AbstractExcessive nitrogen (N) input is one of the major threats for species‐rich grasslands. The ongoing deterioration of habitat quality highlights the necessity to further investigate underlying N turnover processes. Our objectives were (1) to quantify gross and net rates of mineral N production (mineralization and nitrification) and consumption in seminatural grasslands in southwest Germany, with excellent or poor habitat quality, (2) to monitor the temporal variability of these processes, and (3) to investigate differences between calcareous and decalcified soils. In 2016 and 2017, gross N turnover rates were measured using the 15N pool dilution technique in situ on four Arrhenatherion meadows in biweekly cycles between May and November. Simultaneously, net rates of mineralization and nitrification, soil temperature, and moisture were measured. The vegetation was mapped, and basic soil properties were determined. The calcareous soils showed higher gross nitrification rates compared with gross mineralization. In contrast, nitrification was inhibited in the decalcified soils, most likely due to the low pH, and mineralization was the dominant process. Both mineralization and nitrification were characterized by high temporal variability (especially the former) and short residence times of N in the corresponding pools (<2 days) at all sites. This illustrates that high temporal resolution is necessary during the growing season to detect N mineralization patterns and capture variability. Parallel determination of net N turnover rates showed almost no variability, highlighting that net rates are not suitable for drawing conclusions about actual gross turnover rates. During the growing season, the data show no clear relationship between soil temperature/soil moisture and gross N turnover rates. For future experiments, recording of microbial biomass, dissolved organic matter, and root N uptake should be considered.