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  • Nitrogen fixation  (148)
  • Springer  (148)
  • 1980-1984  (116)
  • 1975-1979  (32)
  • 1925-1929
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular evolution 21 (1984), S. 19-32 
    ISSN: 1432-1432
    Keywords: Leghemoglobin ; Gene duplication ; Gene linkage ; Concerted evolution ; Nitrogen fixation ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed the sequences of soybean leghemoglobin genes as an initial step toward understanding their mode of evolution. Alignment of the sequences of plant globin genes with those of animals reveals that (i) based on the proportion of nucleotide substitutions that have occurred at the first, second, and third codon positions, the time of divergence of plant and animal globin gene families appears to be extremely remote (between 900 million and 1.4 billion years ago, if one assumes constancy of evolutionary rate in both the plant and animal lineages) and (ii) in addition to the normal regulatory sequences on the 5′ end, an approximately 30-base-pair sequence, specific to globin genes, that surrounds the cap site is conserved between the plant and animal globin genes. Comparison of the leghemoglobin sequences with one another shows that (i) the relative amount of sequence divergence in various coding and noncoding regions is roughly similar to that found for animal globin genes and (ii) as in animal globin genes, the positions of insertions and deletions in the intervening sequences often coincide with the locations of direct repeats. Thus, the mode of evolution of the plant globin genes appears to resemble, in many ways, that of their animal counterparts. We contrast the overall intergenic organization of the plant globin genes with that of animal genes, and discuss the possibility of the concerted evolution of the leghemoglobin genes.
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  • 2
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    Archives of microbiology 107 (1976), S. 25-31 
    ISSN: 1432-072X
    Keywords: Ferredoxin ; Iron-sulfur proteins ; Nitrogen fixation ; Electron transport to nitrogenase ; Flavodoxin ; Mycobacterium flavum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. Two ferredoxin-type iron-sulfur proteins have been isolated from Mycobacterium flavum 301 grown under nitrogen-fixing, iron-sufficient conditions. No flavodoxin was observed. 2. These ferredoxins are apparently soluble: they were present in the supernatant fraction after disrupting by decompression. Only small amounts were present in particulate fractions. 3. The two ferredoxins were separated by chromatography on DEAE-cellulose, Sephadex or electrophoresis. 4. Both ferredoxins mediated the transfer of electrons from illuminated spinach chloroplasts to a nitrogenase preparation to reduce acetylene. Ferredoxin II was specifically about five times more active than ferredoxin I. Ferredoxin II was also active in the photosynthetic NADP+-reduction whereas ferredoxin I was not. 5. Both ferredoxins were reversibly reduced by either sodium dithionite, illuminated spinach chloroplasts or hydrogen plus hydrogenase from Clostridium pasteurianum. 6. Attempts to determine the primary electron donor for nitrogen fixation in Mycobacterium flavum were unsuccessful. Acetylene reduction in Mycobacterium extracts was obtained only with sodium dithionite or illuminated spinach chloroplasts as electron donors. The reduction of the electron carrier (e. g. ferredoxin) rather than the transfer of electrons from the reduced carrier to nitrogenase was rate-limiting.
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  • 3
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    Archives of microbiology 107 (1976), S. 115-124 
    ISSN: 1432-072X
    Keywords: Alanine dehydrogenase ; Anabaena cylindrica ; Heterocysts ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The l-alanine dehydrogenase (ADH) of Anabaena cylindrica has been purified 700-fold. It has a molecular weight of approximately 270000, has 6 sub-units, each of molecular weight approximately 43000, and shows activity both in the aminating and deaminating directions. The enzyme is NADH/NAD+ specific and oxaloacetate can partially substitute for pyruvate. The K m app for NAD+ is 14 μM and 60 μM at low and high NAD+ concentrations, respectively. The K m app for l-alanine is 0.4 mM, that for pyruvate is 0.11 mM, and that for oxaloacetate is 3.0 mM. The K m app for NH 4 + varies from 8–133 mM depending on the pH, being lowest at high pH levels (pH 8.7 or above). Alanine, serine and glycine inhibit ADH activity in the aminating direction. The enzyme is active both in heterocysts and vegetative cells and activity is higher in nitrogen-starved cultures than in N2-fixing cultures. The data suggest that although alanine is formed by the aminating activity of ADH, entry of newly fixed ammonia into organic combination does not occur primarily via ADH in N2-fixing cultures of A. cylindrica. Ammonia assimilation via ADH may be important in cultures with an excess of available nitrogen. The deaminating activity of the enzyme may be important under conditions of nitrogen-deficiency.
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  • 4
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    Archives of microbiology 107 (1976), S. 263-267 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Root nodule ; Endophyte vesicle ; Alder ; Alnus ; Freeze-etching
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen-fixing root nodules of Alnus crispa var. mollis Fern. were studied by transmission electron microscopy and by freeze-etching technique. Ultrathin sectioning of septate vesicles of the actinomycetal endophyte showed an electron transparent zone, the so-called “void area”, between the vesicle cell wall and its encapsulation material. This void area was not observed in the freeze-etching replicas of cryoprotected nodular tissue. It is suggested that the void area is the result of the coming-off of the vesicle cell wall from the capsule and that its formation reflects difficulty in fixing the voluminous mature vesicle of the root nodule endophyte.
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  • 5
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    Archives of microbiology 114 (1977), S. 197-201 
    ISSN: 1432-072X
    Keywords: Anabaena sp. ; Nitrogen fixation ; Heterocysts ; Cyanobacteria ; Marine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A marine filamentous cyanobacterium capable of rapid growth under N2-fixing conditions has been isolated from the Texas Gulf Coast. This organism appears to be an Anabaena sp. and has been given the strain designation CA. Cultures grown on mineral salts medium bubbled with 1% CO2-enriched air at 42°C show a growth rate of 5.6±0.1 generations per day with molecular nitrogen as the sole nitrogen source. This growth rate is higher than any other reported in the literature to date for heterocystous cyanobacteria growing on N2. Under similar growth conditions, 7.5 mM NH4Cl yields a growth rate of 6.6±0.1 generations per day while 7.5 mM KNO3 allows for a growth rate of 5.8±0.4 generations-day. Nitrogen-fixation rates, as measured by acetylene reduction, show maximum activity values in the range of 50–100 nmoles ethylene produced/minxmg protein. These values compare favorably with those obtained from heterotrophic bacteria and are much higher than values reported for other cyanobacteria. Growth experiments indicate that the organism requires relatively high levels of sodium and grows maximally at 42°C. Because of its high growth rate on N2, this newly isolated organism appears ideal for studying nitrogen metabolism and heterocyst development among the cyanobacteria.
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  • 6
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    Archives of microbiology 118 (1978), S. 177-184 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Knallgas reaction ; Hydrogenase ; Hydrogen utilization ; Nitrogenase ; Nitrogen fixation ; Isolated heterocysts ; Anabaena cylindrica ; Nostoc muscorum ; Anabaena variabilis ; Anacystis nidulans ; Cyanophora paradoxa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several blue-green algae were surveyed for the occurrence of the hydrogenase which was assayed by the oxyhydrogen or Knallgas reaction in the intact organisms. In aerobically grown cultures, the reaction was detectable in Anabaena cylindrica, Nostoc muscorum and in two Anabaena variabilis species, whereas virtually no activity was observed in Anacystis nidulans and Cyanophora paradoxa. In these latter two algae, the reaction was, however, found after growth under molecular hydrogen for several days, which drastically increased the activity levels with all the algae tested. In the nitrogen fixing species, the activity of the Knallgas reaction was enhanced when all combined nitrogen was omitted from the media. H2 and hydrogenase could not significantly support the CO2-fixation in photoreduction experiments with all blue-green algae investigated here. Hydrogenase was assayed by the dithionite and methyl viologen dependent evolution of hydrogen and was found to be present with essentially the same specific activity levels in preparations of both heterocysts and vegetative cells from Anabaena cylindrica. Na2S2O4 as well as H2 supported the C2H2-reduction of the isolated heterocysts. The H2-dependent C2H2-reduction did not require the presence of oxygen but was strictly light-dependent where H2 served as an electron donor to photosystem I of these cells. It is concluded that hydrogen can be utilized by two different pathways in blue-green algae.
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  • 7
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    Archives of microbiology 122 (1979), S. 153-159 
    ISSN: 1432-072X
    Keywords: Rhizobium japonicum ; Nitrogenase activity ; Nitrogen fixation ; Organic acids ; Cell-morphology ; Bacteroids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rhizobium japonicum 61-A-101 grew and fixed nitrogen more effectively on media containing an organic acid and a pentose sugar than on media containing only one of these carbon sources. Peak specific activities in the range 10–15 nmol C2H4 · h-1 · mg protein-1 were found for these organisms in a spot of growth about 1 cm diameter on agar surfaces exposed to air. Increasing concentrations of the organic acids (succinate or malonate) in a medium containing arabinose resulted in longer lasting activity. The inclusion of a third carbon source, glycerol, gave activity which remained at the maximum from about the 8 to the 18 day after inoculation although no growth of the bacteria occurs during the last 8 or 10 days. At low concentration of organic acid l-arabinose was a much better carbon source for supporting nitrogenase activity of these organisms that the d-form. Both organic acids affected the morphology of the bacteria. Higher concentrations, especially of malonate, gave swollen and distorted cells. When bacteria growing on organic acid-containing agar plates were suspended and plated after appropriate dilution on yeast extract — mannitolglycerol agar there was heterogeneity of colony form, with up to 90% microcolonies after growth on high malonate concentrations. The effects of malonate may be correlated with characteristics of the bacteroid form inside the nodule which contains relatively high concentrations of organic acids, especially malonate.
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  • 8
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    Archives of microbiology 123 (1979), S. 37-45 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Blue-green algae ; Hydrogen metabolism ; Nitrogen fixation ; Isolated heterocysts ; Inhibitors ; Plastoquinone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several natural and artificial electron donors were assayed in the C2H2-reduction of heterocysts isolated from the cyanobacterium Anabaena cylindrica. Among these, molecular hydrogen was the most effective one when the assays were performed in the light. The C2H2-reduction and the Knallgas reaction of intact Anabaena filaments as well as the H2-supported C2H2-reduction of isolated heterocysts were compared for their sensitivity towards several inhibitors known to affect the photosynthetic or respiratory electron flow. Among these, dibromothymoquinone (DBMIB) affected all three reactions equally indicating that plastoquinone is a common intermediate of the H2-consumptions by either the respiratory or the photosynthetic electron transport. Metronidazole inhibited the H2-utilization via photosynthesis but did not affect the consumption of this gas by respiration and therefore allows to differentiate between the two pathways of hydrogen utilization. The studies with the inhibitors are suggestive for a segment of electron carriers on the membranes common to both photosynthesis and respiration in heterocysts of Anabaena.
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  • 9
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    Archives of microbiology 123 (1979), S. 259-265 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrogenase ; Gene expression ; Protein synthesis ; Regulation ; Klebsiella pneumoniae ; Microbial ecology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract At growth temperatures above 37°C, Klebsiella pneumoniae does not grow in a medium containing N2 or NO 3 - as nitrogen sources. However, both the growth in the presence of other nitrogen sources as well as the in vitro nitrogenase activity are not affected at this temperature. The inability to fix N2 at high temperature is due to the failure of the cells to synthesize nitrogenase and other nitrogen fixation (nif) gene encoded proteins. When cells grown under nitrogen fixing conditions at 30°C were shifted to 39°C, there was a rapid decrease of the rate of de novo biosynthesis of nitrogenase (component 1), nitrogenase reductase (component 2), and the nifJ gene product. There was no degradation of nitrogenase at the elevated temperature since preformed enzyme remained stable over a period of at least 3 h at 39°C. Thus, temperature seems to represent a third control system, besides NH 4 + and O2, governing the expression of nif genes of K. pneumoniae.
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  • 10
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    Archives of microbiology 129 (1981), S. 238-239 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Disaccharide ; Bacteroid ; Transport ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Slow growing strains of rhizobia appear to lack both uptake systems and catabolic enzymes for disaccharides. In the fast-growing strains of rhizobia there are uptake mechanisms and catabolic enzymes for disaccharide metabolism. In Rhizobium leguminosarum WU 163 and WU235 and R. trifolii WU290, sucrose and maltose uptake appears to be constitutive whereas in R. meliloti WU60 and in cowpea Rhizobium NGR234 uptake of these disaccharides is inducible. There is evidence that there are at least two distinct disaccharide uptake systems in fast-growing rhizobia, one transporting sucrose, maltose and trehalose and the other, lactose. Disaccharide uptake is via an active process since uptake is inhibited by azide, dinitrophenol and carbonyl cyanide m-chlorophenylhydrazone but not by arsenate. Bacteroids of R. leguminosarum WU235 and R. lupini WU8 are unable to accumulate disaccharides.
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  • 11
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    Archives of microbiology 133 (1982), S. 172-177 
    ISSN: 1432-072X
    Keywords: Beggiatoa ; Nitrogen fixation ; Acetylene reduction ; Nitrate assimilation ; Microaerobic ; Isolation of marine strains
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four newly isolated marine strains of Beggiatoa and five freshwater strains were tested for nitrogen fixation in slush agar medium. All strains reduced acetylene when grown microaerobically in media containing a reduced sulfur source and lacking added combined nitrogen. The addition of 2 mmol N, as nitrate or ammonium salts, completely inhibited this reduction. Although not optimized for temperature or cell density, acetylene reduction rates ranged from 3.2 to 12 nmol·mg prot-1 min-1. Two freshwater strains did not grow well or reduce acetylene in medium lacking combined nitrogen if sulfide was replaced by thiosulfate. Two other strains grew well in liquid media lacking both combined nitrogen and reduced sulfur compounds but only under lowered concentrations of air. All freshwater strains grew well in medium containing nitrate as the combined nitrogen source. Since they did not reduce acetylene under these conditions, we infer that they can assimilate nitrate.
