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  • Nitrogen fixation
  • Springer  (346)
  • Cell Press
  • 1990-1994  (88)
  • 1985-1989  (142)
  • 1980-1984  (116)
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  • Springer  (346)
  • Cell Press
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Biology and fertility of soils 18 (1994), S. 231-236 
    ISSN: 1432-0789
    Keywords: A-value ; Bradyrhizobium ; Genotype ; Growth stage ; 15N ; Nitrogen fixation ; Nodulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract TheA-value method, involving the application of a higher15N rate to a reference non-N2-fixing plant, was used to assess the magnitude of N2 fixation in two bambara groundnut cultivars at four growth stages [vegetative, 0–47 days after planting (DAP); early pod-filling, 47–99 DAP; mid-pod-filling, 99–120 DAP; physiological maturity, 120–148 DAP). The cultivars were Ex-Ada, a bunchy type, and CS-88-11, a slightly spreading type. They were grown on a loamy sand. Uninoculated Ex-Ada and CS-88-11 were used as reference plants to measure the N2 fixed in the inoculated bambara groundnuts. In this greenhouse study, soil was the major source of N in bambara groundnuts during vegetative growth, and during this period it accounted for over 80% of the N accumulaed in the plants. However, N2 fixation became the major source of plant N during reproductive growth. There were significant differences between the two cultivars in the ability to fix N2, and at physiological maturity, almost 75% of the N in CS-88-11 was derived from the atmosphere compared to 55% in Ex-Ada. Also, the total N fixed in CS-88-11 at physiological maturity was almost double that in Ex-Ada. Our data indicate that the higher N2 fixation in CS-88-11 was due to two factors, a higher intensity of N2 fixation and a longer active period of N2 fixation. The results also suggest that bambara groundnut genotypes could be selected for higher N2 fixation in farining systems.
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  • 2
    ISSN: 1432-2048
    Keywords: Aspartate aminotransferase (immunocytochemistry, subcellular fractionation) ; Glutamate oxaloacetic transaminase ; Medicago (root nodules, N2 fixation) ; Nitrogen fixation ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aspartate aminotransferase (AAT; EC 2.6.1.1) catalyzes the synthesis of the amino acid aspartate which, in alfalfa root nodules, serves as the immediate precursor of the primary N-transport compound, asparagine. The enzyme AAT may also be important in providing substrates for host-plant and bacteroid respiration. The enzyme occurs as two isoenzymes, AAT-1 and AAT-2, with AAT-1 more abundant in roots and AAT-2 predominant in root nodules. To further elucidate the role of AAT in root-nodule metabolism, subcellular fractionation and immunocytochemical methods were used to determine the intra- and intercellular localization of these two isozymes. Fractionation of nodule subcellular components showed that AAT-2 was localized in amyloplasts. Immunogold labelling with AAT-2 antibodies unequivocally confirmed this, showing that AAT-2 was localized in nodule amyloplasts and leaf chloroplasts. In root nodules, the density of immunogold labelling of infected cell plastids was almost four times that of uninfected cell plastids. The data suggest that aspartate biosynthesis in alfalfa root nodules occurs primarily in the plastids of infected cells.
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  • 3
    ISSN: 1432-0789
    Keywords: Fertilizer use efficiency ; Intercropping ; Natural 15N abundance ; Nitrogen fixation ; Pigeonpea ; Sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A field experiment was conducted to obtain the N balance sheet for sole crops and intercrops of sorghum [Sorghum bicolor (L.) Moench] and pigeonpeas [Cajanus cajan (L.) Millsp.]. Intercropping gave a significant advantage over sole cropping in terms of dry matter production and grain yield, as calculated on the basis of the land equivalent ratio and area-time equivalent ratio. The N fertilizer use efficiency and atmospheric N2 fixation by pigeonpea were estimated using 15N-labeling and natural abundance methods. The N fertilizer use efficiency of sorghum was unaltered by the cropping system, while that of the pigeonpea was greatly reduced by intercropping. Although intercropping increased the fractional contribution of fixed N to the pigeonpeas, no significant difference was observed between the cropping systems in total symbiotically fixed N. There was no evidence of a significant transfer of N from the pigeonpea to the sorghum. This study showed that use of soil N and fertilizer N by pigeonpeas was almost the same as that by sorghum in sole cropping, indicating the potential competence of pigeonpeas to exploit soil N. However, when N was exhausted by a companion crop in intercropping, the pigeonpea crop increased its dependency on atmospheric N2 fixation. We conclude that knowledge of how N from different sources is shared by companion crops is a prerequisite to establishing strategies to increase N use, and consequently land productivity, in intercropping systems.
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  • 4
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    Biology and fertility of soils 17 (1994), S. 1-8 
    ISSN: 1432-0789
    Keywords: Ammonium excretion ; Azospirillum brasilense ; Auxine ; 2,4-Dichlor-phenoxy-acetic acid ; Nitrogen fixation ; Paranodulation ; Maize ; Zea mays ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH 4 + -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N2 fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O2 than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH 4 + -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.
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  • 5
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    Biology and fertility of soils 18 (1994), S. 37-41 
    ISSN: 1432-0789
    Keywords: Available nutrients ; Insecticides ; Microortanisms ; Nitrogen fixation ; Phosphate solubilization ; Rhizosphere soil ; Rice yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A field experiment was conducted to investigate the effects of 1,2,3,4,5,6-hexachlorocyclohexane (BHC), phorate, carbofuran, and fenvalerate, at their recommended doses, on some chemical and microbiological properties of the rhizosphere soil in relation to rice yields. In general, the insecticides had a beneficial effect on rhizosphere soil properties. Carbofuran strongly stimulated the mineralization of organic C. BHC and phorate led to the retention of less total N in the soil. BHC released more NH inf4 sup+ -N than the other insecticides. Phorate, however, liberated the most NO inf3 sup- -N. Phorate and fenvalerate released more available P than BHC and carbofuran did. All the insecticides stimulated the proliferation of aerobic non-symbiotic N2-fixing and phosphate-solubilizing microorganisms, resulting in an overall increase in rice yield. BHC had the greatest effect on rice yields, followed by phorate.
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  • 6
    ISSN: 1432-072X
    Keywords: Rhizobium leguminosarum biovar phaseoli ; nifA ; nifB ; nifH ; Nitrogen fixation ; nif Gene ; fix Gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the isolation, mutational analysis and the nucleotide sequence of the Rhizobium leguminosarum bv. phaseoli nifA gene. Comparison of the deduced amino acid sequence with other NifA sequences indicated the presence of the conserved central activator and the C-terminal DNA-binding domains. Nodules elicited by a R. leguminosarum bv. phaseoli nifA mutant were symbiotically ineffective. The expression of a nifA-gusA fusion was shown to be independent on the oxygen status of the cell. We cloned the three nifH copies of R. leguminosarum bv. phaseoli and determined the nucleotide sequence of their promoter regions. The expression of nifH-gusA fusions is induced under microaerobic conditions and is dependent on the presence of NifA.
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  • 7
    ISSN: 1432-072X
    Keywords: Synechococcus sp. ; Nitrogen fixation ; Photosynthetic oxygen evolution ; Light-dark cycle ; Light-response curve
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract When nitrogen fixing cell cultures of Synechococcus RF-1 were subjected to an alternating lightdark regime (12 h:12 h), a cyclic decrease in the photosynthetic oxygen evolution potential was observed during the dark periods. This rhythm of net photosynthesis rate was maintained for at least two days after transition to continuous light. The decrease in net photosynthesis was accompanied by a stimulation of dark respiration. However, the magnitude of oxygen uptake was considerably smaller than the observed decrease in oxygen evolution. The photosynthetic activity of cells taken from the dark period was characterized by (i) a significantly lower quantum yield and (ii) a strong reduction in the light-saturated rate of photosynthesis. Growing the cultures on nitrate or under continuous light completely suppressed this rhythm. Protein synthesis was not necessary for the recovery of the light-saturated rate of photosynthesis during the light period. The cellular content of chlorophyll a and of phycobiliproteins did not vary between light and dark period, indicating that quantitative changes in the composition of the photosynthetic apparatus are not the basis for the observed oscillations. Regulatory modifications of the photosynthetic efficiency are proposed as an adaptation mechanism to adjust the intracellular oxygen concentration to the needs for nitrogenase activity.
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  • 8
    ISSN: 1432-072X
    Keywords: Key words     Synechococcus sp. ; Nitrogen fixation ; Photosynthetic oxygen evolution ; Light-dark cycle ; Light-response curve
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract      When nitrogen fixing cell cultures of Synechococcus RF-1 were subjected to an alternating light-dark regime (12 h : 12 h), a cyclic decrease in the photosynthetic oxygen evolution potential was observed during the dark periods. This rhythm of net photosynthesis rate was maintained for at least two days after transition to continuous light. The decrease in net photosynthesis was accompanied by a stimulation of dark respiration. However, the magnitude of oxygen uptake was considerably smaller than the observed decrease in oxygen evolution. The photosynthetic activity of cells taken from the dark period was characterized by (i) a significantly lower quantum yield and (ii) a strong reduction in the light-saturated rate of photosynthesis. Growing the cultures on nitrate or under continuous light completely suppressed this rhythm. Protein synthesis was not necessary for the recovery of the light-saturated rate of photosynthesis during the light period. The cellular content of chlorophyll a and of phycobiliproteins did not vary between light and dark period, indicating that quantitative changes in the composition of the photosynthetic apparatus are not the basis for the observed oscillations. Regulatory modifications of the photosynthetic efficiency are proposed as an adaptation mechanism to adjust the intracellular oxygen concentration to the needs for nitrogenase activity.
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  • 9
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Vanadium ; anf Azotobacter ; nif ; Sigma(σ)54 ; Sigma(σ)N ; Molybdenum ; vnf ; ntrC ; rpoN
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several regulatory gene mutants of Azotobacter vinelandii were tested for ability to synthesize functional nitrogenase-1 (Nif phenotype), nitrogenase-2 (Vnf), or nitrogenase-3 (Anf). While nifA mutants were Nif-, Vnf+, and Anf+/-, and ntrC mutants were Nif+, Vnf+, and Anf+, nifA ntrC double mutants were Nif-, Vnf-, and Anf-. A vnfA mutant was Nif+, Vnf+/-, and Anf+/-, and an anfA strain was Nif+, Vnf+, and Anf-. lacZ fusions in the nifH, vnfH, vnfD, anfH, and nifM genes of Azotobacter vinelandii were constructed and introduced into wild-type and regulatory mutants of A. vinelandii. Expression of these operons correlated with the growth phenotype of the regulatory mutants. Apparently either NifA or NtrC can activate expression of nifM. Also, expression of the anf operon required the NifA transcriptional activator, although there are no NifA binding sites at appropriate locations upstream of anfH (or anfA). The results confirm previous reports that VnfA and AnfA are required for expression of vnf and anf genes, respectively, and that VnfA is involved in repression of the nifHDK operon in the absence of molybdenum and of the anfHDGK operon in the presence of vanadium.
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  • 10
    ISSN: 1432-2048
    Keywords: Aspartate aminotransferase (immunocytochemistry, subcellular fractionation) ; Glutamate oxaloacetic transaminase ; Medicago (root nodules, N2 fixation) ; Nitrogen fixation ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aspartate aminotransferase (AAT; EC 2.6.1.1) catalyzes the synthesis of the amino acid aspartate which, in alfalfa root nodules, serves as the immediate precursor of the primary N-transport compound, asparagine. The enzyme AAT may also be important in providing substrates for host-plant and bacteroid respiration. The enzyme occurs as two isoenzymes, AAT-1 and AAT-2, with AAT-1 more abundant in roots and AAT-2 predominant in root nodules. To further elucidate the role of AAT in root-nodule metabolism, subcellular fractionation and immunocytochemical methods were used to determine the intra- and intercellular localization of these two isozymes. Fractionation of nodule subcellular components showed that AAT-2 was localized in amyloplasts. Immunogold labelling with AAT-2 antibodies unequivocally confirmed this, showing that AAT-2 was localized in nodule amyloplasts and leaf chloroplasts. In root nodules, the density of immunogold labelling of infected cell plastids was almost four times that of uninfected cell plastids. The data suggest that aspartate biosynthesis in alfalfa root nodules occurs primarily in the plastids of infected cells.
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  • 11
    ISSN: 1432-2048
    Keywords: Blue-green alga ; Cyanobacterium ; Nitrate (reduction, uptake) ; Nitrogen fixation ; Nodularia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation and nitrate-reduction activities were determined in photoautotrophic cultures of two wild-type strains of cyanobacterium Nodularia, spp. M1 and M2. Air could support growth of the two strains at a similar rate in the presence or absence of exogenous nitrate, ammonium and/or bicarbonate. Nitrogenase activity in air-grown cultures varied with culture age, and totally disappeared after 6 h of darkness. Recovery took place upon culture re-illumination. Ammonium at a concentration of 1 mM resulted in the total disappearance of nitrogenase activity and of heterocysts. In contrast, 20 mM nitrate hardly affected nitrogenase activity and heterocyst formation after ten generations. Under the same conditions, either ammonium or nitrate completely abolished nitrogenase activity and heterocyst formation in Anabaena sp. PCC 7119, a typical heterocystous strain. The inefficiency of nitrate in inhibiting nitrogen fixation in Nodularia M1 and M2 seemed to be caused by a low nitrate-reductase activity, and not by an impairment of nitrate-uptake activity. On the other hand, the presence of nitrate was not required for uptake activity to be expressed in Nodularia.
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  • 12
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    Protoplasma 182 (1994), S. 32-38 
    ISSN: 1615-6102
    Keywords: Rhizobium sp. (Cicer) CC 1192 ; Cicer arietinum ; Nitrogen fixation ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Developing and senescing chickpea (Cicer arietinum L.) nodules formed byRhizobium sp. (Cicer) CC 1192 have been shown by light and electron microscopy to have general morphological and ultrastructural features that are characteristic of indeterminate nodules. These features included the presence of persistent meristematic tissue at the distal ends of the multi-lobed nodules, and a gradient of cells at different stages of development towards the proximal point of attachment of the nodules to the parent root. The cytoplasm of infected cells in the nitrogen-fixing region of the nodules was densely packed with symbiosomes, most of which contained a single bacteroid. Infection threads containing bacteria were noted in invaded cells from the nitrogen-fixing region of the nodules. Other features that were observed in chickpea nodules included the presence of electron-dense occlusions in intercellular spaces in the nitrogen-fixing region, and plasmodesmata that connected infected cells with other infected cells and with uninfected cells. No poly-β-hydroxybutyrate granules were noted in the bacteroids. In later stages of development, infected cells became enlarged and highly vacuolated, and eventually lost their contents. Uninfected cells in the central region were smaller than infected cells and were also highly vacuolated. Some of the degenerative processes that take place in senescing bacteroids were noted.
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  • 13
    ISSN: 1615-6102
    Keywords: Alnus incana ; Frankia ; Hopanoids ; Nitrogen fixation ; Oxygen ; Symbiosis ; Vesicle envelope
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The role ofFrankia vesicle envelope lipids in regulating oxygen diffusion of symbiotic nitrogen fixation inAlnus incana was examined. Total lipids of symbioticFrankia (vesicle clusters) that had been adapted to oxygen tensions of 5,21, or 40 kPa were analyzed with a normal phase HPLC system. During the oxygen treatment, nitrogenase activity was measured as hydrogen evolution in an open flow-through system. When plants were transferred to low oxygen (5 kPa) or high oxygen (40 kPa), nitrogenase activity dropped initially. Activity recovered in both treatments with a rate comparable to the controls (21 kPa O2). Both lipid content and lipid composition of vesicle clusters were affected by the oxygen treatments. With increasing oxygen tension, the vesicle cluster lipid content increased. This correlated with structural data (fluorescence microscopy and TEM) which showed a thicker vesicle envelope at higher oxygen tension. Three hopanoid lipids, bacteriohopanetetrol (bht) and two isomers of phenylacetyl monoester of bht, made up approximately 80% of the vesicle cluster lipids. With changing oxygen concentrations, the ratio of the two bht esters changed whereas the relative proportion of bht remained fairly constant. Therefore, in theFrankia-Alnus incana symbiosis, adaptation to different ambient oxygen tensions occurs at least partly by increasing the thickness of theFrankia vesicle envelope and by changing its lipid composition.
