Optimal conditions for induction of competence in nitrogen-fixing Azotobacter vinelandii

1982 ◽  
Vol 28 (4) ◽  
pp. 389-397 ◽  
Author(s):  
William J. Page
Keyword(s):  
Dissolved Oxygen ◽  
Sodium Acetate ◽  
Azotobacter Vinelandii ◽  
Nitrogenase Activity ◽  
Cell Mass ◽  
Apparent Competition ◽  
Competence Development ◽  
Optimal Conditions ◽  
Nitrogen Fixing ◽  
Ph Range

Competence development in nitrogen-fixing Azotobacter vinelandii cells was optimal at pH 7.2–7.4 which necessitated additional buffering of the iron-limited nitrogen-free competence medium or the addition of a suitable organic acid salt, e.g., sodium acetate. An autolysin was active in this pH range and competent cells were more susceptible to autolysis than the general cell population. Competence development also required restricted aeration of the culture, and only those cultures that attained zero dissolved oxygen became competent. Restricted aeration served to protect the iron-limited cell nitrogenase from oxygen inactivation thus allowing the culture to reach zero dissolved oxygen. The inclusion of additional sources of reductant, e.g., malate, in buffered competence medium resulted in increased respiration and protection of nitrogenase, increased cell mass, and poly-β-hydroxybutyrate synthesis, but decreased competence. A possible explanation for the apparent competition between competence development and nitrogenase activity is discussed.

Transformation of Azotobacter vinelandii strains unable to fix nitrogen with Rhizobium spp. DNA

1978 ◽  
Vol 24 (3) ◽  
pp. 209-214 ◽  
Author(s):  
William J. Page
Keyword(s):  
Transcriptional Control ◽  
Deoxyribonucleic Acid ◽  
Azotobacter Vinelandii ◽  
Nitrogenase Activity ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
Spontaneous Reversion ◽  
Mutant Strains ◽  
Rhizobium Spp ◽  
Rhizobium Sp

The phenotypes of Azotobacter vinelandii ATCC 12837 strains defective in nitrogen fixation (Nif−) were characterized by intrageneric transformation with known Nif− strains of A. vinelandii OP. These former mutant strains were used as recipients for intergeneric transformation by deoxyribonucleic acid (DNA) prepared from Rhizobium spp. to determine if the rhizobia would transform the Azotobacter Nif− phenotypes to Nif+. The frequency of Nif+ transformants using Rhizobium DNA was always less than the frequency using Azotobacter wild-type DNA but was greater than the spontaneous reversion frequency. The Azotobacter Nif+ recombinants also were stable. DNA from all of the Rhizobium spp. transformed to Nif+Azotobacter mutants defective in the nitrogenase component I (molybdoferredoxin); however, some recombinants had a lower nitrogenase activity and a delayed nitrogenase depression time. Mutants defective in the pleiotrophic transcriptional control of both nitrogenase components were transformed to Nif+ by the asymbiotic nitrogen fixing Rhizobium sp. 32H1 and 41A1, but not the symbiotic nitrogen-fixing species. The significance of these results and the possible future applications of this system are discussed.

Molybdenum enhancement of nitrogen fixation in a Mo-starved Azotobacter vinelandii Nif− mutant

1982 ◽  
Vol 28 (10) ◽  
pp. 1173-1180 ◽  
Author(s):  
William J. Page ◽  
S. Karen Collinson
Keyword(s):  
Molecular Weight ◽  
Nitrogen Fixation ◽  
Azotobacter Vinelandii ◽  
Nitrogenase Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Nitrogen Fixing ◽  
Wild Type ◽  
Cell Extracts ◽  
In The Wild ◽  
Dinitrogenase Reductase

