Nitrous oxide production and mRNA expression analysis of nitrifying and denitrifying bacterial genes under floodwater disappearance and fertilizer application

Abstract

A pulse of nitrous oxide (N₂O) emission has been observed following the disappearance of floodwater by drainage. However, its mechanism is not well understood. We conducted a column study to clarify the mechanism for N₂O production during floodwater disappearance by using a microsensor and determining the bacterial gene expression. An increase in N₂O flux was observed following floodwater disappearance after the addition of NH₄⁺, with a corresponding increase in the concentrations of NO₃⁻ and dissolved N₂O in the oxic and anoxic soil layers, respectively. The transcription level of the bacterial amoA mRNA did not change, while that of nirK mRNA increased sharply after an hour of floodwater disappearance. An additional anoxic soil slurry experiment demonstrated that the addition of NO₃⁻ induced the expression of nirK gene and caused a concomitant increase in N₂O production. These findings suggest that NO₃⁻ production in the oxic layers is important as it provides a substrate and induces the synthesis of denitrification enzymes in the anoxic layer during N₂O production.

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