Preventing soil degradation is an important task of modern agrarian science. Preservation and increase in beneficial soil microflora are a precondition for satisfactory ecological functioning of soil and its fertility. The purpose of this study was to investigate the dynamics of beneficial soil microflora in winter wheat crops under the impact of mineral fertilisers and Agrobacterium radiobacter on the lands of the south of Ukraine. The study was conducted in threefold replication in the conditions of the Kherson region in 2016- 2020 in the experimental fields of the Institute of Climate-Smart Agriculture of the National Academy of Agrarian Sciences of Ukraine. The study used a systematic design, and the following factors were investigated: no fertiliser and Agrobacterium radiobacter; N120P90; N90P60 + Agrobacterium radiobacter; N120P90 + Agrobacterium radiobacter. The study on the microflora composition in the soil layer at 0-30 cm was carried out in the main stages of winter wheat growth and development through the inoculation of nutritious environment. Statistical analysis was performed using the methods of analysis of variance, correlation, and regression analysis. Dynamic changes in the soil biota composition under the influence of the studied factor were established. The slightest response to mineral fertilisers and bacterial preparation application was in ammonifying bacteria, while the strongest response was in cellulosedegrading microorganisms. The winter wheat harvesting revealed a decrease in the number of ammonifying bacteria in the soil, while the number of nitrifying and cellulose-degrading microflora increased significantly. Regression models for the prediction of the number of nitrifying bacteria and cellulose-degrading microorganisms, developed based on experimental data, have great accuracy (the error is 3.78% and 7.79%), and allow determining the adverse effect of phosphorus fertiliser on the microflora of dark-chestnut soil. The study has no analogues in Ukraine and expands theoretical knowledge about the influence of mineral fertilisers and bacterial preparation, containing Agrobacterium radiobacter, on the composition of beneficial microflora of the dark-chestnut soil