BIOLOGICAL PROPERTIES OF DIAZOTROPH AZOSPIRILLUM BRASILENSE ISOLATED FROM SPRING TRITICALE ROOTS

Objective. Study the biological properties of the diazotroph Azospirillum brasilense 10/1, promising for improving the nitrogen nutrition of spring triticale and obtaining a high quality crop. Methods. A strain of nitrogen-fixing bacteria A. brasilense 10/1 isolated from washed roots of spring triticale Oberih Kharkivskyi by accumulation cultures method using Dobreiner semi-liquid nitrogen-free medium. Nitrogen-fixing microorganisms were isolated on potato agar with succinic acid by the Dryhalsky method. Potential nitrogenase activity on washed roots of spring triticale plants and nitrogen-fixing activity of azospirilla in pure culture were measured by gas chromatography. Electron microscopic studies of bacterial cells were performed by the method of negative contrast with uranyl acetate. Identification of azospirilla was carried out on the basis of the study of morphological, cultural, physiological and biochemical characteristics and using molecular genetic methods (16S rRNA sequence analysis). The nucleotide sequences were compared with the corresponding sequences from the international database GenBank NCBI using BLAST software. The sensitivity of bacteria to antibiotics and cereal seed pesticides was tested by disk diffusion method. Results. The active strain of nitrogen-fixing bacteria, identified as Azospirillum brasilense 10/1, was obtained by analytical selection methods. The identity of the sequences of 16S rRNA of A. brasilense 10/1 with reference strains of A. brasilense in the GenBank NCBI database is 99.5 % to 99.6 %. Diazotroph A. brasilense 10/1 is sensitive to cefotaxime, norfloxacin, chloramphenicol, gentamicin, erythromycin, kanamycin, furadonin, resistant to polymyxin, ampicillin, oxacillin, amoxicillin, ciprofloxacin, ceftriaxone. Vitavax 200FF and Fundazole dressers do not affect the development of A. brasilense 10/1, Maxim Star 025 FS somewhat inhibits the development of bacteria. Conclusion. The active strain of nitrogen-fixing bacteria A. brasilense 10/1 isolated from washed roots of triticale by methods of analytical selection, is a promising inoculant to increase yields and improve grain quality of this crop. A. brasilense 10/1 is deposited in the Depository of the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine under number B- 7317 and is protected by the patent of Ukraine No. 104212.

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THE INFLUENCE OF BACTERIA OF GENUS AZOSPIRILLUM ON POTENTIAL NITROGENASE ACTIVITY AND BIOSYNTETIC PROCESSES OF SPRING WHEAT AND SPRING TRITICALE

Agriciltural microbiology ◽

10.35868/1997-3004.9.138-146 ◽

2009 ◽

Vol 9 ◽

pp. 138-146

Author(s):

O.O. Shahovnina ◽

O.V. Nadkernichna ◽

Y.O. Vorobey ◽

V.V. Krivopisha

Keyword(s):

Glutamine Synthetase ◽

Spring Wheat ◽

Root Zone ◽

Nitrogenase Activity ◽

Synthetase Activity ◽

Glutamine Synthetase Activity ◽

Nitrogen Fixing ◽

Nitrogen Fixing Bacteria ◽

Spring Triticale ◽

Reliable Increase

Use of new strain of nitrogen fixing bacteria Azospirillum sp. 77 for inoculation of spring wheat and spring triticale permitted to form an effective associative system diasotroph - plant. The bacterization promoted the reliable increase of potential nitrogenase activity in root zone of plant by 38-220 %, activated the biosynthetic processes, in particular, glutamine synthetase activity increased by 57,0-71,9 %, content of protein in leaves - by 9,7-16,3 %, top of the plants - by 11,9- 18,9 %, weight of the roots - by 7,2-7,3 %.

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Characterization of Azospirilla isolated from seeds and roots of turf grass

Canadian Journal of Microbiology ◽

10.1139/m88-040 ◽

1988 ◽

Vol 34 (3) ◽

pp. 212-217 ◽

Cited By ~ 10

Author(s):

Subramaniam Sundaram ◽

Alahari Arunakumari ◽

Robert V. Klucas

Keyword(s):

