Reduction of N2O emission by biochar and/or 3,4-dimethylpyrazole phosphate (DMPP) is closely linked to soil ammonia oxidizing bacteria and nosZI-N2O reducer populations

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.