Chromatographic profiles of antifungal exo- and endometabolites of Bacillus velezensis

The antifungal activity of the Bacillus bacteria is based on their ability to produce metabolites. Therefore, when selecting a strain that produces an effective biofungicide, it is necessary to assess the metabolism of bacteria. The aim of this work is to isolate exo- and endometabolites of the promising B. velezensis BZR 336g and B. velezensis BZR 517 strains and assess their antifungal activity. Studies were carried out in 2020–2021. The object of the study is a liquid culture of the B. velezensis BZR 336g and B. velezensis BZR 517 strains. Methods of liquid extraction, ascending thin layer chromatography (TLC), bioautography with a test-culture of Fusarium oxysporum var. orthoceras and Alternaria sp. fungi were used to analyze metabolites. The ability of the strains to accumulate a complex of active metabolites showing antifungal effect from fungistatic to fungicidal action was revealed. On the bioautogram of exometabolites, we found two most pronounced zones (Rf 0.18 and 0.29) of Fusarium oxysporum var. orthoceras BZR P1 growth inhibition (fungicide). Zones with Rf 0.58 for B. velezensis BZR 336g and Rf 0.70 for B. velezensis BZR 517 correspond to the test fungus growth retardation (fungistatic). Significant suppression of Alternaria sp. BZR P8 growth was also observed in two zones (Rf 0.18 and 0.29). The use of surfactin, iturin A, fengycin (Sigma-Aldrich®) in the TLC analysis made it possible to detect similar lipopeptides in the composition of metabolite complexes produced by the studied bacteria. It should be noted that the studied strains differed both in their ability to produce metabolites of different structure (can be found when analyzing chromatograms under ultraviolet light) and in their effect on phytopathogenic fungi in vitro. This may indicate possible differences in the mechanism of antagonistic activity of bacteria against phytopathogenic fungi. Thus, B. velezensis BZR 336g and B. velezensis BZR 517 produce a significant set of antifungal metabolites and can be used as strains to produce effective biofungicides.