؊ as a relevant modulator of NO 3 ؊ fate in S. loihica strain PV-4, and, by extension, suggest that NO 2 ؊ is a relevant determinant for N retention (i.e., ammonification) versus N loss and greenhouse gas emission (i.e., denitrification).
T wo major dissimilatory pathways determine the fate of nitrate (NO 3Ϫ ) in anoxic environments: denitrification and respiratory ammonification (1, 2). In denitrification, NO 3 Ϫ is stepwise reduced via nitrite (NO 2 Ϫ ), nitric oxide (NO), and nitrous oxide (N 2 O) to dinitrogen (N 2 ). In respiratory ammonification, NO 3 Ϫ is reduced via NO 2 Ϫ to ammonium (NH 4 ϩ ). Nitrite is the common intermediate of the two pathways and the branching point for both dissimilatory pathways. In the presence of NH 4 ϩ , NO 2 Ϫ also can serve as the electron acceptor for anaerobic ammonium oxidation (anammox) (3); however, in soil environments this process appears to be less relevant than the other two processes (4). The fate of NO 2 Ϫ via denitrification or respiratory ammonification has great environmental impact. Denitrification forms gaseous products (N 2 O, N 2 ), which are emitted from the soil, resulting in N loss, whereas respiratory ammonification generates NH 4 ϩ leading to N retention (5, 6). Atmospheric N 2 O is a potent greenhouse gas and an ozone-depleting agent (7,8). Therefore, knowledge of the environmental factors that control these NO 2 Ϫ reduction pathways is needed to estimate, predict, and possibly manipulate N loss versus N retention.The major sources of NO 2 Ϫ in the environment include NH 4 ϩ oxidation performed by nitrosifiers and NO 3 Ϫ reduction. In these conversions, NO 2 Ϫ accumulates when its production is kinetically faster than NO 2 Ϫ consumption (9, 10). Nitrite accumulation had been thought to occur rarely in the environment (11); however, recent observations suggested that NO 2 Ϫ formation can occur as a result of NO 3 Ϫ reduction and NH 4 ϩ oxidation under conditions that favor NO 2 Ϫ production over consumption (12)(13)(14)(15)(16). For example, high pH and abundance of NH 4 ϩ and hydroxylamine (NH 2 OH) affect NO 2 Ϫ oxidizers, causing NO 2 Ϫ accumulation from nitrosification (15), while oxygen intrusion and/or electron donor limitations may cause NO 2 Ϫ accumulation from denitrification (12). In pure-culture studies, several denitrifiers and respiratory ammonifiers were found to reduce NO 3 Ϫ at a higher rate than NO 2 Ϫ , causing dynamic changes of the NO 2 Ϫ :NO 3 Ϫ ratios in the medium (10). Additional NO 2 Ϫ may be generated by NO 3 Ϫ -to-NO 2 Ϫ reducers sensu stricto, which generate NO 2 Ϫ as an end product (12). For example, in activated sludge, the activity of NO 3 Ϫ -to-NO 2 Ϫ reducers leads to NO 2 Ϫ formation; however, the contribution of NO 3 Ϫ -to-NO 2 Ϫ reducers to NO 2 Ϫ accumulation in natural environments is uncertain (17,18).Even though NO 2 Ϫ formation occurs in diverse environments (12,19), information regarding the impact of NO 2 Ϫ on dissimilatory NO 3 Ϫ /NO 2 Ϫ reduction pathways is scarce. Increased NO 2 Ϫ concentrations and respi...