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The aim of the study was to examine the effects of management practice on microbial properties of soil under soybean production. The study included 180 samples of soil under certified organic soybean production and 80 samples in conventional production system. An abundance of the examined microbial groups was assessed using the indirect dilution method, followed by plating of soil suspension on different selective media, while dehydrogenase and β-glucosidase activity was measured spectrophotometrically. Our data indicated that the management practice affected the structure and activity of microbial com­munities. A significant positive effect of organic farming on Azotobacter spp., free N-fixers and abundance of actinomycetes was identified. The influence of management system for the total number of bacteria, ammonifiers and fungi was not observed. Significantly higher dehydrogenase and β-glucosidase activity was recorded in the soils under organic farming compared to the conventional farming. The obtained results showed an increase in organic matter content, associated with organic soil management, and its positive correlation with soil microbial properties. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR 30122]

The objective of this study was to investigate the possibility of using microbial strains as residue decomposers and to determine the effect of these strains on chemical and microbial properties in the residue-amended soil. Greenhouse experiment consisted of eight Bacillus treatments, three Trichoderma treatments, and their combination, all applied to non-sterile chernozem soil amended with wheat straw. Incorporation of wheat straw improved soil chemical and microbial properties, while the extent of residue decom­position under microbial strains was intensified. Microbial treatments significantly affected the soil pH, the content of carbonate, total carbon, soil organic carbon, humus, and available phosphorus and potassium. Bacterial and fungal treatments also significantly influenced the total microbial number, ammonifiers, N2-fixers, fungi, actinomycetes, oligotrophs, copiotrophs, and cellulolytic microorganisms. The effect of microbial treatments varied depending on the applied strains and examined properties, with Bacillus strains being more promising residue decomposers compared to Trichoderma strains. The most effective microbial strains could be used as potential decomposers of crop residues. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR31072]

The objective of this study was to assess the effect of biochar soil amendment (BSA) on chemical and microbial properties in different agricultural soils in Vojvodina Prov­ince. Short-term pot experiment consisted of five biochar application doses (0, 0.5, 1, 2, and 3%) and five contrasting soil types (Mollic Gleysol, Eutric Cambisol, Calcaric Fluvisol, Gleyic Chernozem, and Haplic Chernozem), planted with sunflower (Helianthus annuus L.) and winter wheat (Triticum aestivum L.). The examined chemical and microbial properties were significantly influenced by soil type and interaction of experimental factors. Significant influence of biochar on the contents of calcium carbonate (CaCO3), total nitrogen (N), total carbon (C), soil organic carbon (SOC), humus and potassium (K) of the tested soils was observed. Biochar also significantly affected the number of azotobacters (AZB), fungi (FNG), actino­mycetes (ACT) and copiotrophic bacteria (CB). The effect of BSA varied depending on the applied dose, with higher values of the examined chemical and microbial parameters at higher doses of application. Further studies on using biochar in soils with low fertility will be neces­sary to establish its efficiency as an enhancer for agricultural production in Serbia. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR31072]

The study monitored microbial abundance and dehydrogenase activity in chernozem soil under different cropping systems. Soil samples were taken from the long-term trials Crop Rotations and IOSDV, at the Rimski Šančevi experimental field of the Institute of Field and Vegetable Crops. The soil samples were collected in two years from ten cropping systems at three sampling depths. Number of the examined microbial groups was assessed by the indirect dilution method, while dehydrogenase enzyme activity was measured spectrophotometrically. Number of the tested microbial groups and dehydrogenase activity varied significantly depending on the cropping system and year of study, while sampling depth significantly affected enzyme activity. The highest number of microorganisms was obtained in non-agricultural soil and unfertilized soil under 2-year and 3-year rotation, while the highest dehydrogenase activity was recorded in non-agricultural soil and soil under wheat grown in monoculture.

Aim of the study was to examine the effects of soil use on soil microbial properties at five locations in Central Serbia. The study included 42 samples of soil under certified organic system, 8 samples under conventional system, and 30 samples of abandoned soils. The number of examined microbial groups was assessed using the indirect dilution method followed by plating of soil suspension on different selective media, while dehydrogenase activity was measured spectrophotometrically. The number and activity of microorganisms at the tested plots primarily depended on the soil pH. The acidic soil reaction caused more rapid fungal growth and lower presence of actinomycetes and Azotobacter sp. There were no significant alterations in the number of microorganisms, ammonifiers and fungi under the specific soil use system. Significantly higher dehydrogenase activity was recorded in the abandoned soils and soils under organic systems compared to the conventional farming.

