ANTAGONISTIC, LYTIC AND CELLULASE ACTIVITY BACILLUS, ISOLATED FROM SOIL

Grishko R. V., Kirichenko A. I., Sokolova I. Ye., Dregval’ O. A.,Sklyar T. V.

MICROBIOLOGY

Scentific article

The different groups of agents are used for biological protection of agricultural crops. Some of the most promising and widely used to create microbial preparations are bacteria of the genus Bacillus, which have pathogenic properties regarding harmful insects and phytopathogens. Representatives of the genus Bacillus are one of the main groups of the microbial community of soil and rhizosphere of plants that can produce a number of biologically active compounds (antibiotics, siderophors, lytic enzymes, toxins, phytohormones and vitamins) and have high levels of antagonistic activity to many phytopathogenic fungi and microorganisms. A complex bioinseticidal preparation Bactofungin with a wide spectrum of action against insects of various systematic groups was developed on basis of Bacillus thuringiensis and Beauveria bassiana on the Microbiology, virology and biotechnology department of the Dnipro National University, O. Honchar. The aim of the work was to compare the antagonistic, lytic and cellulase activities of bacilli isolated from the soil, as well as the strain Bacillus thuringiensis IMV-7186, which is part of the complex preparation Bactofungin. The 15 strains of bacilli isolated from the soils of the Poltava region (Bacillus sp. 1 – 15) and deposited strain Bacillus thuringiensis, which was isolated from body of the deceased insect and included in the complex preparation Bactofungin, were investigated. Experimental strains were grown by deep cultivation in synthetic nutrient medium with the addition of 0.5% glucose or Na-CMC as a carbon source. The lytic activity of the culture fluid filtrate of bacilli was determined by turbidimetric method by decreasing of suspensions optical density of live or warmed (100°C, 10 min) test cultures of Pseudomonas syringae pv. atrofaciens, Erwinia amylovora in the process of incubation with an enzyme solution. A chromatographic paper (Watman No. 1) was used as a substrate to determine the cellulase activity. The method is based on quantitative determination of reducing sugars, which are formed as a result of cellulose hydrolysis under the influence of cellulolytic enzymes. In 15 strains of bacteria of the genus Bacillus, isolated from the soil, antagonistic activity was detected in relation to bacterial phytopathogens: X. sampestris, P. syringae pv. atrophaciens, P. syringae pv. lachrуmans, P. carotovorum, as well as with respect to Bacillus thuringiensis, which is part of the bioinsecticide preparation Bactofungin. As a result, 4 strains of bacilli (3rd, 8th, 9th and 10th) were selected, in which the liquid culture was checked for the presence of lytic activity in relation to phytopathogens (Pseudomonas syringae pv atrofaciens and Erwinia amylovora) and cellulase activity. In parallel, the same fermentative activity was also determined in B. thuringiensis. It was found that the greatest lytic activity was shown by B. thuringiensis (Bactofungin component), with the highest rates of specific lytic activity being obtained when glucose was added to the nutritional medium. In strains of bacilli isolated from the soil, the activity indicators on the contrary were higher adding to the medium Na-CMC, but their absolute values were 2-20 times lower than that of B. thuringiensis. The cellulase activity was detected only in two soil strains of Bacillus sp. 3 and Bacillus sp. 8 and ranged from 0.012 to 0.14 units/ml. Moreover, contrary to the expected, the level of cellulase activity in growing strains in a synthetic medium with glucose was significantly greater than that of the Na-CMC medium. The inverse correlation between the accumulation of biomass and the activity of cellulolytic enzymes is revealed, namely, if the cultures are in the stage of active growth, the extracellular enzyme biosynthesis is inhibited. It is planned to create a complex microbial preparation with a wide spectrum of action against insects and phytopathogenic bacteria and fungi.

bacilli, microbial preparation, phytopathogens, antagonism, lytic enzymes, cellulases

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«Bulletin of problems biology and medicine» Issue 4 Part 3 (141), 2017 year, 264-268 pages, index UDK 577.151.52:579.222.3:632.93

10.29254/2077–4214–2017–4–3–141–264-268