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  • 12
    ISSN: 1432-072X
    Keywords: Ammonia assimilation ; Lichen symbioses ; Nitrogen fixation ; 15N kinetics ; Peltigera canina
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract On following N2-incorporation and subsequent metabolism in the lichen Peltigera canina using 15N as tracer, it was found, over a 30 min period, that greatest initial labelling was into NH 4 + followed by glutamate and the amide-N of glutamine. Labelling of the amino-N of glutamine, aspartate and alanine increased slowly. Pulse-chase experiments using 15N confirmed this pattern. On inhibiting the GS-GOGAT pathway using l-methionine-dl-sulphoximine and azaserine, 15N enrichment of glutamate, alanine and aspartate continued although labelling of glutamine was undetectable. From this and enzymic data, NH 4 + assimilation in the P. canina thallus appears to proceed via GS-GOGAT in the cyanobacterium and via GDH in the fungus; aminotransferases were present in both partners. The cyanobacterium assimilated 44% of the 15N2 fixed; the remainder was liberated almost exclusively as NH 4 + and then assimilated by fungal GDH.
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  • 13
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    Archives of microbiology 136 (1983), S. 81-83 
    ISSN: 1432-072X
    Keywords: Ammonia production ; Anabaena ; Cyanobacteria ; Nitrate reductase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the filamentous heterocystous cyanobacterium Anabaena sp. strain ATCC 33047 dinitrogen fixation and nitrate reduction are mutually exclusive processes. Nitrate promotes nitrate reductase synthesis and represses nitrogenase formation. Inhibition of ammonium assimilation by l-methionine-d,l-sulfoximine (MSX) alleviates the repressive effect of nitrate on nitrogenase synthesis, thus indicating that the nitrate effect is indirect through metabolites generated from the ammonium derived from nitrate reduction. In MSX-treated cells both nitrate reduction and dinitrogen fixation take place simultaneously, although at different sites of the filament, without any apparent competition for the required reducing power. The MSX-treated Anabaena cells generate ammonium from both nitrate and dinitrogen, simultaneously.
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  • 14
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    Archives of microbiology 130 (1981), S. 38-43 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Gene expression ; Regulation ; Messenger RNA ; Transcription ; Klebsiella pneumoniae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogenase messenger RNA synthesis in Klebsiella pneumoniae was determined by labelling cells with (3H)uracil and isolating total RnA, which was then hybridized to filterbound recombinant plasmid pSA30 DNA carrying the nitrogenase structural genes nifH, D, and K. Derepression of nitrogenase mRNA starts 1.5 h before the onset of nitrogenase activity (as measured by acetylene reduction). Exposure of nif-derepressed cultures to either NH 4 + , air, or high temperatures (39° C) results in a rapid decrease of the synthesis rates both of nitrogenase mRNA and nitrogenase polypeptides. Nitrogenase mRNA is remarkably stable. After blocking transcription with rifampicin, hybridizable and actively translatable nitrogenase mRNA survives with an average half-life of 18 min. Half-lives are considerably shorter when rifampicin-inhibited cultures are simultaneously shifted to conditions which are non-permissive for nitrogenase synthesis, pointing to some posttranscriptional influence on nitrogenase mRNA stability. In all experiments performed there was no evidence for uncoupling of nitrogenase mRNA synthesis from nitrogenase mRNA translation, indicating that nitrogenase synthesis is regulated solely by transcriptional control.
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  • 15
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    Archives of microbiology 138 (1984), S. 212-216 
    ISSN: 1432-072X
    Keywords: Denitrification ; Nitrate respiration ; Nitrous oxide reduction ; Nitrogen fixation ; Azospirillum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrous oxide reduction can consistently be demonstrated with high activities in cells of Azospirillum brasilense Sp 7 which are grown anaerobically in the presence of low amounts of nitrite. Azospirillum can even grow anaerobically with nitrous oxide in the absence of any other respiratory electron acceptor. Nitrous oxide reduction by Azospirillum is inhibited by acetylene, amytal and weakly by carbon monoxide. Azospirillum converts nitrous oxide to molecular nitrogen without the formation of ammonia. The cells must, therefore, be supplied with ammonia from nitrogen fixation during anaerobic growth with nitrous oxide. When no other nitrogen compound besides nitrous oxide is available in the medium, the bacteria synthesize nitrogenase from protein reserves in about 2 h. Nitrogenase synthesis is blocked by chloramphenicol under these conditions. In contrast, the addition of nitrate or nitrite to the medium represses the synthesis of nitrogenase. Nitrous oxide reduction by Azospirillum and other microorganisms is possibly of ecological significance, because the reaction performed by the bacteria may remove nitrous oxide from soils.
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  • 16
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    Archives of microbiology 136 (1983), S. 219-221 
    ISSN: 1432-072X
    Keywords: Xanthobacter ; Nitrogen fixation ; Oxygen sensitivity ; Nitrogen metabolism ; Glutamine synthetase ; Glutamate synthase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract N2-fixation was investigated in the chemoautotrophic hydrogen bacterium Xanthobacter H4-14. N2-fixing batch cultures of this organism could only be grown at pO2 values of around 0.02 bar, and in continuous culture dissolved oxygen tensions above 16 μM were found to inhibit N2-fixation. Xanthobacter H4-14 utilized a variety of amino acids, nitrate and ammonia as nitrogen sources. Cell-free extracts from steady-state continuous cultures of ammonia grown, nitrate grown and N2-fixing Xanthobacter were assayed for the presence of ammonia assimilation enzymes. No alanine dehydrogenase or glutamate dehydrogenase activity was detected. Ammonia was assimilated exclusively via the glutamine synthetase/glutamate synthase pathway, irrespective of the extracellular concentration of ammonia.
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  • 17
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    Archives of microbiology 107 (1976), S. 235-240 
    ISSN: 1432-072X
    Keywords: Hydrogen bacteria ; Nitrogen fixation ; Acetylene reduction ; Mycobacterium flavum 301 ; Corynebacterium autotrophicum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Seventeen strains of nitrogen-fixing bacteria, isolated from different habitats on hydrogen and carbon dioxide as well as on other substrates, morphologically resembled each other. All strains, including Mycobacterium flavum 301, grew autotrophically with hydrogen. The isolate strain 6 was sensitive to oxygen when dependent on N2 as nitrogen source, a consequence of the sensitivity of its nitrogenase towards oxygen. At the same time, strain 6 was sensitive to hydrogen when growing autotrophically on N2 as nitrogen source, but hydrogen did not affect acetylene reduction by these cells.
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  • 18
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    Archives of microbiology 111 (1976), S. 195-196 
    ISSN: 1432-072X
    Keywords: Mutation ; Blue-green alga ; Anabaenopsis raciborskii ; Gas vacuole ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract From a bloom forming blue-green alga, Anabaenopsis raciborskii, spontaneous mutants, which had lost the ability to form gas vacuoles have been isolated; the mutant frequency was 4.8×10-3. The filaments of gas vacuole-less mutants settled at the bottom of flasks in liquid culture media unlike the parent alga. The growth and nitrogen fixation were comparatively poor in the mutants.
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  • 19
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    Archives of microbiology 114 (1977), S. 43-49 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Anabaena ; Hydrogenase ; Hydrogen-uptake ; Nitrogenase ; Nitrogen fixation ; Protection mechanism ; Inhibitors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The blue-green alga Anabaena cylindrica is found to consume molecular hydrogen in a hydrogenase dependent reaction. This hydrogen uptake proceeds in the dark and is strictly dependent on oxygen, thus representing a Knallgas reactions. Its rate is almost as high as that of the endogenous respiration in Anabaena. Studies with inhibitors reveal that hydrogen is utilized via the complete respiratory chain providing additional energy for the alga. CO plus C2H2 completely block the Knallgas reaction which explains the previously reported considerable increase in the total H2 formation representing the difference between the nitrogenase-dependent H2-evolution and the reutilization of the gas catalysed by the hydrogenase in intact Anabaena. H2 is able to support the C2H2-reduction in the dark in a reaction again strictly dependent on oxygen. Moreover, H2 is also consumed in experiments carried out under far red light and in the presence of dichlorophenyl-dimenthyl-urea (DCMU) where the energy for nitrogen fixation is no longer provided by respiration but by cyclic photophosphorylation. Under these conditions, H2 is found to supply electrons for the formation of C2H4 from C2H2 in a reaction no longer dependent on the presence of oxygen. Moreover, in these experiments, the presence of H2 stabilizes the C2H2-reduction activity against the deleterious effect of oxygen. Thus, this communication provides evidence for a triplicate function of the H2-uptake catalysed by hydrogenase in intact Anabaena which is (a) to provide energy by the Knallgas reaction, (b) to supply reducing equivalents for nitrogenase, (c) to protect nitrogenase from damage by oxygen.
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  • 20
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    Archives of microbiology 120 (1979), S. 161-165 
    ISSN: 1432-072X
    Keywords: Azolla ; Cyanobacteria ; Blue-green algae ; Nitrogen fixation ; Symbiont ; Photoheterotroph ; Algal isolation ; Photosynthesis ; Plant metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A procedure has been developed to isolate cyanobacteria from the aquatic fern Azolla. The method is based upon recovery of cyanobacterial “bundles” from digests of plants and use of this material as a massive inoculum for nitrogen-free media, followed by prolonged incubation in light. The procedure appears to select for those cells capable of growth in vitro. Isolated cyanobacteria were found to resemble Anabaena sp. morphologically but were capable of heterotrophic growth and had high nitrogenase activity when grown on fructose in the dark.
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  • 21
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    Archives of microbiology 123 (1979), S. 227-232 
    ISSN: 1432-072X
    Keywords: Hydrogen production ; Nitrogen fixation ; Photosynthesis ; Cyanobacteria ; Enrichment cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Seventy-one cyanobacteria containing cultures were enriched from various soil and water locations either under aerobic and/or anaerobic conditions on agar medium selective for nitrogen fixation. Kept under argon containing 1% CO2 for 24 and 48 h most of these cultures evolved hydrogen at very variable rates up to 116 μl per mg chlorophyll and hour as a mean value over a time period of 24h. Several samples evolved hydrogen more efficiently compared with known hydrogen producing pure strains from culture collections. Thirty-one of the investigated cultures showed a hydrogen formation higher than 10 μl per mg chlorophyll and hour measured over 24 or 48 h. Among these all the morphological forms of cyanobacteria i.e. unicellular and filamentous with or without heterocysts are found. Hence, selecting for nitrogen fixing cyanobacteria seems to be a practical method to find efficient hydrogen producers.
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  • 22
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    Archives of microbiology 132 (1982), S. 219-224 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Nitrogen fixation ; Nodules ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several mutants defective in nodulation were isolated from Rhizobium japonicum strains 3I1b110 and 61A 76. Mutants of class I do not form nodules after incubation with soybean [Glycine max (L.) Merrill] for 17 days, but will do so by 28 days. When host plants other than G. max are infected with several of these strains, there is no detectable difference in the time of nodulation or size of nodules as compared to the wild type. Two mutants of class I (i. e., SM1 and SM2) have been shown previously to be altered in the lipopolysaccharide portion of their cell wall. Mutants of class II are not slow to nodulate but form fewer nodules than the wild type on all the host plants tested. Mutants of class III are unable to form nodules. Some bacteriophage-resistant mutants, altered in cell surface structure, fall into this class. Two mutants of class III do not bind to soybean roots.
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  • 23
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrogenase regulation ; Glutamine synthetase ; Methionine suofoximine ; Rhodospirillaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methionine sulfoximine (MSX), an irreversible inhibitor of glutamine synthetase of Rhodopseudomonas palustris restored nitrogenase activity to cells in which nitrogenase had been completely inhibited by ammonia switch-off. After addition of MSX, there was a lag period before nitrogenase activity was fully restored. During this lag, glutamine synthetase activity progressively decreased, and near the time of its complete inhibition, nitrogenase activity resumed. Nitrogenase switch-off by ammonia thus required active glutamine synthetase. Glutamine itself caused nitrogenase inhibition whose reversal by MSX depended on the relative ratio of MSX to glutamine. Unlike ammonia, glutamine inhibited nitrogenase under conditions where glutamine synthetase activity was absent. This indicates that glutamine is the effector molecule in nitrogenase switch-off, for instance by interacting with the enzymatic system for Fe protein inactivation. The effects of glutamine and MSX were also dependent on the culture age. Possible explanation for this and for the competitive effects are a common binding site within the regulatory apparatus for nitrogenase, or, in part, within a common transport system. Some observations with MSX were extended to Rhodopseudomonas capsulata and agreed with those in R. palustris.
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  • 24
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    Archives of microbiology 135 (1983), S. 287-292 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Respiration ; Nitrogen fixation ; Heterocysts ; K m for O2 ; Anabaena variabilis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Simultaneous measurements of acetylene reduction by Anabaena variabilis and the concentration of dissolved oxygen in the suspension were made using a specially designed vessel which allowed measurements under steady-state conditions. The rate of acetylene reduction in the dark increased with increasing oxygen concentrations until a maximum value was reached at 300 μM O2 (corresponding to 30% O2 in the gas phase at 35°C). This presumably results from a requirement for energy provided by respiration. Measurements of the dependence of respiration rate on dissolved oxygen concentration were made under comparable conditions using an open system to allow conditions close to steady-state to be obtained. The respiration rate of diazotrophically grown Anabaena variabilis had a dependence on oxygen concentration corresponding to the sum of two activities. These had K m values of 1.0 μM and 69 μM and values of V max of similar magnitude. Only the high affinity activity was observed in nitrate-grown cyanobacteria lacking heterocysts, and this presumably represent activity in the vegetative cells. The oxygen concentration dependence of the low affinity activity resembled that for the stimulation of acetylene reduction. We interpret this as the result of oxygen uptake by the heterocysts. The results are consistent with the idea that in intact filaments of cyanobacteria O2 enters heterocysts much more slowly than it enters the vegetative cells.
