Bacteria Azotobacter Chroococcum Research Articles

BIOLOGICAL ACTIVITY OF RH IZOSPHERE SOIL SPRING WHEAT IN ASSOCIATION WITH BACTERIA AZOTOBACTER CHROOCOCCUMKZT79. MODIFIED N-ACETYL-D-GLUCOSAMINE

Aim. The study of the biological activity of rhizosphere soil of the plants of spring wheat in association with bacteria Azotobacter chroococcum T79, modified N-acetyl-D-glucosamine. Methods. The biological activity of the rhizosphere soil such as the growth-activating ability (method of phytotest), number (by quantitative dilutions of the soil suspension, plating on selective medium growth Eshbi, and counting bacterial colonies) and nitrogenase activity (acetylene method) of nitrogen-fixing microorganisms in the functioning of the association of spring wheat with bacteria Azotobacter chroococcum T79 in combination with N-acetyl-D-glucosamine (0.1 M) in the greenhouse experiments were study. Results. It was shown the significant increase of the number (in 1.4–3.9 times) and nitrogenase activity (in 2.1–5.6 times) of nitrogen-fixing microorganisms during the vegetation of wheat plants as well as the growth-activating ability of the soil (in 1.2 times) in the early stage of formation phytobacteriology association. Conclusion. The treatment of wheat seeds by Azotobacter chroococcum T79, modified N-acetyl-D-glucosamine vs. azotobacter monoinoculant was promoted activation of the functioning of the bacterial nitrogenase (by 10–38 %) and increasing populations of nitrogen-fixing microorganisms (in 1.2–2.2 times) more than their ability to accumulate biologically active substances in the rhizosphere soil of plants.

The Gram-negative bacterium Azotobacter chroococcum NCIMB 8003 employs a new glycoside hydrolase family 70 4,6-α-glucanotransferase enzyme (GtfD) to synthesize a reuteran like polymer from maltodextrins and starch

BackgroundOriginally the glycoside hydrolase (GH) family 70 only comprised glucansucrases of lactic acid bacteria which synthesize α-glucan polymers from sucrose. Recently we have identified 2 novel subfamilies of GH70 enzymes represented by the Lactobacillus reuteri 121 GtfB and the Exiguobacterium sibiricum 255-15 GtfC enzymes. Both enzymes catalyze the cleavage of (α1→4) linkages in maltodextrin/starch and the synthesis of consecutive (α1→6) linkages. Here we describe a novel GH70 enzyme from the nitrogen-fixing Gram-negative bacterium Azotobacter chroococcum, designated as GtfD. MethodsThe purified recombinant GtfD enzyme was biochemically characterized using the amylose-staining assay and its products were identified using profiling chromatographic techniques (TLC and HPAEC-PAD). Glucans produced by the GtfD enzyme were analyzed by HPSEC-MALLS-RI, methylation analysis, 1D/2D [1]H/[13]C NMR spectroscopy and enzymatic degradation studies. ResultsThe A. chroococcum GtfD is closely related to GtfC enzymes, sharing the same non-permuted domain organization also found in GH13 enzymes and displaying 4,6-α-glucanotransferase activity. However, the GtfD enzyme is unable to synthesize consecutive (α1→6) glucosidic bonds. Instead, it forms a high molecular mass and branched α-glucan with alternating (α1→4) and (α1→6) linkages from amylose/starch, highly similar to the reuteran polymer synthesized by the L. reuteri GtfA glucansucrase from sucrose. ConclusionsIn view of its origin and specificity, the GtfD enzyme represents a unique evolutionary intermediate between family GH13 (α-amylase) and GH70 (glucansucrase) enzymes. General significanceThis study expands the natural repertoire of starch-converting enzymes providing the first characterization of an enzyme that converts starch into a reuteran-like α-glucan polymer, regarded as a health promoting food ingredient.

