Phases of denitrification following oxygen depletion in soil

The short-term response of soil denitrification to reduced aeration was studied using the acetylene inhibition method for the assay of denitrification. Two distinct phases of denitrification rate were observed. An initial constant rate, termed phase I, was not decreased by chloramphenicol, was increased slightly or not at all by organic carbon amendment, and lasted for 1–3 h. Phase I was attributed to the activity of pre-existing denitrifying enzymes in the soil microflora. Following phase I the denitrification rate increased; chloramphenicol inhibited this increase. In soils without organic-C amendment a second linear phase, termed phase II, was attained after 4–8 h of anaerobic incubation. The linearity of this phase was attributed to the full derepression of denitrifying enzyme synthesis by the indigenous population and to the lack of significant growth of denitrifiers. Phase I rate was dependent on the initial or in situ aeration state of the soil sample; phase II was not. Therefore, phase I may be more directly related to field denitrification rates. Denitrification rate changes following water saturation of soils in aerobic atmospheres were also examined. Rates were greatly increased by wetting but only after a lag of several hours. Our interpretation is that following wetting of natural soils, anaerobic or partially anaerobic conditions are established by respiration and reduced O2 diffusion rate; this first eliminates O2 inhibition then derepresses the synthesis of denitrifying enzymes. Although denitrifying enzymes are apparently present even in relatively dry soils, their activity is low until O2 inhibition is eliminated. From this evidence we reason that most N is lost from soils during brief periods beginning a few hours after irrigation or a rainfall.