especially under the lower oxygen conditions. Higher N₂O emissions from AO and NiD were associated with the compositional proportion of some dominant AOB species. Denitrification contributed more N₂O (63.6%-69.3%) in the later period during incubation, coinciding with the following characteristics for denitrifiers: a) lower nosZ/(nirS + nirK) ratio, b) more diversity in nirS, and c) different proportions of some dominant species in nirK. Our results demonstrated that higher AO and successive NiD were the main N₂O emission pathways, suggesting that controlling the ammonium content and weakening the AO are critical in decreasing N₂O emissions.