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  • 14
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    Protoplasma 183 (1994), S. 49-61 
    ISSN: 1615-6102
    Keywords: Diffusion ; Intercellular spaces ; Mitochondria ; Nitrogen fixation ; Oxyleghaemoglobin ; Simulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A simulation model is presented for the distribution and consumption of O2 in infected cells of soybean root nodule central tissue. It differs from earlier models in closer adherence to observed structure and embodies new morphometric data about the distribution of 〉 12,000 mitochondria per cell and about the geometry of the gas-filled intercellular spaces near which the mitochondria are located. The model cell is a rhombic dodecahedron and O2 enters only through interfaces (totalling 26% of the cell surface) with 24 gas-filled intercellular spaces. These spaces are located at the edges of each rhombic face of the cell, forming an interconnected network over the cell suface. Next, O2 is distributed through the cytoplasm by a leghaemoglobin-facilitated diffusive process, initially between the mitochondria and amyloplasts in the outer layers of the cell and then between 〉 6,000 symbiosomes (each containing 6 bacteroids) towards the central nucleus. The symbiosomes and mitochondria consume O2, but impede its diffusion; all O2 entering symbiosomes is considered to be consumed there. For the calculations, the cell is considered to consist of 24 structural units, each beneath one of the intercellular spaces, and each is divided into 126 layers, 0.2 μm thick, in and through which O2 is consumed and diffused. Rates of consumption of O2 and of N2 fixation in each diffusion layer were calculated from previously-established kinetics of respiration by mitochondria and bacteroids isolated from soybean nodules and from established relationships between bacteroid respiration and N2 fixation. The effects of varying the O2-supply concentration and the concentration and type of energy-yielding substrates were included in the simulations. When the model cell was supplied with 0.5 mM malate, mitochondria accounted for a minimum of 50% of the respiration of the model cell and this percentage increased with increased concentration of the O2 supply. Gradients of concentrations of free O2 dissolved in the cytoplasm were steepest near the cell surface and in this location respiration by mitochondria appeared to exert a marked protective effect for nitrogen fixation in layers deeper within the cell. Estimates of N2 fixation per nodule, calculated from the model cell, were similar to those calculated from field measurements.
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  • 15
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; nifA ; Metal ions ; Oxygen regulation ; Transcriptional activation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The NIFA protein from Rhizobium leguminosarum biovar trifolii (R. trifolii) strain ANU843 lacks an N-terminal domain present in homologous NIFA proteins from other diazotrophs. The R. trifolii nifA gene product is unstable when expressed in Escherichia coli under both aerobic and microaerobic conditions. Stability is increased by fusion of additional amino acids to the N-terminus of the protein or by expression of nifA in sno mutant (presumed protease deficient) strains of E. coli. Transcriptional activation in vivo by R. trifolii NIFA decreases under aerobic growth conditions, or when cultures are depleted of metal ions. In sno mutant strains this decrease in activity reflects a loss of specific activity rather than proteolytic degradation, implying that R. trifolii NIFA requires metal ions for activity and is oxygen sensitive. The addition of 30 amino acids to the amino-terminus of R. trifolii NIFA results in an oxygen-tolerant protein, with metal ion-dependent activity. Metal ions are therefore not only required for oxygen sensing by R. trifolii NIFA but may play an additional role in determining NIFA structure or activity.
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  • 16
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    Trees 8 (1993), S. 99-103 
    ISSN: 1432-2285
    Keywords: Robinia pseudoacacia L. ; Hydrogen uptake ; Hydrogenase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Hydrogen uptake is thought to increase the efficiency of nitrogen fixation by recycling H2 produced by nitrogenase that would otherwise be lost by diffusion. Here we demonstrate the capacity of eight Rhizobium strains to take up molecular hydrogen. Uptake by nodule homogenates from Robinia pseudoacacia was measured amperometrically under nitrogenase repression. Markedly lower activities were found than in soybean nodules. In addition hydrogenase activity was detected by the ability of bacteroids to reduce methylene blue in the presence of hydrogen. It was demonstrated that hydrogenase structural genes are present in the black locust symbiont, Rhizobium sp. strain R1, using hybridization with a plasmid, which contained hydrogenase genes from R. leguminosarum bv. viceae.
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  • 17
    ISSN: 1432-2048
    Keywords: Carbohydrate and nitrogen distribution ; Nitrate ; Nitrogenase activity ; Nitrogen fixation ; Respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study was undertaken to determine how carbon utilization of fruit production might affect symbiotic activity in hydroponically cultured white bean (Phaseolus vulgaris L.) cv. Rico and its supernodulating genotype R32-BS15 (abbreviated as RBS15). Total plant biomass production of both genotypes was similar. Nodule dry weight of RBS15 consistently scored approximately twice the amount recorded for Rico, while nodule numbers in the mutant were almost six times as high. Nodule respiration on a per-plant basis and specific respiration was initially (day 37) disproportionally higher in the mutant, reaching up to three times the values recorded for Rico, while plant N2 fixation estimated by 15N dilution was almost identical. This indicates a lower fixation efficiency of RBS15 nodules, which we suggest to be a result of the larger number of smaller nodules with a higher proportion of growth and maintenance respiration per unit nodule mass. During reproductive development, specific respiration of the mutant dropped below that of Rico without a reduction in fixation, indicating a change in relative efficiency of fixation. Continuous removal of fruits from day 37 onwards stimulated respiration of nodules in both genotypes with highest values per plant being documented for RBS15, while specific activity was higher for Rico. The results indicate that symbiotic activity was not detrimentally affected by competition for carbohydrates between fruits and nodules. It appeared that nodules did not possess excess N2-fixation capacity which could be stimulated by additional provision of photosynthate. Hence, the early onset of reproduction during the life cycle of P. vulgaris is unlikely to be responsible for inadequate fixation performance in the field.
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  • 18
    ISSN: 1432-1939
    Keywords: Plant defenses ; Polyphenols ; Herbivory ; Savanna ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In East African savanna we found that leaves of mature tree species with symbiotic N2 fixation contained lower concentrations of polyphenols than leaves of species without this symbiosis. We suggest that the root symbiosis is costly to the plant in terms of photosynthate that otherwise could be used in chemical defense. Further, a negative relationship between concentration of polyphenols and the height of the species was found, independent of their ability to fix N2. These findings suggest that root symbioses and apparency to herbivory are important factors mediating the production of chemical defenses in plants.
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  • 19
    ISSN: 1432-1939
    Keywords: Actinorhizal plants ; Evolutionary tradeoffs ; Nitrogen fixation ; Nutrient resorption ; Phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nutrient resorption was measured in an actinorhizal nitrogen-fixing shrub,Comptonia peregrina, for five years in the understory of a deciduous oak forest in Rhode Island, USA. Mean resorption of nitrogen was extremely inefficient (11%) compared to most deciduous species (50%+), yet resorption of phosphorus was efficient (53%) and comparable to other species. Of the seven additional nutrients studied, only copper (6%) and zinc (10%) were resorbed from senescing leaves. Resorption of nitrogen (5%–20%) and phosphorus (40%–71%) varied significantly among years. Copper was resorbed from leaves in three years and accreted into leaves in two years. Five-year resorption means differed among individual genets by as much as a factor of 2.5 for nitrogen, and 1.3 for phosphorus. Resorption of nitrogen, copper, and zinc were highly correlated, yet resorption of phosphorus remained autonomous from other nutrients. The ecophysiological tradeoffs inComptonia which have resulted in the cooccurence of actinorhizal nitrogen fixation, inefficient nitrogen resorption, and efficient phosphorus resorption suggest that plant nutrient status does have an impact on resorption efficiency and that the evolution of nutrient conservation strategies is nutrient-specific.
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  • 20
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    Biology and fertility of soils 15 (1993), S. 35-38 
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; Eucalyptus saligna ; Albizia falcataria ; Pontoscolex corethrurus ; Litterfall
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Tree species differ in the quantity and quality of litter produced, and these differences may significantly affect ecosystem structure and function. I examined the importance of tree species in determining earthworm densities in replicated stands of Eucalyptus saligna Sm. and Albizia falcataria (L.) Fosberg, and in mixed stands (25% albizia and 75% eucalyptus). Mean earthworm densities ranged from 92 m-2 in the pure eucalyptus, to 281 m-2 in the mixture, and a maximum of 469 m-2 in the pure albizia stands. Only two earthworm species were present, Pontoscolex corethrurus and Amynthas gracilis. Leaf biomass on the forest floor was highest in the pure eucalyptus and lowest in the pure albizia stands, whereas the annual fine litterfall production was lowest in the pure eucalyptus and highest in the albizia stands. The N content of fine litterfall was correlated positively with earthworm density, and the fine litterfall biomass: N ratio was correlated negatively with earthworm density. Greater leaf biomass on the forest floor under eucalyptus stands despite lower rates of litterfall suggests that litter quality, rather than litter quantity, was primarily responsible for the greater earthworm density in the albizia stands. Some biogeochemical effects of tree species in the tropics may be mediated through effects on earthworm populations.
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  • 21
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    Biology and fertility of soils 15 (1993), S. 73-78 
    ISSN: 1432-0789
    Keywords: Firewood crops ; Green-leaf manure ; Macronutrients ; Nodulation ; Nitrogen fixation ; Sesbania spp. ; Acetylen reduction assay ; ARA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In three field trials conducted during the summer season of 1986, 1987 and 1989 in an alkaline soil, 17 accessions of annual Sesbania spp. were evaluated for nodulation, N2 fixation (acetylene reduction assay), dry weight of roots and shoots, woody biomass production, and nutrient uptake. At 50 days after sowing all the accessions were effectively nodulated (average 36.4 root nodules plant-1) with a high nodule score (3.4). There was a lot of variation in nodule volume and mass and in acetylene reduction activity but not in N content (5.2%). N uptake in shoots, roots and nodules averaged 639, 31, and 13 mg plant-1, respectively, and much of the fixed N remained in shoots. Accessions of ‘S. cannabina’ complex performed better than others. S. rostrata had poor root nodulation but exhibited excellent stem nodulation (300 nodules plant-1) even though not inoculated with Azorhizobium sp. Average concentrations of N, P, K, S, Ca, and Mg in the shoots were high, at 3.2, 0.28, 1.5, 0.28, 1.5, and 0.4% respectively, and Na was low (0.15%), reflecting the usefulness of Sesbania spp. as an integrated biofertilizer source. Green matter production was 26.0 Mg ha-1 (5.9 Mg dry matter) and N uptake was 158 kg ha-1, 54 days after sowing. Average woody biomass of six accessions at maturity, 200 days after sowing, was high (19.9 Mg ha-1), showing its potential for shortterm firewood production. Total nutrient uptake for production of woody biomass (200 days of growth) was no more demanding than growing the plant to the green-manuring stage of 50–60 days' growth.
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  • 22
    ISSN: 1432-0789
    Keywords: δ 15N ; Elevation ; Nitrogen fixation ; Non-nodulating ; Glycine max ; Soybeans ; Isolines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Dissimilarities in soil N uptake between N2-fixing and reference non-N2-fixing plants can lead to inaccurate N2 fixation estimates by N difference and 15N enrichment methods. The natural 15N abundance (δ 15N) method relies on a stabilized soil 15N pool and may provide reliable estimates of N2 fixation. Estimates based on the δ 15N and differences in N yield of nodulating and non-nodulating isolines of soybean were compared in this study. Five soybeans from maturity groups 00, IV, VI, and VIII and their respective non-nodulating isolines were grown at three elevations differing in ambient temperature and soil N availability. Despite large differences in phenological development and N yield between the non-nodulating isolines, the δ 15N values measured on seeds were relatively constant within a site. The δ 15N method consistently produced lower N2 fixation estimates than the N difference method, but only in three of the 15 observations did they differ significantly. The average crop N derived from N2 fixation across sites and maturity groups was 81% by N difference compared to 71% by δ 15N. The magnitude of difference between the two methods increased with increasing proportions of N derived from N2 fixation. These differences between the two methods were not related to differences in total N across sites or genotypes. The low N2 fixation estimates based on δ 15N might indicate that the nodulating isolines had assimilated more soil N than the non-nodulating ones. A lower variance indicated that the estimates by N difference using non-nodulating isolines were more precise than those by δ 15N. Since the differences between the estimates were large only at high N2 fixation levels (low soil N availability), either method may be used in most situations when a non-nodulating isoline is used as the reference plant. The δ 15N method may have a comparative advantage over N difference and 15N enrichment methods in the absence of a suitable non-N2-fixing reference plant such as a non-nodulating isoline.
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  • 23
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    Biology and fertility of soils 15 (1993), S. 275-278 
    ISSN: 1432-0789
    Keywords: Peanuts ; Arachis hypogaea ; Continuous cropping ; Nitrogen fixation ; Bradyrhizobium spp. ; Effectiveness of rhizobia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The objective of this study was to assess the number and effectiveness of peanut rhizobia in soils of the major peanut-growing areas of Thailand. Three cropping areas, (1) continuously cropped with peanuts, (2) continuously cropped with non-legumes, and (3) non-cultivated fields, were chosen in each region. Peanut rhizobia were found in the soil at 38 to 55 sites sampled. Cultivated fields with a peanut cultivation history contained (as estimated by most probable numbers) an average of 1.6×103 cells g-1 of soil. The numbers of peanut rhizobia in most of the fallow fields and some of the noncultivated shrub or forest locations were much the same as at the sites where Arachis hypogaea was cultivated. In contrast, there were no or few (28–46 cells g-1 soil) peanut rhizobia in the majority of fields continuously cultivated with sugarcane, cassava, corn, and pineapple. It appears that in these areas the indigenous peanut rhizobial populations are not adequate in number for a maximal nodulation of peanuts. A total of 343 Bradyrhizobium isolates were tested for effectiveness and were found to vary widely in their ability to fix N2. In some areas the majority of rhizobia were quite effective while in others they were less effective than the inoculum strain THA 205 recommended in Thailand.
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  • 24
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    Biology and fertility of soils 16 (1993), S. 299-301 
    ISSN: 1432-0789
    Keywords: A N value ; 15N ; Nitrogen fixation ; Glycine max ; Hordeum vulgare
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Pot experiments were conducted with two soils, from Rottenhaus and Seibersdorf in Austria, to ascertain whether the rate of fertilizer N application and the test crop would influence the amount of N available in the soil as assessed by the A-value method. 15N-labelled fertilizer was applied at rates of 10, 25, 40, 60, and 100 mg N kg-1 soil, corresponding approximately to 20, 50, 80, 120 and 200 kg N ha-1 respectively, and two crop species, barley (Hordeum vulgareL.) and non-nodulating soybean (Glycine max L.) were used to determine the soil A N value under the various fertilizer regimes. The results showed that the Rottenhaus soil had a higher A N value than the Seibersdorf soil, suggesting that the former was more fertile than the latter. The A N values of both soils were significantly affected by the level of N application. When grown in the same soil, the two test crops showed significantly different fertilizer use efficiency and per cent N derived from fertilizer when the rate of N application exceeded 20 kg ha-1. Thus, the A N value as determined by the two test crops differed significantly for the same soil when the rate of N application was greater than 20 kg/ha. The difference was greater when the soil fertility level was high. The dependence of the A N value on the level of N application and the species of crop seriously compromises the suitability of this method for determining plant-associated N2 fixation. Hence, considerable caution is required when using this method to estimate plant-associated N2 fixation.
