Chemical formation of hybrid di-nitrogen calls fungal codenitrification into question

Abstract

Abstract Removal of excess nitrogen (N) can best be achieved through denitrification processes that transform N in water and terrestrial ecosystems to di-nitrogen (N 2 ) gas. The greenhouse gas nitrous oxide (N 2 O) is considered an intermediate or end-product in denitrification pathways. Both abiotic and biotic denitrification processes use a single N source to form N 2 O. However, N 2 can be formed from two distinct N sources (known as hybrid N 2 ) through biologically mediated processes of anammox and codenitrification. We questioned if hybrid N 2 produced during fungal incubation at neutral pH could be attributed to abiotic nitrosation and if N 2 O was consumed during N 2 formation. Experiments with gas chromatography indicated N 2 was formed in the presence of live and dead fungi and in the absence of fungi, while N 2 O steadily increased. We used isotope pairing techniques and confirmed abiotic production of hybrid N 2 under both anoxic and 20% O 2 atmosphere conditions. Our findings question the assumptions that (1) N 2 O is an intermediate required for N 2 formation, (2) production of N 2 and N 2 O requires anaerobiosis, and (3) hybrid N 2 is evidence of codenitrification and/or anammox. The N cycle framework should include abiotic production of N 2 .