Azospirillum Brasilense ◽

Deoxyribonucleic Acid ◽

Reference Strain ◽

Nitrogenase Activity ◽

Nitrogen Fixing ◽

Dna Homology ◽

Nitrogen Fixing Bacteria ◽

Physiological Characterization ◽

Azospirillum Amazonense

Nitrogen-fixing bacteria were isolated from surface-sterilized seeds and roots of turf grasses growing in Nebraska. The percentage of successful isolations from surface-sterilized seeds ranged from 0 to 100 depending upon the source of the seeds. Based upon morphological and physiological characterization, some of the root and seed isolates appeared to be Azospirillum spp. Deoxyribonucleic acid homology studies on four selected isolates indicated that two root and one seed isolates were related to Azospirillum brasilense SP-7 with 70% or greater DNA homology, and one seed isolate was related to reference strain Azospirillum amazonense Y1 with 100% DNA homology. After repeated culturing on semisolid malate medium, some of the isolates lost their capacity to fix nitrogen. However, when certain non-nitrogen-fixing isolates were used as inoculants on turf grasses grown from surface-sterilized seeds which possessed no detectable indigenous nitrogen-fixing bacteria, nitrogenase activity as measured by acetylene reduction was detected.

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New marine nitrogen-fixing bacteria isolated from an eelgrass (Zostera marina) bed

Canadian Journal of Microbiology ◽

10.1139/m88-153 ◽

1988 ◽

Vol 34 (7) ◽

pp. 886-890 ◽

Cited By ~ 6

Author(s):

Wung Yang Shieh ◽

Usio Simidu ◽

Yoshiharu Maruyama

Keyword(s):

Zostera Marina ◽

Nitrogenase Activity ◽

Molecular Nitrogen ◽

Exponential Growth Phase ◽

Nitrogen Fixing ◽

Low Oxygen ◽

Nitrogen Fixing Bacteria ◽

Nitrogen Fixation Activity ◽

The Family ◽

Ph Decrease

Four strains of marine nitrogen-fixing bacteria were isolated from the roots of eelgrass (Zostera marina) and from sediments in an eelgrass bed in Aburatsubo Inlet, Kanagawa Prefecture, Japan. Significant levels of nitrogenase activity were detected in all four strains after a few hours of incubation under anaerobic conditions. Nitrogenase activity in all cases was Na+ dependent. These strains grew anaerobically or under conditions of low oxygen, using molecular nitrogen as the sole nitrogen source. Bacterial growth in liquid nitrogen-free medium was accompanied by a marked pH decrease during the exponential growth phase. Neither yeast extract nor vitamins were required for the nitrogen fixation activity of these strains. Taxonomically, all strains were facultatively anaerobic, Gram-negative rods. They were motile in liquid medium by means of a single polar flagellum and required NaCl for their growth. These characteristics, as well as the guanine + cytosine content of their DNA (43.5 – 44.8 mol%), placed them in the family Vibrionaceae. These strains, however, could not be identified to the genus level because they were distinct from the two halophilic genera Vibrio and Photobacterium of the family Vibrionaceae by a variety of characteristics.

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Research on Improvement of Nitrogenase Activity of Free-Living Nitrogen-Fixing Bacteria Using DBD Plasma

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.671-674.2674 ◽

2013 ◽

Vol 671-674 ◽

pp. 2674-2678 ◽

Cited By ~ 1

Author(s):

Yan Yun Zhu ◽

Xiao Li Zhu ◽

Fang She Yang

Keyword(s):

Rhizosphere Soil ◽

Barrier Discharge ◽

Nitrogenase Activity ◽

Nitrogen Fixing ◽

Bacterial Strains ◽

Dbd Plasma ◽

Nitrogen Fixing Bacteria ◽

Free Living ◽

Reduction Assay ◽

Rrna Sequencing

Nitrogen-fixing bacteria were screened from the rhizosphere soil of plants in Shaanxi in China. 36 free-living nitrogen-fixing bacterial strains were isolated and their nitrogenase activity were determined by acetylene reduction assay (ARA), two strains named FLNB03 and FLNB09 with higher nitrogenase activity were isolated and identified by 16S rRNA sequencing. The datum showed that FLNB03 was similar to Acinetobacter and their similarity reached 99%, FLNB09 was similar to Agrobacterium sp. and their similarity reached 99%. Then both of them were treated using Dielectric Barrier Discharge (DBD) plasma for mutation and their mutants called FLNB03-2 and FLNB09-3 were obtained. The nitrogenase activity of FLNB03-2 was 0.61±0.10 nmol•107cfu-1•h-1, and that of FLNB09-3 was 0.40±0.05 nmol•107cfu-1•h-1, their nitrogenase activity increased by 22.00% and 14.29% than their original bacteria respectively. FLNB03-2 and FLNB09-3 might be used as microbial fertilizer.