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  • 25
    ISSN: 1432-072X
    Keywords: Azotobacter vinelandii ; Continuous culture ; Oxygen control ; Nitrogen fixation ; Respiratory protection
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    Notes: Abstract Azotobacter vinelandii strain OP was grown in continuous culture at various dissolved oxygen concentrations of air (100% air saturation of the medium=225 ±14 μM O2). Sucrose was added as carbon source and either dinitrogen or ammonia as nitrogen sources. Irrespective of the nitrogen source steady state cultures showed the following general responses with dissolved oxygen concentrations increasing from about 1% to 30% air saturation: (i) cell protein levels, (ii) the amount of cell protein formed per sucrose consumed as well as (iii) nitrogenase activity decreased by at least a factor of two while (iv) cellular respiration increased. At higher oxygen concentrations the parameters changed only slightly, if at all. Increasing the sucrose concentration in the inflowing medium (s R) from 3 g/l to 15 g/l increased the total level of cellular respiration with nitrogen-fixing cultures but was more pronounced with ammonium-assimilating cultures. With nitrogen-fixing cultures cell protein levels increased five-fold while the ratio of protein formed per sucrose consumed as well as cellular nitrogenase activity remained unaffected. With ammonium-assimilating cultures the cell protein level was only doubled and the level of cell protein formed per sucrose consumed was decreased at the higher s R. Increasing the dilution rate at a constant oxygen concentration of 45% air saturation resulted in an almost parallel increase of both cellular respiratory and nitrogenase activity at low and moderate dilution rates. At high dilution rates nitrogenase activity increased steeply over the respiratory activity. Nitrogen-fixing cultures adapted to various oxygen concentrations were subjected to oxygen stress by increasing the oxygen concentration for 7 min. In all cases, this resulted in a complete inhibition (‘switch-off’) of nitrogenase activity. Upon restoration of the original oxygen concentration nitrogenase activity returned to a decreased level. The discussion arrives at the conclusion that some of the results are incompatible with the concept of respiratory protection of nitrogenase.
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  • 26
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    Archives of microbiology 140 (1984), S. 215-217 
    ISSN: 1432-072X
    Keywords: Glutathione reductase ; Cyanobacteria ; Nostoc muscorum ; O2 protection ; Glutathione ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Glutathione reductase activity was detected and characterized in heterocysts and vegetative cells of the cyanobacterium Nostoc muscorum. The activity of the enzyme varied between 50 and 150 nmol reduced glutathione· min-1·mg protein-1, and the apparent Km for NADPH was 0.125 and 0.200 mM for heterocysts and vegetative cells, respectively. The enzyme was found to be sensitive to Zn+2 ions, however, preincubation with oxidized glutathione rendered its resistance to Zn+2 inhibition. Nostoc muscorum filaments were found to contain 0.6–0.7mM glutathione, and it is suggested that glutathione reductase can regenerate reduced glutathione in both cell types. The combined activity of glutathione reductase and isocitrate dehydrogenase in heterocysts was as high as 18 nmol reduced glutathione·min-1·mg protein-1. A relatively high superoxide dismutase activity was found in the two cell types; 34.2 and 64.3 enzyme units·min-1·mg protein-1 in heterocysts and vegetative cells, respectively. We suggest that glutathione reductase plays a role in the protection mechanism which removes oxygen radicals in the N2-fixing cyanobacterium Nostoc muscorum.
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  • 27
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    Archives of microbiology 110 (1976), S. 207-213 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; NH 4 + excretion ; Photosynthesis ; Rhodospirillum rubrum ; Photosynthetic bacteria ; Enzymes of ammonia assimilation
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    Topics: Biology
    Notes: Abstract NH 4 + excretion was undetectable in N2-fixing cultures ofRhodospirillum rubrum (S-1) and nitrogenase activity in these cultures was repressed by the addition of 10 mM NH 4 + to the medium. The glutamate analog,l-methionine-dl-sulfoximine (MSX), derepressed N2 fixation even in the presence of 10 mM extracellular NH 4 + . When 10 mg MSX/ml was added to cultures just prior to nitrogenase induction they developed nitrogenase activity (20% of the control activities) and excreted most of their fixed N2 as NH 4 + . Nitrogenase activities and NH 4 + production from fixed N2 were increased considerably when a combined nitrogen source, NH 4 + (〉40 μmoles NH 4 + /mg cell protein in 6 days) orl-glutamate (〉60 μmoles NH 4 + /mg cell protein in 6 days) was added to the cultures together with MSX. Biochemical analysis revealed thatR. rubrum produced glutamine synthetase and glutamate synthase (NADP-dependent) but no detectable NADP-dependent glutamate dehydrogenase. The specific activity of glutamine synthetase was observed to be maximal when nitrogenase activity was also maximal. Nitrogenase and glutamine synthetase activities were repressed by NH 4 + as well as by glutamate. The results demonstrate that utilization of solar energy to photoproduce large quantities of NH 4 + from N2 is possible with photosynthetic bacteria by interfering with their regulatory control of N2 fixation.
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  • 28
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Aerobic hydrogen bacteria ; Oxygen sensitivity ; Efficiency ; Aerobic and anaerobic acetylene reduction ; Corynebacterium autotrophicum
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    Topics: Biology
    Notes: Abstract The coryneform hydrogen bacterium strain GZ 29, assigned to Corynebacterium autotrophicum fixed molecular nitrogen under autotrophic (H2, CO2) as well as under heterotrophic (sucrose) conditions. Physiological parameters of nitrogen fixation were measured under heterotrophic conditions. The optimal dissolved oxygen concentration for cells grown in a fermenter with N2 was rather low (0.14 mg O2/l) compared with cells grown in the presence of NH 4 + (4.45 mg O2/l). C. autotrophicum GZ 29 had a doubling time of 3.7 h at 30°C with N2 as N-source and sucrose as carbon source and at optimal pO2. Acetylene reduction reached values of 12 nmoles of ethylene produced/minxmg protein. Although the oxygen concentration in the growing culture was kept constant, the optimal dissolved oxygen tension for the acetylene reduction assay shifted to higher pO2-values. The overall efficiency of nitrogen fixation amounted to 22 mg N fixed/g sucrose consumed; it reached a maximal value of 65 mg N fixed/g sucrose consumed at the beginning of the exponential growth phase. Intact cells reduced acetylene even under anaerobic test conditions; further anaerobic metabolic activity could not be ascertained so far.
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  • 29
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    Archives of microbiology 116 (1978), S. 113-118 
    ISSN: 1432-072X
    Keywords: Hydrogen evolution ; Hydrogenase ; Nodules ; Bacteroids ; Nitrogenase ; Rhizobium leguminosarum ; Acetylene reduction ; Nitrogen fixation ; Pisum sativum ; Legumes
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    Topics: Biology
    Notes: Abstract Hydrogen evolution from root nodules has been reported to make N2 fixation by some legume-Rhizobium symbiotic systems inefficient. We have surveyed the extent of H2 evolution and estimated relative efficiencies of nodules of Austrian winter peas formed by 15 strains of R. leguminosarum. Their rates of H2 evolution in air were about 30% of the rates of H2 evolution under an atmosphere in which N2 was replaced by Ar. Relative efficiency values based on C2H2 reduction rates ranged from 0.55 to 0.80. With some of the strains, hydrogenase activities were demonstrated in intact nodules and in bacteroids, but the levels of activity were insufficient to recycle all the H2 evolved by the nitrogenase system. In both intact nodules and bacteroids the hydrogenase is less sensitive to O2 damage than the nitrogenase system, so H2 uptake capacity was observed in intact nodules by suppressing the nitrogenase-dependent H2 evolution with an atmosphere containing a high O2 concentration, and in bacteroids by using aerobically prepared bacteroid suspensions. The hydrogenase activity of both was dependent on O2 consumption. A K mfor H2 of near 4 μM was determined in suspension of bacteroids from nodules formed by strains 128C53 and 128C56.
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  • 30
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    Archives of microbiology 121 (1979), S. 155-159 
    ISSN: 1432-072X
    Keywords: Blue-green algae ; Nitrogen fixation ; Anabaena ; Cyanobacteria ; Marine
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    Notes: Abstract Five strains of heterocystous blue-green algae capable of high rates of growth and nitrogenase activity were isolated from shallow coastal environments. Growth of the organisms was characterized with respect to temperature, NaCl concentration in the medium, and nitrogen source. The temperature optima ranged from 35–42°C, and all but one of the strains displayed a requirement for added NaCl. The generation times under N2-fixing conditions were 5.1–5.9 h, and were as low as 3.4 h for growth on NH4Cl. Nitrogenase activity (C2H2 reduction) was high throughout the logarithmic growth phase of each strain. The maximum value observed for one strain was 65.5 nmoles C2H4 produced/mg protein x min, and the average values for the five strains ranged from 24.5–46.7 nmoles C2H4/mg protein x min. The organisms all belong to the genusAnabaena. The growth and nitrogenase activity of these strains are much higher than those of the heterocystous blue-green algae commonly used for investigation of nitrogen metabolism, and they thus should prove to be useful physiological tools. Their prevalence, as judged by the ease of their enrichment and isolation, in bay and estuarine environments suggests that they are important contributors of combined nitrogen.
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  • 31
    ISSN: 1432-072X
    Keywords: Fluorescent antibody staining ; Nitrogen fixation ; Symbiosis ; Anabaena azollae ; Azolla caroliniana
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    Notes: Abstract Fluorescent antibody staining indicated differences in surface antigenicity in Anabaena azollae cells fresh from the leaf cavities of the fern, Azolla caroliniana, and algae which were isolated and subcultured from this fern. Such results suggest that either changes in antigenicity occur in this phycobiont during culturing or that isolation selects for an antigenically different mutant strain capable of in vitro growth.
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  • 32
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    Archives of microbiology 135 (1983), S. 103-109 
    ISSN: 1432-072X
    Keywords: RNA polymerase ; Transcription ; Nitrogen fixation ; Symbiosis ; Rhizobium
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    Topics: Biology
    Notes: Abstract DNA-dependend RNA polymerase (EC 2.7.7.6) from Rhizobium japonicum was purified. The subunit structure was found to be ββ′α2σ, with the following apparent molecular weights determined by electrophoresis: M r (β and β') 150,000 each, M r (σ) 96,000, M r (α) 40,000, M r (holoenzyme) 490,000, M r (core enzyme) 380,000. The recovery of σ was 28%. RNA polymerase from aerobically grown R. japonicum cells and from nitrogen-fixing cells have the same electrophoretic properties suggesting that no chemical modification of the enzyme occurs when cells undergo this metabolic differentiation. The enzyme is Mg2+-dependent, rifampicin-sensitive, and has optimal activity at alkaline pH (8–10) and at 35–40° C. It binds strongly to bacteriophage T7 promoters, weakly to antibiotic resistance genes, and not at all to cloned R. japonicum nif DNA. Preliminary in vitro transcription experiments, including nif DNA as template, revealed that additional factors may be required for selective transcription from promoters.
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  • 33
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    Archives of microbiology 130 (1981), S. 96-100 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrate respiration ; Denitrification ; Assimilatory nitrate reduction ; Dissimilatory nitrate reduction ; Acetylene reduction ; Azospirillum
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    Topics: Biology
    Notes: Abstract Azospirillum spp. participate in all steps of the nitrogen cycle except nitrification. They can fix molecular nitrogen and perform assimilatory nitrate reduction and nitrate respiration. Culture conditions have been defined under which nitrate is used both as terminal respiratory electron acceptor and as nitrogen source for growth. Nitrate and, possibly to a very limited extent, nitrite, but not sulfate, iron or fumarate support anaerobic respiration. Under anaerobic conditions, nitrate can also supply energy for nitrogen fixation but without supporting growth. Nitrate-dependent nitrogenase activity lasts only for 3–4 h until the enzymes of assimilatory nitrate reduction are synthesized. Nitrite accumulates during this period and inhibits nitrogenase activity at concentrations of about 1 mM.
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  • 34
    ISSN: 1432-072X
    Keywords: Rhizobium trifolii ; Symbiosis ; Nodulation ; Nitrogen fixation ; Symbiotic genes ; Reiterated sequences ; Plasmid
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    Notes: Abstract A Rhizobium trifolii symbiotic plasmid specific gene library was constructed and the physical organisation of regions homologous to nifHDK, nifA and nod genes was determined. These symbiotic gene regions were localised to u 25 kb region on the sym-plasmid, pPN1. In addition four copies of a reiterated sequence were identified on this plasmid, with one copy adjacent to nifH. No rearrangement of these reiterated sequences was observed between R. trifolii bacterial and bacteroid DNA. Analysis of a deletion derivative of pPN1 showed that these sequences were spread over a 110 kb region to the left of nifA.
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  • 35
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    Archives of microbiology 121 (1979), S. 141-145 
    ISSN: 1432-072X
    Keywords: Azospirillum ; Denitrification ; Nitrogen fixation
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    Notes: Abstract The interaction between nitrate respiration and nitrogen fixation inAzospirillum lipoferum andA. brasilense was studied. All strains examined were capable of nitrogen fixation (acetylene reduction) under conditions of severe oxygen limitation in the presence of nitrate. A lag phase of about 1 h was observed for both nitrate reduction and nitrogenase activity corresponding to the period of induction of the dissimilatory nitrate reductase. Nitrogenase activity ceased when nitrate was exhausted suggesting that the reduction of nitrate to nitrite, rather than denitrification (the further reduction of nitrite to gas) is coupled to nitrogen fixation. The addition of nitrate to nitrate reductase negative mutants (nr-) ofAzospirillum did not stimulate nitrogenase activity. Under oxygen-limited conditionsA. brasilense andA. lipoferum were also shown to reduce nitrate to ammonia, which accumulated in the medium. Both species, including strains ofA. brasilense which do not possess a dissimilatory nitrite reductase (nir-) were also capable of reducing nitrous oxide to N2.
