The aim of this research was to evaluate the efficacy of biological fungicides against winter barley root rot pathogens for their subsequent practical application in ecological protection technologies within the Krasnodar Territory. The experiments were conducted at the Federal Scientific Center for Biological Plant Protection (FRCBPP) under field infection nursery conditions during 2023–2025. The object of study comprised winter barley root rot pathogens of Fusarium and Helminthosporium etiology. Selected for the study were preparations containing various agents of biological origin and their metabolites: Geostim Fit, A, L (2 l/t); Alirin B, L (2 l/ha); Pseudobacterin-2, L (1 l/ha); Sterniphag, WP (80 g/t); Trichocin, WP (20 g/t); an experimental sample from the FR-CBPP based on Bacillus velezensis BZR336 g, L (3 l/t); an experimental sample from the FRCB-PP based on Pseudomonas chlororaphis BZR245-F, L (3 l/t); Orgamica F, L (0.7 l/t); and Scarlet, ME (0.4 l/t) (chemical standard). The maximum efficacy against Fusarium seed infection was observed after treatment with Trichocin, WP (94.5 %), Geostim Fit, A, L (92.4 %), Alirin-B, L (92.3 %), and Organica F, L (91.1 %). Laboratory and field germination rates were highest after seed treatment with preparations based on Trichoderma asperellum – 96 % (Organica F, L), Bacillus velezensis – 95 % (experimental sample from the FRCBPP based on Bacillus velezensis BZR336 g, L), and Pseudomonas chlororaphis – 95 % (experimental sample from the FRCBPP based on Pseudomonas chlororaphis BZR245-F, L). The highest plant stand densities were recorded on experimental plots treated with Alirin-B, L, Geostim Fit, A, L (488 plants/m 2 ), Sterniphag, WP (485 plants/m 2 ), Pseudobacterin-2, L, and the Scarlet, ME standard (484 plants/m 2 ). High efficacy rates against root and radical rots were determined for preparations based on Bacillus subtilis Alirin-B, L (68.3 %), Pseudomonas chlororaphis BZR245-F, L (65.2 %), and Trichoderma harzianum Sterniphag, WP (60.3 %). Research into the efficacy of biological preparations for protecting barley from root rot is crucial for practical application within the agro-industrial sector of both the region and the country.