Inoculation effect of endophytic fungus Piriformospora indica and bacteria Azotobacter chroococcum on activity of antioxidant enzymes and wheat tolerance to zinc deficiency (Niknejad cultivar) in greenhouse conditions

Inoculation effect of endophytic fungus Piriformospora indica and bacteria Azotobacter chroococcum on activity of antioxidant enzymes and wheat tolerance to zinc deficiency (Niknejad cultivar) in greenhouse conditions

Interrelated effects of mycorrhiza and free-living nitrogen fixers cascade up to aboveground herbivores.

Aboveground plant performance is strongly influenced by belowground microorganisms, some of which are pathogenic and have negative effects, while others, such as nitrogen-fixing bacteria and arbuscular mycorrhizal fungi, usually have positive effects. Recent research revealed that belowground interactions between plants and functionally distinct groups of microorganisms cascade up to aboveground plant associates such as herbivores and their natural enemies. However, while functionally distinct belowground microorganisms commonly co-occur in the rhizosphere, their combined effects, and relative contributions, respectively, on performance of aboveground plant-associated organisms are virtually unexplored. Here, we scrutinized and disentangled the effects of free-living nitrogen-fixing (diazotrophic) bacteria Azotobacter chroococcum (DB) and arbuscular mycorrhizal fungi Glomus mosseae (AMF) on host plant choice and reproduction of the herbivorous two-spotted spider mite Tetranychus urticae on common bean plants Phaseolus vulgaris. Additionally, we assessed plant growth, and AMF and DB occurrence and density as affected by each other. Both AMF alone and DB alone increased spider mite reproduction to similar levels, as compared to the control, and exerted additive effects under co-occurrence. These effects were similarly apparent in host plant choice, that is, the mites preferred leaves from plants with both AMF and DB to plants with AMF or DB to plants grown without AMF and DB. DB, which also act as AMF helper bacteria, enhanced root colonization by AMF, whereas AMF did not affect DB abundance. AMF but not DB increased growth of reproductive plant tissue and seed production, respectively. Both AMF and DB increased the biomass of vegetative aboveground plant tissue. Our study breaks new ground in multitrophic belowground–aboveground research by providing first insights into the fitness implications of plant-mediated interactions between interrelated belowground fungi–bacteria and aboveground herbivores.

Combined effects of Piriformospora indica and Azotobacter chroococcum enhance plant growth, antioxidant potential and steviol glycoside content in Stevia rebaudiana

The herb Stevia rebaudiana Bertoni is an important sugar substitute due to the presence of steviol glycosides-mainly stevioside and rebaudioside-A. These compounds possess antioxidant activity with industrial importance. The fungus Piriformospora indica has been used to improve medicinally important biomolecules, growth and secondary metabolite content in plants. The soil bacterium Azotobacter chroococcum has also been reported to possess plant growth promoting ability. The present study was undertaken to test the ability of P. indica (Pi) and A. chroococcum (Az), alone and in combination to improve the growth, antioxidant activity and steviol glycoside (SG) content of in vitro S. rebaudiana plantlets. Inoculation with Pi or Az alone showed enhancement in all these characters compared with non-inoculated plantlets. Combined inoculation with Pi and Az further enhanced these traits.

Azotobacter Genomes: The Genome of Azotobacter chroococcum NCIMB 8003 (ATCC 4412).