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  • 36
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    Archives of microbiology 124 (1980), S. 161-167 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Photosynthesis ; Green alga ; Chl a and b ; DCMU ; Light and O2 dependency
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    Topics: Biology
    Notes: Abstract A unicellular alga which can grow in the light without a combined nitrogen source was isolated from a hot spring. The cells were almost spherical, usually 5–10 μm in diameter. Absorption spectra of the watersoluble pigments and of the acetone-extracted ones revealed the existence of chlorophyll a and b and the absence of phycobilins. Thin sections examined by electron microscopy revealed an eukaryotic organization with features typical of the coccoid green algae (the Chlorococcales). Cells divided by internal cytokinesis and subsequent liberation of daughter cells from the parental wall, in a way similar to Chlorella. The alga reduced acetylene to ethylene and incorporated 15N2 into cell protoplasm when incubated in a low oxygen atmosphere. Nitrogenase activity was light-dependent, microaerophilic and thermophilic. Although the association of symbiotic nitrogen fixing prokaryotes with the cells may still be possible, any such organisms have not so far been detected.
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  • 37
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    Archives of microbiology 127 (1980), S. 163-165 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Arthrobacter ; Corynebacterium ; Anabaena azollae
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    Topics: Biology
    Notes: Abstract Coryneform bacteria were found associated with the nitrogen fixing blue-green alga, Anabaena azollae in the leaf cavity of Azolla caroliniana. Plate counts indicated ca. 7,400±1,900 bacterial cells per mature leaf cavity or approximately 1 bacterial cell for every algal cell. No other type of bacterium was found in these cavities.
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  • 38
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    Archives of microbiology 127 (1980), S. 115-118 
    ISSN: 1432-072X
    Keywords: Proteus mirabilis ; Serratia marcescens ; Erwinia herbicola ; Nitrogen fixation ; nif genes ; his genes ; Plasmids
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    Notes: Abstract Plasmid pRD1, an R plasmid of the P incompatibility group which carries his and nif genes from Klebsiella pneumoniae in addition to drug resistance markers derived from RP4, was transferred to His- mutants of Serratia marcescens, Erwinia herbicola and Proteus mirabilis. His+ transconjugants were obtained at low but different frequencies according to recipient genus. Transconjugants all acquired the drug resistance, and were Nif+ in S. marcescens and E. herbicola, having acetylene-reducing activities of the same order of magnitude as the parent K. pneumoniae and fixing 15N2. No evidence for nif expression in P. mirabilis transconjugants was obtained though the nif genes were present.
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  • 39
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Regulation ; Guanosine 5′-diphosphate 3′-diphosphate (ppGpp)
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    Notes: Abstract Addition of ammonium to N2 fixing cultures of Azotobacter vinelandii, Klebsiella pneumoniae and Clostridium pasteurianum rapidly reduced the intracellular levels of guanosine 5′-diphosphate 3′-diphosphate (ppGpp) by 70–90%. This change might reflect a regulatory role of ppGpp in nitrogen metabolism.
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  • 40
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    Archives of microbiology 132 (1982), S. 333-337 
    ISSN: 1432-072X
    Keywords: Anabaena variabilis Kütz ; 14C-prelabeled blue-green algae ; Interaction respiration/photosynthesis ; CO2 exchange ; Nitrogen fixation
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    Topics: Biology
    Notes: Abstract Prelabeled Anabaena variabilis Kütz. evolves 14CO2 in the light with KCN and DCMU (2,4-dichlorophenyl-1,1-dimethylurea) present, comparable to the dark control without inhibitors added. Double-reciprocal plots of CO2 release vs. light intensity with either KCN or KCN+DCMU present result in two straight lines intersecting at the ordinate. Apparently, reducing equivalents originating from carbohydrate catabolism are channeled into the photosynthetic electron-transport chain, competing for electrons from photosystem II. Under these conditions, the CO2 release is accompanied by a light-dependent oxygen uptake, presumably due to oxygen-reducing photosystem-I activity while ribulose-bisphosphate carboxylase is inhibited by KCN. Comparing nine blue-green algae it was shown that only nitrogen-fixing species release substantial amounts of CO2 in the light with KCN or KCN+DCMU present. This release is particularly obvious with Anabaena variabilis Kütz. under nitrogen-fixing conditions, but small when the alga is grown with combined nitrogen. We conclude that nitrogen-fixing species share a common link between respiratory and photosynthetic electron transport. The physiological role may be electron supply of nitrogenase by photosystem I.
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  • 41
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    Archives of microbiology 133 (1982), S. 6-10 
    ISSN: 1432-072X
    Keywords: Rhodospirillaceae ; Rhodopseudomonas globiformis ; Nitrogen metabolism ; Nitrogen fixation ; Glutamine synthetase
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    Topics: Biology
    Notes: Abstract Rhodopseudomonas globiformis strain 7950 grew with a variety of amino acids, urea, or N2 as sole nitrogen sources. Cultures grown on N2 reduced acetylene to ethylene; this activity was absent from cells grown on nonlimiting NH 4 + . Glutamate dehydrogenase could not be detected in extracts of cells of strain 7950, although low levels of an alanine dehydrogenase were present. Growth ofR. globiformis on NH 4 + was severely inhibited by the glutamate analogue and glutamine synthetase inhibitor, methionine sulfoximine. High levels of glutamine synthetase (as measured in the γ-glutamyl transferase assay) were observed in cell extracts of strain 7950 regardless of the nitrogen source, although N2 and amino acid grown cells contained somewhat higher glutamine synthetase contents than cells grown on excess NH 4 + . Levels of glutamate synthase inR. globiformis were consistent with that reported from other phototrophic bacteria. Both glutamate synthase and alanine dehydrogenase were linked to NADH as coenzyme. We conclude thatR. globiformis is capable of fixing N2, and assimilates NH 4 + primarily via the glutamine synthetase/glutamate synthase pathway.
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  • 42
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    Archives of microbiology 133 (1982), S. 312-317 
    ISSN: 1432-072X
    Keywords: Rhizobium japonicum ; Rhizobium leguminosarum ; Formate metabolism ; Formate dehydrogenase ; Nitrogen fixation ; Nitrogenase ; Bacteroids ; Symbiosis
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    Topics: Biology
    Notes: Abstract Formate metabolism supported nitrogen-fixation activity in free-living cultures of Rhizobium japonicum. However, formate0dependent nitrogense activity was observed only in the presence of carbon sources such as glutamate, ribose or aspartate which by themselves were unable to support nitrogenase activity. Formate-dependent nitrogenase activity was not detected in the presence of carbon sources such as malate, gluconate or glycerol which by themselves supported nitrogenase activity. A mutant strain of R. japonicum was isolated that was unable to utilise formate and was shown to lack formate dehydrogenase activity. This mutant strain exhibited no formate-dependent nitrogenase activity. Both the wild-type and mutant strains nodulated soybean plants effectively and there were no significant differences in the plant dry weight or total nitrogen content of the respective plants. Furthermore pea bacteroids lacked formate dehydrogenase activity and exogenously added formate had no stimulatory effect on the endogenous oxygen uptake rate. The role of formate metabolism in symbiotic nitrogen fixation is discussed.
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  • 43
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Ultrastructure ; Nitrogen fixation ; Water stress ; Taxonomy ; DNA ; Plasmids
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    Notes: Abstract Two strains of desiccation-tolerant coccoid cyanobacteria, Chroococcus S24, a marine form, and Chroococcus N41, a cryptoendolith isolated from a hot-desert rock, have been characterized. The mol % DNA base compositions of the strains are 47.1 and 48.9% respectively. Plasmid DNA was not detected in either strain. The pigment contents and nutritional characteristics of the strains are identical. Both lack phycoerythrinoid pigments and, in culture, behave as slow-growing halotolerant marine forms with elevated requirements for Na+, Cl−, Mg2+ and Ca2+. Sucrose was the only carbon source of those tested that supported photoheterotrophic growth. Each strain synthesizes nitrogenase under anaerobic conditions but not in air. Morphologically the two strains are indistinguishable. They are considered to be independent isolates of the same cyanobacterial species. Chroococcus N41 was studied in detail with the electron microscope. When brought to equilibrium at matric water potentials of-168 MPa and lower (to-673 MPa=c0.12a w) the protoplast shrinks, but the cells maintain the same size and diameter as those at-2,156 kPa (MN medium; control); the sheath expands and remains attached to the cell wall outer membrane by fibrils. The cell wall, cell membrane, thylakoid membranes, cyanophycin granules and carboxysomes appeared intact in desiccated cells.
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  • 44
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    Archives of microbiology 136 (1983), S. 20-25 
    ISSN: 1432-072X
    Keywords: Hydrogen production ; Nitrogen fixation ; Hydrogen recycling ; Hydrogenase
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    Topics: Biology
    Notes: Abstract Mutants of Rhodopseudomonas capsulata lacking uptake hydrogenase activity have been isolated among those unable to grow photoautotrophically. Studies with these mutants showed increases in nitrogenase mediated H2 production from all substrates tested. In addition, photosynthetic synthetic growth on N2 with malate as carbon source was not affeced by the block in H2 uptake even under low light. Under these growth conditions hydrogen was observed to accumulate in mutant but not in wild-type cultures. This finding suggested that H2 was evolved by nitrogenase during N2 fixation by this photosynthetic bacterium and was efficiently recycled in the wild type.
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  • 45
    ISSN: 1432-072X
    Keywords: Ammonium export ; Ammonium assimilation ; Glutamine synthetase ; Nitrogen fixation ; Rhizobium sp. 32H1
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    Topics: Biology
    Notes: Abstract The relationship between ammonium assimilation and ammonium export has been studied in free-living, N2-fixing Rhizobium sp. 32H1. After 55 to 67 h of microaerobic growth under a gas phase of 0.2% O2 – 1.0% CO2 – 98.8% Ar high levels of nitrogenase were observed concomitant with a slightly adenylylated glutamine synthetase (GSI) and some glutamine synthetase (GSII) activity. However, after growth of 89 h, or longer, GSI became adenylylated and the level of GSII had decreased. When the gas phase was shifted to 0.2% O2 – 1.0% CO2 – 98.8% N2, a lag was observed before ammonium export could be detected in the 55 to 67 h cultures. No lag in ammonium export was observed in the cultures previously grown for 89 h. The onset of ammonium export in the 55 to 67 h cultures was found to correlate with the adenylylation state of GSI. There appeared to be no correlation between the level of GSII and the export of ammonium. Neither an increase in the adenylylation level of GSI nor ammonium export was observed when the 55 to 67 h cultures were maintained under the Ar gas mixture.
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  • 46
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    Environmental management 7 (1983), S. 177-187 
    ISSN: 1432-1009
    Keywords: Nitrogen ; Model ; Agriculture ; Mass balance ; Ground-water ; Denitrification ; Immobilization ; Dry deposition ; Nitrogen fixation ; Nitrate ; Florida
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract A detailed nitrogen budget was devised for agricultural activities in the Florida peninsula, based on routine data published by state agricultural agencies. The model demonstrates that important unmonitored fluxes of nitrogen can often be calculated by mass balance on individual model compartments, and that the reasonability of poorly quantified fluxes can be assessed. The results of such models can be very useful in designing and assessing the results of field experiments and in prioritizing environmental monitoring programs.
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  • 47
    ISSN: 1432-2048
    Keywords: Nitrogen fixation ; Nodule development ; Senescence (nodules) ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cowpea (Vigna unguiculata (L.) Walp cv. Vita 3) seedlings inoculated with Rhizobium strain CB756 were cultured with their root systems maintained in air or in Ar: O2 (80:20, v/v) during early nodule development (up to 24 d after sowing). Compared with those in air, seedlings in Ar:O2 showed progressive N deficiency with inhibited shoot growth, reduced ribulose-1,5-bisphosphate carboxylase and total protein levels and loss of chlorophyll in the leaves. Nodule initiation, differentiation of infected and uninfected nodule tissues and the ultrastructure of bacteriod-containing cells were similar in the air and Ar: O2 treatments up to 16 d after sowing. Thereafter the Ar: O2 treatment caused cessation of growth and development of nodules, reduced protein levels in bacteroids and nodule plant cells, and progressive degeneration of nodule ultrastructure leading to premature senescence of these organs. Provision of NO 3 - (0.1–0.2 mM) to Ar: O2-grown seedlings overcame the abovementioned consequences of N2 deficiency on nodule and plant growth, but merely delayed the degenerative effects of Ar: O2 treatment on nodule structure and senescence. Treatment of Ar: O2-grown seedlings with NO 3 - greatly increased the protein level of nodules but the increase was largely restricted to the plant cell fraction as opposed to the bacteroids. By contrast, NO 3 - treatment of air-grown seedlings increased protein of bacteroid and host nodule fractions to the same relative extents when compared with air-grown plants not supplemented with NO 3 - . These findings, taken together with studies of the distribution of N in nodules of symbiotically effective plants grown from 15N-labeled seed, indicate that direct incorporation of fixation products by bacteroids may be a critical feature in the establishment and continued growth of an effective symbiosis in the cowpea seedling.
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  • 48
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    Planta 152 (1981), S. 544-552 
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Lichens ; Nitrogen fixation ; Peltigera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Nostoc in the cephalodia of the lichen Peltigera aphthosa Willd. fixed 15N2 and the bulk of the nitrogen fixed was continuously transferred from it to its eukaryotic partners (a fungus and a green alga, Coccomyxa sp.). Kinetic studies carried out over the first 30 min, after exposure of isolated cephalodia to 15N2, showed that highest initial 15N2-labelling was into NH 4 + . After 12 min little further increase in the NH 4 + label occurred while that in the amide group of glutamine and in glutamate continued to increase. The 15N-labelling of the amino group of glutamine and of aspartate increased more slowly, followed by an increase in the labelling of alanine. When total incorporation of 15N-label was calculated, the overall pattern was found to be rather similar except that, throughout the experiment, the total 15N incorporated into glutamate was about six times greater than that into the amide group of glutamine. Pulse chase experiments, in which 14N2 was added to cephalodia previously exposed to 15N2, showed that the NH 4 + pool rapidly became depleted of 15N-label, followed by decreases in the labelling of glutamate, the amide group of glutamine and aspartate. The 15N-labelling of alanine, however, continued to increase for a period. When isolated cephalodia were treated with L-methionine-SR-sulphoximine, an inhibitor of glutamine synthetase (EC 6.3.1.2), and azaserine, an inhibitor of glutamate synthase (EC 2.6.1.53), there was no detectable labelling in glutamine although the 15N-labelling of glutamate increased unimpaired. On treating the cephalodia with amino-oxyacetate, an inhibitor of aminotransferase activity, the alanine pool decreased. Evidence was obtained that glutamine synthetase and glutamate synthase were located in the Nostoc, and that glutamate dehydrogenase (EC 1.4.1.4) and various amino-transferases were located in the cephalodial fungus. Possible implications of these findings are discussed.