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  • 25
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; Photosynthetic bacteria ; Stem nodules ; Aeschynomene scabra ; Sesbania rostrata ; Azorhizobium caulinodans ; Erythrobacter sp. ; Roseobacter denitrificans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Bradyrhizobial strain BTAi 1 nodulates both stems and roots of Aeschynomene spp. Previous work has shown that it contains bacteriochlorophyll a and forms photosynthetic reaction centers, and has provided indirect evidence of photosynthesis by bacteroids within stem nodules. Here we report physiological and biochemical characteristics of BTAi 1 ex planta, which also suggest the presence of photosynthetic activity. Light-stimulated uptake of 14CO2 by BTAi 1 was detected at all stages of growth. Inhibitors of photosynthesis, 1,10-orthophenanthroline and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and the uncoupler NH4Cl, immediately suppressed light-driven 14CO2 uptake and increased O2 uptake. BTAi 1 is strictly aerobic and was unable to grow without organic C even in the light; also, it was unable to grow chemoautotrophically in an atmosphere enriched with H2 and CO2. In micro-aerobic conditions, strain BTAi 1 expressed acetylene reducing activity ex planta in an N-free medium. The highest rates of light-stimulated 14CO2 uptake and acetylene-reducing activity occurred during the exponential and early stationary phases of growth. Acetylene-reducing rates at a low glucose concentration were increased following a light-dark cycle in comparison with continuous dark conditions.
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  • 26
    ISSN: 1432-072X
    Keywords: Klebsiella pneumoniae ; Nitrogen fixation ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nifL gene product of Klebsiella pneumoniae inhibits the activity of the positive activator protein NifA in response to increased levels either of fixed nitrogen or of oxygen in the medium. In order to demonstrate that the responses to these two effectors are discrete we have subjected nifL to hydroxylamine mutagenesis and isolated nifL mutants that are impaired in their ability to respond to oxygen but not to fixed nitrogen. Two such mutations were sequenced and shown to be single base pair changes located in different parts of nifL. The amino acid sequence of NifL shows limited homology to the histidine protein kinases which comprise the sensing component of bacterial two-component regulatory systems. In the light of the location of one of the oxygen-insensitive mutations (Leu294Phe) we have reassessed this homology and we suggest that the Gln273-Leu317 region of NifL may facilitate interactions between NifL and NifA.
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  • 27
    ISSN: 1432-072X
    Keywords: Aromatic catabolism ; Free-living diazotrophs ; Nitrogen fixation ; catechol dioxygenases ; Protocatechuate dioxygenases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Six species of free-living nitrogen fixing bacteria, Azomonas agilis, Azospirillum brasilense, Azospirillum lipoferum, Azotobacter chroococcum, Azotobacter vinelandii, and Beijerinckia mobilis, were surveyed for their ability to grow and fix N2 using aromatic compounds as sole carbon and energy source. All six species grew and expressed nitrogenase activity on benzoate, catechol, 4-hydroxybenzoate, naphthalene, protocatechuate, and 4-toluate. In many cases, growth rates on one or more aromatic compounds were comparable to or greater than those on the non-aromatic substrates routinely used for cultivation of the organisms. Specific activity of nitrogenase in extracts of aromatic-grown cells often exceeded that in cells grown on non-aromatic substrates. All six species growing on substrates typically converted to catechol expressed inducible catechol 1,2-dioxygenase and/or catechol 2,3-dioxygenase. When grown on substrates typically converted to protocatechuate, inducible protocatechuate 3,4-dioxygenase and/or protocatechuate 4,5-dioxygenase was expressed. A. chroococcum expressed only ortho cleavage dioxygenases during growth on naphthalene and 4-toluate and only meta cleavage dioxygenases on the other aromatics. B. mobilis expressed only ortho cleavage dioxygenases. The other four species examined expressed both ortho and meta cleavage enzymes.
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  • 28
    ISSN: 1432-072X
    Keywords: Rhodobacter capsulatus ; Nitrogen fixation ; Nitrogenase activity ; Nitrogenase expression ; Aerobiosis ; Oxygen ; Light ; Modification of the Fe protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rhodobacter capsulatus was grown chemotrophically in the dark in oxygen-regulated chemostat culture and in the presence of limiting amounts of fixed N. When the oxygen partial pressure was varied, in situ nitrogen fixation occurred only at 1% of air saturation of the medium. By contrast, nitrogenase proteins and their activity measured in the absence of oxygen could be detected up to 30% of air saturation. This revealed that expression of nitrogenase is much less sensitive toward oxygen than the in situ function of the enzyme. At oxygen partial pressures 〉 1% of air saturation, the degree of modification of the Fe protein of nitrogenase was increased. Light was of no stimulatory effect on both the activity and the expression of nitrogenase. This holds true for growth at 1% or 5% of air saturation. At 5% of air saturation, however, high illumination enhanced the inhibitory effect of oxygen on nitrogenase formation.
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  • 29
    ISSN: 1432-072X
    Keywords: Calcium ; Nitrogen fixation ; Nodulation ; Periplasmic proteins ; Rhizobium fredii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Calcium is essential for the growth of rhizobia and the formation of nitrogen-fixing root-nodules on legumes, but its precise role in these processes remains unknown. We have found that Rhizobium fredii USDA208 accumulates a major 38 kDa protein when grown in media supplemented with 0.3–2 μmM CaCl2. We have purified this protein and raised polyclonal antibodies against it. The protein initially is synthesized as a 40 kDa precursor which subsequently undergoes calcium-dependent processing to give rise to the mature polypeptide. Subcellular and immunocytochemical localization studies indicate that the 38 kDa protein accumulates preferentially in the periplasmic space. Its N-terminal sequence, AETIKIGVAGPMTG, shows significant homology to the N-termini of amino acid binding proteins from the periplasm, including leucine-, isoleucine-, and valine-specific binding proteins of Pseudomonas aeruginosa and Escherichia coli and a leucine-specific binding protein of E. coli. The R. fredii protein does not, however, bind [3H]-leucine. The 38 kDa protein is encoded by the bacterial chromosome. It is absent in several rhizobia other than R. fredii, but antigenically related polypeptides are present in Escherichia coli and Erwinia carotovora subsp. carotovora.
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  • 30
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    Archives of microbiology 159 (1993), S. 423-427 
    ISSN: 1432-072X
    Keywords: Chlorogloeopsis ; Continuous culture ; Cyanobacterium ; Nitrogen fixation ; Oxygen effects ; Photosynthesis ; Respiration ; Temperature effects ; Thermophile
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of temperature and oxygen on diazotrophic growth of the thermophilic cyanobacterium HTF (High Temperature Form) Chlorogloeopsis was investigated using cells grown in light-limited continuous culture at a dilution rate of 0.02 h-1. Diazotrophy was more sensitive to elevated temperatures than growth with combined nitrogen. The maximum temperature for growth of cultures gassed with CO2-enriched air was more than 55 °C but less than 60 °C with N2 as the sole nitrogen source, but between 60°C and 65°C when nitrate was present in the medium. The effect of temperature on nitrogenase activity, photosynthesis and respiration in the dark was determined using cells grown at 55°C. Maximal rates of all three processes were observed at 55°C and rates at 60°C during shortterm incubations were not less than 75% of the maximum. However, nitrogenase activity at 60°C was unstable and decayed at a rate of 2.2 h-1 under air and at 0.3 h-1 under argon. Photosynthesis and respiration were more stable at 60°C than anoxic nitrogen fixation. The upper temperature limits for diazotrophic growth thus seem to be set by the stability of nitrogenase.
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  • 31
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    Molecular genetics and genomics 239 (1993), S. 435-440 
    ISSN: 1617-4623
    Keywords: Enterobacter agglomerans ; nifQ gene ; FeMo-cofactor ; Nitrogen fixation ; Overexpression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nifQ gene, involved in early stages of iron-molybdenum cofactor (FeMo-co) biosynthesis, was identified downstream of the nijB and nijF genes of Enterobacter agglomerans. This gene was cloned and its nucleotide sequence determined. The amino acid sequence, as deduced from the nucleotide sequence, revealed an accumulation of cysteine amino acid residues at the C-terminal end of the protein. The cysteine cluster showed the following consensus sequence Cys-X4-Cys-X2-Cys-X5-Cys, which is a typical characteristic of metal-binding proteins. Further, the nifQ gene was cloned downstream of strong transcriptional (bacteriophage λpLpR) and translational (atpE) signals of the expression vector pCYTEXP1 and expressed as an unfused, soluble protein in Escherichia coli. The molecular mass of 19 kDa, as deduced by SIDS-PAGE, is in good agreement with the molecular mass deduced from the nucleotide sequence. The availability of high-level expression clones should facilitate purification of large quantities of the recombinant NifQ protein and elucidation of its properties.
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  • 32
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; Novel type of NifU ; Ethane formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA sequence analysis of a 3494-bp HindIII-Bc1I fragment of the Rhodobacter capsulatus nif region A revealed genes that are homologous to ORF6, nifU, nifS, nifV and nifW from Azotobacter vinelandii and Klebsiella pneumoniae. R. capsulatus nifU, which is present in two copies, encodes a novel type of NifU protein. The deduced amino acid sequences of NifUI and NifUII share homology only with the C-terminal domain of NifU from A. vinelandii and K. pneurnoniae. In contrast to nifA andnifB which are almost perfectly duplicated, the predicted amino acid sequences of the two NifU proteins showed only 39% sequence identity. Expression of the ORF6-nifU ISVW operon, which is preceded by a putative σ54-dependent promoter, required the function of NifA and the nif-specific rpoN gene product encoded by nifR4. Analysis of defined insertion and deletion mutants demonstrated that only nifS was absolutely essential for nitrogen fixation in R. capsulatus. Strains carrying mutations in nifV were capable of very slow diazotrophic growth, whereas ORF6, nifU I and nifW mutants as well as a nifU I/nifUII, double mutant exhibited a Nif+ phenotype. Interestingly, R. capsulatus nifV mutants were able to reduce acetylene not only to ethylene but also to ethane under conditions preventing the expression of the alternative nitrogenase system. Homocitrate added to the growth medium repressed ethane formation and cured the NifV phenotype in R. capsulatus. Higher concentrations of homocitrate were necessary to complement the NifV phenotype of a polar nifV mutant (NifV−NifW−), indicating a possible role of NifW either in homocitrate transport or in the incorporation of this compound into the iron-molybdenum cofactor of nitrogenase.
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  • 33
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    Molecular genetics and genomics 241 (1993), S. 124-128 
    ISSN: 1617-4623
    Keywords: Aspartate aminotransferase ; Alleles ; Alfalfa ; Nitrogen fixation ; Transit peptide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Aspartate aminotransferase (AAT) plays a key enzymatic role in the assimilation of symbiotically fixed nitrogen in legume root nodules. In alfalfa, two distinct genetic loci encode dimeric AAT enzymes: AAT1, which predominates in roots, and AAT2, which is expressed at high levels in nodules. Three allozymes of AAT2 (AAT2a, −2b and −2c), differing in net charge, result from the expression of two alleles, AAT2A and AAT2C, at this locus. Utilizing antiserum to alfalfa AAT2, we have previously isolated from an expression library one AAT2 cDNA clone. This clone was used as a hybridization probe to screen cDNA libraries for additional AAT2 cDNAs. Four different clones were obtained, two each that encode the AAT2a and AAT2c enyzme subunits. These two sets of cDNAs encode polypeptides that differ in net charge depending upon the amino acid at position 296 (valine or glutamic acid). Within each set of alleles, the two members differ from each other by the presence or absence of a 30 by (ten amino acid) sequence. The presence or absence of this ten amino acid sequence has no effect on the size or charge of the mature AAT2 protein because it is located within the region encoding the protein's transit peptide, which is proteolytically removed upon transport into plastids. The data suggest that a deletion event has occurred independently in two AAT2 progenitor alleles, resulting in the four allelic cDNA variants observed. The deletion of this ten amino acid sequence does not appear to impair the normal maturation of the enzyme.
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  • 34
    ISSN: 1432-0789
    Keywords: Typic cryoboroll ; N yield ; 15N ; Root length ; Grass-legume intercrop ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Barley-field pea intercrops have been shown to increase N yield when grown under cryoboreal subhumid conditions. In this study, we extended previous research by testing the hypotheses that (1) the intercropped field pea fixes a greater proportion of its shoot and root N than does sole-cropped field pea; (2) N is transferred from the annual legume to the cereal during the growing season; and (3) root production is greater under intercropped than sole-cropped conditions. Unconfined microplots seeded to barley, field peas, or a barley-field pea intercrop were fertilized with N at 10 kg ha-1 as (NH4)2SO4 (5.21 atom % 15N excess). Both the intercropped and sole-cropped barley derived more than 93% of their N from the soil. In contrast, 40% of N in the intercropped field pea was derived from soil. This study provided no evidence for transfer of N from the legume to the cereal. On average, the proportion of N derived from air by both pea intercrops was 39% higher than that derived by the sole-cropped pea. Root length determined by a grid intersection method following digitization using an image analyzer tended to be higher under intercropping than in sole crops. We conclude that even on fertile soils benefits may accrue from annual intercropping that includes a legume. The benefits arise from (1) increased N production, (2) greater N-fixation efficiency, and/or (3) more shoot and root residue-N mineralization for subsequent crops.
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  • 35
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    Biology and fertility of soils 13 (1992), S. 165-172 
    ISSN: 1432-0789
    Keywords: Azospirillum brasilense ; 15N-isotope enrichment ; Nitrogen fixation ; Auxine ; 2,4-dichlorphenoxy acetic acid ; 3,5-dichlor-phenoxy acetic acid ; Acetylene reduction assay ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Wheat seedlings, treated with the auxine 2,4-dichlor-phenoxy acetic acid (2,4-D) during germination developed only a residual root system. Root elongation was extremely restricted and root tips were deformed to thick club-shaped tumours. When 2,4-D was added in a later stage of plant growth the plants developed additional nodule-like knots along primary roots. Root and shoot dry-matter production was slightly repressed in all 2,4-D treatments and N translocation from roots to shoots was repressed as well. When transferred to an auxine-free growth medium, the 2,4-D-affected roots were not capable of complete recovery. In plants inoculated gnotobiotically with Azospirillum brasilense, either with the wild type or with the NH 4 + -excreting mutant strain C3, a 2,4-D addition increased rhizosphere acetylene-reduction activity at pO2 1.5 kPa. The O2 sensitivity of root-associated nitrogenase activity tended to be reduced. The number of root-colonizing bacteria, at approximately 108 colony-forming units (cfu) per g dry root, was similar in the 2,4-D treatments and untreated controls. Plant treatment with high concentrations of the chemical isomer 3,5-dichlor-phenoxy acetic acid (3,5-D) did not have comparable effects, either on plant development or on rhizosphere-associated nitrogenase activity. Root-tumour tissue inhabited by A. brasilense showed purple staining when subjected to a tetrazolium chloride solution, which may indicate intensive local nitrogenase activity in this tissue. Exposed to an 15N2-enriched atmosphere, plants treated with 2,4-D and with A. brasilense incorporated significantly higher amounts of 15N than untreated controls. In all cases the highest values of 15N enrichment were found following inoculation with the NH 4 + -excreting mutant strain C3.
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  • 36
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    Archives of microbiology 158 (1992), S. 256-261 
    ISSN: 1432-072X
    Keywords: Frankia ; Amino acids ; Glutamine synthetase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The utilization of some amino acids, added at 1 mM and 10 mM concentrations, as the sole combined nitrogen sources by Frankia sp. strain CpI1, has been investigated. Glutamine, like NH 4 + , provided rapid growth without N2 fixation. Histidine at 1 mM yielded poor N2-fixing activity but better cell growth than N2. Aspartate, glutamate, alanine, proline, each at 1 mM concentration, supported similar levels of N2 fixation and growth. Growth on 10 mM glutamate, proline, or histidine resulted in poor N2-fixing activity and poor cell growth. Cells grown on 10 mM alanine had about half the N2-fixing activity of cells grown on N2 but growth was good. Aspartate at 10 mM concentration, however, stimulated N2-fixing activity dramatically and promoted faster growth. Enzyme analysis suggested that asparate is catabolized by glutamate-oxaloacetate transaminase (GOT), since GOT specific activity was induced, and aspartase activity was not detected, in cells grown on aspartate as the sole combined nitrogen source. Thinlayer chromatography (TLC) of metabolites extracted from N2-grown cells fed with [14C]-aspartate showed that label was rapidly accumulated mainly on aspartate and/or glutamate, depending on the cells' physiological state, without detectable labeling on fumarate or oxaloacetate (OAA). These findings provide evidence that aspartate is catabolized by GOT to OAA which, in turn, is rapidly converted to α-ketoglutarate through the TCA cycle and then to glutamate by GOT or by glutamate synthase (GOGAT). The stimulation of N2 fixation and growth by aspartate is probably caused by an increased intracellular glutamate pool.