Molybdenum (Mo)-starved wild-type and Nif− strains of Azotobacter vinelandii reduced acetylene (fixed nitrogen) in Mo-limited nitrogen-free medium. Vanadate enhanced this activity in all of the strains. Molybdate caused repression of nitrogenase activity in the Nif− mutants and enhanced the nitrogenase activity in the wild type. The nitrogenase activity in the Nif− mutant UW3, however, was enhanced by Mo, became maximal after 3 h, and then declined to zero after 10 h of incubation. The activation of nitrogenase by Mo followed a 5- to 10-min lag and was inhibited when streptomycin or rifampin was added with Mo. Examination of Mo-starved nitrogen-fixing UW3 cell extracts by two-dimensional polyacrylamide gel electrophoresis revealed molecular weight 57 000, 50 000, and 30 000 proteins that were Mo and NH4+ repressive. The molecular weight 30 000 protein appeared in the same position on the gel as the wild-type dinitrogenase reductase, although UW3 did not produce this protein under Mo-sufficient nitrogen-fixing conditions. Cell extracts prepared 3 h after Mo addition lacked the molecular weight 57 000 and 50 000 proteins but contained a new protein corresponding to the β subunit of dinitrogenase. When UW3 nitrogenase activity was lost, the dinitrogenase reductase-like protein also was absent. The results suggest that a complex active in nitrogen fixation may form between components of the traditional Mo-sufficient and alternative Mo-starved cell nitrogen fixation systems.

Formation of the nitrogen-fixing enzyme system in Azotobacter vinelandii

1968 ◽  
Vol 14 (1) ◽  
pp. 25-31 ◽  
Author(s):  
G. W. Strandberg ◽  
P. W. Wilson
Keyword(s):  
Color Change ◽  
Azotobacter Vinelandii ◽  
Nitrogenase Activity ◽  
Color Difference ◽  
Potassium Nitrate ◽  
Whole Cells ◽  
System A ◽  
Growth System ◽  
A Cell ◽  
Enzyme Formation

The formation and activity of nitrogenase2 in Azotobacter vinelandii OP was examined using a cell-free assay system. A lag period of about 30 min occurred between the exhaustion of the combined nitrogen source and growth on N2. Cells grown on ammonium acetate or potassium nitrate had no detectable nitrogenase activity. Nitrogenase activity appeared in cells, grown under a flowing gas phase of 20% O2 – 60% He, about 45 min after the exhaustion of ammonia. Nitrogenase formation was inhibited in a closed system with an atmosphere containing 40% O2 but not by one containing 20% O2. Hydrogen did not inhibit enzyme formation. The question of whether N2 is required for the formation of the enzyme could not be answered as this gas could not be completely eliminated from the growth system. Chloramphenicol prevented the formation of the enzyme and inhibited nitrogen fixation in whole cells, but had no effect on cell-free enzyme activity. A brief rise in turbidity which occurred during nitrogenase formation appeared to be due to a color change in the cells from reddish brown to dark brown. Spectrophotometric examination of extracts from ammonia- and N2-grown cells did not reveal any components responsible for this color difference, but this result may reflect only the presence of interfering substances in the crude extract.

scholarly journals Rhizobium tropici Genes Involved in Free-Living Salt Tolerance are Required for the Establishment of Efficient Nitrogen-Fixing Symbiosis with Phaseolus vulgaris

2002 ◽  
Vol 15 (3) ◽  
pp. 225-232 ◽  
Author(s):  
Joaquina Nogales ◽  
Rosario Campos ◽  
Hanaa BenAbdelkhalek ◽  
José Olivares ◽  
Carmen Lluch ◽  
...  
Keyword(s):  
Salt Tolerance ◽  
Phaseolus Vulgaris ◽  
Environmental Changes ◽  
Nitrogenase Activity ◽  
Regulation Of Gene Expression ◽  
Elongation Factor ◽  
Regulatory System ◽  
Trna Synthetase ◽  
Nitrogen Fixing ◽  
Rhizobium Tropici