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  • 49
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Glycine ; Nitrogen fixation ; Proplastid ; Purine synthesis ; Root nodule ; Ureide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Subcellular organelle fractionation of nitrogen-fixing nodules of soybean (Glycine max (L.) Merr.) indicates that a number of enzymes involved in the assimilation of ammonia into amino acids and purines are located in the proplastids. These include asparagine synthetase (EC 6.3.1.1), phosphoribosyl amidotransferase (EC 2.4.2.14), phosphoglycerate dehydrogenase (EC 1.1.1.95), serine hydroxymethylase (EC 2.1.2.1), and methylene-tetrahydrofolate dehydrogenase (EC 1.5.1.5). Of the two isoenzymes of asparate aminotransferase (EC 2.6.1.1) in the nodule, only one was located in the proplastid fraction. Both glutamate synthase (EC 1.4.1.14) and triosephosphate isomerase (EC 5.3.1.1) were associated at least in part with the proplastids. Glutamine synthetase (EC 6.3.1.2) and xanthine dehydrogenase (EC 1.2.1.37) were found in significant quantities only in the soluble fraction. Phosphoribosylpyrophosphate synthetase (EC 2.7.6.1) was found mostly in the soluble fraction, although small amounts of it were detected in other organelle fractions. These results together with recent organelle fractionation and electron microscopic studies form the basis for a model of the subcellular distribution of ammonium assimilation, amide synthesis and uredie biogenesis in the nodule.
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  • 50
    ISSN: 1432-2048
    Keywords: Alnus ; Ammonium ; Carbon translocation ; Endophyte damage ; Nitrogen fixation ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cloned plants of Alnus incana (L.) Moench were inoculated and grown without combined nitrogen for seven weeks. The effects of ammonium on the function and structure of the root nodules were studied by adding 20 mM NH4Cl (20 mM KCl=control) for four days. Nitrogenase activity decreased to ca. 50% after one day and to less than 10% after two days in ammonium treated plants, but was unaffected in control plants. The results were similar at photon flux densities of 200 and 50 μmol m-2 s-1. At the higher light level the effect was concentration dependent between 2 and 20 mM NH4Cl. The recovery was slow, and more than 11 d were needed for plants treated with 20 mM ammonium to reach initial activity. The distribution of 14C to the root nodules after assimilation of 14CO2 by the plants was not changed by the ammonium treatment. Microscopical studies of root nodules showed high frequencies of endophyte vesicles being visually damaged in nodules from ammonium-treated plants, but not in nodules from control plants. When nitrogenase activity was restored, visually damaged vesicles were again few, whereas young developing vesicles were numerous. The slow recovery, the 14C-translocation pattern, and the structural changes of the endophyte indicate a more complex mechanism of ammonium influence than simply a short-term reduction in supply of carbon compounds to the nodules.
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  • 51
    ISSN: 1432-2048
    Keywords: Leguminosae ; Nitrate-reductase mutants ; Nitrogen fixation ; Nodulation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of nitrate on the symbiotic properties of nitrate-reductase-deficient mutants of a strain of cowpea rhizobia (32H1), and of a strain of Rhizobium trifolii (TA1), were examined; the host species were Macroptilium atropurpureum (DC.) Urb. and Trifolium subterraneum L. Nitrate retarded initial nodulation by the mutant strains to an extent similar to that found with the parent strains. It is therefore unlikely that nitrite produced from nitrate by the rhizobia, plays a significant role in the inhibition of nodulation by nitrate. Nitrite is an inhibitor of nitrogenase, and its possible production in the nodule tissue by the action of nitrate reductase could be responsible for the observed inhibition of nitrogen fixation when nodulated plants are exposed to nitrate. However, the results of this investigation show that nitrogen fixation by the plants nodulated by parent or mutant strains was depressed by similar amounts in the presence of nitrate. No nitrite was detected in the nodules. Nodule growth, and to a lesser extent, the nitrogenase specific activity of the nodules (μmol C2H4g−1 nodule fr. wt. h−1), were both affected by the added nitrate.
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  • 52
    ISSN: 1432-2048
    Keywords: Centrosema leghaemoglobin ; Nitrate reductase ; Nitrogen fixation ; Nitrogenase ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence of events leading up to the establishment of symbiotic nitrogen-fixation were studied in two tropical legumes, Centrosema pubescens Benth, and Vigna unguiculata L. Walp. Parameters measured included fresh and dry weights, chlorophyll and leghaemoglobin contents, as well as the activities of NADH-nitrate reductase (EC 1.6.6.1), and nitrogenase (nitric-oxide reductase-EC 1.7.99.2) in plants that were inoculated with suitable rhizobia or which were watered with potassium nitrate. Dry weight and photosynthetic activity of both species followed the sigmoidal pattern which is characteristic of most plants. Growth was little different in either a qualitative or quantitative sense whether nitrogen was supplied as nitrate or through dinitrogen fixation. Although the biochemical sequence of events was dependent on the limiting sensitivities of the individual assays used, the data suggest that nitrate reductase is the first measurable enzymatic activity in the nodules (and roots), followed by acetylene reduction and leghaemoglobin in that order. It is possible therefore, that low levels of symbiotic nitrogen fixation occur in the nodules in the absence of leghaemoglobin. Nitrate reductase activity in C. pubescens nodules was negatively exponentially correlated with nitrogenase activity of the same nodules, suggesting a changing metabolism in old nodules. These data are discussed in terms of environmental and physical factors known to control nitrogen fixation.
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  • 53
    ISSN: 1432-2048
    Keywords: Allantoin ; Amino acids ; Bleeding sap ; Nitrogen fixation ; Phaseolus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dwarf french beans, Phaseolus vulgaris L., were grown with or without inoculation with rhizobia (strain 3644), and with or without a combined nitrogen source (nitrate or ammonium ions). The distribution of radioactivity into products of dark 14CO2 assimilation was studied in roots or nodules from these plants. A detailed study was also made of the distribution and rates of excretion of nitrogen in xylem bleeding sap in 28 day old plants grown on the various sources of nitrogen. Whereas detached nodules accumulated radioactive glycine, serine and glutamate when incubated with 14CO2, bleeding sap from plants root fed 14CO2 contained low levels of radioactivity in these compounds but higher levels in allantoin. Chemical analysis showed allantoin to be the major compound transported in the xylem of nodulated plants, whether or not they were fed on combined nitrogen. In contrast uninoculated plants accumulated mainly amino acids in the bleeding sap, the amount and chemical composition of which depended on the combined nitrogen source.
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  • 54
    ISSN: 1432-2048
    Keywords: Glyoxylate ; Isonicotinic acid hydracide ; Medicago ; Nitrogen fixation ; Photorespiration ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation (measured as acetylene reduction) by whole nodulated alfalfa plants was stimulated when the plants were treated with isonicotinic acid hydracide (INH) and glyoxylate, both inhibitors of the glycolate pathway of carbohydrate metabolism, at concentrations of 300 and 100 mM, respectively. Reducing energetic loses caused by photorespiration results in an increase in the symbiotic nitrogen fixation.
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  • 55
    ISSN: 1432-2048
    Keywords: Glutamine synthetase ; Leghaemoglobin ; Nitrogenase ; Nitrogen fixation ; Phaseolus ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activities of glutamine synthetase (GS), nitrogenase and leghaemoglobin were measured during nodule development in Phaseolus vulgaris infected with wild-type or two non-fixing (Fix-) mutants of Rhizobium phaseoli. The large increase in GS activity which was observed during nodulation with the wild-type rhizobial strain occurred concomitantly with the detection and increase in activity of nitrogenase and the amount of leghaemoglobin. Moreover, this increase in GS was found to be due entirely to the appearance of a novel form of the enzyme (GSn1) in the nodule. The activity of the form (GSn2) similar to the root enzyme (GSr) remained constant throughout the experiment. In nodules produced by infection with the two mutant strains of Rhizobium phaseoli (JL15 and JL19) only trace amounts of GSn1 and leghaemoglobin were detected.
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  • 56
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    Theoretical chemistry accounts 60 (1982), S. 579-587 
    ISSN: 1432-2234
    Keywords: Nitrogen fixation ; Nitrogen complexes ; Carbon monoxide complexes ; Electronic effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract MNDO method is used to study the interaction of nitrogen and carbon monoxide molecules with a proton, hydrogen atom, hydride ion, hydrogen molecule ion and hydrogen molecule. Predicted geometries and heats of reaction of different complexes are presented. The wave functions are analyzed in terms of ground state charge distributions and overlap populations. Electronic effects accompanying complexation are also discussed.
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  • 57
    ISSN: 1432-2048
    Keywords: Ammonia ; Nitrogen fixation ; Nodule ; Senescence (root nodules) ; Ureide ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract During early development (up to 18 d after sowing) of nodules of an “effective” cowpea symbiosis (Vigna unguiculata (L.) Walp cv. Vita 3: Rhizobium strain CB756), rapidly increasing nitrogenase (EC 1.7.99.2) activity and leghaemoglobin content were accompanied by rapid increases in activities of glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 2.6.1.53), enzymes of denovo purine synthesis (forming inosine monophosphate) xanthine oxidoreductase (EC 1.2.3.2), urate oxidase (EC 1.7.3.3), phosphoenolpyruvate carboxylase (EC 4.1.1.31) and led to increased export of ureides (allantoin and allantoic acid) to the shoot of the host plant in the xylem. Culturing plants with the nodulated root systems maintained in the absence of N2 (in 80 Ar: 20 O2, v/v) had little effect on the rates of induction and increase in nitrogenase activity and leghaemoglobin content but, in the absence of N2 fixation and consequent ammonia production by bacteroids, there was no stimulation of activity of enzymes of ammonia assimilation or of the synthesis of purines or ureides. Addition of NO 3 - (0.1–0.2 mM) relieved host-plant nitrogen deficiency caused by the Ar: O2 treatment but failed to increase levels of enzymes of N metabolism in either the bacteroid or the plant-cell fractions of the nodule. Premature senescence in Ar: O2-grown nodules occurred at 18–20 d after sowing, and resulted in reduced levels of nitrogenase activity and leghaemoglobin but increased the activity of hydroxybutyrate oxidoreductase (EC 1.1.1.30).
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  • 58
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    Hydrobiologia 49 (1976), S. 53-57 
    ISSN: 1573-5117
    Keywords: Nitrogen fixation ; Acetylene reduction ; Lake Mendota
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The contribution of nitrogen fixation to the nitrogen budget of Lake Mendota has been calculated. On average, the equivalent of 1.28 × 105 kg of NH3 (as determined by the acetylene reduction technique) was added to this eutrophic lake during June, July and August. Diurnal variation (approximately two-thirds of the day's fixation occurs prior to noon) in algal nitrogen fixation, and variation of fixation with depth (3.6% of the fixation in the column occurs in the top decimeter) were characterized as prerequisites to this calculation.
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  • 59
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    Hydrobiologia 56 (1977), S. 273-279 
    ISSN: 1573-5117
    Keywords: Nitrogen fixation ; nutrient enrichment ; limnocorral
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation was measured in situ by the 15N tracer technique in the Bay of Quinte, Lake Ontario, and three lake enclosures with different nutrient enrichment. The fixation rates in the Bay were low but detectable during the summer season. The fixation activities were found to be correlated with the presence of nitrogen-fixing blue-green algae and the distribution of the algal species in the water was affected by nitrate enrichment. The study showed that, with the addition of nitrate, species not able to fix atmospheric nitrogen became predominant. However, in the absence of external nitrogen (i.e., nitrate), species able to fix nitrogen became dominant. Phosphorus enrichment alone did result in higher N-fixation rates in the water and, without the addition of phosphorus, the fixation rates are lower and fluctuate throughout the season, presumably dependent on the availability of phosphorus in the water. A comparison between the 15N-isotopic method and the acetylene reduction method is reported and the factors involved in the variations between these two methods are discussed.
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  • 60
    ISSN: 1573-5036
    Keywords: Biofuel ; Leguminous trees ; Nitrogen fixation ; Semi-arid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sand culture pot experiments were carried out with Proposis seedlings in the greenhouse on a nitrogen free nutrient solution with increasing levels of sodium chloride. All species tolerated a 6,000 mg/l salinity with no reduction in growth.P. velutina was the only species that poorly tolerated the 12,000 mg/l salinity level.P. articulata, P. pallida, andP. tamarugo tolerated 18,000 mg/l NaCl with little reduction in growth and grew slightly in a salinity (36,000 mg/l NaCl) greater than seawater. This is the first legume known to grown in salinities equivalent to seawater.
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  • 61
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    Plant and soil 51 (1979), S. 163-165 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Actinomycete symbioses ; Datisca cannabina ; Nitrogen fixation ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The occurrence of root nodules inDatisca cannabina is reported. The nodules are typically of the Alnus type, forming dichotomously-branched coralloid clusters. The enlarged cortical cells contain vesicle clusters of the endophyte. The nodules reduced acetylene to ethylene at a rate of 5.5 μmoles ethylene per g fresh wt of nodules per hour.