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  • 37
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    Archives of microbiology 158 (1992), S. 155-161 
    ISSN: 1432-072X
    Keywords: Anoxygenic phototrophic bacteria ; Photosynthesis ; Heliobacteria ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three species of anoxygenic phototrophic heliobacteria, Heliobacterium chlorum, Heliobacterium gestii, and Heliobacillus mobilis, were studied for comparative nitrogen-fixing abilities and regulation of nitrogenase. Significant nitrogenase activity (acetylene reduction) was detected in all species grown photoheterotrophically on N2, although cells of H. mobilis consistently had higher nitrogenase activity than did cells of either H. chlorum or H. gestii. Nitrogen-fixing cultures of all three species of heliobacteria were subject to “switch-off” of nitrogenase activity by ammonia; glutamine also served to “switch-off” nitrogenase activity but only in cells of H. mobilis and H. gestii. Placing photosynthetically grown heliobacterial cultures in darkness also served to “switch-off” nitrogenase activity. Dark-mediated “switch-off” was complete in lactate-grown heliobacteria but in pyruvate-grown cells substantial rates of nitrogenase activity continued in darkness. In all heliobacteria examined ammonia was assimilated primarily through the glutamine synthetase/glutamate synthase (GS/GOGAT) pathway although significant levels of alanine dehydrogenase were present in extracts of cells of H. gestii, but not in the other species. The results suggest that heliobacteria, like phototrophic purple bacteria, are active N2-fixing bacteria and that despite their gram-positive phylogenetic roots, heliobacteria retain the capacity to control nitrogenase activity by a “switch-off” type of mechanism. Because of their ability to fix N2 both photosynthetically and in darkness, it is possible that heliobacteria are significant contributors of fixed nitrogen in their paddy soil habitat.
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  • 38
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    Archives of microbiology 157 (1992), S. 361-366 
    ISSN: 1432-072X
    Keywords: Rhodobacter capsulatus ; Ammonium assimilation ; Nitrogen fixation ; Glutamate synthase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen assimilation in Rhodobacter capsulatus has been shown to proceed via the coupled action of glutamine synthetase (GS) and glutamate synthase (GOGAT) with no measurable glutamate dehydrogenase (GDH) present. We have recently isolated a novel class of mutants of R. capsulatus strain B100 that lacks a detectable GOGAT activity but is able to grow at wild type rates under nitrogen-fixing conditions. While NH 4 + -supported growth in the mutants was normal under anaerobic/photosynthetic conditions, the growth rate was decreased under aerobic conditions. Ammonium and methylammonium uptake experiments indicated that there was a clear difference in the ammonium assimilatory capabilities in these mutants under aerobic versus anaerobic growth. Regulation of expression of a nifH : : lacZ fusion in these mutants was not impaired. The possible existence of alternative ammonium assimilatory pathways is discussed.
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  • 39
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    Archives of microbiology 157 (1992), S. 431-435 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrogenase ; “Switchoff” ; Metabolic regulation ; Rhodospirillum rubrum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of NAD(P) and analogs of this nucleotide on nitrogenase activity in Rhodospirillum rubrum has been studied. Addition of NAD+ to nitrogen fixing Rsp. rubrum leads to inhibition of nitrogenase. NADP+ has the same effect but NADH or analogs modified in the nicotinamide portion do not cause inhibition. In contrast to ammonium ions, addition of NAD+ leads to inhibition of nitrogenase in cells that have been N-starved under argon. The inhibitory effect of NAD+ is more pronounced at lower light intensities. Addition of NAD+ also leads to inhibition of glutamine synthetase, a phenomenon also occurring when “switchoff” is produced by the addition of effectors such as ammonium ions or glutamine. It is also shown that NAD+ is taken up by Rsp. rubrum cells.
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  • 40
    ISSN: 1432-072X
    Keywords: Carbohydrate ; Cell cycle ; Hydrogen production ; Marine ; Synechococcus sp. ; Nitrogen fixation ; Synchronous culture ; Unicellular cyanobacterium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Synchronously growing cells of nitrogen-fixing Synechococcus sp. Miami BG 043511 were harvested periodically and the capability for hydrogen photoproduction in closed vessels was measured under hydrogen production conditions. The capability for hydrogen photoproduction in cells was correlated with that of cellular carbohydrate content. Cells with a high carbohydrate content exhibited a high capacity for hydrogen production and those with low carbohydrate content exhibited low capacity for hydrogen production. Nitrogenase activity at the onset of incubation did not coincide with a capability for the cells to produce hydrogen during the subsequent incubation period. Interestingly, when cells with a high capacity for hydrogen photoaccumulation were incubated, alternate periods of hydrogen and oxygen accumulation were observed at 12 hour intervals. About 0.5 ml of hydrogen per ml of cell suspension was accumulated in flasks during the initial 12-h incubation period. These observations indicate that the use of synchronous culture can be one of the ways of provide materials suitable not only for basic studies but also for applied aspects of hydrogen photoproduction.
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  • 41
    ISSN: 1573-1561
    Keywords: Nitrogen fixation ; glyceollin ; phytoalexin ; Bradyrhizobium japonicum ; nodulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Nitrogen fixation in root nodules formed by strain 2143 ofBradyrhizobium japonicum andGlycine max (L.) Merr. cv Williams 82 reaches a maximum at 21 to 28 days postinoculation and then begins to decline. The phytoalexin, glyceollin, accumulates in nodules coincident with the decline in nitrogen fixation. Nodules formed by strain 3122, which are unable to fix nitrogen, accumulate even higher levels of glyceollin and do so beginning 21 days postinoculation even though these nodules contain no recoverable bacteria. The typical phytoalexin response occurs within days of infection. The mechanism by which this response in theBradyrhizobium japonicum-soybean combination is delayed 2 to 3 weeks after infection is presently unknown but phytoalexin accumulation could contribute to the inability of the soybean-Bradyrhizobium japonicum combination to maintain high levels of nitrogen fixation throughout the growing season.
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  • 42
    ISSN: 1432-2242
    Keywords: Glutamate oxalate transaminase ; Isozymes ; Nitrogen fixation ; Medicago sativa L. ; Rhizobium meliloti
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The enzyme aspartate aminotransferase (AAT) plays a key role in the assimilation of fixed-N in alfalfa (Medicago sativa L.) root nodules. AAT activity in alfalfa nodules is due to the activity of two dimeric isozymes, AAT-1 and AAT-2, that are products of two distinct genes. Three forms of AAT-2 (AAT-2a, -2b, and-2c) have been identified. It was hypothesized that two alleles occur at the AAT-2 locus, giving rise to the three AAT-2 enzymes. In a prior study bidirectional selection for root nodule AAT and asparagine synthetase (AS) activities on a nodule fresh weight basis in two diverse alfalfa germ plasms resulted in high nodule enzyme activity subpopulations with about 20% more nodule AAT activity than low enzyme activity subpopulations. The objectives of the study presented here were to determine the inheritance of nodule AAT-2 production and to evaluate the effect of bidirectional selection for AAT and AS on AAT-2 allelic frequencies, the relative contributions of AAT-1 and AAT-2 to total nodule activity, nodule enzyme concentration, and correlated traits. Two alleles at the AAT-2 locus were verified by evaluating segregation of isozyme phenotypes among F1 and S1 progeny of crosses or selfs. Characterization of subpopulations for responses associated with selection was conducted using immunoprecipitation of in vitro nodule AAT activity, quantification of AAT enzyme protein by ELISA, and AAT activity staining of native isozymes on PAGE. Results indicate that selection for total AAT activity specifically altered the expression of the nodule AAT-2 isozyme. AAT-2 activity was significantly greater in high compared to low activity subpopulations, and high AAT subpopulations from both germ plasms had about 18% more AAT-2 enzyme (on a nodule fresh weight basis). No significant or consistent changes in AAT-2 genotypic frequencies in subpopulations were caused by selection for AAT activity. Since changes in AAT activity were not associated with changes in AAT-2 genotype, selection must have affected a change(s) at another locus (or loci), which indirectly effects the expression of nodule AAT.
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  • 43
    ISSN: 1432-2242
    Keywords: Medicago sativa ; Nodulation (spontaneous) ; Nitrogen fixation ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rhizobium nodulation genes can produce active extracellular signals for legume nodulation. The R. meliloti host-range nodH gene has been postulated to mediate the transfer of a sulfate to a modified lipo-oligosaccharide, which in its sulfated form is a specific nodulation factor for alfalfa (Medicago sativa L.). We found that alfalfa was capable of effective nodulation with signal-defective and non-nodulating nodH mutants (Nnr) defining a novel gene-for-gene interaction that conditions nodulation. Bacteria-free nodules that formed spontaneously at about a 3–5% rate in unselected seed populations of alfalfa cv ‘Vernal’ in the total absence of Rhizobium (Nar) exhibited all the histological, regulatory and ontogenetic characteristics of alfalfa nodules. Inoculation of such populations with nodH mutants, but not with nodA or nodC mutants, produced a four- to five-fold increase in the percentage of nodulated plants. Some 10–25% of these nodulated plants formed normal pink nitrogen-fixing nodules instead of white empty nodules. About 70% of the S1 progeny of such Nnr+ plants retained the parental phenotype; these plants were also able to form nodules in the absence of Rhizobium. If selected Nar+ plants were self-pollinated almost the entire progeny exhibited the parental Nar+ phenotype. Segregation analysis of S1 and S2 progeny from selected Nar+ plants suggests that the Nar character is monogenic dominant and that the nodulation phenotype is controlled by a gene dose effect. The inoculation of different S1 Nar+ progeny with nodH mutant bacteria gave only empty non-fixing nodules. Our results indicate that certain alfalfa genotypes can be selected for suppression of the non-nodulation phenotype of nodH mutants. The fact that the Nnr plant phenotype behaved as a dominant genetic trait and that it directly correlated with the ability of the selected plants to form nodules in the absence of Rhizobium suggests that the interaction of plant and bacterial alleles occurs early during signal transduction through the alteration of a signal reception component of the plant so that it responds to putative signal precursors.
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  • 44
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    Molecular genetics and genomics 235 (1992), S. 49-54 
    ISSN: 1617-4623
    Keywords: Rhizobium meliloti ; Two-component sensor protein ; FixL ; Nitrogen fixation ; Oxygen control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary FixL protein of Rhizobium meliloti is a haemoprotein kinase which activates the transcription of nifA and fixK genes via the transcriptional activator protein FixJ under microaerobic conditions. FixL and FixJ proteins belong to the family of two-component regulatory systems for which primary sequence data predicts a modular structure. We showed, using Escherichia coli as heterologous host, that FixL indeed has a modular structure. The amino-terminal hydrophobic domain is dispensable for the oxygen-regulated activity of FixL in vivo. The central cytoplasmic non-conserved domain is necessary for the oxygen-sensing function of FixL whereas it is not necessary for the activation of FixJ by FixL. We propose that, under aerobic conditions, the central domain represses the activating function associated with the carboxy-terminal conserved domain.
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  • 45
    ISSN: 1617-4623
    Keywords: Azorhizobium caulinodans ; Nitrogen fixation ; fixABCX genes ; Gene regulation ; mRNA processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report here the transcriptional analysis of the fixABCXORF1 region of Azorhizobium caulinodans. This led to the identification of a 0.9 kb transcript covering fixX and ORF1, which was synthesized only under conditions of nitrogen fixation. The 5′ end of this transcript was mapped by primer extension and S 1 nuclease protection analyses and shown to be located 70 ± 1 nucleotides upstream of the fixX start codon. By means of transcriptional fixX- and ORF1-lacZ fusions, it was shown that fixX and ORF1 were most probably transcribed from the fixA promoter and that expression of fixX and ORF1 was dependent on NifA activation. This suggests that the 0.9 kb mRNA results from post-transcriptional processing of a large mRNA covering fix-A,B,C,X and ORF1. In addition, ORF1 mutants were constructed and were shown not to be impaired in nitrogenase activity.
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  • 46
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; Nucleotide binding site ; Oxygen regulation ; Random mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Rhizobium meliloti the NifA protein plays a central role in the expression of genes involved in nitrogen fixation. The R. meliloti NifA protein has been found to be oxygen sensitive and therefore acts as a transcriptional activator only under microaerobic conditions. In order to generate oxygen-tolerant variants of the NifA protein a plasmid carrying the R. meliloti nifA gene was mutagenized in vitro with hydroxylamine. About 70 mutated nifA genes were isolated which mediated up to 12-fold increased NifA activity at high oxygen concentrations. A cloning procedure involving the combination of DNA fragments from mutated and wild-type nifA genes allowed mapping of the mutation sites within the central part of the nifA gene. For 17 mutated nifA genes the exact mutation sites were determined by DNA sequence analysis. It was found that all 17 mutated nifA genes carried identical guanosine — adenosine mutations resulting in a methionine — isoleucine exchange (M217I) near the putative nucleotide binding site within the central domain. Secondary structure predictions indicated that the conformation of the putative nucleotide binding site may be altered in the oxygen-tolerant NifA proteins. A model is proposed which assumes that at high oxygen concentrations the loss of activity of the R. meliloti NifA protein is due to a conformational change in the nucleotide binding site that may abolish binding or hydrolysis of the nucleotide. Such a conformational change may be blocked in the oxygen-tolerant NifA protein, thus allowing interaction with the nucleotide at high oxygen concentrations.
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  • 47
    ISSN: 1617-4623
    Keywords: Archaea ; Nitrogen fixation ; nifH promoter ; Cell-free transcription ; Methanococcus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nifH1 gene of Methanococcus thermolithotrophicus, which encodes the putative dinitrogenase reductase of an archaeon, was accurately transcribed in a homologous cell-free transcription system. Extracts of cells grown with N2 or ammonia as nitrogen source initiated transcription at the nifH1 promoter with similar efficiencies. We confirmed that cells grown under non-N2-fixing conditions do not contain significant amounts of nifH1-specific mRNA. The levels of cell-free transcription initiation at the nifH1 promoter wer similar to those observed at a tRNA promoter. The DNA sequence from −40 to +5 relative to the initiator nucleotide of nifH1 mRNA contained all the information required for promoter activity. A mutational analysis of this section of DNA demonstrated that a TATA box at −25 and the TTGT motif (initiator element) at the transcription start site are essential for cell-free transcription. These elements are similar to the structural determinants of a known tRNA promoter of Methanococcus. Mutation of a sequence, showing homology to the bacterial NifA site, which overlaps the transcription start site, did not affect promoter activity. Hence, cell-free transcription of the Methanococcus nifH1 gene is independent of upstream activator elements and does not require alternate cis-acting sequences that differ from the methanogen consensus promoter. These findings suggest that the activation of nif promoters is brought about by fundamentally different mechanisms in Archaea and bacteria.
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  • 48
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    Molecular genetics and genomics 231 (1992), S. 494-498 
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; Azotobacter vinelandii ; Molybdenum ; Vanadium ; Metalloenzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Strains with mutations in 23 of the 30 genes and open reading frames in the major nif gene cluster of A. vinelandii were tested for ability to grow on N-free medium with molybdenum (Nif phenotype), with vanadium (Vnf phenotype), or with neither metal present (Anf phenotype). As reported previously, nifE, nifty, nifU, nifS and nifV mutants were Nif− (failed to grow on molybdenum) while nifM mutants were Nif−, Vnf− and Anf−. nifV, nifS, and nifU mutants were found to be unable to grow on medium with or without vanadium, i.e. were Vnf− Anf−. Therefore neither vnf nor anf analogoues of nifU, nifS, nifV or nifM are expected to be present in A. vinelandii.