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Polyhydroxyalkanoate analysis inAzospirillum brasilense

Canadian Journal of Microbiology ◽

10.1139/m95-171 ◽

1995 ◽

Vol 41 (13) ◽

pp. 73-76 ◽

Cited By ~ 19

Author(s):

Robin Itzigsohn ◽

Oded Yarden ◽

Yaacov Okon

Keyword(s):

Azospirillum Brasilense ◽

Bacterial Species ◽

Carbon Sources ◽

Dry Mass ◽

Nitrogen Fixing ◽

Nitrogen Fixing Bacteria ◽

Qualitative And Quantitative ◽

Alkanoic Acids ◽

Quantitative Analyses

The considerable industrial interest in the qualitative and quantitative production of polyhydroxyalkanoates in microorganisms has led to the characterization of those synthesized in the nitrogen-fixing bacteria Azospirillum brasilense and Azotobacter paspali. In contrast to some other bacterial species, Azospirillum brasilense does not produce copolymers of hydroxyalkanoates when grown under the different carbon sources assayed, namely n-alkanoic acids, hydroxyalkanoates, and sugars with varying C:N ratios. Rather, only homopolymers of polyhydroxybutyrate were detected, comprising up to 70% of the cell dry mass. No copolymers were detected in Azotobacter paspali. Quantitative analyses of poly(β-hydroxybutyrate) are also presented.Key words: Azospirillum spp., Azotobacter paspali, polyhydroxyalkanoate analysis, PHA, PHB.

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Influence of ammonium and moisture on survival and nifH transcription in the diazotrophic Pseudomonas mendocina S10

Notulae Scientia Biologicae ◽

10.15835/nsb12110545 ◽

2020 ◽

Vol 12 (1) ◽

pp. 114-123

Author(s):

Sushma SHARMA ◽

Dileep K. SINGH

Keyword(s):

Nitrogenase Activity ◽

Physiological State ◽

Bacterial Count ◽

Crop Productivity ◽

Soil Conditions ◽

Pseudomonas Mendocina ◽

Nitrogen Fixing ◽

Nitrogen Fixing Bacteria ◽

Real Time Quantitative Pcr ◽

Moisture Conditions

Nitrogen is important for crop productivity and usually added in form of urea into the soil which negatively affects the environment. It is important to utilize nitrogen fixing bacteria for improving the nitrogen content of soil in India. Here, we have isolated nitrogen fixing-bacteria Pseudomonas mendocina S10 from rhizospheric soil and studied its nitrogenase activity along with its survival under sterile soil conditions. Enterobacterial Repetitive Intergenic Consensus (ERIC) PCR and Real-time quantitative PCR were employed to investigate the population and nifH transcripts level respectively in presence of ammonium and moisture additives. Strain S10 was capable of growth and expressing nifH transcripts in the presence of 2.5 mM ammonium and 20 percent water availability. Population of isolated strain and it’s nifH mRNA was found at low levels when exposed to 5 mM ammonium for 60 days of incubation period. However, viable bacterial count and nifH transcript levels remained low in the presence of 1.25 mM ammonium and zero percent water content. These findings indicate that isolated strain could tolerate ammonium up to 5 mM for 60 days and can maintained their cell viability in low moisture conditions. Results revealed the advantage of using gene expression to evaluate the physiological state of microorganism’s population in soil.

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THE INFLUENCE OF BRADYRHIZOBIUM JAPONICUM AND AZOSPIRILLUM BRASILENSE ON THE VERTICAL MIGRATION OF NUTRIENTS AT SOYBEAN CULTIVATION

Agriciltural microbiology ◽

10.35868/1997-3004.21.39-43 ◽

2015 ◽

Vol 21 ◽

pp. 39-43

Author(s):

S. F. Kozar ◽

I. M. Pyschur ◽

V. M. Nesterenko

Keyword(s):

Chlorophyll Content ◽

Vertical Migration ◽

Bradyrhizobium Japonicum ◽

Azospirillum Brasilense ◽

Water Soluble ◽

Nitrogen Fixing ◽

Nitrogen Fixing Bacteria ◽

Joint Application ◽

Calcium And Magnesium ◽

Soybean Plants

The paper presents the research results of pre-sowing seeds bacterization with nitrogen fixing bacteria Bradyrhizobium japonicum and Azospirillum brasilense influence on the loss of moisture, water soluble humus and nutrients. It was shown that seeds bacterization reduces the leaching intensity of nitrates, phosphorus, potassium, calcium and magnesium. Moreover, the least losses of nutrients were observed in a variants with joint application of both studied nitrogen fixing microorganisms. It was noted that seeds bacterization with B. japonicum and A. brasilense had promoted increase of chlorophyll content in the leaves of soybean plants. The highest yield was observed in the variant with the joint use of rhizobia and azospirillum.

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