During the last decades, intensive application of synthetic fungicides in the agricultural crop protection practice caused growing concern for the existence of toxic chemical residues in food as well as in the whole environment. Instead of using synthetic fungicides, it is suggested that crop protection be carried out by using preparations based on compounds of natural origin (secondary metabolites of plants or microorganisms, including macrofungi from Basidiomycota) as biological control agents. The potential of macrofungal species as biocontrol agents was analyzed in this investigation of eight autochthonous species from different locations in Serbia. Both the terricolous species: Coprinus comatus, Coprinellus truncorum, Amanita strobiliformis, Hydnum repandum and the lignicolous species: Flammulina velutipes, Stereum subtomentosum, Trametes versicolor and Bjerkandera adusta were examined, with an aim to detect some novel sources of antifungal agents. This study surveyed antifungal activity of selected macrofungal extracts (MeOH, EtOH and CHCl3) against phytopathogenic Fusarium and Alternaria strains isolated from garlic, soybean and rice: F. proliferatum, F. verticillioides, F. proliferatum, F. graminearum and A. padwickii. Microdilution method in 96 well microplates was applied for the estimation of antifungal effects of macrofungi extracts in the range from 24.75 to 198.00 mg/ml and determination of minimal inhibitory (MIC) and minimal fungicidal concentration (MFC). EtOH extract of mychorhizal species H. repandum showed antifungal activity against all analyzed phytopathogenic strains, with the strongest effect on Fusarium strains (MIC 24.75 mg/ml; MFC 24.75 mg/ml). Among others, MeOH extracts of S. subtomentosum and C. micaceus showed similar effects while only B. adusta showed slight effect on Fusarium strains (MIC 24.75-99.00 mg/ml; MFC 24.75-99.00 mg/ml) and none effect on A. padwickii. The obtained results indicate the possibility of using examined extracts as efficient antifungal agents and provide the basis for the new formulations of biocontrol agents against phytopathogenic fungi in the future.

The aim of this study was to isolate and identify Bradyrhizobium japonicum strains on the basis of molecular characteristics. From root nodules of different soybean cultivars were obtained 56 isolates, characterized according to morphological, cultural, and biochemical properties. Among these isolates, 33 isolates showing resemblance with Bradyrhizobium sp. were further subjected to molecular identification. Following DNA extraction, a partial 16S rDNA gene sequence from the isolates was amplified by PCR using universal primers fD1 (27F) and rP3 (1492R). Purification and sequencing of the amplified fragments were done in the bio­technology company Macrogen, Seoul, South Korea. Sequences were analyzed using the pro­gram FinchTV and BLAST (Basic Local Alignment Search Tool) and compared to sequences in GenBank and the Bradyrhizobium ID-database for identification. Comparison of the se­quences with the Bradyrhizobium ID-database showed that all tested isolates were identified as Bradyrhizobium japonicum. Each isolate was deposited in the NCBI GenBank database under a unique accession number. Identification of Bradyrhizobium species from root nodules of soybean is of great importance because the symbiosis between rhizobia and legumes are a cheaper and usually more effective agronomic practice for ensuring an adequate supply of nitrogen for legumes, while preserving and improving fertility and productivity of soils. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 31022]

Biocontrol using plant growth-promoting rhizobacteria (PGPR) represents an alternative approach to disease management, since PGPR are known to promote growth and reduce diseases in various crops. Among the different PGPR, members of the genus Bacillus are prefered for most biotechnological uses due to their capability to form extremely resistant spores and produce a wide variety of metabolites with antimicrobial activity. The objective of this research was to identify antagonistic bacteria for management of the plant diseases. Eleven isolates of Bacillus spp. were obtained from the soil samples collected from different localities in the Province of Vojvodina. The antifungal activity of bacterial isolates against five fungal species was examined using a dual plate assay. Bacillus isolates exhibited the highest antifungal activity against Fusarium proliferatum, Fusarium oxysporum f. sp. cepae and Alternaria padwickii, while they had the least antagonistic effect on Fusarium verticillioides and Fusarium graminearum. Molecular identification showed that effective bacterial isolates were identified as Bacillus safensis (B2), Bacillus pumilus (B3, B11), Bacillus subtilis (B5, B7) and Bacillus megaterium (B8, B9). The highest antagonistic activity was exhibited by isolates B5 (from 39% to 62% reduction in fungal growth) and B7 (from 40% to 71% reduction in fungal growth). These isolates of B. subtilis could be used as potential biocontrol agents of plant diseases. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR-31073]