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  • 49
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    Hydrobiologia 234 (1992), S. 59-64 
    ISSN: 1573-5117
    Keywords: Nitrogen fixation ; cyanobacteria ; Lyngbya wollei
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The ability of the benthic cyanobacterium Lyngbya wollei to fix nitrogen was studied using field samples and axenic cultures. L. wollei was collected and isolated from Lake Okeechobee, Florida, where it forms extensive mats. Rates of acetylene reduction up to 39.1 nmol mg dry wt−1 h−1 were observed for field samples. The maximum observed rate of acetylene reduction in axenic laboratory cultures was 200 nmol mg dry wt−1 h−1. Aerobic conditions limited nitrogen fixation activity, but dark/light cycles promoted the development of activity. Reduced oxygen levels appeared to be required for the development of significant levels of nitrogenase activity. The level of irradiance also had a significant impact on the level of activity. The potential significance of nitrogen fixation to Lyngbya production is discussed.
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  • 50
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    World journal of microbiology and biotechnology 8 (1992), S. 171-174 
    ISSN: 1573-0972
    Keywords: Nitrogen fixation ; nodulation ; rhizobia ; soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Eighty soybean cultivars were assessed for their potential for nodulation and nitrogen fixation with indigenous rhizobia in a Nigerian soil. Seventy-six days after planting (DAP) 87%, 3% and 10% of the soybean cultivars had from 0 to 30, 31 to 60 and over 61 nodules/plant, respectively. Only 8% had a nodule dry weight of 600 to 1100 mg/plant. At 84 DAP the proportion of nitrogen derived from the atmosphere (Ndfa) ranged from 0 to 65% 16% of the cultivars derived 51 to 65% of their N2 from the atmosphere. The diversity of soybean germplasm and the variation in nodulation and N2 fixation permitted the selection of the five best cultivars in terms of their compatibility with indigenous rhizobia, % Ndfa and the amount of N2 which they fixed.
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  • 51
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    Oecologia 88 (1991), S. 451-455 
    ISSN: 1432-1939
    Keywords: Nitrogen fixation ; Carbon isotope ratio ; Nitrogen isotope ratio ; Acacia ; Namibia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nitrogen (N2) fixation was estimated along an aridity gradient in Namibia from the natural abundance of 15N (δ15N value) in 11 woody species of the Mimosacease which were compared with the δ15N values in 11 woody non-Mimosaceae. Averaging all species and habitats the calculated contribution of N2 fixation (N f ) to leaf nitrogen (N) concentration of Mimosaceae averaged about 30%, with large variation between and within species. While in Acacia albida N f was only 2%, it was 49% in Acacia hereroensis and Dichrostachys cinerea, and reached 71% in Acacia melifera. In the majority of species N f was 10–30%. There was a marked variation in background δ15N values along the aridity gradient, with the highest δ15N values in the lowland savanna. The difference between δ15N values of Mimosaceae and non-Mimosaceae, which is assumed to result mainly from N2 fixation, was also largest in the lowland savanna. Variations in δ15N of Mimosaceae did not affect N concentrations, but higher δ15N-values of Mimosaeae are associated with lower carbon isotope ratios (δ13C value). N2 fixation was associated with reduced intrinsic water use efficiency. The opposite trends were found in non-Mimosaceae, in which N-concentration increased with δ15N, but δ13C was unaffected. The large variation among species and sites is discussed.
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  • 52
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    Biology and fertility of soils 11 (1991), S. 210-215 
    ISSN: 1432-0789
    Keywords: Rhizosphere ; Maize ; Bacillus circulans ; Enterobacteriaceae ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary We studied the dominant diazotrophs associated with maize roots and rhizosphere soil originating from three different locations in France. An aseptically grown maize plantlet, the “spermosphere model”, was used to isolate N2-fixing (acetylene-reducing) bacteria. Bacillus circulans was the dominant N2-fixing bacterium in the rhizosphere of maize-growing soils from Ramonville and Trogny, but was not found in maize-growing sandy soil from Pissos. In the latter soil, Enterobacter cloacae, Klebsiella terrigena, and Pseudomonas sp. were the most abundant diazotrophs. Azospirillum sp., which has been frequently reported as an important diazotroph accociated with the maize rhizosphere, was not isolated from any of these soils. The strains were compared for their acetylene-reducing activity in the spermosphere model. The Bacillus circulans strains, which were more frequently isolated, also exhibited significantly greater acetylene-reducing activity (3100 nmol ethylene day-1 plant-1) than the Enterobacteriaceae strains (180 nmol ethylene day-1 plant-1). This work indicates for the first time that Bacillus circulans is an important maizerhizosphere-associated bacterium and a potential plant growth-promoting rhizobacterium.
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  • 53
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    Biology and fertility of soils 11 (1991), S. 273-278 
    ISSN: 1432-0789
    Keywords: Genetic variability ; N-15 methods ; Nitrogen fixation ; Provenances ; Rhizobium strains ; Gliricidia sepium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Variation in nodulation and N2 fixation by the Gliricidia sepium/Rhizobium spp. symbiosis was studied in two greenhouse experiments. The first included 25 provenances of G. sepium inoculated with a mixture of three strains of Rhizobium spp. N2 fixation was measured using the 15N isotope dilution method 12 weeks after planting. On average, G. sepium derived 45% of its total N from atmospheric N2. Significant differences in fixation were observed between provenances. The percentage of N derived from atmospheric N2 ranged from 26 to 68% (equivalent to 18–62 mg N plant-1) and was correlated with total N in the plant (r=0.70; P=0.05). The second experiment included six strains of Rhizobium spp. and two methods of inoculation and the plants were harvested 14,35 and 53 weeks after planting. In the first harvest significant differences were found between the number of nodules and the percentage and amount of N2 fixed. There was also a significant correlation between the number of nodules and the amount of N2 fixed (r=0.92; P=0.05). In the final harvest no correlation was observed, although there were significant differences between the number of nodules and the percentage of N derived from the atmosphere. The amount of N2 fixed increased with time (from an average of 27% at the first harvest to 58% at the final harvest) and was influenced by the Rhizobium spp. strain and the method of inoculation. It ranged from 36% for Rhizobium sp. strain SP 14 to 71% for Rhizobium SP 44 at the last harvest. Values for the percentage of atmosphere derived N2 obtained by soil inoculation were slightly higher than those obtained by seed inoculation.
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  • 54
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    Biology and fertility of soils 11 (1991), S. 306-312 
    ISSN: 1432-0789
    Keywords: Alkali soil ; Blue-green algae ; Calcium carbonate ; Gypsum ; Nitrogen fixation ; Organic matter ; Soil reclamation ; Sodic soil ; Waterlogged soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Virgin alkali (sodic) soils have a high pH and high exchangeable Na and are often barren. Blue-green algae, however, tolerate excess Na and grow extensively on the soil surface in wet seasons. Experiments using a highly degraded alkali soil (silt loam, pH 10.3, electrical conductivity 3.5 dS m-1, 90% exchangeable Na) were conducted in soil columns, with or without gypsum, in order to study the influence of waterlogging on the growth of indigenous and inoculated blue-green algae and hence, soil reclamation. The growth of indigenous blue-green algae was initially slow in alkali soil, due to the high pH and exchangeable Na, and depressed in gypsum-amended soil, due to excess Ca. Inoculation hastened the establishment of blue-green algae in both the unamended alkali soil and the gypsum-amended soil, overcoming the adverse influence of excess Na in the former and excess Ca in the latter. Gypsum was effective in amelioration (pH 9.05, electrical conductivity 1.2 dS m-1, 41% exchangeable Na after 11 weeks) but blue-green algae were ineffective even after 17 weeks. In combination with gypsum, blue-green algae had no additional effect, and the C and N increases due to the growth of indigenous or inoculated blue-green algae were insignificant. Alkali soil reclamation by biological methods requires mobilization of Ca from native soil calcite and the exchange of Ca for Na in the exchange complex. The ineffectiveness of blue-green algae was ascribed to their inability to mobilize Ca. It is argued that current theories favouring blue-green algae as a biological amendment to bring about alkali soil reclamation are untenable and are not comparable with an effective chemical amendment such as gypsum.
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  • 55
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    Biology and fertility of soils 12 (1991), S. 100-106 
    ISSN: 1432-0789
    Keywords: Azospirillum ; 15N-isotope dilution ; Nitrogen fixation ; Acetylene reduction activity ; ARA ; Rhizosphere ; Mineral nitrogen ; Oxygen tension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Acetylene reduction activity by Azospirillum brasilense, either free-living in soils or associated with wheat roots, was determined in a sterilised root environment at controlled levels of O2 tension and with different concentrations of mineral N. In an unplanted, inoculated soil nitrogenase activity remained low, at approximately 40 nmol C2H4 h-1 per 2kg fresh soil, increasing to 300 nmol C2H4 h-1 when malic acid was added as a C source via a dialyse tubing system. The N2 fixation by A. brasilense in the rhizosphere of an actively growing plant was much less sensitive to the repressing influence of free O2 than the free-living bacteria were. An optimum nitrogenase activity was observed at 10 kPa O2, with a relatively high level of activity remaining even at an O2 concentration of 20 kPa. Both NO inf3 sup- and NH inf4 sup+ repressed nitrogenase activity, which was less pronounced in the presence than in the absence of plants. The highest survival rates of inoculated A. brasilense and the highest rates of acetylene reduction were found in plants treated with azospirilli immediately after seedling emergence. Plants inoculated at a later stage of growth showed a lower bacterial density in the rhizosphere and, as a consequence, a lower N2-fixing potential. Subsequent inoculations with A. brasilense during plant development did not increase root colonisation and did not stimulate the associated acetylene reduction. By using the 15N dilution method, the affect of inoculation with A. brasilense in terms of plant N was calculated as 0.067 mg N2 fixed per plant, i.e., 3.3% of the N in the root and 1.6% in the plant shoot were of atmospheric origin. This 15N dilution was comparable to that seen in plants inoculated with non-N2-fixing Psudomonas fluorescens.
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  • 56
    ISSN: 1432-0789
    Keywords: Ulex gallii ; Legume ; Nitrogenase activity ; Nitrogen fixation ; Acetylene reduction activity ; Phosphorus fertilizer ; Forest soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary N2(C2H2) fixation by Ulex gallii Planchon (dwarf or autumn flowering gorse/furze) seedlings was determined following 8 months of growth (December-August) in the glasshouse in a very acid, N- and P-deficient forest soil. Application of Na2HPO4·12H2O or North African ground rock phosphate fertilizer was essential for growth, nodulation and C2H2 reduction activity. Overall, both the sodium phosphate and the rock phosphate were equally effective P sources and the maximum acetylene reduction by intact roots was measured as 4.09 and 4.69 μmol C2H4g-1 fresh weight nodule h-1, respectively. Applied NH4Cl severely inhibited nodulation and restricted acetylene reduction activity but not seedling growth. The results are discussed in relation to the spread of U. gallii in the south of Ireland and its potential as a leguminous nurse crop for Sitka spruce on the very impoverished forest soils of the region.
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  • 57
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    Archives of microbiology 156 (1991), S. 270-276 
    ISSN: 1432-072X
    Keywords: Coding regions ; Codon frequency table ; IS-like elements ; Nitrogen fixation ; Nodulation ; Repeated sequence ; Bradyrhizobium japonicum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To date, the sequences of 45 Bradyrhizobium japonicum genes are known. This provides sufficient information to determine their codon usage and G+C content. Surprisingly, B. japonicum nodulation and NifA-regulated genes were found to have a less biased codon usage and a lower G+C content than genes not belonging to these two groups. Thus, the coding regions of nodulation genes and NifA-regulated genes could hardly be identified in codon preference plots whereas this was not difficult with other genes. The codon frequency table of the highly biased genes was used in a codon preference plot to analyze the RSRjα9 sequence which is an insertion sequence (IS)-like element. The plot helped identify a new open reading frame (ORF355) that escaped previous detection because of two sequencing errors. These were now corrected. The deduced gene product of ORF355 in RSRjα9 showed extensive similarity to a putative protein encoded by an ORF in the T-DNA of Agrobacterium rhizogenes. The DNA sequences bordering both ORFs showed inverted repeats and potential target site duplications which supported the assumption that they were IS-like elements.
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  • 58
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    Archives of microbiology 156 (1991), S. 362-366 
    ISSN: 1432-072X
    Keywords: Peribacteroid membrane ; Symbiosomes ; Grycine max ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Addition of ATP to intact symbiosomes isolated from soybean nodules, resulted in generation of a membrane potential (positive inside) across the peribacteroid membrane (PBM). This energisation was monitored as oxonol fluorescence quenching. The rate of fluorescence quenching was inhibited by the inclusion of permeant anions in the reaction medium. Using this inhibition as a measure of anion uptake across the PBM, the presence of a phthalonate-sensitive dicarboxylate carrier on the PBM was confirmed. Following dissipation of the membrane potential by a permeant anion, a pH gradient, measured using [14C]methylamine uptake, was slowly established across the PBM. This ΔpH was abolished by addition of an uncoupler but was insensitive to inhibitors of bacteroid respiration. The difference in pH between the external medium and the symbiosome interior was estimated to be in the range of 1–1.6 pH units. The magnitude in planta will depend on the concentrations of ATP and permeant anions in the cytosol of the host cell.
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  • 59
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    Trees 5 (1991), S. 227-231 
    ISSN: 1432-2285
    Keywords: Robinia pseudoacacia L. ; Nitrate ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Experiments with black locust (Robinia pseudoacacia L.) seedlings grown under strictly controlled laboratory conditions indicated that the availability of nitrate has a marked impact on nitrogen fixation. When nitrate concentrations were very low, both nodulation and seedling growth were impaired, whereas nitrate concentrations high enough to promote plant growth strongly inhibited symbiotic nitrogen fixation. When nitrate was added to the growth medium after infection, nodulation and nitrogen fixation of the seedlings decreased. This effect was even more marked when nitrate was applied before infection with rhizobia. Higher nitrogen concentrations also reduced nodule number and nodule mass when applied simultaneously with the infecting bacteria. The contribution of symbiotic nitrogen fixation to black locust shoot mass by far exceeded its effects on shoot length and root mass. When nitrate availability was very low, specific nitrogen fixation (i. e. nitrogenase activity per nodule wet weight) was improved with increasing nitrogen supply, but rapidly decreased with higher nitrogen concentrations.
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  • 60
    ISSN: 1432-2048
    Keywords: Flavin ; Glycine (nitrogen fixation) ; Leghemoglobin ; Nitrogen fixation ; Oxygen (activated) ; Pyridine nucleotides ; Symbiosis (legume-Rhizobium)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reduction of ferric leghemoglobin (Lb3+) from soybean (Glycine max (L.) Merr.) nodules by riboflavin, FMN and FAD in the presence of NAD(P)H was studied in vitro. The system NAD(P)H + flavin reduced Lb3+ to oxyferrous (Lb2+ · O2) or deoxyferrous (Lb2+) leghemoglobin in aerobic or anaerobic conditions, respectively. In the absence of O2 the reaction was faster and more effective (i.e. less NAD(P)H oxidized per mole Lb3+ reduced) than in the presence of O2; this phenomenon was probably because O2 competes with Lb3+ for reductant, thus generating activated O2 species. The flavin-mediated reduction of Lb3+ did not entail production of superoxide or peroxide, indicating that NAD(P)H-reduced flavins were able to reduce Lb3+ directly. The NAD(P)H + flavin system also reduced the complexes Lb3+ · nicotinate and Lb3+ · acetate to Lb2+ · O2, Lb2+ or Lb2+ · nicotinate, depending on the concentrations of ligands and of O2. In the presence of 200 μM nitrite most Lb remained as Lb3+ in aerobic conditions but the nitrosyl complex (Lb2+ · NO) was generated in anaerobic conditions. The above-mentioned characteristics of the NAD(P)H + flavin system, coupled with its effectiveness in reducing Lb3+ at physiological levels of NAD(P)H and flavins in soybean nodules, indicate that this mechanism may be especially important for reducing Lb3+ in vivo.
