The importance of heterotrophic nitrification was studied in soil from a mixed-conifer forest. Three sites in the forest were sampled: a clear cut area, a young stand and a mature stand. In the mature stand, the mineral soil (0-10 cm) and the organic layer were sampled separately. Gross rates of N mineralization and nitrification were measured by (NH4+)-N-15 (NO3-)-N-15 isotopic pool dilution, respectively. The rates of autotrophic and heterotrophic nitrification were distinguished by use of acetylene as a specific inhibitor of autotrophic nitrification. In samples supplemented with (NH4+)-N-15 and treated with acetylene, no (NO3-)-N-15 was detectable showing that the acetylene treatment effectively blocked the autotrophic nitrification, and that NH4+ was not a substrate for heterotrophic nitrification. In the clear cut area, autotrophic nitrification was the most important NO3- generating process with total nitrification (45 ug N kg(-1) h(-1)) accounting for about one-third of gross N mineralization (140 ug N kg(-1) h(-1)). In the young and mature forested sites, gross nitrification rates were largely unaffected by acetylene treatment indicating that heterotrophic nitrification dominated the NO3- generating process in these areas. In the mature forest mineral and organic soil, nitrification (heterotrophic) was equal to only about 5% of gross mineralization (gross mineralization rates of 90 ug N kg(-1) h(-1) mineral; 550 ug N kg(-1) h(-1) organic). The gross nitrification rate decreased from the clear cut area to the young forest area to the mineral soil of the mature forest (45; 17; 4.5 ug kg(-1) h(-1) respectively). The N-15 isotope pool dilution method, combined with acetylene as an inhibitor of autotrophic nitrification provided an effective technique for assessing the importance of heterotrophic nitrification in the N-cycle of this mixed-conifer ecosystem.