The article presents the results of research on the development of parameters for practical application of phage biopreparation Pseudomonas syringae in order to indicate the data of phytopathogenic bacteria in sanitary control objects. The introduction of the article describes pathogens of Pseudomonas syringae and plants that they affect, which proves research actuality aimed at developing speed-up methods of indication and identification of bacteria-pathogens, which will allow specialists to develop measures to combat above-mentioned phytopathogens in a short time. It was established that cultivation of the "bacteriophage-test material" system at a temperature of 28 ± 1 0C during 3.5 hours allows detecting Pseudomonas syringae bacteria in soil samples, river water, and cucumber seeds by the RSF method at a concentration of 103 MK/ml., increase in the exposure time does not improve the quality of the reaction. The bacteriophages Ps. s-7 UlGAU and Ps. s-27 UlGAU in monoculture were used in experiments at a concentration of 103 BFU/ml. When testing a polyphage biological product in RSF, including all the above-mentioned bacteriophages, it was found that the results obtained in the studies of the river water sample were similar to the experimental data on cucumber seed sample. Concentration of detected Pseudomonas syringae bacteria was 103 m.k./ml. It was experimentally established that reducing the initial titer of polyphage biopreparation to a concentration of 102 BFU / ml allowed for the indication of Pseudomonas syringae bacteria in a soil sample at a concentration of 103 mk/g.
The article presents the results of studies on isolation and study of biological properties of bacteriophages specific for phytopathogenic bacteria Pseudomonas syringae. 8 new virulent bacteriophages were isolated and selected from environmental objects. The main biological properties of isolated phages, including lytic activity, were studied (according to Appelman, the indicator was equal to from 10-5 to 10-8; by diffusion method in «soft agar» - from 1.0±0.1*106 to 2.0±0.1*109 (PFU/ml), «spot test» - spot test confirmed the results obtained by diffusion method in «soft agar» - n6 – n9) and its diapason, which ranged from 21.4% (Ps.s-13 UlGAU) to 85.7% (Ps.s-7 UlGAU, Ps.s-27 UlGAU), morphology of plaque-forming units (morphology of negative colonies), specificity within genus and species, changes in lytic activity index during storage, features of impact of physical and chemical factors (trichloromethane at the ratio of 1:10, the impact of which during 5-85 minutes does not influence on biological activity of phages when determining by indirect method; urea, whose 30% solution reduces the yield of plaque-forming units of each bacteriophage studied by about 77.6-85.5 %; temperature in the range of -5 to 10°C and from 50 to 100°C). The study of biological properties allowed us to systematize biological features of each of isolated clones of virulent bacteriophages and select the 4 most promising bacteriophages for further research. The research is carried out with the support of Russian Foundation for Basic Research, the project “Fundamental foundations for the development of phage preparation specific to Pseudomonas syringae, and applied aspects of its application for phagoidentification and bioprocessing of food and agricultural raw materials” No 19-44-730014.
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