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  • 61
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    Protoplasma 163 (1991), S. 82-92 
    ISSN: 1615-6102
    Keywords: Root nodule ; Infection process ; Extracellular matrix ; Nitrogen fixation ; Ceanothus ; Frankia ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Root nodules are induced in actinorhizal plants by the nitrogen-fixing actinomyceteFrankia. Nodules may be initiated by root hair infection or by intercellular penetration. InCeanothus spp. (Rhamnaceae),Frankia colonized the host root tissue by intercellular infection, in spite of the occurrence of root hairs in the infected region. The intercellular infection pathway was characterized by an extensive darkly-staining matrix which filled prominent intercellular spaces of the root cortex, gradually decreasing through a transition zone into the nodule cortex. At the ultrastructural level, most of the matrix was composed of fibrillar electron dense material. Holes or spaces occurred in the electron dense matrix, often in conjunction with apparent loosening of wall layers. Secondary cell division was observed within the root cortical cells embedded in the intercellular matrix. Unusually high levels of pectic compounds and proteins were identified histochemically in the matrix.
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  • 62
    ISSN: 1615-6102
    Keywords: Rhizobium meliloti ; Development ; Symbiosis ; Nitrogen fixation ; Ultrastructure ; Spontaneous nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The development of spontaneous nodules, formed in the absence ofRhizobium and combined nitrogen, on alfalfa (Medicago sativa L. cv. Vernal) was investigated at the light and electron microscopic level and compared to that ofRhizobium-induced normal nodules. Spontaneous nodules were initiated from cortical cell divisions in the inner cortex next to the endodermis, i.e., the site of normal nodule development. These nodules, on uninoculated roots, were white multilobed structures, histologically composed of nodule meristems, cortex, endodermis, central zone and vascular strands. Nodules were devoid of intercellular or intracellular bacteria confirming microbiological tests. Early development of spontaneous nodules was initiated by series of anticlinal followed by periclinal divisions of dedifferentiated cells in the inner cortex of the root. These cells formed the nodular meristem from which the nodule developed. The cells in the nodule meristems divided unequally and differentiated into two distinct cell types, one larger type being filled with numerous membrane-bound starch grains, and the other smaller type with very few starch grains. There were no infection threads or bacteria in the spontaneous nodules at any stage of development. This size differentiation is suggestive of the different cell sizes seen inRhizobium-induced nodules, where the larger cell type harbours the invading bacteria and the smaller type is essential in supportive metabolic roles. The ontogenic studies further support the claim that these structures are nodules rather than aberrant lateral roots, and that plant possess all the genetic information needed to develop a nodule with distinct cell types. Our results suggest that bacteria and therefore theirnod genes are not necessarily involved in the ontogeny and morphogenesis of spontaneous and normal nodules in alfalfa.
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  • 63
    ISSN: 1617-4623
    Keywords: Leghemoglobin ; Symbiosis ; Nitrogen fixation ; Nodulins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Heme-deficient mutants of Rhizobium and Bradyrhizobium have been found to exhibit diverse phenotypes with respect to symbiotic interactions with plant hosts. We observed that R. meliloti hemA mutants elicit nodules that do not contain intracellular bacteria; the nodules contain either no infection threads (“empty” nodule phenotype) or aberrant infection threads that failed to release bacteria (Bar− phenotype). These mutant nodules expressed nodulin genes associated with nodules arrested at an early stage of development, including ENOD2, Nms-30, and four previously undescribed nodulin genes. These nodules also failed to express any of six late nodulin genes tested by hybridization, including leghemoglobin, and twelve tested by in vitro translation product analysis which are not yet correlated with specific cloned genes. We observed that R. meliloti leucine and adenosine auxotrophs induced invaded Fix− nodules that expressed late nodulin genes, suggesting that it is not auxotrophy per se that causes the hemA mutants to elicit Bar− or empty nodules. Because R. meliloti hemA mutants elicit nodules that do not contain intracellular bacteria, it is not possible to decide whether or not the Fix− phenotype of these nodules is a direct consequence of the failure of R. meliloti to supply the heme moiety of hololeghemoglobin. Our results demonstrate the importance of establishing the stage in development at which a mutant nodule is arrested before conclusions are drawn about the role of small metabolite exchange in the symbiosis.
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  • 64
    ISSN: 1617-4623
    Keywords: Gene regulation ; Nitrogen fixation ; Oxygen control ; Symbiosis ; Two-component systems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The cloning, sequencing and mutational analysis of the Bradyrhizobium japonicum symbiotic nitrogen fixation genes fixL and fixJ are reported here. The two genes were adjacent and probably formed an operon, fixLJ. The predicted FixL and FixJ proteins, members of the two-component sensor/regulator family, were homologous over almost their entire lengths to the corresponding Rhizobium meliloti proteins (approx. 50% identity). Downstream of the B. japonicum fixJ gene was found an open reading frame with 138 codons (ORF138) whose product shared 36% homology with the N-terminal part of FixJ. Deletion and insertion mutations within fixL and fixJ led to a loss of approximately 90% wildtype symbiotic nitrogen fixation (Fix) activity, whereas an ORF138 mutant was Fix+. In fixL, fixJ and ORF138 mutant backgrounds, the aerobic expression of the fixR-nifA operon was not affected. NifA itself did not regulate the expression of the fixJ gene. Thus, the B. japonicum FixL and FixJ proteins were neither involved in the regulation of aerobic nifA gene expression nor in the anaerobic NifA-dependent autoregulation of the fixRnifA operon; rather they appeared to control symbiotically important genes other than those whose expression was dependent on the NifA protein. The fixL and fixJ mutant strains were unable to grow anaerobically with nitrate as the terminal electron acceptor. Therefore, some of the FixJ-dependent genes in B. japonicum may be concerned with anaerobic respiration.
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  • 65
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    Molecular genetics and genomics 227 (1991), S. 86-90 
    ISSN: 1617-4623
    Keywords: Azospirillum lipoferum ; Nitrogen fixation ; nifH promoter ; NifA ; σ54
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Southern hybridization experiments strongly indicate that the regulatory region of theAzospirillum lipoferum nifH gene is located on a cloned 1.1 kbBamHI-XhoI restriction fragment. By cloning this fragment into a promoter-probe plasmid inEscherichia coli, a promoter was identified oriented towards thenifH gene. Using a set of several bacterial strains and plasmids, both NifA and the alternative σ factor, σ54, fromKlebsiella pneumoniae were shown to be required for the induction of the assumednifH promoter in this particular heterologous system. However, NtrC fromK. pneumoniae did not stimulate this promoter. No other promoter activity was detected in the direction opposite to the identified promoter, indicating that the transcription of the adjacentnifJ gene cannot be initiated from the 1.1 kbBamHI-XhoI fragment. Thus, the genesnifH andnifJ inA. lipoferum cannot be oriented divergently, in contrast to the situation in several other nitrogen-fixing bacteria.
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  • 66
    ISSN: 1617-4623
    Keywords: Azorhizobium caulinodans ; Electron transfer flavoprotein ; nifOfixABCX genes ; nifW homologue ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of a 4.1 kb DNA fragment containing the fixABC region of Azorhizobium caulinodans was established. The three gene products were very similar to the corresponding polypeptides of Rhizobium meliloti. The C-terminal domains of both fixB products displayed a high degree of similarity with the α-subunits of rat and human electron transfer flavoproteins, suggesting a role for the FixB protein in a redox reaction. Two open reading frames (ORF) were found downstream of fixC. The first ORF was identified as fixX on the basis of sequence homology with fixX from several Rhizobium and Bradyrhizobium strains. The second ORF potentially encoded a 69 amino acid product and was found to be homologous to a DNA region in the Rhodobacter capsulatus nif cluster I. Insertion mutagenesis of the A. caulinodans fixX gene conferred a Nif− phenotype to bacteria grown in the free-living state and a Fix− phenotype in symbiotic association with the host plant Sesbania rostrata. A crude extract from the fixX mutant had no nitrogenase activity. Furthermore, data presented in this paper also indicate that the previously identified nifO gene located upstream of fixA was probably a homologue of the nifW gene of Klebsiella pneumoniae and Azotobacter vinelandii.
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  • 67
    ISSN: 1617-4623
    Keywords: Soybean ; Symbiosis ; Nitrogen fixation ; RFLP ; Plant genome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The genetic locus (nts) controlling nitrate-tolerant nodulation, supernodulation, and diminished autoregulation of nodulation of soybean (Glycine max (L.) Merill) was mapped tightly to the pA-132 molecular marker using a restriction fragment length polymorphism (RFLP) detected by subclone pUTG-132a. The nts (nitrate-tolerant symbiotic) locus of soybean was previously detected after its inactivation by chemical mutagenesis. Mutant plant lines were characterized by abundant nodulation (supernodulation) and tolerance to the inhibitory effects of nitrate on nodule cell proliferation and nitrogen fixation. The large number of RFLPs between G. max line nts382 (homozygous for the recessive nts allele) and the more primitive soybean G. soja (P1468.397) allowed the detection of co-segregation of several diagnostic markers with the supernodulation locus in F2 families. We located the nts locus on the tentative RFLP linkage group E about 10 cM distal to pA-36 and directly next to marker pA-132. This very close linkage of the molecular marker and the nts locus may allow the application of this clone as a diagnostic probe in breeding programs as well as an entry point for the isolation of the nts gene.
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  • 68
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    Molecular genetics and genomics 231 (1991), S. 97-105 
    ISSN: 1617-4623
    Keywords: Aspartate aminotransferase ; Alfalfa ; Nitrogen fixation ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated an alfalfa leaf cDNA clone that encodes aspartate aminotransferase (AAT, EC 2.6.1.1) by direct complementation of an Escherichia coli aspartate auxotroph with a plasmid cDNA library. DNA sequence analysis of the recombinant plasmid, pMU1, revealed that a 1514 by cDNA was inserted in the correct orientation and in-frame with the start of the lacZ coding sequence in the vector, pUC18. The resulting fusion protein is predicted to be 424 amino acids in length with a molecular weight of 46387 Daltons. The cDNA-encoded protein has a characteristic pyridoxal phosphate attachment site motif and has substantial amino acid sequence homology to both animal and bacterial AATs. Plasmid pMUl encodes an AAT with a Km for aspartate of 3.3 mM, a Km for 2-oxoglutarate of 0.28 mM, and a pH optimum between 8.0 and 8.5. Several lines of evidence including Western blot analysis, the isoelectric point of the encoded protein, and the effect of pH on the activity of the fusion protein, suggest that the cDNA encodes the isozyme AAT-1 rather than AAT-2. Northern blot analysis showed that the aat-1 clone hybridized to a 1.6 kb transcript present in alfalfa leaves, roots and nodules. The relative concentrations of aat-1 mRNA in these tissues were 1: 2: 5, respectively. Thus, transcription of aat-1 appears to be induced during nodule development. Southern blot analysis suggested that AAT-1 in alfalfa is encoded by either a single-copy gene or a small, multigene family.
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  • 69
    ISSN: 1432-0789
    Keywords: Acacia mangium ; Acacia auriculiformis ; Bradyrhizobium spp. ; Rhizobium spp. ; Nitrogen fixation ; Nodule efficiency ; Tree legumes ; Agroforestry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Two Australian Acacia species, A. mangium and A. auriculiformis were inoculated in vitro with eight strains of Bradyrhizobium spp. and two strains of Rhizobium spp. On the two plant species, only Bradyrhizobium spp. strains formed effective N2-fixing nodules. A. mangium, which nodulates effectively with a restricted range of Bradyrhizobium spp. strains, is a specific host compared to A. auriculiformis. A. auriculiformis is assumed to be a promiscuous host because it nodulates effectively with a wide range of Bradyrhizobium spp. strains. Nodule efficiency as expressed by the ratio of N2 fixed to nodule dry weight appeared to be higher in A. auriculiformis (0.44–0.81) than in A. mangium (0.23–0.55).
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  • 70
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; Atriplex spp. ; Root-associated diazotrops ; Acetylene reduction assy (ARA) ; Saline sodic soils ; Enterobacter agglomerans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary TwoAtriplex spp. growing in low-fertility saline sodic soils were assayed for root-associated nitrogenase activity. The excised washed and unwashed root of the two species.A. lentiformis andA. amnicola, showed high root-associated nitrogenase activity. Acetylene-reducing activity seemed to be directly influenced by moisture. The highest number of diazotrophs, enumerated using a most probable number technique was observed on the root surface. Most of the isolated diazotrophs were identified asEnterobacter agglomerans. Root-associated nitrogenase activity inAtriplex spp. may explain the high protein and biomass content of these plants growing in low-fertility saline sodic soils.
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  • 71
    ISSN: 1432-2048
    Keywords: β-1,4-Glucan ; Nitrogen fixation ; Phaseolus (root nodules) ; Rhizobium (mutant) ; Root nodules ; produced by mutantRhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Root nodules induced inPhaseolus vulgaris L. by the wild-type (WT) and a C4-dicarboxylic-acid mutant strain ofRhizobium leguminosarum biovar.phaseoli were compared on the basis of ultrastructure and cytochemistry of cellulose subunits. The mutant bacteroids failed to colonize infected host cells in a normal manner, and presented a premature degenerative appearance. Starch granules, rough endoplasmic reticulum and mitochondria were found to accumulate in the ineffective nodules. The most striking difference between effective and ineffective nodules was the presence of unusual spherical, laminated structures in plastids of mutant-infected host cells only. Cytochemical observations showed that these structures containβ-1,4-glucans. The presence ofβ-1,4-glucans within such structures may be caused by the activity of a cellulase which is produced by either the bacteroids or the host cell and is locally hydrolyzing the host cell-wall, thus releasing cellulose subunits into the cytoplasm. Another possibility is denovo synthesis ofβ-1,4-glucans in the host cell.
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  • 72
    ISSN: 1432-2048
    Keywords: Cyanobacteria ; Fe-protein (immuno-gold localization) ; Nitrogenase ; Nitrogen fixation ; Oscillatoria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The filamentous non-heterocystous cyanobacterium Oscillatoria limosa was subjected to Western blot analyses using two antisera raised against the small subunit (Fe-protein) of the nitrogenase complex. Two polypeptides were recognized in nitrogen-fixing cultures irrespective of the antiserum used while no bands were detectable in nitrate-grown cultures. The apparent molecular weights of the two polypeptides were approximately 40.5 and 39.5 kDa respectively, with the former, probably an inactive form, dominating. In situ immunogold electron microscopy was used to reveal the cellular and subcellular localization on the Fe-protein. All cells of the trichomes of nitrogen-fixing O. limosa showed a dense label. The label was homogeneously distributed throughout the cytoplasm including the thylakoid area. Nitrate-grown cultures contained a very low label.
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  • 73
    ISSN: 1432-2048
    Keywords: Bradyrhizobium ; Dicarboxylate transport ; Glycine ; Nitrogen fixation ; Peribacteroid membrane ; Zhizobium ; Root nodule ; Symbiosis (soybean-Bradyrhizobium)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Malate and succinate were taken up rapidly by isolated, intact peribacteroid units (PBUs) from soybean (Glycine max (L.) Merr.) root nodules and inhibited each other in a competitive manner. Malonate uptake was slower and was severely inhibited by equimolar malate in the reaction medium. The apparent Km for malonate uptake was higher than that for malate and succinate uptake. Malate uptake by PBUs was inhibited by (in diminishing order of severity) oxaloacetate, fumarate, succinate, phthalonate and oxoglutarate. Malonate and butylmalonate inhibited only slightly and pyruvate,isocitrate and glutamate not at all. Of these compounds, only oxaloacetate, fumarate and succinate inhibited malate uptake by free bacteroids. Malate uptake by PBUs was inhibited severely by the uncoupler carbonylcyanidem-chlorophenyl hydrazone and the respiratory poison KCN, and was stimulated by ATP. We conclude that the peribacteroid membrane contains a dicarboxylate transport system which is distinct from that on the bacteroid membrane and other plant membranes. This system can catalyse the rapid uptake of a range of dicarboxylates into PBUs, with malate and succinate preferred substrates, and is likely to play an important role in symbiotic nitrogen fixation. Energization of both the bacteroid and peribacteroid membranes controls the rate of dicarboxylate transport into peribacteroid units.
