Bacteria isolated from different layers of four soils of the Cologne area were analyzed for denitrifying, nitrifying and N2-fixing isolates by colony hybridization using gene probes. In the soils tested, the percentage of denitrifying bacteria among the total population isolated was 3-8% (in one case exceptionally 15%) and thus small. Denitrifying bacteria were particularly enriched in the upper layer (depth approximately 5 cm) and were present only in low amounts at 25 cm depth in two gleysol soils. Nitrate apparently did not determine the distribution of denitrifying bacteria in these soils. The potential denitrification activity of different soil layers coincided with the distribution pattern of isolates assessed by DNA-probing. The total number of bacteria and of denitrifying isolates was considerably higher in or at the roots of plants than in the bulk, root-free soil adjacent to the plants. The percentage of the isolated aerobic N2-fixing bacteria varied between 0-3%, and these bacteria could be isolated mainly from the upper 5 cm layer. A small portion of the isolates hybridized with the probe coding for part of one subunit of ammonia monooxygenase from Nitrosomonas europaea. The investigation showed that DNA-probing can provide useful information about the relative distribution of denitrifying and N2-fixing bacteria in different soils and their layers.