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  • 62
    ISSN: 1573-5036
    Keywords: Aldicarb ; 14C-carbon metabolism ; Cowpea ; Growth ; Nitrogen fixation ; Nodulation ; Pesticide ; Respiration ; Rhizobium ; Soil ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Addition of aldicarb (2 methyl-2(methyl thio) propionaldehyde-0-methyl carbamoyl oxime) in the growth medium enhanced the growth ofRhizobium sp. (cowpea group) at 2 ppm level while an inhibition was observed at the normal (5 ppm) and higher (10 ppm) concentrations. Respiration of the cells was also inhibited by 5 and 10 ppm levels of the chemical eventhough a stimulation was observed at 2 ppm (lower) concentration. The insecticide, when incorporated at 5 and 10 ppm levels in the medium increased the14C-glucose incorporation and considerably altered the assimilation of the radioactive carbon in different fractions of rhizobium cells. Soil application of this insecticide (Temik 10 G) reduced the number of nodules formed and the total nitrogen content in cowpea plants inoculated with theRhizobium sp. but enhanced the dry matter production of cowpea plants.
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  • 63
    ISSN: 1573-5036
    Keywords: acetylene reduction assay ; Nitrogen fixation ; 15N natural abundance ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The use of variations in natural abundance of15N between nitrogen fixing and non nitrogen fixing soybeans was investigated for quantitative estimate of symbiotic nitrogen fixation. Isotopic analysis of 4 varieties of inoculated and non-inoculated soybeans growing under field conditions, with and without N-fertilizer was determined. It was found that inoculated soybeans had a significantly lower15N content than non-inoculated ones. Estimates of the participation of fixed N to the total nitrogen content of inoculated soybeans were calculated from these differences. They were compared to estimates calculated from differences in N yield between inoculated and noninoculated plants and to the nitrogenase activity, measured by the C2H2 reduction assay over the growing season. Estimates given by the15N measurements were correlated with the C2H2 reducing activity but not with the differences in the N yield. This shows that the isotopic composition was dependent on the amount of fixed nitrogen and consequently that the estimates of fixed nitrogen based on natural15N abundance should be reliable. The absence of correlation between estimates based on15N content and estimates based on N yield was explained by differences in the uptake of soil nitrogen between inoculated and non inoculated soybeans.
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  • 64
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    Plant and soil 53 (1979), S. 319-328 
    ISSN: 1573-5036
    Keywords: Dolichos ; Growth ; Greenhouse ; Lablab ; Leaf area ; Nitrogen fixation ; Nodulation ; Sand culture ; Sugars ; Sulfur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In order to explore interrelations between S nutrition, soluble sugars, leaf area, nodulation and N2 fixation, greenhouse experiments were done with several levels of S added to perlite-sand cultures or to a moderately S-deficient soil. Sulfur had indirect effects on nodulation and N2 fixation, possibly by improving sugars supply and N metabolism. In perlite-sand culture, leaf area increased with concentrations of supplied S up to 50 and 200 μM for symbiotic and N-treated plants respectively, then decreased at higher concentrations. Plant yield and total sugars content (mg per plant) for the N-treated plants behaved similar to leaf area in response to added S but in the symbiotic plants maximum values were obtained at 100 μM S. In soil, Mo had no effect on growth but interacted significantly with S in affecting total sugars content. High levels of S depressed sugars content at low Mo but raised it at high Mo. Sulfur increased the N content of soil-grown plants. It increased the N content of plants grown in perlite-sand culture except at very high levels of S. There was little effect on concentration of N in the shoots. Nitrogen content correlated significantly with leaf area and sugar content, and highly significantly with S concentration in the shoots.
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  • 65
    ISSN: 1573-5036
    Keywords: Drought stress ; Nitrogen fixation ; Nodulation ; Prosopis spp ; Semi-arid ; Tree legumes ; Water potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The literature does not contain a field report of nodulation for the xerophytic tree legumes of the genus Prosopis despite their widespread occurrence in semi-arid regions of the world. A phraetophytically simulated greenhouse experiment was conducted with mesquite in a 3m deep soil column to determine if nodules would form in deper depths where moisture is more abundant. The upper 0.5m in the soil column was allowed to dry to 2200 kPa but the 3.2 m depth was maintained at soil moistures more positive than 70 kPa by water additions to the bottom of the soil column. Over 100 nodules and an acetylene reduction rate of 1.9 mg/h were observed at the 3.2m depth. Nodulation or acetylene reduction were not observed closer than 2.7m from the surface. Air temperatures during these assays exceeded 45°C. Leaf xylem water potentials were in the 2800–3500 kPa range.
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  • 66
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    Plant and soil 65 (1982), S. 383-396 
    ISSN: 1573-5036
    Keywords: Frankia ; Microbial ecology ; Nitrogen fixation ; Purshia ; Revegetation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nitrogen fixing trees and shrubs may be useful in revegetation efforts. Speculation that insufficient endophyte populations in surface soils may limit non-leguminous symbiotic nitrogen fixation in marginal land was explored.Purshia tridentata andP. glandulosa seedlings were grown in greenhouse trials using ten soils from nativePurshia sites. Treatments include a control, an inoculated treatment, and six mmole nitrogen amendment. When inoculated with aP.tridentata crushed nodule inoculum, two of five non-nodulating soils and three sparsely nodulating soils produced well nodulated plants. Inoculation also increased nodule mass, total nitrogen, nitrogen content and shoot dry mass in plants from some of the soils. Of the three soils failing to produce nodulated plants when inoculated, one produced plants that responded well to nitrogen additions but failed to nodulate under low nitrogen conditions; another produced severely stunted plants indicating nutritional limitations on the host; and the third produced plants that were not nitrogen deficient. An application of nitrogen completely suppressed nodulation in all but one soil. The twoPurshia species were similar in nodulation, nitrogen fixation and growth, although important exceptions exist that indicate species may differ in adaptability to certain soil conditions.
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  • 67
    ISSN: 1573-5036
    Keywords: Isotope techniques ; Nitrogen-15 ; Nitrogen fixation ; Nitrogen utilization ; Phaseolus vulgaris ; Rhizobium ; Varietal comparison
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Differences in N2-fixation byPhaseolus vulgaris bean cultivars were successfully evaluated in the field using15N isotope dilution technique with a non-fixing test crop of a different species (wheat). The Phaseolus cultivars could have been similarly ranked for N2-fixation capacity from either seed yield or total nitrogen yield, but the isotope method provided a direct measure of N2-fixation and made it possible to estimate the proportion of fixed to total nitrogen in the crop and in plant parts. Amounts of nitrogen fixed varied between 24.59 kg N/ha for the 60-day cultivar Goiano precoce to 64.91 kg N/ha for the 90-day cultivar Carioca. The per cent of plant nitrogen due to fixation was 57–68% for the 90-day cultivars and 37% for Goiano precoce (60-day cultivar). Fertilizer utilization was 17–30% of a 20 kg N/ha fertilizer application. 100 kg N/ha fertilizer application decreased N2-fixation without suppressing it totally. Differences in yield between the highest yielding (Carioca) and the lowest (Moruna) 90-day cultivars were also due apparently to varietal differences in efficiency of conversion of nitrogen to economic matteri.e. seed, as well as to differences in capacity of genotypes for N2-fixation.
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  • 68
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    Plant and soil 66 (1982), S. 21-28 
    ISSN: 1573-5036
    Keywords: Agro-forestry ; Acetylene reduction ; Leucaena leucocephala ; Nematode root knots ; Nitrogen fixation ; Nodule biomass ; Woody legume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The nitrogen fixation rate in a 4-year-old stand of the woody legumeLeucaena leucocephala (Lam.) de Wit. was estimated in the field at a rather dry site in Tanzania by use of an acetylene reduction technique. The diurnal mean value during April–May was 35 nmol C2H4 mg−1 (dry weight) nodules h−1, with a variation between 22±8 and 48±12 nmol C2H4 mg−1 (dry weight) nodules h−1 in early morning and at midday, respectively. The nodule biomass was determined by auger sampling to be 51±16 kg (dry weight) ha−1. Most of the nodules were found at the 10–30 cm soil depth level. A rough calculation of the amount of nitrogen fixed annually arrived at 110±30 kg ha−1. The results give strong support for the use ofL. leucocephala for soil enrichment in less humid areas of tropical Africa.
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  • 69
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Actinomycete-nodulated ; Legume ; Lime ; Mine spoil ; Nitrogen fixation ; Reclamation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary An acid mine spoil in Southern Indiana was amended with lime (CaCO3) (0.0, 12.5, 25 and 39t/ha) and planted withElaegnus umbellata Thunb.,Alnus glutinosa Gaertn.,Robinia pseudoacacia L.,Robinia fertilis Ashe, ‘Arnot’,Myrica pensylvania Lois,Caragana arborescens L. andShepherdia argentea Nutt. Survival and soil data were collected periodically and plants were harvested 15 months after planting. Nodule and top dry weights were determined and acetylene reduction assays performed on the nodules. Addition of lime caused significant increases in pH, and 39 t/ha of lime were required to maintain a pH above 5.5. Survival of plant material was greatest at the highest lime addition, although response of individual species varied.Elaeagnus umbellata, R. pseudoacacia, R. fertilis ‘Arnot’, andA. glutinosa appeared more tolerant of the harsh conditions. OnlyC. arborescens showed a linear increase in top dry weight due to lime addition.Alnus glutinosa andS. argentea achieved statistically the same growth regardless of pH, andR. fertilis ‘Arnot’ andE. umbellata did not increase in top dry weight above an addition of 25 t/ha.Robinia pseudoacacia achieved maximum top dry weight at 25 t/ha, whereasM. pensylvanica growth declined with increasing pH. Nodule dry weights increased with increasing pH; however,S. argentea showed greater nodule dry weights at lower lime levels. Acetylene reduction rates increased with lime addition.Elaegnus umbellata did not respond above 25 t/ha lime, whereasA. glutinosa did not show an increase until this point.
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  • 70
    ISSN: 1573-5036
    Keywords: Actinorhizae ; Datisca cannabina ; Frankia ; Nitrogen fixation ; Root nodules ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The fine structures of the microsymbiont inside the root nodules ofDatisca cannabina have been studied by light, by transmission- and by scanning-electron microscopy. The endophyte is prokaryotic and actinomycetal in nature. The hyphae are septate and branched, diameter 0.3–0.5 μm. The tips of hyphae are swollen to form electron-dense, clubshaped to filamentous vesicles, ranging in diameter: 0.4–1.4 μm. The endophyte penetrates through walls of the cortial cells. The infected zone is kidney shaped and confined to one side of the acentric stele. The orientation of infection is reversed from other actinorhizae exceptCoriaria. The hyphae are near the host cell wall and vesicles are directed towards the central vacuole. Vesicles are aseptate and no collapsing of the vesicle cell wall (void area) has been observed. Vesicle clusters structures are globular with an opening at one side of the cluster. The host cell is multinucleate or contains a lobed nucleus. Groups of mitochondria are located in between the hyphae, suggesting a strong association between the host and the endophyte for energy supply and amino acid production. The consequences of the inability to separate the mitochondria from the vesicle clusters in nodule homogenates in physiological studies have been discussed. Isolated vesicles clusters showed dehydrogenase activity, indicated by the presence of formazan crystals, after incubation with NADH and NBT. Strongest reducing activity was found within the vesicles. The possible role of filamentous vesicles in nitrogen fixation has been discussed.
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  • 71
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    Plant and soil 68 (1982), S. 321-329 
    ISSN: 1573-5036
    Keywords: Alginate ; A value ; Endomycorrhiza ; Glomus mosseae ; Inoculant ; Nitrogen fixation ; Rhizobium japonicum ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary This field study was undertaken to determine the effect of inoculation withGlomus mosseae on N2 fixation and P uptake by soybean. The inoculation withGlomus mosseae was achieved using a new type of inoculant, alginate-entrapped (AE) endomycorrhizal fungus. N2 fixation was assessed using the A value method. In P-fertilized plots, inoculation with AEGlomus mosseae increased the harvest index based on dry weight (+20%) and N content of seeds (+17%), the A value (+31%) and %N derived from fixation (+75%). Inoculation with AEGlomus mosseae decreased the coefficient of variation for the A value and for the dry weights of the different plant parts.
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  • 72
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    Plant and soil 69 (1982), S. 233-238 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; Azospirillum spp. ; Azospirillum lipoferum ; Wheat yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Seven isolates belonged toA. brasilense and 3 belonged toA. lipoferum. Isolates having more denitrifying capacity fixed less nitrogen in nitrogen free semi-solid malate medium. One strain ofA. lipoferum having high nitrogen fixing capacity with negative test for denitrification was tested as inoculant to supplement the nitrogen need of a wheat crop in field condition with different doses of N with and without the inoculant. While control without nitrogen yielded 1260 kg/ha the yield in inoculated treatment was 2070 kg/ha resulting in significant increase. In a treatment receiving 40 kg N/ha the grain yield was 2370 kg/ha as against yield of 3110 kg/ha in a similar treatment receiving fertiliser plus inoculant. Thus increase in yield was about 30%. Further the treatment receiving 80 kg N/ha yielded 2970 kg/ha as against yield of 4150 kg/ha in a treatment receiving inoculant alongwith the above dose of the fertiliser. Thus increase in yield due to application of inoculant was about 36%. Similarly, the uptake of N in different treatments was augmented due to inoculation of seeds with the culture.
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  • 73
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    Plant and soil 60 (1981), S. 309-315 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Aeschynomene americana ; Flooding ; Leaf water potential ; Nitrogen fixation ; Soil moisture deficits
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Studies on the tolerance ofAeschynomene americana L. to periods of flooding or soil moisture deficit were conducted in an attempt to elucidate nitrogen fixation as affected by soil moisture. Nitrogenase activity was not reduced significantly in pot-grown Aeschynomene plants subjected to flooding in greenhouse conditions. After 20 days of withholding water from the soil, nitrogenase activities of the drought-stressed plants were much lower than those of either the well-watered or flooded plants. Leaf water potentials were similar in flooded and control plants; however, the droughted plants had leaf water potentials that were 4 bars lower than those of the control plants. Aeschynomene plants were tolerant to long-term periods of flooding, but exhibited a reduction in nitrogenase activity and leaf water status when subjected to soil moisture deficits.
