In agroecosystems, microorganisms are the main factor in the soil formation process, plant nutrition and phytosanitary condition of the soil. Therefore, all measures aimed at restoring soil fertility and increasing productivity, environmental safety of agricultural production are closely linked to the activities of microorganisms. Increased local activity, biomass and diversity of microbiota are one of the most important characteristics that distinguish the rhizosphere from the total soil volume. The aim of the research was to study the number and composition of the microbial complex of the rhizosphere of cereals (winter wheat of different varieties of domestic selection) in the process of ontogenesis. To conduct a set of microbiological studies, soil samples were taken and prepared (typical chernozem) in compliance with the standard requirements for sample preparation and storage of samples in the laboratory. The analysis of rhizosphere soil samples was performed according to the variants of different varieties of winter wheat of domestic selection. The number of soil microorganisms was determined by the method of inoculation of suspensions on nutrient media according to generally accepted methods in microbiology. Determination of the total microbial biomass in the soil was carried out by the rehydration method, and the intensity of "respiration" of the soil, respectively, by the release of CO2 and O2 absorption by the manometric method (Warburg apparatus). It is established that varietal specificity is significantly related to the peculiarities of microbiome formation in different phases of plant growth and development, which is an integral indicator of functional and metabolic activity of soil microorganisms. The number and composition of the microbial complex of winter wheat rhizosphere in the process of ontogenesis changes significantly, especially in the ratio of the number of spore-forming and non-spore forms of microorganisms under the same conditions of crop cultivation. The total pool of saprotrophic microorganisms of the rhizosphere demonstrates the variability of biomass and changes in favor of ecoplastic Bacillus. An increase in the number of spore-forming bacteria to 4.2 x 107 CFU/g in the variants of growing certain varieties was found. It is shown that in the cultivation of different varieties of winter wheat there are stable indicators of the intensity of CO2 emissions - from 5.2 to 7.0. A similar trend can be traced to the absorption of O2 (not more than 5.3-6.8).
From an agrocoenosis of winter wheat (Triticum aestivum L.; phylloplane and rhizosphere of the root system; typical chernozem, soil column measuring up to 40 cm), using the classical microbiological methods, we had isolated soil bacteria and characterized them according to the morphological features as representatives of Gram-positive and spore-forming bacteria of Bacillus sp. genus. In the earing-swelling phase of grain, the screening studies found non-pigmented forms of colonies of bacterial isolates, 19 of which were classified to colonial-morphological diversity of R-type with the diameter of 7 to 13 mm. The analysis of physiological condition of cells of populations of soil isolates revealed technologic specificity according to parameters of spore formation in different conditions and incubation time (up to 48–72 h). We observed 90.0% of free spores in axenic cultures as early as after 72 h of cultivation and no more than 10.0% of prospores in the studied monoisolates with stable morphologic traits. Isolates Н10 and Н45 demonstrated the ability to grow in higher cultivation temperatures (+37…+40 °С). According to environmental рН, isolates were able to grow in рН ranging 4.5–8.0. Differential diagnostic testing revealed that as the source of carbon, with formation of acid, soil isolates used arabinose, xylose, mannitol, glucose, galactose, fructose, maltose, sorbitol, glycerin, dextrin, starch, rhamnose and dulcite (with development of alkaline). There was observed active use of mineral forms of nitrogen: ammonium salt and nitrates, aminoacids and proteins. The isolates hydrolyzed casein, gelatin, starch, and litmus was being reduced in the young during growth in milk with litmus. They also exerted catalase activity and were oxidase-positive. Biochemical testing using API test system determined that the studied isolated bacteria differed by a range of fermentation carbohydrates, reduction of nitrates. In the conditions of submerged fermentation, isolates Н38 and Н40 grew in heightened temperature ranges of cultivation (40 °С) for 48 h (according to fact of spore development). Therefore, according to the key morphologic and biochemical traits, strains Н3, Н10, Н13, Н36, Н38, Н40, Н43, Н45 were similar to such of reference strain B. subtilis 8A, and were identified to Bacillus sp., species B. subtilis.
Objective. To assess the performance of individual strains of Bacillus subtilis and the efficiency of their action on the condition and activity of the photosynthetic system of juvenile winter wheat plants (Triticum aestivum L.). Methods. Microbiological, instrumental (microscopic), biophysical (determination of photochemical activity of winter wheat sprouts in model conditions of the induction of chlorophyll fluorescence). Results. The technological stability of B. subtilis H38, H40 and H45 strains was shown both during fermentation (formation of viable spore titre in the range of 1.9‒2.4 billion/mL) and during storage of bacterial suspensions (BS) for 60 days (1.8‒2.3 billion spores/mL). Analysis of the influence of B. subtilis H38, H40, H45 on the activity of the photosynthetic system of winter wheat (Triticum aestivum L.) plants revealed a high informative value of induced changes in chlorophyll fluorescence (ICF), which was recorded in the structural organization of chloroplasts of wheat sprouts at the initial, maximum, and stationary levels of fluorescence and viability index (Rfd after 1:50 dilution of BS of B. subtilis strains is 1.56–1.69; Rfd after 1:100 dilution corresponds to the normal quantum efficiency of photosynthesis in the range of 1.20‒1.40). Conclusion. The studied strains of B. subtilis are characterized by high performance (a significant number of spores are formed during the cultivation of bacteria; their number remains relatively stable during a 60-day research period), which implies the possibility of their successful use in production and the possibility of long-term storage of preparations based on them. The prospective use of the evaluated strains for the intensification of the photochemical activity of winter wheat (Triticum aestivum L.) plants in the process of ontogenesis has been proven. This means scientific and practical importance for ecological monitoring, assessment of plant resistance, and the introduction of biological agents in the technology of growing agricultural crops.
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