Recent studies indicate that soil soluble organic nitrogen (SON) plays an important regulatory role in the soil-plant N cycle. The aims of this study were to identify the vertical distribution of SON and its correlation with N mineralization, nitrification, and amidohydrolase activities, in a soil repeatedly amended with cow manure or chemical fertilizer. For this purpose, soil samples were collected from 0-20, 20-40, 40-60, 60-80, and 80-100 cm depths of a calcareous soil, which has been annually amended for 5 y with cow manure (CM) at two rates of 50 (CM 50 ) and 100 (CM 100 ) Mg CM ha -1 y -1 . Treatments with chemical fertilizer (CF) and a control (CT) were also included. Soluble organic N, N mineralization, nitrification rates, as well as L-glutaminase and L-asparaginase activities were determined. Both CM 50 and CM 100 enhanced SON content throughout the soil profile. Nitrogen-mineralization rate (N m ) was increased at the 0-20 cm depth of the CM 100 treatment and remained unaffected at the deeper depths. Nitrification rate (N n ) was significantly higher at the 0-60 cm depth of CM 100 compared to CF and CT. L-glutaminase and L-asparaginase activities were significantly increased at the 0-40 cm depth in both CM 50 and CM 100 compared to CF and CT. The amidohydrolase activities could not be detected below 40 cm, regardless of the fertilizer treatments. Our results suggest that SON makes a minor contribution to N mineralization in deep soil layers. It was also concluded that changes in the SON throughout the soil profile were not associated with changes in the N-transformation rates (N m and N n ) and amidohydrolase activities. While we conclude that SON is a major N pool in the whole profile of the manure applied soil further investigation is required to characterize SON and to investigate the bioavailability of SON for microbial activity in different soil depths.