The European Commission’s “Farm to Folk” (F2F) strategy recommends reducing fertilizers by at least 20% by 2030. In this aspect, the main goal of our study was to verify whether a 20 and even 40% reduction in nitrogen (N) fertilization rate would be sufficient to maintain soil fertility (expressed as dehydrogenase activity—DHA, humic acid substances—HA substances, E4/E6 ratio), and yields of wheat and rapeseed cultivated in strip-till technology. Two fields (10 ha each) were established for wheat and rapeseed cultivation, and soils (0–20 cm) were sampled before sowing and after harvesting. It was found that a 20% fertilization reduction does not adversely affect the biological activity expressed by DHA. Nevertheless, the reduction in nitrogen (N) fertilizer rates led to a slight decrease in the E4/E6 ratio, which could serve as a potential indicator of fertilization reduction. DHA, E4/E6 ratio and content of HA substances were also dependent on the crop. Following harvesting, both rapeseed and wheat exhibited a positive correlation between the E4/E6 ratio and the degree of fertilization rate (FR), as well as yield.