N2O emissions and product ratios of nitrification and denitrification as affected by freezing and thawing

Agricultural soils contribute significantly to atmospheric nitrous oxide (N2O). A considerable part of the annual N2O emission may occur during the cold season, possibly supported by high product ratios in denitrification (N2O/(N2+N2O)) and nitrification (N2O-N/(NO3−-N+NO2−-N)) at low temperatures and/or in response to freeze–thaw perturbation. Water-soluble organic materials released from frost-sensitive catch crops and green manure may further increase winter emissions. We conducted short-term laboratory incubations under standardized moisture and oxygen (O2) conditions, using nitrogen (N) tracers (15N) to determine process rates and sources of emitted N2O after freeze–thaw treatment of soil or after addition of freeze–thaw extract from clover. Soil respiration and N2O production was stimulated by freeze–thaw or addition of plant extract. The N2O emission response was inversely related to O2 concentration, indicating denitrification as the quantitatively prevailing process. Denitrification product ratios in the two studied soils (pH 4.5 and 7.0) remained largely unaltered by freeze–thaw or freeze–thaw-released plant material, refuting the hypothesis that high winter emissions are due to frost damage of N2O reductase activity. Nitrification rates estimated by nitrate (NO3−) pool enrichment were 1.5–1.8 μg NO3-N g−1 dw soil d−1 in freeze–thaw-treated soil when incubated at O2 concentrations above 2.3 vol% and one order of magnitude lower at 0.8 vol% O2. Thus, the experiments captured a situation with severely O2-limited nitrification. As expected, the O2 stress at 0.8 vol% resulted in a high nitrification product ratio (0.3 g g−1). Despite this high product ratio, only 4.4% of the measured N2O accumulation originated from nitrification, reaffirming that denitrification was the main N2O source at the various tested O2 concentrations in freeze–thaw-affected soil. N2O emission response to both freeze–thaw and plant extract addition appeared strongly linked to stimulation of carbon (C) respiration, suggesting that freeze–thaw-induced release of decomposable organic C was the major driving force for N2O emissions in our soils, both by fuelling denitrifiers and by depleting O2. The soluble C (applied as plant extract) necessary to induce a CO2 and N2O production rate comparable with that of freeze–thaw was 20–30 μg C g−1 soil dw. This is in the range of estimates for over-winter soluble C loss from catch crops and green manure plots reported in the literature. Thus, freeze–thaw-released organic C from plants may play a significant role in freeze–thaw-related N2O emissions.