Purpose. To study the effect of the experimental microbiological preparation “Naturost”, created on the basis of Bacillus subtilis No. 111, on the growth and productivity of barley in the Vologda region. Materials and methods. The research was carried out during the growing seasons of 2019, 2020 and 2022 at the experimental field of RAS Vologda Research Center and under production conditions in the fields of the agricultural production cooperative (collective farm) “Plemzavod Prigorodnyi” (Vologda region). The preparation “Naturost” was introduced twice: while soaking the seeds and spraying the phyllosphere of the plants in the tillering phase. Whole-genome sequencing of the strain of the bacterium B. subtilis-111 was carried out at the molecular genetic laboratory of “Biotrof” company with the use of MiSeq platform (Illumina, Inc.) Results. During the whole-genome sequencing of B. subtilis-111 strains, it was found that it has auxin biosynthesis genes; a range of various protective mechanisms contributing to the survival of the strain in the natural environment was also detected in the genome. In addition, the genome of the strain contains an array of groups of genes that are responsible for the production of bacteriocins with pronounced antimicrobial activity. The treatment of barley plants with the experimental preparation “Naturost” contributed to an increase in the growth parameters of the crop: dry mass increased to 33%, average daily increments – up to 48%, which occurred against the background of an increase in the content of photosynthetic pigments. Grain productivity in the conditions of small-scale experiments with the use of the preparation increased by 7–19%, and in the actual field conditions – by 14% relative to the control. Conclusion. The experimental preparation based on B. subtilis-111 had a stimulating effect on the growth and productivity of barley of the breed Sonet in the Vologda region.