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  • 74
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    Oecologia 84 (1990), S. 176-185 
    ISSN: 1432-1939
    Keywords: Prosopis glandulosa ; Mesquite ; Nodulation ; Nitrogen fixation ; Vegetation change
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We report the recovery of root nodules from P. glandulosa var. glandulosa in the eastern portion of its range, where the species reaches its greatest vegetational development. Single cores 4.7 cm in diameter and up to 250 cm deep yielded from 0 to over 250 nodules. Nodules were found at all depths below 10 cm, with the highest concentration often around 100 cm. Detailed studies of three trees revealed relatively small volume densities of about 0.02 nodules cm−3, high surface area densities of 2–4 nodules cm−2, and high nodule biomass of 8–23 g m−2, when compared to cultivated legumes. Nodules are small, weakly attached to roots that are seldom over 0.5 mm in diameter, and not easily observed under field conditions. No nodules were recovered from cores from the more arid western portion of P.glandulosa's range, although seedlings nodulated readily in these soils in the glasshouse as well as in most unamended soils from throughout mesquite's geographical range. Local differences in nodulating potential of soils included a negative association with mesquite canopies and a positive association with depth. These results suggest a significant role for biological fixation in the nitrogen regime and vegetation dynamics of Prosopis-dominated ecosystems.
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  • 75
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    Oecologia 85 (1990), S. 241-246 
    ISSN: 1432-1939
    Keywords: Plant-soil interactions ; Nutrient cycling ; Nitrogen fixation ; Decomposition ; Primary succession
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plant nutrient status and physiological processes were examined in relation to soil nutrient characteristics under individuals of five species colonizing a young cinder deposit in Hawaii Volcanoes National Park. Two exotic species, Buddeleja asiatica and Myrica faya, had high photosynthetic rates and high nitrogen concentrations and relatively easily decomposed leaves; soils under them had high concentrations of nitrogen, cations, and organic matter and high rates of net nitrogen mineralization. At the other extreme, the natives Metrosideros polymorpha and Vaccinium reticulatum had low plant concentrations and photosynthetic rates, and low concentrations and turnover rates of N in the soil. Thus, a strong correlation exists between soil processes and plant processes, suggesting a positive feedback cycle.
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  • 76
    ISSN: 1432-0789
    Keywords: Phosphorus fertilizer ; vesicular-arbuscular mycorrhizal fungus ; Azospirillum brasilense ; Glomus versiforme ; Barley ; Nitrogen fixation ; Nitrogen-15 ; Hordeum vulgare
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Pot-culture studies were carried out to examine the response of barley (Hordeum vulgare L.) to inoculation with Azospirillum brasilense and Glomus versiforme, singly and/or in combination, under varying levels of nitrogenous [(15NH4)2SO4] and soluble phosphatic (single superphosphate) fertilizers. The interaction between both the endophytes led to increased growth and nutrition of the barley plants. Roots from plants inoculated with Azospirillum brasilense and Glomus versiforme exhibited very low acetylene reduction activity. N2 fixation in the plants increased with the increase in plant growth but the mycorrhiza alone gave a low level of N2 fixation in the plants compared to combined inoculation with both the endophytes.
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  • 77
    ISSN: 1432-072X
    Keywords: Propionic fermentation ; Fermentation of aspartate ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract From estuarine mud a rod-shaped, motile, gram-negative, anaerobic bacterium was isolated (strain asp 66). Asp 66 fermented several substrates including glucose, fructose, malate, fumarate, citrate and aspartate. Fermentation products were acetate, propionate and presumably CO2. Hydrogen was never formed nor utilized. Succinate conversion to propionate was catalyzed by cell suspensions but did not support growth. Asp 66 did not require vitamins and grew well in mineral media with a fermentable substrate. The pH range for growth was from 6.5 to 8.5. Temperature optimum was 27 to 30°C. The strain was able to fix N2 as evidenced by its growth with N2 as sole nitrogen source and its ability to reduce acetylene to ethylene. Cell-free extracts of cultures grown under air without shaking contained cytochrome(s) with absorption peaks at 523 nm and at 553 nm. The G+C content of the DNA was 60.8+-1 mol%. The taxonomic position of strain asp 66 is discussed.
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  • 78
    ISSN: 1432-072X
    Keywords: Selenium ; Hydrogenase ; Bradyrhizobium japonicum ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It has been established that the hydrogenase from autotrophically cultured Bradyrhizobium japonicum contains selenium as a bound constituent. About 80% of the enzyme selenium remains bound during precipitation with 5% trichloroacetic acid (TCA). However, 85% of the selenium bound to the enzyme is released by a combined treatment of urea, heat and TCA. Neither selenomethionine nor selenocysteine could be detected on analysis of anaerobically hydrolyzed enzyme. These results are consistent with the report showing that the structural genes for this enzyme do not contain a TGA codon (Sayavedra-Soto et al. 1988) which has been reported to code for selenocysteine incorporation into several proteins (Chambers et al. 1986; Zinoni et al. 1986; Stadtman 1987). We have demonstrated that 75Se from the labeled hydrolyzed enzyme forms the derivative' selenodicysteine. The form of selenium resulting in the synthesis of this derivative apparently is SeO inf3 sup= or a compound such as Se= which is easily oxidized to SeO inf3 sup= . In a separate approach it was established that 12–16% of the total 75Se in the native enzyme reacted with 2,3-diaminonaphthalene indicating that this fraction was present as SeO inf3 sup= . The remaining 75Se was bound to the enzyme protein. From this research, we concluded that Se in Bradyrhizobium japonicum hydrogenase is present in a labile bound form. In this respect, this enzyme is similar to xanthine dehydrogenase and nicotinic acid hydroxylase, both of which contain labile Se constituents that have not been defined.
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  • 79
    ISSN: 1432-072X
    Keywords: Plectonema ; Nitrogen fixation ; Nitrogen starvation ; Cyanobacteria ; Glycogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen-starved cells of the filamentous, nonheterocystous cyanobacterium, Plectonema boryanum 581, were cultured microaerophilically in the presence and absence of exogenous, organic substrates (fructose, glucose, sucrose, and CO2) to determine the effect of an external carbon source on nitrogenase activity. In unsupplemented and in fructose-, glucose-, and sucrose-enriched cultures, nitrogenase activity was detected 75 min after the exclusion of oxygen, compared to 90 min for the CO2-supplemented cultures. Nitrogenase activity increased for more than 40 h at levels unique to each carbon-supplemented and unsupplemented culture. The relationship between nitrogenase activity, carbon source, and glycogen content is discussed.
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  • 80
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    Archives of microbiology 154 (1990), S. 510-513 
    ISSN: 1432-072X
    Keywords: Frankia ; Nitrogen fixation ; Glutamine synthetase ; Ammonium assimilation ; Serine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen-starved cells of Frankia strain HFPArl3 incorporated [13N]-labeled ammonium into glutamine ≫ serine ≫ (glutamate, alanine, aspartate), after five-minute radioisotope exposures. High initial endogenous pools of glutamate were reduced, while total glutamine increased, during short term NH inf4 sup+ incubation. Preincubation of cells in methionine sulfoximine (MSX) resulted in [13N]glutamine reduced by more than 80%, while [13N]glutamate and [13N]alanine levels increased. The results suggest that glutamine synthetase is the primary enzyme of ammonium assimilation, and that glutamate dehydrogenase and alanine dehydrogenase may also function in ammonium assimilation at low levels. Efflux of [13N]serine and lesser amounts of [13N]glutamine was detected from the Frankia cells. The identity of both Ser and Gln in the extracellular compartment was confirmed with gas chromatography/mass spectrometry. Serine efflux may be of significance in nitrogen transfer in Frankia.
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  • 81
    ISSN: 1432-2048
    Keywords: β-1,4-Glucan ; Nitrogen fixation ; Phaseolus (root nodules) ; Rhizobium (mutant) ; Root nodules ; produced by mutant Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Root nodules induced in Phaseolus vulgaris L. by the wild-type (WT) and a C4-dicarboxylic-acid mutant strain of Rhizobium leguminosarum biovar. phaseoli were compared on the basis of ultrastructure and cytochemistry of cellulose subunits. The mutant bacteroids failed to colonize infected host cells in a normal manner, and presented a premature degenerative appearance. Starch granules, rough endoplasmic reticulum and mitochondria were found to accumulate in the ineffective nodules. The most striking difference between effective and ineffective nodules was the presence of unusual spherical, laminated structures in plastids of mutant-infected host cells only. Cytochemical observations showed that these structures contain β-1,4-glucans. The presence of β-1,4-glucans within such structures may be caused by the activity of a cellulase which is produced by either the bacteroids or the host cell and is locally hydrolyzing the host cell-wall, thus releasing cellulose subunits into the cytoplasm. Another possibility is denovo synthesis of β-1,4-glucans in the host cell.
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  • 82
    ISSN: 1432-2242
    Keywords: Genetics ; Symbiosis ; Nitrogen fixation ; Coevolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary To determine the relationship between nodulation restriction by the Rj4 allele of soybean, rhizobitoxine-induced chlorosis, and taxonomic grouping of bradyrhizobia, 119 bradyrhizobial isolates were tested in Leonard jar culture for nodulation response and chlorosis induction. In addition to strain USDA 61, the strain originally reported as defining the Rj4 response, eight other isolates (i.e., USDA 62, 83, 94, 238, 252, 259, 260, and 340) were discovered to elicit the nodulation interdiction of the Rj4 allele. Only 16% of all the bradyrhizobial strains tested induced chlorosis, but seven of the nine strains (78%) interdicted by the Rj4 allele were chlorosis-inducing strains. Furthermore, in tests for antibiotic resistance profile, eight of the nine interdicted strains (89%) were classed in DNA homology group II. This evidence suggests that the Rj4 allele has a positive value to the host plant in shielding it from nodulation by certain chlorosis-inducing bradyrhizobia of a DNA homology group with impaired efficiency of nitrogen fixation with soybean.
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  • 83
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    Theoretical and applied genetics 80 (1990), S. 223-227 
    ISSN: 1432-2242
    Keywords: Selection index ; Nitrogen fixation ; Peanut ; Nonparametric method ; Stepwise regression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Eight characters related to nitrogen fixation and pod development measured 30 days after flowering were evaluated for their correct grading of the relative yield performance of 17 genetically diverse lines of groundnut (Arachis hypogaea L.). Each line was assigned a high or low yield status based on its pod yield, shelling percentage, and 100-kernel weight. Seventeen character combinations were examined for their relative merit in correct identification of the yield status of lines. The character sets, nitrogenase activity alone or in combination with nitrogen percent or shoot weight identified the status of 77% of lines correctly. The extent to which various characters accounted for the variation in pod yield was also checked by multiple regression analysis. While the character combination, nitrogen percent plus leaf area explained 75% of variation in pod yield, nodule mass, nitrogenase activity, and leaf area occurred in some other combinations that explained yield variation to a lesser extent. These analyses point to the profitability of involving crop physiological traits such as leaf area and nitrogen percent in selecting for relative yield performance in groundnut.
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  • 84
    ISSN: 1617-4623
    Keywords: Cyanobacteria ; Anabaena ; Heterocyst ; Nitrogen fixation ; DNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A 1.8 kb transcript corresponding to a region of the Anabaena 7120 chromosome 4 kb downstream of the nifHDK operon appears 12–18 h after heterocyst induction. The DNA corresponding to this transcript was sequenced and found to contain two open reading frames, designated ORF 1 and ORF 2. Two polypeptides, of 30 kDa and 13 kDa, encoded by these ORFs were expressed in Escherichia coli. An apparent start site for the transcript, detected by S1 nuclease protection, was located 42 bp upstream of the ATG start codon of ORF 1. ORF 2 shows strong sequence similarity to ORF 6 in the nif gene region of Azotobacter vinelandii. ORF 1 was interrupted using a 1.4 kb neomycin resistance cassette and the resulting mutant grew very slowly on medium lacking combined nitrogen. The mutant had 45% of wild-type acetylene reduction activity, which could be complemented by a 2.8 kb EcoRI fragment of wild-type Anabaena DNA containing only ORF 1 and ORF 2. Thus, one or both of these ORFs is required for efficient nitrogen fixation in Anabaena.
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  • 85
    ISSN: 1617-4623
    Keywords: Proline biosynthesis ; Nitrogen fixation ; Osmoregulation ; proC gene ; Glycine max
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated several cDNA clones encoding Δ1 1-pyrroline-5-carboxylate reductase (P5CR, l-proline: NAD(P)+ 5-oxidoreductase, EC 1.5.1.2) which catalyzes the terminal step in proline biosynthesis, by direct complementation of a proC mutation in Escherichia coli with an expression library of soybean root nodule cDNA. The library was constructed in the λ ZapII vector, converted to a plasmid library by in vivo excision of recombinant pBluescript phagemids, and used for transformation of the E. coli mutant. Complementing plasmids contained inserts of about 1.2 kb which hybridized to a 1.3 kb RNA transcript in nodules, uninfected roots and leaves. DNA sequence analysis of one full length cDNA clone showed that it encoded a 28 586 Mr polypeptide with 39% amino acid identity to the E. coli P5CR sequence. Genomic analysis showed that there are two to three copies of the P5CR gene in the soybean genome. The steady-state level of P5CR mRNA in root nodules was twice as high as in uninfected roots and about five times higher than in leaves. Subjecting young seedlings to osmotic stress by watering with 400 mM NaCl resulted in an almost six-fold increase in the level of root P5CR mRNA, suggesting that this gene may be osmoregulated.
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  • 86
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; nifDKEN operon ; Nucleotide sequence ; Protein sequence conservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequences of genes homologous to the Klebsiella pneumoniae nifEN genes have been determined in Bradyrhizobium japonicum 110. The coding regions for the nifE and nifN consist, respectively, of 1641 and 1407 nucleotides. The nifD gene (coding for the β-subunit of dinitrogenase) and nifE are linked, and separated by 95 nucleotides. In the region of 12 nucleotides that separates nifE from nifN the stop codon for nifE overlaps the putative ribosome binding site for nifN. In contrast to Klebsiella and Azotobacter vinelandii, the B. japonicum nifEN genes are linked to the nifDK genes in the same operon. Comparison of dinitrogenase polypeptides (nifDK products) and the polypeptides of the nifE and nifN genes reveals considerable homology between nifD and nifE, and between nifK and nifN. Several protein domains, containing highly conserved cysteine residues, are conserved among the gene products of nifD, nifK, nifE and nifN. This result allows us to propose a probable evolutionary pathway for the common origin of these genes.
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  • 87
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; Plant gene regulation ; Nodulin genes ; 5′ promoter analysis ; DNA regulatory motifs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The qualitative and quantitative contributions of four separate cis-acting DNA elements controlling the root nodule-specific soybean leghemoglobin lbc 3 gene were analyzed in transgenic Lotus corniculatus plants. Expression from internal deletions in the 5′ region between positions −49 and −1956 was monitored from a CAT reporter gene. The strong positive element (SPE; −1090, −947) responsible for high-level expression was demonstrated to be an organ-specific element by deleting proximal nodule-specific control elements. Deletion of the downstream qualitative organ-specific element (OSE; −139, −102) containing the putative nodulin consensus sequences 5′AAAGAT and 5′CTCTT resulted in a low expression level. Efficient SPE enhancement is therefore dependent on the organ-specific element, which by itself does not enhance expression. This quantitative effect of the immediate upstream region carrying the consensus sequences was also found in hybrid promoter studies using the soybean nodulin N23 gene promoter, suggesting the involvement of these motifs in a regulatory mechanism for nodulin genes. Deletion of the lbc 3 negative element (NE, −102, t-49) linking the SPE and OSE onto the TATA box did not lead to unregulated expression. These results indicate that interaction between positive, negative and neutral qualitative elements controls lbc 3 expression. Binding of the nuclear protein NAT2 at the lbc 3 weak positive element (WPE; −230, −170) is probably not directly required for this mechanism.