Characterization of nine transposon-induced mutants of Rhizobium tropici with decreased salt tolerance (DST) allowed the identification of eight gene loci required for adaptation to high external NaCl. Most of the genes also were involved in adaptation to hyperosmotic media and were required to overcome the toxicity of LiCl. According to their possible functions, genes identified could be classified into three groups. The first group included two genes involved in regulation of gene expression, such as ntrY, the sensor element of the bacterial ntrY/ntrX two-component regulatory system involved in regulation of nitrogen metabolism, and greA, which encodes a transcription elongation factor. The second group included genes related to synthesis, assembly, or maturation of proteins, such as alaS coding for alanine-tRNA synthetase, dnaJ, which encodes a molecular chaperone, and a nifS homolog probably encoding a cysteine desulfurase involved in the maturation of Fe-S proteins. Genes related with cellular build-up and maintenance were in the third group, such as a noeJ-homolog, encoding a mannose-1-phosphate guanylyltransferase likely involved in lipopolysaccharide biosynthesis, and kup, specifying an inner-membrane protein involved in potassium uptake. Another gene was identified that had no homology to known genes but that could be conserved in other rhizobia. When inoculated on Phaseolus vulgaris growing under nonsaline conditions, all DST mutants displayed severe symbiotic defects: ntrY and noeJ mutants were impaired in nodulation, and the remaining mutants formed symbiosis with very reduced nitrogenase activity. The results suggest that bacterial ability to adapt to hyper-osmotic and salt stress is important for the bacteroid nitrogen-fixing function inside the legume nodule and provide genetic evidence supporting the suggestion that rhizobia face severe environmental changes after their release into plant cells.

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

1988 ◽  
Vol 34 (7) ◽  
pp. 886-890 ◽  
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.

scholarly journals TEMPO-Functionalized Nanoporous Au Nanocomposite for the Electrochemical Detection of H2O2

2018 ◽  
Vol 2018 ◽  
pp. 1-11
Author(s):  
Dongxiao Wen ◽  
Qianrui Liu ◽  
Ying Cui ◽  
Huaixia Yang ◽  
Jinming Kong
Keyword(s):  
Electrochemical Detection ◽  
Nanoporous Gold ◽  
Carboxyl Groups ◽  
Time Interval ◽  
Optimal Conditions ◽  
Sem Images ◽  
One Step ◽  
Ph Range ◽  
Synthesis Approach ◽  
Higher Sensitivity

A novel nanocomposite of nanoporous gold nanoparticles (np-AuNPs) functionalized with 2,2,6,6-tetramethyl-1-piperidinyloxy radical (TEMPO) was prepared; assembled carboxyl groups on gold nanoporous nanoparticles surface were combined with TEMPO by the “bridge” of carboxylate-zirconium-carboxylate chemistry. SEM images and UV-Vis spectroscopies of np-AuNPs indicated that a safe, sustainable, and simplified one-step dealloying synthesis approach is successful. The TEMPO-np-AuNPs exhibited a good performance for the electrochemical detection of H2O2 due to its higher number of electrochemical activity sites and surface area of 7.49 m2g-1 for load bigger amount of TEMPO radicals. The TEMPO-functionalized np-AuNPs have a broad pH range and shorter response time for H2O2 catalysis verified by the response of amperometric signal under different pH and time interval. A wide linear range with a detection limit of 7.8 × 10-7 M and a higher sensitivity of 110.403 μA mM-1cm-2 were obtained for detecting H2O2 at optimal conditions.

