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Abstract Nitrous oxide (N₂O) is a climate-active gas with emissions predicted to increase due to agricultural intensification. Microbial reduction of N₂O to dinitrogen (N₂) is the major consumption process but microbial N₂O reduction under acidic conditions is considered negligible, albeit strongly acidic soils harbornosZgenes encoding N₂O reductase. Here, we study a co-culture derived from acidic tropical forest soil that reduces N₂O at pH 4.5. The co-culture exhibits bimodal growth with aSerratiasp. fermenting pyruvate followed by hydrogenotrophic N₂O reduction by aDesulfosporosinussp. Integrated omics and physiological characterization revealed interspecies nutritional interactions, with the pyruvate fermentingSerratiasp. supplying amino acids as essential growth factors to the N₂O-reducingDesulfosporosinussp. Thus, we demonstrate growth-linked N₂O reduction between pH 4.5 and 6, highlighting microbial N₂O reduction potential in acidic soils. 

Abstract Nitrous oxide (N2O), a greenhouse gas with ozone destruction potential, is mitigated by the microbial reduction to dinitrogen catalyzed by N2O reductase (NosZ). Bacteria with NosZ activity have been studied at circumneutral pH but the microbiology of low pH N2O reduction has remained elusive. Acidic (pH < 5) tropical forest soils were collected in the Luquillo Experimental Forest in Puerto Rico, and microcosms maintained with low (0.02 mM) and high (2 mM) N2O assessed N2O reduction at pH 4.5 and 7.3. All microcosms consumed N2O, with lag times of up to 7 months observed in microcosms with 2 mM N2O. Comparative metagenome analysis revealed that Rhodocyclaceae dominated in circumneutral microcosms under both N2O feeding regimes. At pH 4.5, Peptococcaceae dominated in high-N2O, and Hyphomicrobiaceae in low-N2O microcosms. Seventeen high-quality metagenome-assembled genomes (MAGs) recovered from the N2O-reducing microcosms harbored nos operons, with all eight MAGs derived from acidic microcosms carrying the Clade II type nosZ and lacking nitrite reductase genes (nirS/K). Five of the eight MAGs recovered from pH 4.5 microcosms represent novel taxa indicating an unexplored N2O-reducing diversity exists in acidic tropical soils. A survey of pH 3.5–5.7 soil metagenome datasets revealed that nosZ genes commonly occur, suggesting broad distribution of N2O reduction potential in acidic soils.