The genome of the soil-dwelling heterotrophic N2-fixing Gram-negative bacterium Azotobacter chroococcum NCIMB 8003 (ATCC 4412) (Ac-8003) has been determined. It consists of 7 circular replicons totalling 5,192,291 bp comprising a circular chromosome of 4,591,803 bp and six plasmids pAcX50a, b, c, d, e, f of 10,435 bp, 13,852, 62,783, 69,713, 132,724, and 311,724 bp respectively. The chromosome has a G+C content of 66.27% and the six plasmids have G+C contents of 58.1, 55.3, 56.7, 59.2, 61.9, and 62.6% respectively. The methylome has also been determined and 5 methylation motifs have been identified. The genome also contains a very high number of transposase/inactivated transposase genes from at least 12 of the 17 recognised insertion sequence families. The Ac-8003 genome has been compared with that of Azotobacter vinelandii ATCC BAA-1303 (Av-DJ), a derivative of strain O, the only other member of the Azotobacteraceae determined so far which has a single chromosome of 5,365,318 bp and no plasmids. The chromosomes show significant stretches of synteny throughout but also reveal a history of many deletion/insertion events. The Ac-8003 genome encodes 4628 predicted protein-encoding genes of which 568 (12.2%) are plasmid borne. 3048 (65%) of these show > 85% identity to the 5050 protein-encoding genes identified in Av-DJ, and of these 99 are plasmid-borne. The core biosynthetic and metabolic pathways and macromolecular architectures and machineries of these organisms appear largely conserved including genes for CO-dehydrogenase, formate dehydrogenase and a soluble NiFe-hydrogenase. The genetic bases for many of the detailed phenotypic differences reported for these organisms have also been identified. Also many other potential phenotypic differences have been uncovered. Properties endowed by the plasmids are described including the presence of an entire aerobic corrin synthesis pathway in pAcX50f and the presence of genes for retro-conjugation in pAcX50c. All these findings are related to the potentially different environmental niches from which these organisms were isolated and to emerging theories about how microbes contribute to their communities.

Metal ion induced dissolution of melanin extracted from the bacterium Azotobacter chroococcum

Metal ion induced dissolution of melanin extracted from the bacterium Azotobacter chroococcum

Melanin from the Nitrogen-Fixing Bacterium Azotobacter chroococcum: A Spectroscopic Characterization

Melanins, the ubiquitous hetero-polymer pigments found widely dispersed among various life forms, are usually dark brown/black in colour. Although melanins have variety of biological functions, including protection against ultraviolet radiation of sunlight and are used in medicine, cosmetics, extraction of melanin from the animal and plant kingdoms is not an easy task. Using complementary physicochemical techniques (i.e. MALDI-TOF, FTIR absorption and cross-polarization magic angle spinning solid-state 13C NMR), we report here the characterization of melanins extracted from the nitrogen-fixing non-virulent bacterium Azotobacter chroococcum, a safe viable source. Moreover, considering dihydroxyindole moiety as the main constituent, an effort is made to propose the putative molecular structure of the melanin hetero-polymer extracted from the bacterium. Characterization of the melanin obtained from Azotobacter chroococcum would provide an inspiration in extending research activities on these hetero-polymers and their use as protective agent against UV radiation.

Plant Growth Promotion Rhizobacteria in Onion Production

The aim of the research was to examine the effect of rhizospheric bacteria Azotobacter chroococcum, Pseudomonas fluorescens (strains 1 and 2) and Bacillus subtilis on the growth and yield of onion and on the microorganisms in the rhizosphere of onion. The ability of microorganisms to produce indole-acetic acid (IAA), siderophores and to solubilize tricalcium phosphate (TCP) was also assessed. The experiment was conducted in field conditions, in chernozem type of soil. Bacillus subtilis was the best producer of IAA, whereas Pseudomonas fluorescens strains were better at producing siderophores and solubilizing phosphates. The longest seedling was observed with the application of Azotobacter chroococcum. The height of the plants sixty days after sowing was greater in all the inoculated variants than in the control. The highest onion yield was observed in Bacillus subtilis and Azotobacter chroococcum variants. The total number of bacteria and the number of Azotobacter chroococcum were larger in all the inoculated variants then in the control. The number of fungi decreased in most of the inoculated variants, whereas the number of actinomycetes decreased or remained the same.

