Climate change-related warming and increased precipitation may alter winter snow cover and thawing events, and therefore, may carry significant consequences for nitrous oxide (N2O) production pathways such as denitrification, and the abundance and expression of denitrifying microorganisms. We used a soil microcosm study to investigate the combined effect of soil thaw temperature, initial water filled pore space (WFPS) prior to soil freezing, and snowmelt infiltration simulated by the addition of water on N2O emission and denitrification rates, soil respiration rate, and the abundance and transcription of denitrifying (nirK, nirS, and nosZ) bacteria during a single freeze-thaw event. Soil respiration rate was primarily controlled by an increase in soil thaw temperature, whereas soil N2O emission and denitrification rates were generally greater in soils with a higher initial WFPS and soil thaw temperature. In contrast, snowmelt infiltration generally had a negligible effect on these rates, which may be related to pre-existing soil conditions that were already conducive to denitrification. Unexpectedly, the nosZ transcript/nosZ gene abundance ratio was lower in soils thawed at 8.0 °C compared to 1.5 °C; however, this may have resulted in a lower N2O reduction, thus explaining the greater levels of N2O emitted from soils thawed at 8.0 °C. Overall, this study demonstrated that increased N2O production during a single freeze-thaw event was primarily linked to antecedent conditions of high initial WFPS, soil thaw temperature, and a synergistic interplay between these two environmental parameters, and provides evidence that an increase in annual temperature and precipitation, along with the timing of precipitation, may further stimulate N2O production pathways.
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