The substrate and especially its reaction is the most important aspect affecting the success of highbush blueberry cultivation. The limited availability of suitable soils forces producers to look for alternative cultivation methods. The substrates can be used in a variety of growing systems, including forming substrates in the mound form or filling with them trenches. Both systems require different outlays to set up a plantation. They also create different conditions for plant growth. The experiment evaluated the yield, chemical composition (N, P, K, Ca, Mg, Cu, Fe, Mn and Zn) of highbush blueberry leaves and fruits and changes in chemical composition of soil and substrate during cultivation (pH, general and available mineral nutrients). The research materials were collected at a research station of the West Pomeranian University of Technology Szczecin in Poland in 2014-2016. Two-year-old bushes of the 'Brigitta Blue' were planted in 2012. In the field experiment, sawdust, peat and cocoa husk were used in the cultivation in the flat (trenches with a depth of 35 cm and a width of 100 cm) and mound systems (with 35 cm in height and 100 cm in width). The blueberries used for growing sawdust, peat and cocoa husk substrates had a different chemical composition. The most suitable conditions for the growth of blueberries were provided by sawdust and peat substrates and the worst of cocoa husk. The cocoa husk substrate had a high pH and was resistant to changes in reaction (start 6.58; end 6.32–6.61 in KCl). It was also characterized by excessive abundance in macroelements. The growth of bushes was the weakest and yielding at a low level (0.39–0.53 kg per bush). The substrate of sawdust was more prone to lowering the pH (6.12; end 4.15–4.53 in KCl). Peat pH was stable during the experiment. Better grew and yielded bushes resulted when planted in peat in the flat system, or in sawdust in the mound system. The mound cultivation system had a more favorable effect on the substrate's chemical properties than the flat soil system. In general, in the flat cultivation system, the plants took better off the macro and micronutrients from the tested substrates. Due to the lower workload, it is recommended to prepare the plantations for growing highbush blueberries in the mound system. The mound cultivation system shaped the chemical properties of substrates more advantageously than the flat system. Sawdust and peat affected the soil negatively - mainly for lowering pH and mineral composition. Larger changes were found in the mounds cultivation system.