English

The biomass of the bacterium Azotobacter chroococcum can be used as a biofertilizer due to its ability to fix nitrogen from the atmosphere. In order to optimize the production of bacterial biomass for this purpose, a cultivation of A. chroococcum was done by using different media and cultivation techniques (batch, fed batch and repeated batch). Chemically defined and complex media with 20 g/l of sugar were selected as the most appropriate media for batch cultivation in stirred tank bioreactor. In order to obtain higher fed batch and repeated batch techniques were examined. During these cultivations, increase of bioprocess efficiency parameters (yield coefficient and productivity) were observed compared with the batch cultivation. On the basis of the obtained results, repeated batch technique appeared to be the most suitable for the bacterial biomass production at industrial scale.   Key words: Azotobacter chroococcum, chemically defined and complex media, different cultivation techniques, biofertilizers.

A new phenol oxidase produced during melanogenesis and encystment stage in the nitrogen-fixing soil bacterium Azotobacter chroococcum

Laccases are copper-containing phenol oxidases that are commonly found in many types of plant, insect, fungi and bacteria. Whilst phenol oxidases have been well characterized in fungal species, laccase-type enzymes originating from bacteria have been much less well defined. Bacteria belonging to the family Azotobacteraceae share many morphological characteristics with strains already known to exhibit polyphenol and phenol oxidase activity; and hence the aim of this work was to identify and characterize a novel laccase from the isolated strain Azotobacter chroococcum SBUG 1484 in an attempt to provide further understanding of the roles such enzymes play in physiological development. Laccase activity was clearly observed through oxidation of 2,6-dimethoxyphenol, other typical substrates including: methoxy-monophenols, ortho- and para-diphenols, 4-hydroxyindole, and the non-phenolic compound para-phenylenediamine. A. chroococcum SBUG 1484 showed production of a cell-associated phenol oxidase when grown under nitrogen-fixing conditions, and was also observed when cells enter the melanogenic and encystment stages of growth. Catechol which is structurally related to melanin compounds was also released from Azotobacter cells into the surrounding culture medium during nitrogen-fixing growth. From our results we propose that a membrane-bound laccase plays an important role in the formation of melanin, which was monitored to correlate with progression of A. chroococcum SBUG 1484 cells into the encystment stage of growth.

Enhancement of wood waste decomposition by microbial inoculation prior to vermicomposting

To investigate the feasibility of microbial pre-decomposition of timber wastes to quality production of vermicompost with higher agronomic value, timber wastes were inoculated with different combinations of the fungi Phanerochete chrysosporium, Trichoderma reesei, Aspergillus niger and the bacteria Azotobacter chroococcum (MTCC 3853) and Bacillus cereus (MTCC 4079) and incubated at 28-30 °C in a mechanical composter. The inoculation enhanced the degradation of timber wastes, increased total nitrogen and improved the quality and enhanced production of vermicompost generated with the native earthworm Drawida willsi Michelsen. Total nitrogen increased from 0.16% to 1.52% and total organic carbon (TOC) decreased from 42% to 13%. Out of 10 microbial combinations tested for pre-decomposition, the combination of P. chrysosporium+T. reesei was found best in terms of ligno-cellulosic decomposition, and P. chrysosporium+A. niger+B. cereus with respect of cast output. The study shows that microbial pre-decomposition of timber wastes to produce quality vermicompost is a feasible technology.

Gardian and Pivot Herbicide Effect on Some Non-Symbiotic Nitrogen Fixing Bacteria

ABSTRACTStudying the ecological risk of the herbicides used in modern agriculture is very important for environment protection from pollution. Therefore investigations of herbicide effect on important for soil fertility and plant nutrition microorganisms is actual and necessary. An ecological assessment of two herbicides/gardian and pivot/was done by measuring the bacterial growth of non-symbiotic nitrogen fixing bacteria—Azotobacter chroococcum 18 and two strains of Azospirillum brasilense—Sp. 7 and Sp. 107. Inhibition of bacterial growth is observed and it depends on the herbicide used, its concentration and the bacterial strain. Three rates of herbicides were studied. Gardian inhibition on the bacterial growth varies from 22 % to 75%. The decrease of the size of bacterial colonies of A. brasilense Sp. 7, when the highest concentration is used is 75%; in case of A. brasilense Sp. 107 - it is 50%. In case of pivot application A. brasilense Sp. 107 is more resistant again/inhibition varies from 17% to 50%/; for A. brasilense Sp. 7 it is respectively 33% and 56%. In case of A. chrococcum the inhibition of the highest concentration of gardian is 56% and of pivot - 50%. The results show that gardian is more toxic than pivot as concerning the growth of the bacterial species studied.

