[en] Highlights: • Synthetic inhibitors mixed with urea and manure significantly lowed N₂O fluxes. • The reduction of N₂O emission closely linked with AOB community, but not AOA. • Soil AOB abundance were markedly inhibited by the nitrapyrin added treatments. • The changes in AOB community ascribed to edaphic factors of pH, NH₄⁺ and SOC. • Elevated temperature notably increased AOA, but decreased AOB abundance. Synthetic inhibitors and organic amendment have been proposed for mitigating greenhouse gas N₂O emissions. However, their combined effect on the N₂O emissions and ammonia-oxidizer (ammonia-oxidizing bacteria and archaea, AOB and AOA) communities remains unclear in calcareous soils under climate warming. We conducted two incubation experiments (25 and 35 °C) to examine how N₂O emissions and AOA and AOB communities responded to organic amendment (urea plus cattle manure, UCM), and in combination with urease (N-(n-butyl) thiophosphoric triamide, NBPT) and nitrification inhibitor (nitrapyrin). The treatments of UCM + nitrapyrin and UCM + nitrapyrin + NBPT significantly lowered total N₂O emissions by average 64.5 and 71.05% at 25 and 35 °C, respectively, compared with UCM treatment. AOB gene abundance and α-diversity (Chao1 and Shannon indices) were significantly increased by the application of urea and manure (P < 0.05). However, relative to UCM treatment, nitrapyrin addition treatments decreased AOB gene abundance and Chao 1 index by average 115.4 and 30.4% at 25 and 35 °C, respectively. PCA analysis showed that UCM or UCM plus nitrapyrin notably shifted AOB structure at both temperatures. However, fertilization had little effects on AOA community (P > 0.05). Potential nitrification rate (PNR) was greatly decreased by nitrapyrin addition, and PNR significantly positively correlated with AOB gene abundance (P = 0.0179 at 25 °C and P = 0.0029 at 35 °C) rather than AOA (P > 0.05). Structural equation model analysis showed that temperature directly increased AOA abundance but decrease AOB abundance, while fertilization indirectly influenced AOB community by altering soil NH₄⁺, pH and SOC. In conclusion, the combined application of organic amendment, NBPT and nitrapyrin significantly lowered N₂O emissions via reducing AOB community in calcareous soil even at high temperature. Our findings provide a solid theoretical basis in mitigating N₂O emissions from calcareous soil under climate warming.

[en] Highlights: • Applying biochar with DMPP could reduce more N₂O emissions than biochar alone. • Biochar could adsorb DMPP when applied together in soil. • The response pattern of diverse denitrifier clades to biochar and/or DMPP was different. • Dynamics of AOB and nosZI-N₂O reducers were positively and negatively correlated with N₂O emission rate, respectively. -- Abstract: Biochar has been demonstrated to reduce nitrous oxide (N₂O) emissions from soils, but its effect is highly soil-dependent. In particular, in soils with strong nitrification potential, biochar addition may increase N₂O emissions. Thus, in soils with strong nitrification potential, the combination of biochar with the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) may be more effective in reducing N₂O emissions than biochar alone. However, the combined use of biochar and DMPP on soil N₂O emissions is relatively unexplored, and underlying microbial mechanisms of how biochar and/or DMPP amendment affect N₂O emissions is still largely unknown. Here, a 30-day incubation experiment was established with four treatments: CK (control), BC (biochar), DMPP, and BD (biochar and DMPP), all at agronomically recommended rates, and N cycling assessed following addition of urea. Treatment of soil with BC, DMPP and BD reduced N₂O emissions (compared with urea alone) by 59.1%, 95.5% and 74.1%, respectively. Quantification of N cycling genes (amoA, nirS, nirK, and nosZ) indicated that biochar stimulated growth of ammonia oxidizing archaea (AOA) and bacteria (AOB), while DMPP alone inhibited the activity and growth of AOB. In the BD treatment, DMPP was absorbed onto biochar reducing its efficacy in inhibiting AOB growth. The response patterns of nirS/nirK nitrite-reducing denitrifiers to biochar and/or DMPP addition varied among clades. Notably, biochar and/or DMPP increased the abundance of nosZI and nosZII-N₂O reducers, but nosZI-clade taxa were more closely associated with reducing N₂O emission than nosZII taxa. Overall, our findings proved that the dynamics of AOB and nosZI-N₂O reducers resulting from the addition of biochar and/or DMPP played a key role in governing soil N₂O emissions.

[en] Highlights: • NirS and nosZ communities were affected by organic but not chemical fertilizer. • SOC, NO₃⁻, NH₄⁻ and pH were main drivers for the alteration of denitrifier community abundance. • The significant factors influencing soil N₂O emissions were the nirS community, SOC and nitrate • N₂O emission significantly decreased when biofertilizer was applied with chemical fertilizer. The effects of consecutive application of chemical fertilizer with or without organic fertilizer on soil N₂O emissions and denitrifying community structure in a drip-irrigated field were determined. The four fertilizer treatments were (i) unfertilized, (ii) chemical fertilizer, (iii) 60% chemical fertilizer plus cattle manure, and (iv) 60% chemical fertilizer plus biofertilizer. The treatments with organic amendments (i.e. cattle manure and biofertilizer) reduced cumulative N₂O emissions by 4.9–9.9%, reduced the N₂O emission factor by 1.3–42%, and increased denitrifying enzyme activities by 14.3–56.2%. The nirK gene copy numbers were greatest in soil which received only chemical fertilizer. In contrast, nirS- and nosZ-copy numbers were greatest in soil amended with chemical fertilizer plus biofertilizer. Chemical fertilizer application with or without organic fertilizer significantly changed the community structure of nirK-type denitrifiers relative to the unfertilized soil. In comparison, the nirS- and nosZ-type denitrifier genotypes varied in treatments receiving organic fertilizer but not chemical fertilizer alone. The changes in the denitrifier communities were closely associated with soil organic carbon (SOC), NO₃⁻, NH₄⁺, water holding capacity, and soil pH. Modeling indicated that N₂O emissions in this soil were primarily associated with the abundance of nirS type denitrifying bacteria, SOC, and NO₃⁻. Overall, our findings indicate that (i) the organic fertilizers increased denitrifying enzyme activity, increased denitrifying-bacteria gene copy numbers, but reduced N₂O emissions, and (ii) nirS- and nosZ-type denitrifiers were more sensitive than nirK-type denitrifiers to the organic fertilizers.