Garlic (Allium sativum L.) is considered to be one of the oldest crops in the world. During 2016, infected garlic bulbs occurred in storages on several localities of the Province of Vojvodina. Symptomatic cloves showed typical rot symptoms such as softened and spongy areas covered with white fungal growth with deep lesions formed on the cloves which became dry over time. A total of 36 isolates of Fusarium species were obtained from diseased cloves of garlic. Colony morphology and microscopic properties of isolated Fusarium species were recorded from the cultures grown on PDA and CLA, respectively. Identification of two chosen isolates was performed by sequencing the EF-1α gene. The TEF sequence of isolate JBL12 showed 100% similarity with several F. tricinctum sequences and sequence of JBL539 showed 99% identity with several F. acuminatum sequences and they were deposited in the NCBI GenBank. Based on the results of the morphological and molecular identification, isolates JBL12 and JBL539 were identified as F. tricinctum and F. acuminatum, respectively, as new causal agents of garlic bulbs rot in Serbia. Specific primers were designed for the PCR identification of the F. tricinctum. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. TR31030]

The objective of this study was to compare the effects of conventional and organic production system on microbial activity in the soil cultivated with bean and maize crops. The trial in Đurđevo was set up according to the conventional farming system, while organic farming system was used in Futog. Two maize hybrids and two bean cultivars were used in the trial. Soil samples were collected in two periods during 2014 (before sowing, at flowering stage of bean crops, and at 9-11 leaf stage of maize) at two depths, at both locations. The following microbiological parameters were tested: the total number of micro­organisms, number of ammonifiers, Azotobacter sp., free nitrogen fixing bacteria, fungi, actinomycetes, and activity of dehydrogenase enzyme. The results showed that the total number of microorganisms, number of free N-fixers and dehydrogenase activity were higher within organic production, while Azotobacter sp. was more abundant in conventional production. Variations in the number of ammonifiers, fungi and actinomycetes in relation to the type of production were not obtained. Significant differences in microbial activity were also obtained between period and depths of sampling.

The effect on sugar beet yield parameters and microbiological soil status was studied using two techniques of sugar beet inoculation with strains of Azotobacter chroococcum. Cultivar “Drena” was used in the study, and field trial was set under the conditions of organic farming system in Bački Petrovac. A mixture of three strains of Azotobacter chroococcum was used as microbial fertilizer. Inoculation was performed by: (A) incorporation of strains into soil before sowing; and (B) repeated incorporation of strains into soil two weeks after sowing. PGP characterization of the strains confirmed the ability of producing indole-3-acetic acid (IAA) from 12.63 μg ml-1 to 14.95 μg ml-1, nitrogen fixation, and P-solubilization. Positive effects on the number of azotobacter and free nitrogen fixers in rhizosphere were obtained by inoculation, as well as positive effects on the tested sugar beet yield parameters. The largest increase in root yield, yield of crystal sugar, and yield of polarised sugar compared with the control was obtained by repeated soil inoculation, ranging from 22 to 23%.

Plant growth promoting rhizobacteria (PGPR) represent a wide variety of bacteria inhabiting the root surface and root vicinity. They are directly or indirectly involved in promoting plant growth and development via production and secretion of various regulatory compounds in the rhizosphere. PGPR are generally beneficial to a wide range of crops including maize. The main objective of this study was to determine the effect of PGPR strains on the microbial abundance in maize rhizosphere. The trial was conducted at Rimski Šančevi experimental field of Institute of Field and Vegetable Crops in Novi Sad, Serbia. The trial included five maize hybrids (NS 3014, NS 4015, NS 5043, NS 6010 and NS 6030) developed at the Institute and three treatments with microorganisms. Non-inoculated treatment was control. Inoculation was performed with Bacillus Q7, Pseudomonas PS2 and their mixture with Azotobacter chroococcum (Q7 + PS2 + AC). Application method was incorporation immediately before planting with liquid culture of strains (density 109 CFU/ml). It was observed that the certain treatment with rhizobacteria increased the number of all three studied microbial groups in the rhizosphere of the same hybrid. The best effect on the number of microorganisms in the rhizosphere of NS 6010 was achieved with Q7 + PS2 + AC, in NS 5043 with Q7 and with PS2 in NS 4015. The results obtained in this study indicate the importance of PGPR in crop production, which requires a constant expansion of knowledge about the mutual interactions of plants and microorganisms.

Аmong 50 bacterial isolates obtained from maize rhizospherе, 13 isolates belonged to the genus Azotobacter. Isolates were biochemically characterized and estimated for pH and halo tolerance ability and antibiotic resistance. According to characterization, the six representative isolates were selected and further screened in vitro for plant growth promoting properties: production of indole-3-acetic acid (IAA), siderophores, hydrogen cyanide (HCN), exopolysaccharides, phosphate solubilization and antifungal activity (vs. Helminthosporium sp., Macrophomina sp., Fusarium sp.). Beside HCN production, PGP properties were detected for all isolates except Azt7. All isolates produced IAA in the medium without L-tryptophan and the amount of produced IAA increased with concentration of precursor in medium. The highest amount of IAA was produced by isolates Azt4 (37.69 and 45.86 μg ml-1) and Azt5 (29.44 and 50.38 μg ml-1) in the medium with addition of L-tryptophan (2.5 and 5 mM). The isolates showed the highest antifungal activity against Helminthosporium sp. and the smallest antagonistic effect on Macrophomina sp. Radial Growth Inhibition (RGI) obtained by the confrontation of isolates with tested phytopathogenic fungi, ranged from 10 to 48%. [Projekat Ministarstva nauke Republike Srbije, br. TR 31073]