Compost production from Municipal Solid Waste (MSW) employing bioinoculants

The effect of antibiotic producer fungi (Penicillium chrysogenum, P. funiculosum) and free living nitrogen fixing bacteria (Azotobacter chroococcum) on the quality of vermicompost prepared from MSW was observed in the present study. Prior to vermicomposting of MSW the interaction studies among bioinoculants and E. fetida, in Farm Yard Manure (FYM), was also carried out to see the effect of bioinoculants on the growth and fecundity of earthworms. This combination of bioinoculants (A. chroococcum, P. chrysogenum and P. funiculosum) proved to be better, in terms of quality of vermicompost, than all other combinations tried.

Contribution of Biofertilizers and Fertilizer Nitrogen to Nutrient Uptake and Yield of Egyptian Winter Wheat

ABSTRACT Nutrient uptake and grain and straw yield of Egyptian winter wheat (Triticum aestivum L. Merr.) were evaluated for two site-years after the seed inoculation with two biofertilizer products, Phosphorien, containing the phosphorus (P)-solubilizing bacteria Bacillus megatherium, and Nitrobien, containing a combination of nitrogen (N)-fixing bacteria Azotobacter chroococcum and Azospirillum liposerum. Ammonium nitrate and polymer-coated urea fertilizers were applied to plots alone and together with the biofertilizers at rates of either 83 kg N ha−1 or 186 kg N ha−1 for comparison. The highest grain yield (5.76–6.74 Mg ha−1) and straw yield (11.49–13.32 Mg ha−1) occurred at the highest fertilizer rates with N fertilizer. There was a slight additional increase in grain and straw yields when a biofertilizer was applied along with N fertilizer. A slightly higher grain and straw yield was measured with the polymer-coated urea treatment than with the ammonium nitrate treatment. The biofertilizer materials were not as effective as N fertilizers in producing grain (4.02–4.09 Mg ha−1) or straw (7.71–8.11 Mg ha−1) for either year, although the Nitrobien + Phosphorien combination increased these parameters over the N-fertilizer control. The effect of the Nitrobien biofertilizer in increasing grain yields was equivalent to a urea application rate of about 13 kg N ha−1. Biofertilizer inoculations increased iron (Fe), manganese (Mn), zinc (Zn), and copper (Cu) concentrations in wheat tissue (at boot stage), but these higher levels did not influence grain or straw yield.

5-n-Alkylresorcinols from the Nitrogen-fixing Soil Bacterium Azotobacter chroococcum Az12

A mixture of five saturated 5-n-alkylresorcinol homologues was isolated from vegetative cells of the nitrogen-fixing soil bacterium Azotobacter chroococcum Az12. Their structures were established by spectrometry (1H NMR, EI-MS, FAB-MS, FAB-MS/MS) and chromatography (GC, TLC) means.