Nitrogen fixation

1976 ◽  
Vol 274 (934) ◽  
pp. 341-358 ◽  
Keyword(s):  
Nitrogen Fixation ◽  
Metabolic Regulation ◽  
Higher Plants ◽  
Focal Point ◽  
Nitrogenase Activity ◽  
Nitrogen Fixing ◽  
Blue Green Algae ◽  
International Biological Programme ◽  
The 1960S ◽  
Biological Programme

The International Biological Programme served as a focal point for studies on biological nitrogen fixation during the 1960s. The introduction of the acetylene reduction technique for measuring nitrogenase activity in the field led to estimates becoming available of the contribution of lichens, blue-green algae, nodulated non-legumes and bacterial-grass associations, as well as of legumes. Other studies carried out on the physiology and biochemistry of the process led to the eventual purification and characterization of the nitrogenase enzyme. These studies, collectively, provided the springboard for current work, so essential in view of the present energy crisis, on how to increase the use and efficiency of nitrogen-fixing plants, on the metabolic regulation of the nitrogenase enzyme and on the genetics of the nitrogen-fixing process, both in higher plants and in free-living micro-organisms.

scholarly journals SCREENING OF SOIL BACTERIA FOR PLANT GROWTH PROMOTION ACTIVITIES IN VITRO

2016 ◽  
Vol 4 (1) ◽  
pp. 27 ◽  
Author(s):  
Edi Husen
Keyword(s):  
Plant Growth ◽  
Soil Bacteria ◽  
Phosphate Solubilization ◽  
Azotobacter Vinelandii ◽  
Growth Promotion ◽  
Nitrogenase Activity ◽  
Chelating Agent ◽  
Plant Growth Promoting Rhizobacteria ◽  
Iaa Production

Fourteen isolates of soil bacteria, including two known plant growth promoting rhizobacteria (PGPR) strains, Azotobacter vinelandii Mac 259 and Bacillus cereus UW 85, were tested in vitro. Parameters assessed were indoleacetic acid (IAA) production, phosphate solubilization, dinitrogen fixation, and siderophore (Fe-III chelating agent) production. IAA production was assayed colorimetrically using ferric chlorideperchloric acid reagent. Phosphate-solubilization and siderophore production were tested qualitatively by plating the bacteria in Pikovskaya and chrome azurol S agar, respectively. The ability to fix dinitrogen was measured based on nitrogenase activity of the bacteria by gas chromatography. The results showed that twelve isolates produced IAA, ranged from 2.09 to 33.28 µmol ml-1. The ability to solubilize precipitated phosphate was positively exhibited by four isolates (BS 58, BTS, TCaR 61, and BTCaRe 65). Seven isolates including Mac 259 positively produced siderophore. None of the isolates showed nitrogenase activity. Only one isolate (TS 3) did not exhibit any of the traits tested. Isolate TCeRe 60 and reference strain Mac 259 were found to have IAA- and siderophore-producing traits. Four P-solubilizing bacteria (BS 58, BTS, TCaR 61, and BTCaRe 65) were also IAA- and siderophore-producing bacteria. Potential use of these PGPR isolates needs further test in enhancing plant growth.

scholarly journals Influence of hydrotalcite preparation conditions on its physico-chemical properties

2019 ◽  
Vol 12 (1) ◽  
pp. 119-126
Author(s):  
Miroslava Mališová ◽  
Michal Horňáček ◽  
Pavol Hudec ◽  
Jozef Mikulec ◽  
Vladimír Jorík ◽  
...  
Keyword(s):  
Catalytic Properties ◽  
Chemical Properties ◽  
Synthesis Temperature ◽  
Optimal Synthesis ◽  
Preparation Process ◽  
Optimal Conditions ◽  
Temperature Range ◽  
Preparation Conditions ◽  
Physico Chemical ◽  
Ph Range

Abstract The aim of the research was to prepare and characterize hydrotalcite synthesized under different preparation conditions. The most common hydrotalcite preparation is the co-precipiaton method. The preparation process strongly influences the catalytic properties of hydrotalcite; therefore, optimal conditions have to be determined. During the study, seven samples of the catalyst were prepared in the pH range from 8 to 12 and the synthesis temperature range from 25 to 55 °C. Based on several catalyst properties, optimal synthesis pH 10 was found. Ideal temperature of the preparation was determined to be 35 °C, but the temperature does not have a significant effect on the catalyst properties.

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