The size of the microbial biomass and the activity of soil enzymes are among the most sensitive indicators of agricultural land quality. The aim of this study was to determine the spatial and temporal variability of microbial biomass, the activity of dehydrogenase (DHA) enzyme and the concentration of micro- (Na, Mg and Ca) and macroelements (Cu, Zn, Mn and Fe) in the soil, collected from 37 measurement sites (depth 0–30 cm) in a 40-hectare field during two growing seasons (wheat and oilseed rape). The percentage of nitrogen (%N) in the wheat grain and rapeseeds was also determined. Mapping the spatial distribution of the microbial biomass, the level of DHA activity and the concentration of the selected elements was used to assess the soil productivity. All tested soil parameters exhibited temporal and spatial variability. The creation of raster maps showing the distribution of the tested parameters allowed the observation of a higher nitrogen content in wheat grains in the south-western part of the field, with high values of DHA activity, bacterial biomass and soil pH. In the case of oilseed rape, plants grown in the northern part of the field were characterized by a higher nitrogen content in the grain, where the soil was characterized by a higher content of fungal biomass. On the basis of the obtained research results, a positive, statistically significant correlation was also shown between the biomass of bacteria and the level of DHA in the soil under the cultivation of both wheat and rape. The cultivation of both crops had a significant impact on the size of the microbial biomass pool and on the DHA activity level but did not affect the concentration of the nutrients in the soil. High concentrations of the analyzed elements at the measuring points correlated with a greater %N content in the grain/seeds of the crops harvested at those locations in the field. The results conclude that the mapping of the physicochemical parameters, microbial biomass and activity on the field permits the development of an effective strategy for maintaining sustainable soil productivity through the appropriate management of agricultural practices and the better approximation of mineral fertilization.