EFFECTS OF TRIFLURALIN ON SOIL MICROBIAL POPULATIONS AND THE NITROGEN FIXATION ACTIVITIES

Effects of trifluralin on soil microbial populations and the nitrogen fixation activity of nitrogen-fixing bacteria Azotobacter chroococcum and Bradyrhizobium japonicum and the decomposition of trifluralin by soil microorganisms were studied. Trifluralin at lower concentrations from 0.5mg μg−1 dry soil to lower than 10.0mg μg−1 dry soil appeared to stimulate the growth of soil bacteria, actinomycetes, mould, and the pure cultures of Br.japonicum and A. chroococcum. Not only the colony amounts of these two species of nitrogen fixing bacteria increased, grown on agar medium containing lower concentrations of trifluralin, but also these colonies also enlarged in size and appeared obviously in shorter formation time. However, trifluralin at higher concentrations would inhibit the development of microbial colonies both in amount and size. Trifluralin inhibited the activity of acetylene reduction of A.chroococcum when it was added at the same time of inoculation with A. chroococcum, but it showed a noteworthy stimulation to nitrogen fixation of A.chroococcum when it was put into culture after the cells of the nitrogen fixing bacterium had grown well. The observation that soil microorganisms could use trifluralin as sole carbon and nitrogen resources for their growth, indicated that microorganisms could decompose trifluralin well.

Effect of aeration and carbon/nitrogen ratio on the molecular mass of the biodegradable polymer poly-β-hydroxybutyrate obtained from Azotobacter chroococcum 6B

The bacterial polyester poly-β-hydroxybutyrate (PHB) was quantified and characterized on an isolate␣of the nitrogen-fixing bacteria Azotobacter chroococcum 6B on the basis of its average molecular mass, determined from the relative viscosity at different aeration rates and carbon/nitrogen ratios during culture in fermentors. A higher value for the molecular mass (1100 kDa) was obtained with the lower aeration rates investigated, which diminished, significantly at the highest aeration rate of 2.5 vvm (a 100-fold decrease). The yield of PHB relative to the amount of glucose consumed increased with the C/N ratio (a maximum of 0.16 g PHB/g glucose consumed with a carbon/nitrogen ratio of 137.7), but the molecular mass was lowered from 800 kDa to nearly 100 kDa. The maximum PHB content was 63.5% (on a cellular dry-weight basis) after 47 h in fed-batch culture with an initial C/N ratio of 68.9 and aeration at a rate of 0.5 vvm. Calorimetric measurements on the isolated PHB showed a melting point near 175 °C.

In vivo modification of Azotobacter chroococcum glutamine synthetase.

A monospecific anti-(glutamine synthetase) antibody raised against glutamine synthetase of the unicellular cyanobacterium Synechocystis sp. strain PCC 6803 immunoreacted with glutamine synthetase from the N2-fixing heterotrophic bacterium Azotobacter chroococcum. In Western-blotting experiments this antibody recognized a single protein of a molecular mass of 59 kDa corresponding to glutamine synthetase subunit. This protein was in vivo-labelled in response to addition of ammonium, both [3H]adenine and H(3)32PO4 preincubation of the cells being equally effective. Nevertheless, the amount of glutamine synthetase present in A. chroococcum was independent of the available nitrogen source. Modified, inactive glutamine synthetase was re-activated by treatment with snake-venom phosphodiesterase but not by alkaline phosphatase. L-Methionine-DL-sulphoximine, an inhibitor of glutamine synthetase, prevented the enzyme from being covalently modified. We conclude that, in A. chroococcum, glutamine synthetase is adenylylated in response to ammonium and that for the modification to take place ammonium must be metabolized.

Nitrate permease from Azotobacter chroococcum

Active transport systems in bacteria can be divided into two groups: those that are osmotic shock‐resistant with one single membrane protein, and those that are shock‐sensitive and have a membrane‐bound protein complex plus a soluble periplasmic protein. Whether the bacterial assimilatory nitrate transport falls into the one or the other of these two groups has not been studied before. We report that nitrate uptake by the strictly aerobic, N2‐fixing heterotrophic bacterium Azotobacter chroococcum is sensitive to osmotic shock. The polypeptide composition of cytoplasmic membranes changes in response to the nitrogen source available to the cells. Incorporation of [35S]‐methionine into proteins as well as use of the A. chroococcum TRI mutant, which is defective in nitrate transport, and the A. choococcum MCD1 strain, a mutant unable to use nitrate as a nitrogen source, suggest that nitrate transport into A. chroococcum cells is mediated by a multicomponent system tightly bound to the cytoplasmic membrane.