Bacteria of the genus Bacillus play an important role in biological methods of plants protection. The bacteria, as producers of active substances of biopesticides, favourably differ from other microorganisms. They grow well and quickly on artificial media with relatively simple chemical composition. The bacteria of the genus Bacillus show an antagonistic activity against a large spectrum of plant pathogens that infect a wide range of crops. Live sporulating bacterial culture inhibits the proliferation of pathogenic fungi and bacteria by their metabolic products. Currently, in the Republic of Moldova any biological product based on Bacillus is not available. In this connection, the search and development of new biological agents that would become the producents of new biological products are indispensable. In this regard, the scope of the study was determination of the cultural and morphological features of the growth of bacteria of genus Bacillus, isolated from the rhizosphere of tomato, having the ability to inhibit the widespread in republic pathogenic fungi. On the results of extensive screening of bacterial agents with the antifungal activity against phytopathogenic fungi, the bacteria were selected, which were identified according to manual Bergey’s, as Bacillus subtilis. Cultural morphological characteristics of individual colonies were performed and described using three media: PDA, MPA and M9 minial medium. Comparing the growth features of two studied bacteria it was noticed that the culture of the B. subtilis strain S4 grew as single distinct colonies on all of the presented media. Whereas the B. subtilis strain S2 created well-defined colony only on MPA medium, but on PDA and M9 media – culture spreads, blurring the margins of the colonies. The number of colonies on these media could be counted only by the central areas of the colony. The most significant growth features of microorganisms on solid cultivation medium is the colony characters. Different bacterial cultures behave differently when spreading on cultivation media. In our study the bacterial strain Bacillus subtilis S4 have presented the same growth characteristics on the all three cultivation media, giving individual clear colonies, while the strain Bacillus subtilis S2 – only on the MPA medium. This suggests that for determination the titer of the cells of the Bacillus subtilis S2 it is desirable to use MPA medium. We tried to inoculate the selected strain of Bacillus subtilis on PDA cultivation medium for best storage. It was obtained a good strong growth and biomass accumulation of both bacterial strains on this medium. This is consistent with the classic conditions for storage of this type of cultures. Preliminary experiments on the selective media to ensure high titer cultures revealed that cultures such highest titer observed on the medium MPA (Bacillus subtilis S2 – 4.1×109 CFU/ml, Bacillus subtilis S4 – 9.4×108 CFU/ml), on PDA medium (Bacillus subtilis S2 – 1.2×109 CFU/ ml, Bacillus subtilis S4 – 3.3×109 CFU/ml), on M9 medium titer was lower (Bacillus subtilis S2 – 3.0×108 CFU/ml, Bacillus subtilis S4 – 4.3×107 CFU/ml). Given that the medium of the PDA and the MPA have a complex composition and are quite expensive, it is appropriate to continue to use the M9 medium (that is easy to prepare and low cost) for the development of new media that contribute to maitaining and increasing the antifungal properties of the studied strains. PDA and MPA are the natural media, whereas M9 – is a synthetic medium. As are known the natural media are not easy to use for production of culture, because they are very expensive and required the use of food products and a long time for their preparation. The synthetic medium M9 is cheaper and easier to prepare. Thus, the M9 medium can serve as a basis for further development of the best cultivation medium for the studied bacteria.