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  • 74
    ISSN: 1573-5036
    Keywords: Bacteroids ; Cowpea ; Nitrogen fixation ; Peanut ; Rhizobium ; Siratro
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Acetylene reduction activity and nitrogen accumulation in the plant top per unit nodule mass were compared among peanut, cowpea and siratro plants nodulated by six different strains of Rhizobium. Peanut was found to have several fold higher values than cowpea and siratro for both parameters for all strains of Rhizobium which nodulated it effectively, but the bacteroid content of the peanut nodules was similar to those of cowpea and siratro.
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  • 75
    ISSN: 1573-5036
    Keywords: Azospirillum brasilense ; Cereals ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The potential of the nitrogen fixing bacteriumAzospirillum brasilense to enhance development and increase growth of several gramineae was investigated. In both sterilized and non-sterilized systems heading and flowering occurred earlier in the inoculated plants as compared to the noninoculated ones. Total shoot and root weights, total-N content, plant height and leaf length were significantly increased by inoculation.
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  • 76
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    Plant and soil 73 (1983), S. 431-434 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; 15N2 diffusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary The kinetic of15N2 diffusion has been measured in a system similar to that for the estimation of N2 fixation in plant microorganism associations cultivated in soil. The15N2 enrichment of the soil atmosphere reached an homogenous value one hour after injection of15N2 and is identical to that obtained by calculation, indicating that no adsorption occurs in the soil particles.
    Notes: Résumé La cinétique de diffusion du15N2 est mesurée sur un système identique à ceux pouvant être utilisés pour la mesure de fixation de l'azote chez les associations plantes-microorganismes cultivées sur sol. L'enrichissement homogène de l'atmosphère du sol est obtenu une heure environ après l'injection de15N2 et correspond à l'enrichissement calculé, ce qui indique qu'aucune adsorption n'a lieu dans les particules du sol.
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  • 77
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    Plant and soil 73 (1983), S. 299-305 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; Rhizobium trifolii populations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effectiveness ofRhizobium trifolii isolates from five locations in southern Britain representing contrasting soil types has been examined with five white clover varieties. The average effectiveness of Rhizobium isolates varied considerably as did the average productivity of plant varieties. The largest differences were, however, associated with Rhizobium population × plant variety interactions. These were often large enough to reverse relative yield differences between white clover varieties. The implications of these results for improving clover productivity in nitrogen fixation are discussed.
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  • 78
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    Plant and soil 82 (1984), S. 329-335 
    ISSN: 1573-5036
    Keywords: Legume breeding ; Medicago sativa ; Nitrogen fixation ; Rhizobium ; Trifolium repens ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary This paper examines evidence which quantifies the relative importance of legume and Rhizobium genotypes as determinants of phenotypic variation in symbiotic nitrogen fixation. It demonstrates potentially large and unpredictable effects of the Rhizobium genotype. The likely importance of such effects on crop yield is considered. The information is then used to assess ways in which legume breeding programmes may be altered to encompass the effects of genetic variation in Rhizobium.
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  • 79
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    Plant and soil 75 (1983), S. 309-342 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Amino-acid composition ; Chromosome number ; Cross-inoculation ; Growth various substrates ; Medium composition ; Nitrogen fixation ; Parasponia parviflora ; Root nodule ; Temperate Rhizobium species
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Cross-inoculation experiments withParasponia parviflora plants and a large number of Rhizobium strains isolated from legumes, mainly of temperate origin, showed that strains ofRhizobium trifolii andR. lupini could produce root nodulation in Parasponia. Root nodulation was also obtained with some tropical Rhizobium isolates such as those from Arachis and Albizzia, but not withRhizobium japonicum strains. In addition, strains ofR. leguminosarum, R. phaseoli andR. meliloti produced abnormal root nodules or pseudo-root nodules in Parasponia seedlings. All root nodules induced in Parasponia by foreign Rhizobium species lacked, however, nitrogenase activity. Testing the reverse combination Parasponia-Rhizobium on legumes yielded with regard to root nodulation negative results, except withVigna sinensis and twoMacroptilium species (M. atropurpureum andM. lathyroides). The root nodules produced in the latter legumes showed nitrogenase activity.Trema cannabina seedlings inoculated with Parasponia-Rhizobium invariably did not produce root nodules. The feasibility of Parasponia to accept foreign rhizobia bacteria or to produce pseudo-nodulation is probably genetically determined. In a karyological study it was shown thatParasponia parviflora had the chromosome number 2n=20. Growth experiments revealed thatP. parviflora can be cultivated in water culture, perlite and in soil (pot) culture. It can also be grown in agar tubes, although growth under these conditions is sometimes rather variable. Nitrogenase (C2H2) tests showed that water-culture root nodules were 5–10 times less active than those from perlite culture, soil or agar tubes. On the whole, nitrogenase activity of root nodules grown on the various substrates and receiving different treatments, was rather variable.Vigna sinensis plants inoculated with Parasponia-Rhizobium showed on basis of fresh nodule weight a 3–4 times higher nitrogenase activity as compared with the same strain on Parasponia. However, as shown by acetylene reduction tests of nodule slices, the main activity of mature Parasponia root nodules is situated in the apical part of the root nodule. On fresh weight basis the nitrogenase activity of this nodule section is of the same order as that of leguminous root nodules. Intact Parasponia plants showed higher nitrogenase activities than excised root nodules. The maximal nitrogen fixation rate ofParasponia parviflora plants was 20–25 μmol. C2H4·g−1 fresh weight nodule tissue.h−1, which value is comparable to that of legumes and higher than estimates obtained by Parasponia in the field. No decrease of the nitrogen-fixation rate was observed in Parasponia plants tested in a diurnal cycle during the 12-hours dark period. Amino-acid analyses of Parasponia root nodules showed large quantities of aspartic acid, asparagine, glutamic acid and glutamine suggesting a role of these amino acids in the nitrogen fixation or transport processes. In contrast to actinorhizal root nodules Parasponia root nodules do not contain citrulline. The latter amino acid was, however, found—although in rather low concentration—inVigna sinensis andMacroptilium atropurpureum nodules obtained withParasponia-Rhizobium.
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    Plant and soil 77 (1984), S. 3-14 
    ISSN: 1573-5036
    Keywords: Asian-type soybean ; Fast-growingRhizobium japonicum ; Glycine max ; Nitrogen fixation ; Rhizobium japonicum ; Rhizobium physiology ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Physiological and symbiotic characteristics were identified in fast-growing (FG)Rhizobium japonicum. Carbon nutritional patterns linked these rhizobia to other FG rhizobia. They were able to use hexoses, pentoses, disaccharides, trioses, and organic acids for growth, but they were unable to use dulcitol or citrate. These rhizobia produced acid with all carbon sources except intermediates of the Krebs cycle. FGR. japonicum showed no vitamin requirements and were tolerant to 1% NaCl but not to 2%. They nodulated cowpea, pigeon pea, and mung bean but not peanut. Effective, nitrogen-fixing symbioses were observed only with cowpea and pigeon pea. In addition, FGR. japonicum formed effective symbioses with Asian-type soybeans. We concluded that although the physiological characteristics of FGR. japonicum were similar to other FG rhizobia, their symbiotic properties were similar to slow-growing rhizobia of the cowpea miscellany.
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  • 81
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    Plant and soil 51 (1979), S. 187-196 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Aerobic ; Anaerobic ; Bacillus ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Among 390 isolates from Egytiian soils initially grown on Brown's N-free agar, 15 facultative Bacillus isolates were able to reduce acetylene in Stanier's N-poor broth under both aerobic and anaerobic (N2 atmosphere) conditions. Some of these isolates were Gram-positive, with unswollen sporangia and thin-walled endospores. Other strains were with slightly or definitely bulged sporangia. Yeast extract (0.01%) was essential for growth stimulation and N2[C2H2] fixation by these isolates. Replacing yeast extract with 20 μg/ml (NH4)2SO4 or biotin, thiamine and amino acids (singly or in combination) resulted in stimulation of growth and N2[C2H2] fixation, though at lower rates than in yeast extract. One isolate was able to grow and reduce C2H2 in Stanier's N-free liquid medium. Nitrogenase [C2H2] activity of the anaerobically grown and incubated cultures was greater than aerobic cultures. Addition of 0.1% CaCO3 to the culture media significantly increased and O2 partially inhibited, N2[C2H2] fixation by these Bacillus isolates. Studies of the characteristics and N2[C2H2] fixing activities of these isolates indicate that at least some of them are new nitrogen-fixingBacillus species.
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  • 82
    ISSN: 1573-5036
    Keywords: Arachis hypogaea ; Peanuts ; Rhizobium ; Nitrogen fixation ; Nitrogen fertilizers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The potential of peanuts for symbiotic nitrogen fixation is considerable and under optimal edaphic and climatic conditions it reached 222 kg N2/ha, which was 58% of the nitrogen accumulated in the plants. The effect of the Rhizobium inoculation on crude protein accumulation in the yield (kg/ha) was 3–4 times greater than its effect on the yield of pods and hay. There was an inverse relationship between the protein and oil content in the kernels. Seasonal changes in nitrogenase activity in the nodules were determined by the acetylene reduction method during two growing seasons. Under favorable conditions the specific activity of the nitrogenase reached a very high level (up to 975 μmoles C2H2 g dry wt nod/h) and the total activity (μmoles C2H4/plant/h) was also high in spite of the relatively poor nodulation (weight and number). The high activity was drastically reduced (to 75 μmoles C2H4 g dry wt nod/h) due to exceptionally hot and dry weather, which occurred in the middle of the second half of the growing season. It appears that N2-fixation (nitrogenase activity) is more sensitive to these unfavorable conditions, than is nodule growth. Maximum nitrogenase activity was observed during the podfilling stage; until 50–60 days after planting, nitrogenase activity was very low.
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  • 83
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    Plant and soil 52 (1979), S. 49-57 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Azospirillum ; Dry weight ; Greenhouse ; Inoculation ; Millet ; Nitrogen fixation ; Pennisetum ; Spirillum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Millet plants (Pennisetum glaucum) were grown at three levels of nitrogen fertilization with and without an inoculum of live nitrogen-fixing Azospirillum cells. The highest average rate of nitrogen fixation as estimated from acetylene reduction by excised preincubated roots was only 23g N2 fixed per ha per day and occurred after treatment with low levels of nitrogen amendment. The average rates of acetylene reduction for intact plants at all treatments were also low. The lack of significant nitrogen fixation due to an Azospirillum-millet association in this study was substantiated by plant dry weight analysis, and determination of the nitrogen content of plants, pot leachate, and soil. There was significant correlation between the total nitrogen content of the plants per pot at the termination of the experiment and the amount of nitrogen fertilizer added initially, but there was no effect of inoculum on final total nitrogen content.
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  • 84
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    Plant and soil 52 (1979), S. 571-578 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Actinomycetous symbiosis ; Alnus glutinosa ; Hydrogenase Hydrogen evolution ; Hydrogen uptake ; Nitrogen fixation ; Respiration ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In the growing season no net H2 evolution is detected when root nodules ofAlnus glutinosa are incubated in air or in argon containing 20% O2. Due to the hydrogenase activity, N2-fixing root nodules consume added H2 at a rate of about 1.4 μmoles H2.g fresh nodule−1.h−1. The uptake of H2 is only found in summer. At the end of the season, in autumn, nodules evolve significant quantities of H2 although the nodules still continue to fix nitrogen. In-vitro studies with fractionated homogenates of summer-harvested nodules show that the recovery of the hydrogenase is high when using methylene-blue or phenazine metasulfate as electron acceptors. No hydrogenase activity is detected in homogenates of autumn-harvested nodules. The hydrogenase is localised in the microsymbiont.
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  • 85
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    Plant and soil 54 (1980), S. 33-44 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Harwood forests ; Nitrogen cycle ; Nitrogen fixation ; United States ; Wood litter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nitrogen-fixing activity in hardwood forests of the northeastern United States occurred in wood litter, greater than 2 cm in diameter. Activity in large dead wood was independent of species, in the case of deciduous wood litter, but was restricted to partially decayed wood with a high moisture content. Maximum rates of activity were observed in the summer months, minimum rates in the winter. Evidence from six stands of varying ages showed that fixation in large wood litter occurred in only 25% of the samples assayed. Fixation was highest in the youngest, 4 years, and oldest, over 200 years, stands; being about 2 kg/ha/yr. The quantity of nitrogen fixed appears to be related to the biomass of dead wood. Large amounts of wood litter in the youngest stands were from slash left after cutting. As the supply of slash is exhausted by decay, nitrogen fixation decreases, with a low around year 20. Fixation then gradually increases as natural thinning adds wood to the litter compartment. Apparently, the amount of nitrogen fixed in dead wood the first 20 years following clearcutting can only replace a modest fraction of the amount lost as a result of the cutting and product removal. Finally, the results indicate that nitrogen fixation in wood litter does not equal nitrogen fixation in a northern hardwood forest calculated using a mass balance approach, suggesting that additional nitrogen inputs exist.
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  • 86
    ISSN: 1573-5036
    Keywords: Acid-sensitive step ; Nitrogen content ; Nitrogen fixation ; Pisum sativum ; Rhizobium leguminosarum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Experiments were performed to measure the pH-sensitive steps in nodulation and symbiotic fixation byPisum sativum and isolate RP-212-1 ofRhizobium leguminosarum. An aeroponic system with rigorous pH control was used to obtain numerous effective nodules. After exposure to various pH levels, the following responses were measured: (1) legume root growth and development, (2) survival and growth rate of a single effective bacterial isolate, (3) degree of nodulation, (4) rate of nitrogen fixation, (5) plant biomass, and (6) nitrogen content of plants. Both bacterial growth and root development were adequate at all pH levels from 4.4 to 6.6, but efficient nodulation and nitrogen fixation did not occur at pH 4.8 and below. The processes required for symbiosis were about 10 times as sensitive to acidity as either bacterial growth or root growth alone. Nodulation was the most acid-sensitive step.