The dynamics of phosphorus and sulphur in soil is closely related to the dynamics of the biological cycle in which microorganisms play a central role. There is not much microbiological activity in acid soils because aerobes are scarce, rhizosphere is restricted to the shallow surface layer, and the biomass of microorganisms decreases with higher acidity. The aim of the research was to investigate the number of microorganisms, which decompose organic and inorganic phosphorus compounds and organic sulphur compounds in calcocambisol, luvisol, and pseudogley. The following parameters were determined in the soil samples: pH in H2O and in 1MKCl; the content of CaCO3 (%); humus content (%), nitrogen content (%); the content of physiologically active phosphorus and potassium (mg P2O5/100g of soil; mg K2O/100g of soil). The number of microorganisms was determined by the method of agar plates on appropriate nutrient media: the number of microorganisms solubilizing phosphates on a medium by Muramcov; the number of microorganisms that decompose organic phosphorus compounds on a medium with lecithin; and the number of microorganisms that transform organic sulphur compounds on a medium by Baar. All three types of soil are acid non-carbonate soils with a low level of available phosphorus and a more favorable amount of potassium, nitrogen, and humus. The largest number of bacteria, which transform organic phosphorus compounds, was found in calcocambisol. The largest number of phosphate solubilizing bacteria was recorded in pseudogley, whereas the largest number of phosphate solubilizing fungi was recorded in calcocambisol. The largest number of bacteria, which transform organic sulphur compounds, was recorded in pseudogley.

Free-living soil bacteria beneficial to plant growth are usually referred to as plant growth promoting rhizobacteria (PGPR), capable of promoting plant growth by colonizing the plant root. Application of PGPR to increase the yield is limited by variability among the results obtained in the laboratory, in greenhouse and field. Rhizobacteria that promote plant growth (PGPR) participate in interactions with plants (rice, wheat, maize, sugarcane, sugar beet, cotton) and significantly increase their vegetative growth and yield. Apart from Azotobacter and Azospirillum, PGPR also include Acetobacter, Azoarcus and several species of Enterobacteriaceae (Klebsiella, Enterobacter, Citrobacter, and Pseudomonas). PGPR represent an alternative to plant growth enhancement chemicals.

The aim of this study was to determine the effects of three different inoculation methods with selected Azotobacter chroococcum strains on productive and technological traits of sugar beet, as well as on the total number of microorganisms and azotobacter in rhizosphere. The results of this two-year study showed that effectiveness of the tested inoculation methods in increasing root yield and sugar content varies greatly, depending on year and azotobacter strains. Effectiveness of inoculation methods was not largely impacted by year on granulated sugar. Achieved granulated sugar yield was significantly higher by using pre-sowing azotobacter application, than by using seed inoculation. A significantly increased number of microorganisms in sugar beet rhizosphere was determined, not only by using pre-sowing azotobacter application but also by using sugar beet seed inoculation. Pre-sowing azotobacter application and inter-row cultivation both caused an equal increase in the number of these bacteria in sugar beet rhizosphere (42.2% and 46.9%). Use of sugar beet seed inoculation caused an increase of 33.7% in the number of azotobacter. In order to achieve higher effectiveness in applying azotobacter on productive and technological traits of sugar beet, and considering determined interaction between a certain year, an inoculation method and a strain, it is necessary for future research to focus on determining efficiency of these strains when they are in a mixture.

Plant growth promoting rhizobacteria (PGPR) are a group of diverse soil bacteria which stimulate the growth of the host plant. It has been shown that Azotobacter chroococcum may be used as biofertilizer for increasing the yield and improving technological characteristics of sugar beet. The effect of three different ways of inoculation of sugarbeet with Azotobacter chroococcum on the production features of sugar beet and microbiological status of rhizosphere soil has been tested. One variety of sugar beet, Drena, was included in the tests in the region Rimski Šančevi, Novi Sad, during 2008 and 2009. Five strains of Azotobacter chroococcum (1, 5, 8, 10 and 14) were used as microbiological fertilizers. There were three variations of inoculation: (A) incorporation in the soil, (B) before the fi rst cultivation, and (C) applying the liquid culture of strain on the seed before sowing. The highest increase in yield of sugar beet roots 6.25 t ha-1 and yield of white sugar 0.91 t ha-1 was achieved with strain 10 with variant of inoculation of soil before the first cultivation. In both years, a positive effect of Azotobacter chroococcum was observed on the total number of microorganisms in the rhizosphere.