Biofertilizers are an alternative to mineral fertilizers for increasing soil productivity and plant growth in sustainable agriculture. The objective of this study was to evaluate possible effects of three mineral fertilizers and four plant growth promoting rhizobacteria (PGPR) strains as biofertilizer on soil properties and seedling growth of barley (Hordeum vulgare) at three different soil bulk densities, and in three harvest periods. The application treatments included the control (without bacteria inoculation and mineral fertilizers), mineral fertilizers (N, NP and P) and plant growth promoting rhizobacteria species (Bacillus licheniformis RC04, Paenibacillus polymyxa RC05, Pseudomonas putida RC06, and Bacillus OSU-142) in sterilized soil. The PGPR, fungi, seedling growth, soil pH, organic matter content, available P and mineral nitrogen were determined in soil compacted artificially to three bulk density levels (1.1, 1.25 and 1.40 Mg m⁻³) at 15, 30, and 45 days of plant harvest. The results showed that all the inoculated bacteria contributed to the amount of mineral nitrogen. Seed inoculation significantly increased the count of bacteria and fungi. Data suggest that seed inoculation of barley with PGPR strains tested increased root weight by 9–12.2%, and shoot weight by 29.7–43.3% compared with control. The N, NP and P application, however, increased root weight up to 18.2, 25.0 and 7.4% and shoot weight by 31.6, 43.4 and 26.4%, respectively. Our data show that PGPR stimulate barley growth and could be used as an alternative to chemical fertilizer. Soil compaction hampers the beneficial plant growth promoting properties of PGPR and should be avoided.