Impact of nitrogen compounds on fungal and bacterial contributions to codenitrification in a pasture soil

Ruminant urine patches on grazed grassland are a significant source of agricultural nitrous oxide (N₂O) emissions. Of the many biotic and abiotic N₂O production mechanisms initiated following urine-urea deposition, codenitrification resulting in the formation of hybrid N₂O, is one of the least understood. Codenitrification forms hybrid N₂O via biotic N-nitrosation, co-metabolising organic and inorganic N compounds (N substrates) to produce N₂O. The objective of this study was to assess the relative significance of different N substrates on codenitrification and to determine the contributions of fungi and bacteria to codenitrification. ¹⁵N-labelled ammonium, hydroxylamine (NH₂OH) and two amino acids (phenylalanine or glycine) were applied, separately, to sieved soil mesocosms eight days after a simulated urine event, in the absence or presence of bacterial and fungal inhibitors. Soil chemical variables and N₂O fluxes were monitored and the codenitrified N₂O fluxes determined. Fungal inhibition decreased N₂O fluxes by ca. 40% for both amino acid treatments, while bacterial inhibition only decreased the N₂O flux of the glycine treatment, by 14%. Hydroxylamine (NH₂OH) generated the highest N₂O fluxes which declined with either fungal or bacterial inhibition alone, while combined inhibition resulted in a 60% decrease in the N₂O flux. All the N substrates examined participated to some extent in codenitrification. Trends for codenitrification under the NH₂OH substrate treatment followed those of total N₂O fluxes (85.7% of total N₂O flux). Codenitrification fluxes under non-NH₂OH substrate treatments (0.7-1.2% of total N₂O flux) were two orders of magnitude lower, and significant decreases in these treatments only occurred with fungal inhibition in the amino acid substrate treatments. These results demonstrate that in situ studies are required to better understand the dynamics of codenitrification substrates in grazed pasture soils and the associated role that fungi have with respect to codenitrification.