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    Plant and soil 56 (1980), S. 335-340 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Field method ; Nitrogen fixation ; Non-destructive ; Pisum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Direct injection of acetylene into soil around plant roots, followed by determination of ethylene/acetylene ratios in the soil atmosphere has been tested as a rapid, non-destructive method of estimating acetylene reducing activity. In pots of artificial media as well as in field soil, the ratios determined within 10 min. after injection were significantly correlated with the rates of acetylenedependent ethylene production in detached roots. The method may be useful in preliminary screening of large numbers of plant-bacteria combinations.
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    Plant and soil 64 (1982), S. 251-253 
    ISSN: 1573-5036
    Keywords: Competition of Rhizobium ; Nitrogen fixation ; Nodulation ; Vigna radiata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Competition of five strains of Rhizobium of the cowpea group, onVigna radiata (L) Wilcjeck variety ML 5, was tested in loamy clay and loamy sand soils. Strains RM 6 and RM 5 were effective nodulators in loamy clay soil, and strains MNH, M 20 and RM 6 were effective nodulators in loamy sand soil. Strains RM 6 and MNH predominated nodule formation in loamy clay and loamy sand soils respectively.
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  • 89
    ISSN: 1573-5036
    Keywords: Moisture stress ; Nitrogen fixation ; Tropical forage legume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effect of soil moisture stress on growth, nodulation and nitrogenase activity of two tropical forage legumes,Macroptilium atropurpureum cv. Siratro andDesmodium intortum cv. Greenleaf was studied in a pot experiment. After ten weeks growth, the highest moisture stress (20 per cent water holding capacity) significantly reduced only the top weight of both plants. Moisture stress progressively retarded top growth in the two legumes. Similar trends were also observed in defoliated plants. Moisture stress had little or no effect on the nodulation or nitrogenase activity of the plants.
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    Plant and soil 66 (1982), S. 125-127 
    ISSN: 1573-5036
    Keywords: Charcoal ; Garden pea ; Inoculation ; Nitrogen fertilisation ; Nitrogen fixation ; Nodulation ; Pisum sativum ; Wood charcoal
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In a factorial pot experiment on garden peas investigating the effect of growing medium, inoculation, nitrogen fertilisation and charcoal addition, shoot growth was significantly better in John Innes potting compost than in 50/50 peat/sand compost and when receiving nitrogen fertilisation but was depressed by the presence of wood charcoal. In the peat/sand medium, root systems were smaller but carried more and larger nodules. The number of nodules was increased by inoculation but the addition of carbon decreased root size, nodule number and nodule size.
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    Plant and soil 66 (1982), S. 413-416 
    ISSN: 1573-5036
    Keywords: Growth ; Nematicide ; Nitrogen fixation ; Nodulation ; Peanut ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effect of two granular nematicidesviz. oxamyl and fenamiphos, on the nodulation and growth of Rhizobium inoculatedArachis hypogaea L. was studied in glasshouse and field trials. In the glasshouse trial at the suggested rates of application shoot fresh weight was significantly reduced by oxamyl whilst root fresh weight was similarly affected by fenamiphos. In the field trial vegetative growth and plant emergence were significantly reduced by both nematicides. Nodulation at the higher rates of application was increased by both oxamyl and fenamiphos whilst oxamyl caused a significant increase in pod number at the highest rate of application.
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    Plant and soil 79 (1984), S. 37-49 
    ISSN: 1573-5036
    Keywords: Azotobacter ; Nitrogen fixation ; N2-fixing efficiency ; Respiration ; Soil ; Water potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Respiration and N2-fixation (acetylene reduction) ofAzotobacter vinelandii have been studied at a variety of soil water potentials. Both processes were strictly linked and strongly reduced at water potentials between −0.6 and −1.3 MPa. Complete inhibition occurred below −2.1MPa. Osmotic potentials in soil compared to matric potentials of the same value were less inhibitory to respiration and acetylene reduction by Azotobacter. The N2-fixing efficiency (mg N/g glucose) was not influenced by water potentials ranging from −0.1 to −2.1 MPa.
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  • 93
    ISSN: 1573-5036
    Keywords: Mesquite ; Nitrogen fixation ; 15N natural abundance ; Nodule morphology ; Rhizobium bacteria ; Woody legumes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The relative nitrogen fixation efficiencies (RE 1-[H2 evolved÷C2H2 reduced]·100) of four mesquite (Prosopis glandulosa var.torreyana) rhizobia (Strains WR 1001, WR 1002, L5, L9) and a cowpea rhizobia (Strain 176A32) on mesquite were evaluated in a glasshouse experiment. Plant yield, shoot N accumulation, and the natural15N abundance (δ15N) of nodule tissue were determined. Strain WR 1002 failed to nodulate mesquite and strain L5 produced ineffective nodules. Among the three effective strains (WR 1001, L9, 176A32) the cowpea strain (176A32) and strain L9 had significantly higher RE than strain WR 1001. Differences in RE, however, were not accompanied by significantly higher plant yield and shoot N accumulation. The difference in15N abundance between foliar tissue and nodules (nodules minus leaves) was 0.47 δ15N for the ineffective L5 nodules, while for the effective WR 1001, L9, and 176A32 nodules, respectively, this difference was 8.35, 7.81, and 8.35 δ15N. This indicates a similar relationship between N2-fixing effectiveness and natural15N enrichment of nodules that was previously observed in soybeans (Glycine max, L. Merr.). Strains WR 1001 and L9 produced elongate, indeterminate nodules typical for mesquite. The ineffective L5 nodules had few infected cells and an abundance of cortical amyloplasts. Mesquite nodules produced by the cowpea strain were spherical and were somewhat more similar in internal morphology to determinate nodules typical of cowpea than indeterminate nodules normally associated with mesquite.
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  • 94
    ISSN: 1573-5036
    Keywords: Azotobacter vinelandii ; Gnotobiotic cultures ; Nitrogen fixation ; Oryza sativa Rhodopseudomonas capsulata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary An all glass tight growth chamber, entirely sterilizable, has been constructed to carry out axenic and gnotobiotic cultures of rice plants (Oryza sativa L.). When grown in liquid medium and in the absence of combined nitrogen but in the presence of the diazotrophsAzotobacter vinelandii andRhodopseudomonas capsulata, rice plants exhibited a complete biological cycle from germination up to ear stage, during a period of time similar to the one encountered in french paddy soil of Camargue. In one experiment, mannitol was given to rice culture medium together withAzotobacter vinelandii andRhodopseudomonas capsulata. In another experiment, mannitol was not given together with Rhodopseudomonas, and still positive nitrogen gain was obtained, although it was less than culture with mannitol. When15N labeled cells of Rhodopseudomonas were added in rice culture medium,15N was partly transferred to rice plant. Among the nitrogen substances excreted from the bacteria in the rhizosphere medium, large organic molecules were shown to be the most abundant in our experimental conditions. Moreover, the concentration of free ammonia or aminoacids present in the rice rhizosphere were always compatible with a bacterial nitrogenase activity.
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  • 95
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    Plant and soil 69 (1982), S. 265-273 
    ISSN: 1573-5036
    Keywords: Azotobacter chroococcum ; Fertilizers ; Farmyard manure ; Nitrogen ; Nitrogen fixation ; Potassium ; Phosphorus ; Yields of grain stover ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Field trials were conducted during theKharif seasons of 1979 and 1980 to examine the effect of the nitrogen-fixing bacteriumAzotobacter chroococcum (isolate M4) on yields of maize and N-economy. Different levels of nitrogen and farmyard manure (FYM) were supplied to assess their interaction with Azotobacter inoculation. Seed inoculation without fertilization raised grain and stover yields significantly. FYM coupled with inoculation gave higher yields than either could singly. The financial gain due to an increase in grain yield upon applying per ha 80 kg N, 10×103 kg FYM and Azotobacter over the treatment 40N + FYM + Azotobacter was offset by the cost of the additional N; thus, the higher N-application was not economical. Azotobacter inoculation was economically most efficient at lower doses of fertilizer nitrogen which not only increased yields but resulted in a saving of fertilizer N when applied in combination with FYM.
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  • 96
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    Plant and soil 61 (1981), S. 53-63 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Inoculation ; Klebsiella oxytoca ; Nitrogen fixation ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Of 45 fermentative gram negative bacterial isolates examined from wheat roots, three were capable of fixing atmospheric nitrogen as determined by the acetylene reduction technique and by protein contents of cells. A gram negative non-motile facultatively anaerobic bacterial strain capable of N2 fixation was identified asKlebsiella oxytoca ZMK-2. Optimal growth and N2 fixation occurred at pH 6.5. The optimum temperatures for growth under anaerobic conditions ranged between 30°–37°C. Acetylene reduction by intact cells was strikingly inhibited by 0.1 atm. or greater partial pressure of O2. Furthermore, the accumulation of H2 in the gas phase over cultures ofKlebsiella oxytoca ZMK-2 at partial pressures greater than 0.02 atm. resulted in a striking inhibition in the rate of C2H2 reduction. The addition of suspensions of eitherKlebsiella oxytoca ZMK-2 orAzotobacter vinelandii or a mixed culture of these two organisms to axenic cultures of wheat plants produced no significant increase in plant growth as measured by plant dry weight or nitrogen content of plants.
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  • 97
    ISSN: 1573-5036
    Keywords: Host selection ; Nitrogen fixation ; Nodule structure ; Rhizobium trifolii ; TemperatureTrifolium pratense
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary 1. Nodule structure was examined in the red clover cultivar S123 and in lines (designated H2) separately bred for high yield withRhizobium trifolii strains RCR 0403 and RCR 5. Plants were grown at 16/11, 22/17 and 27/22°C in a 16 h day at 25,000 lux. 2. The larger yields of bred lines compared with S123 were directly correlated with larger aggregate nodule size, larger infected zones and with more bacteroid tissue. Over all treatments the correlation of plant dry weight with aggregate areas of bacteroid tissue, assessed in median longitudinal section, accounted for 79% of the variance. 3. Yields were unrelated to the proportion of uninfected cells and vacuoles within the bacteroid zone. 4. Effects of host type and strain on nodule structure and yield were greatest at the moderate temperature and least and most irregular at high temperature. 5. Bacteriod tissue degenerated more rapidly in S123 than in the bred lines, more rapidly with RCR 0403 than RCR 5 and more rapidly at the high temperature than at moderate or low temperatures. 6. Aggregate nodule size irrespective of treatment was correlated with the aggregate sizes of nodule meristem and differentiating tissues. Nodule comtex comprised a larger proportion of the nodule section at 17 days than subsequently. Other than as specified above, the size relationships of the different tissues of the nodule were unaffected by host selection, temperature, bacterial strain or plant age. 7. At 22/17°C over a 17–25 day period the increment in dry matter per mm2 active nodule tissue was similar for H2 and S123 plants and was less for RCR 0403 (3.33 mg) than for RCR 5 (6.87 mg). At 27/22°C the effeciency of bacteroid tissue was much less,viz. 0.67 and 1.23 mg per mm2 respectively. 8. Nodule tissue areas and volumes were closely related so that unit volumes of active bacteriod tissue in general promoted similar dry matter increment in S123 and bred lines,viz. about 50.5 mg mm−3 at 22/17° for RCR 0403 and 80.8 mg mm−3 for RCR 5. The latter was associated with a fixation rate of 0.157 mg N mm−3 active tissue day−1.
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  • 98
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    Plant and soil 72 (1983), S. 321-334 
    ISSN: 1573-5036
    Keywords: Leguminous plants ; Nitrogen fixation ; Protein ; Seed ; Yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Mineral nitrogen did not increase grain yield and seed protein levels ofVicia faba L. andLupinus luteus L. in field trials and pot experiments. Fixed N2 was substituted by mineral nitrogen in these cases because of inhibition of N2 fixation by mineral nitrogen. Contrary to these results mineral nitrogen increased grain yields and seed protein amounts ofLupinus albus L.,Pisum sativum L., andGlycine max. (L.) Merr. The nitrogen effect was caused at an early stage by saving energy due to inhibition of N2 fixation (measurement of gas exchange by means of IRGA). In case of the N application after flowering grain, yields and seed protein levels increased because the mineral N was an additional nitrogen source for plants. At this stage the plants had ceased fixing atmospheric nitrogen. The high sink activity of growing fruits induced a lack of assimilates in nodules (determined by means of14CO2 application). The N effect was therefore the consequence of the lower assimilate pool for supplying root nodules in these plants in comparison withVicia faba L. andLupinus luteus L. Hence it follows that response to mineral nitrogen can be a criterion for discovering more effective Rhizobium-host combinations.
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  • 99
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    Plant and soil 75 (1983), S. 131-138 
    ISSN: 1573-5036
    Keywords: Hydrogen inhibition ; Nitrogen fixation ; Peas ; Relative efficiency ; Rhizobium leguminosarum ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Experiments were performed to investigate the causes of low relative efficiency, RE, in legume root nodules. Nitrogen fixing activity and RE varied with time of incubation of nodules and with different temperatures and oxygen concentrations. The effects of nitrogen concentration and carbon dioxide concentration were also examined. In each case the RE was inversely related to nitrogen fixing activity; measured by acetylene reduction. Increasing the nitrogen concentration had no effect on either nitrogen fixing activity or RE. Experiments with isolated bacteroids gave higher RE values than the whole nodules from which they were isolated. All the results were consistent with hydrogen inhibition of nitrogenase within the nodule being the cause of low RE.
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  • 100
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    Plant and soil 82 (1984), S. 273-284 
    ISSN: 1573-5036
    Keywords: Legume ; Nitrogen fixation ; Rhizobium ; C and N economy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Studies of the C and N economy of a range of temperate and tropical legume/Rhizobium symbioses indicate considerable variation (up to three-fold) in the cost of N2 fixation. Comparisons between and within symbioses indicate that the proportion of net photosynthate utilized in nodule functioning varies almost ten-fold from as low as 3% to as high as 25%. Factors possibly responsible for variation in efficiency of C use in nodules and in the proportioning of translocated photosynthetic products to nodules are discussed.
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