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  • 88
    ISSN: 1573-5079
    Keywords: Anabaena ; ferredoxin ; flavodoxin ; FPLC determination of ferredoxin and flavodoxin ; Iron deficiency ; Nitrogen fixation ; Photosynthesis ; strain differences in gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Iron-dependent formation of ferredoxin and flavodoxin was determined in Anabaena ATCC 29413 and ATCC 29211 by a FPLC procedure. In the first species ferredoxin is replaced by flavodoxin at low iron levels in the vegetative cells only. In the heterocysts from Anabaena ATCC 29151, however, flavodoxin is constitutively formed regardless of the iron supply. Replacement of ferredoxin by flavodoxin had no effect on photosynthetic electron transport, whereas nitrogen fixation was decreased under low iron conditions. As ferredoxin and flavodoxin exhibited the same Km values as electron donors to nitrogenase, an iron-limited synthesis of active nitrogenase was assumed as the reason for inhibited nitrogen fixation. Anabaena ATCC 29211 generally lacks the potential to synthesize flavodoxin. Under iron-starvation conditions, ferredoxin synthesis is limited, with a negative effect on photosynthetic oxygen evolution.
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  • 89
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    Biology and fertility of soils 8 (1989), S. 356-368 
    ISSN: 1432-0789
    Keywords: Plant-root associations ; Azospirillum spp ; Rhizosphere ; Nitrogen fixation ; Acetylene reduction assay (ARA) ; Phytohormones
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Bacteria of the genus Azospirillum are extensively studied for their plant-growth promoting effect following inoculation. Physiological and biochemical studies of these diazotrophic bacteria are now benefiting from recent breakthroughs in the development of genetic tools for Azospirilum. Moreover, the identification and cloning of Azospirillum genes involved in N2 fixation, plant interaction, and phytohormone production have given new life to many research projects on Azospirillum. The finding that Azospirillum genes can complement specific mutations in other intensively studied rhizosphere bacteria like Rhizobia will certainly trigger the exploration of new areas in rhizosphere biology. Therefore a review of the Azospirillum-plant interactions is particularly timely.
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  • 90
    ISSN: 1432-0789
    Keywords: Nodule damage ; Rivellia angulata ; Nitrogen fixation ; Cajanus cajan ; Pigeonpea ; Vertisol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Damage caused by Rivellia angulata larvae to pigeonpea root nodules at the ICRISAT center in India was greater in the crop grown on Vertisols (up to 86%) compared to that on Alfisols (20%). Attempts to quantify the field effects of nodule damage on growth and yield of pigeonpea in a Vertisol, involving many heavy applications of soil insecticides (aldrin and hexachlorocyclohexane) failed because the insecticides did not control the pest and adversely affected the growth of the pigeonpea and the subsequent crop of sorghum (Sorgorum bicolor L. Moench). The impact of nodule damage on pigeonpea growth, yield and nutrient uptake was successfully studied in greenhouse-grown plants at three N levels. In this pot study, artificial inoculation with Rivellia sp. led to substantial nodule damage (70%). The results of this damage were a significant overall reduction in nodule dry weight (46%), acetylene reduction activity (31%), total leaf area (36%), chlorophyll content of leaves (39%) and shoot dry weight (23%) 68 days after sowing. At maturity, Rivellia sp. infestation caused significant reductions in top dry weight (22%), root and nodule dry weight (27%), seed dry weight (14%), and total N (29%) and P uptake (19%). The problems and prospects of manipulating nodule damage so as to reduce N losses in pigeonpea are discussed.
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  • 91
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    Biology and fertility of soils 7 (1989), S. 269-274 
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; Frankia-Ceanothus spp. association ; Acetylene reduction assay (ARA) ; Microsymbiont population ; Nodules ; Actinomycetes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Wildland shrub improvement is needed for sound range and disturbed land revegetation practice. The possibility of selecting superior N2-fixingFrankia-Ceanothus spp. actinorhizal associations was examined. Greenhouse tests were used to expose various soil-borne microsymbiont andCeanothus sp. population accessions in reciprocal combination. The acetylene reduction rate was used as a measure of N2-fixation capacity. There was no significant interaction between host and microsymbiont regardless of source for all variables measured. The acetylene reduction rate, nodule number and mass, plant biomass, and root: shoot ratio were significantly different among soil sources. The acetylene reduction rate was not significantly different amongCeanothus sp. accessions. Neither was it strongly correlated with other variables. It was concluded that the N2-fixation rate is more a function ofFrankia sp. than the hostCeanothus sp. in actinorhizal associations. It appears possible to select soil sources with superior N2-fixing microsymbiont populations.
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  • 92
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    Oecologia 79 (1989), S. 566-568 
    ISSN: 1432-1939
    Keywords: Carbon dioxide ; Lichen ; Lobaria ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thalli of Lobaria pulmonaria (L.) Hoffm., a nitrogen-fixing epiphyte common in mesic temperate forests, were collected in a Douglas-fir (Pseudotsuga menziesii Franco) forest near Corvallis, Oregon, and maintained for 20 to 40 days in controlled-environment chambers with atmospheric CO2 concentrations of 374 and 700 μll-1. Nitrogenase activity, which was assayed by the acetylene reduction method, was approximately doubled in the lichen maintained in elevated CO2. Increases in nitrogen fixation by lichens may be an important part of the integrated ecosystem response to rising CO2.
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  • 93
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    Archives of microbiology 151 (1989), S. 445-453 
    ISSN: 1432-072X
    Keywords: Denitrification ; Growth yield measurements ; Nitrate respiration ; Nitrogen fixation ; Proton translocations in respirations ; Azospirillum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract For Azospirillum brasilense Sp7, the energy transformation efficiencies were measured in anaerobic respirations with either nitrate, nitrite or nitrous oxide as respiratory electron acceptors by determining the maximal molar growth yields and the H+-translocations using the oxidant pulse method. In continuous cultures grown with malate limiting, the maximal molar growth yields (Y s max -values) were essentially the same with O2 or N2O but were 1/3 and 2/3 lower with NO 2 - or NO 3 - , respectively, as respiratory electron acceptors. Both the maximal molar growth yields and the maintenance energy coefficients were surprisingly high when Azospirillum was grown with nitrite as the sole electron acceptor and source for N-assimilation. Growth under N2-fixing conditions drastically reduced the Y s max -values in the N2O and O2-respiring cells. In the H+-translocation measurements, the $$\vec H^ + $$ /oxidant ratios were 5.6 for O2→H2O, 2.5–2.8 for NO 3 - →NO 2 - , 2.2 for NO 2 - →N2O and 3.1 for N2O→N2 respirations when the cells were preincubated with valinomycin and K+. All the values were enhanced when the experiments were performed with valinomycin plus methyltriphenylphosphonium (=TPMP+) cation. The uncoupler carbonyl cyanide-m-chlorophenyl-hydrazone diminished the H+-excretion indicating that this translocation was due to vectorial flow across the membrane. In the absence of any ionophore, nitrate and nitrite respirations were accompanied by a H+-uptake $$(NO_3^ - \to N_2 = - 2.9 \vec H^ + /NO_3^ - and NO_2^ - \to N_2 = - 2.5 \vec H^ + /NO_2^ - )$$ . Any significant H+-translocation could not be detected in N2O- and O2-respirations under these conditions. It is concluded that nitrate reduction proceeds inside the cytoplasmic membrane, whereas nitrite is reduced extramembraneously. The data are not conclusive for the location of nitrous oxide reductase. The maximal molar growth yield determinations and the absence of any H+-uptake in untreated cells indicate a cytoplasmic orientation of the enzyme similar to the terminal cytochrome oxidase of respiration. The low H+-extrusion values for N2O-respiration compared to O2-respiration in cells treated with valinomycin plus TPMP+ are, however, not in accord with such an interpretation.
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  • 94
    ISSN: 1432-072X
    Keywords: Bradyrhizobium ; Gene cloning ; Heme ; Marker exchange mutagenesis ; Nitrogen fixation ; Respiration ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Random and site-directed Tn5-induced mutagenesis of Bradyrhizobium japonicum yielded two mutations, one in strain 2960 and the other in strain 2606::Tn5-20, which mapped close to each other but in separate genes. The corresponding wild-type genes were cloned, and their approximate location on the cloned DNA was determined. Mutant 2960 was Fix- and formed green nodules on soybean, whereas strain 2606::Tn5-20 had ca. 4% of wild-type Fix activity and formed white nodules. Cytochrome oxidase assays (Nadi tests) showed a negative reaction with both mutants, indicating a functional deficiency of cytochrome c or its terminal oxidase or both. However, the mutants grew well under aerobic conditions on minimal media with different carbon sources. Furthermore, mutant 2960 had a reduced activity in hydrogen uptake, was unable to grow anaerobically with nitrate as the terminal electron acceptor and 2960-infected soybean nodules contained little, if any, functional leghemoglobin. Southern blot analysis showed that a B. japonicum heme biosynthesis mutant [strain LO505: O'Brian MR, Kirshbom PM, Maier RJ (1987) Proc Natl Acad Sci USA 84: 8390–8393] had its mutation close to the Tn5 insertion site of our mutant 2606::Tn5-20. This finding, combined with the observed phenotypes, suggested that the genes affected in mutants 2960 and 2606::Tn5-20 were involved in some steps of heme biosynthesis thus explaining the pleiotropic respiratory deficiencies of the mutants. Similar to strain LO505, the mutant 2606::Tn5-20 (but not 2960) was defective in the activity of protoporphyrinogen IX oxidase which catalyzes the penultimate step in the heme biosynthesis pathway. This suggests that one of the two cloned genes may code for this enzyme.
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  • 95
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    Archives of microbiology 151 (1989), S. 180-182 
    ISSN: 1432-072X
    Keywords: Klebsiella pneumoniae ; Nitrogen fixation ; nifL ; Regulation ; Oxygen control ; Nitrogen control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A number of in-frame deletions have been constructed in the Klebsiella pneumoniae regulatory gene nifL. The effects of each nifL mutation on NifA-mediated expression from the nifH promoter of K. pneumoniae have then been assessed with respect to both nitrogen and oxygen control. These experiments indicate that, in contrast to the situation with the homologous regulatory proteins NtrB and NtrC, NifA activity is not impaired in the absence of NifL. We conclude that the only function of NifL is to inactivate NifA in response to an increase in the nitrogen or oxygen status of the cell.
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  • 96
    ISSN: 1432-2048
    Keywords: Glutamate synthase ; Glutamine synthetase ; Nitrogen fixation ; Phaseolus (glutamate synthase) ; Plastid (glutamate synthase) ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The two isoenzymes of NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14), previously identified in root nodules of Phaseolus vulgaris L., have both been shown to be located in root-nodule plastids. The nodule specific NADH-GOGAT II accounts for the majority of the activity in root nodules, and is present almost exclusively in the central tissue of the nodule. However about 20% of NADH-GOGAT I activity is present in the nodule cortex, at about the same specific activity as this isoenzyme is found in the central tissue. Glutamine synthetase (GS; EC 6.3.1.2) occurs predominantly as the γ polypeptide in the central tissue, whereas in the cortex, the enzyme is represented mainly by the β polypeptide. Over 90% of both GS and NADH-GOGAT activities are located in the central tissue of the nodule and GS activity exceeds NADH-GOGAT activity by about twofold in this region. Using the above information, a model for the subcellular location and stoichiometry of nitrogen metabolism in the central tissue of P. vulgaris root nodules is presented.
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  • 97
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    Theoretical and applied genetics 78 (1989), S. 433-435 
    ISSN: 1432-2242
    Keywords: Rhizobium ; Irradiation ; Nitrogen fixation ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Sumary The objective of this work was to know the behaviour and variability of Rhizobium leguminosarum after irradiation. The induced variation was tested under greenhouse conditions on the variety JV 3 of broad beans (Vicia faba) in six replications. Induced genetic variabilty was observed for strain, parent and mutant versus parent. Out of 24 irradiated strains, strain 93-32 performed better with a greater number of nodules and higher dry weight of nodules per plant and biological yield. Environment played an important role in the expression of characters observed. High heritability and genetic advance of these traits indicated that the nitrogen fixation ability of Rhizobium can easily be improved by selection.
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  • 98
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    Molecular genetics and genomics 216 (1989), S. 484-491 
    ISSN: 1617-4623
    Keywords: Nitrogen fixation ; nifL ; Repression ; Metal ions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ability of the Klebsiella pneumoniae nifL gene product to antagonise NIFA mediated transcriptional activation from the nifH promoter in vivo was inhibited either by metal deprivation, or by the presence of the iron chelators EDDA or Desferal in the growth medium. This inhibition of the repressive activity of NIFL was reversed by the addition of ferrous or manganous ions to the medium but was unaffected by other transition metals. The dependence on metal ions for NIFL activity was observed when NIFL was overexpressed and when cultures were exposed to oxygen or high levels of fixed nitrogen. Immunochemical evidence suggests that NIFL and NIFA associate to form a functional protein complex. Metal ions are apparently not required for the formation of this complex.
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  • 99
    ISSN: 1617-4623
    Keywords: glnB ; Klebsiella pneumoniae ; Nitrogen control ; Glutamine synthetase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The role of theKlebsiella pneumoniae PII protein (encoded byglnB) in nitrogen regulation has been studied using two classes ofglnB mutants. In Class I mutants PII appears not to be uridylylated in nitrogen-limiting conditions and in Class II mutants PII is not synthesised. The effects of these mutations on expression from nitrogen-regulated promoters indicate that PII is not absolutely required for nitrogen control. Furthermore the uridylylated form of PII(PII-UMP) plays a significant role in the response to changes in nitrogen status by counteracting the effect of PII on NtrB-mediated dephosphorylation of NtrC. PII is not involved in thenif-specific response to changes in nitrogen status mediated by NifL.
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  • 100
    ISSN: 1617-4623
    Keywords: Rhodobacter capsulatus ; Nitrogen fixation ; DNA sequence analysis ; nifE, nifN, nifX genes ; Protein comparisons
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Rhodobacter capsulatus genes homologous to Klebsiella pneumoniae nifE, nifN and nifX were identified by DNA sequence analysis of a 4282 bp fragment of nif region A. Four open reading frames coding for a 51188 (NifE), a 49459 (NifN), a 17459 (NifX) and a 17472 (ORF4) dalton protein were detected. A typical NifA activated consensus promoter and two imperfect putative NifA binding sites were located in the 377 bp sequence in front of the nifE coding region. Comparison of the deduced amino acid sequences of R. capsulatus NifE and NifN revealed homologies not only to analogous gene products of other organisms but also to the α and β subunits of the nitrogenase iron-molybdenum protein. In addition, the R. capsulatus nifE and nifN proteins shared considerable homology with each other. The map position of nifX downstream of nifEN corresponded in R. capsulatus and K. pneumoniae and the deduced molecular weights of both proteins were nearly identical. Nevertheless, R. capsulatus NifX was more related to the C-terminal end of NifY from K. pneumoniae than to NifX. A small domain of approximately 33 amino acid residues showing the highest degree of homology between NifY and NifX was also present in all nifB proteins analyzed so far. This homology indicated an evolutionary relationship of nifX, nifY and nifB and also suggested that NifX and NifY might play a role in maturation and/or stability of the iron-molybdenum cofactor. The open reading rame (ORF4) downstream of nifX in R. capsulatus is also present in Azotobacter vinelandii but not in K. pneumoniae. Interposon-induced insertion and deletion mutants proved that nifE and nifN were necessary for nitrogen fixation in R. capsulatus. In contrast, no essential role could be demonstrated for nifX and ORF4 whereas at least one gene downstream of ORF4 appeared to be important for nitrogen fixation.
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