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  • Nitrogen fixation  (76)
  • Springer  (76)
  • International Union of Crystallography
  • Periodicals Archive Online (PAO)
  • 1980-1984  (76)
  • 1984  (32)
  • 1983  (23)
  • 1981  (21)
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  • Springer  (76)
  • International Union of Crystallography
  • Periodicals Archive Online (PAO)
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  • 1980-1984  (76)
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  • 1984  (32)
  • 1983  (23)
  • 1981  (21)
  • 1982  (27)
  • 1980  (13)
  • 1
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    Springer
    Journal of molecular evolution 21 (1984), S. 19-32 
    ISSN: 1432-1432
    Keywords: Leghemoglobin ; Gene duplication ; Gene linkage ; Concerted evolution ; Nitrogen fixation ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have analyzed the sequences of soybean leghemoglobin genes as an initial step toward understanding their mode of evolution. Alignment of the sequences of plant globin genes with those of animals reveals that (i) based on the proportion of nucleotide substitutions that have occurred at the first, second, and third codon positions, the time of divergence of plant and animal globin gene families appears to be extremely remote (between 900 million and 1.4 billion years ago, if one assumes constancy of evolutionary rate in both the plant and animal lineages) and (ii) in addition to the normal regulatory sequences on the 5′ end, an approximately 30-base-pair sequence, specific to globin genes, that surrounds the cap site is conserved between the plant and animal globin genes. Comparison of the leghemoglobin sequences with one another shows that (i) the relative amount of sequence divergence in various coding and noncoding regions is roughly similar to that found for animal globin genes and (ii) as in animal globin genes, the positions of insertions and deletions in the intervening sequences often coincide with the locations of direct repeats. Thus, the mode of evolution of the plant globin genes appears to resemble, in many ways, that of their animal counterparts. We contrast the overall intergenic organization of the plant globin genes with that of animal genes, and discuss the possibility of the concerted evolution of the leghemoglobin genes.
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  • 2
    ISSN: 1432-2048
    Keywords: Ammonia ; Nitrogen fixation ; Nodule ; Senescence (root nodules) ; Ureide ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract During early development (up to 18 d after sowing) of nodules of an “effective” cowpea symbiosis (Vigna unguiculata (L.) Walp cv. Vita 3: Rhizobium strain CB756), rapidly increasing nitrogenase (EC 1.7.99.2) activity and leghaemoglobin content were accompanied by rapid increases in activities of glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 2.6.1.53), enzymes of denovo purine synthesis (forming inosine monophosphate) xanthine oxidoreductase (EC 1.2.3.2), urate oxidase (EC 1.7.3.3), phosphoenolpyruvate carboxylase (EC 4.1.1.31) and led to increased export of ureides (allantoin and allantoic acid) to the shoot of the host plant in the xylem. Culturing plants with the nodulated root systems maintained in the absence of N2 (in 80 Ar: 20 O2, v/v) had little effect on the rates of induction and increase in nitrogenase activity and leghaemoglobin content but, in the absence of N2 fixation and consequent ammonia production by bacteroids, there was no stimulation of activity of enzymes of ammonia assimilation or of the synthesis of purines or ureides. Addition of NO 3 - (0.1–0.2 mM) relieved host-plant nitrogen deficiency caused by the Ar: O2 treatment but failed to increase levels of enzymes of N metabolism in either the bacteroid or the plant-cell fractions of the nodule. Premature senescence in Ar: O2-grown nodules occurred at 18–20 d after sowing, and resulted in reduced levels of nitrogenase activity and leghaemoglobin but increased the activity of hydroxybutyrate oxidoreductase (EC 1.1.1.30).
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  • 3
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    Planta 152 (1981), S. 544-552 
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Lichens ; Nitrogen fixation ; Peltigera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Nostoc in the cephalodia of the lichen Peltigera aphthosa Willd. fixed 15N2 and the bulk of the nitrogen fixed was continuously transferred from it to its eukaryotic partners (a fungus and a green alga, Coccomyxa sp.). Kinetic studies carried out over the first 30 min, after exposure of isolated cephalodia to 15N2, showed that highest initial 15N2-labelling was into NH 4 + . After 12 min little further increase in the NH 4 + label occurred while that in the amide group of glutamine and in glutamate continued to increase. The 15N-labelling of the amino group of glutamine and of aspartate increased more slowly, followed by an increase in the labelling of alanine. When total incorporation of 15N-label was calculated, the overall pattern was found to be rather similar except that, throughout the experiment, the total 15N incorporated into glutamate was about six times greater than that into the amide group of glutamine. Pulse chase experiments, in which 14N2 was added to cephalodia previously exposed to 15N2, showed that the NH 4 + pool rapidly became depleted of 15N-label, followed by decreases in the labelling of glutamate, the amide group of glutamine and aspartate. The 15N-labelling of alanine, however, continued to increase for a period. When isolated cephalodia were treated with L-methionine-SR-sulphoximine, an inhibitor of glutamine synthetase (EC 6.3.1.2), and azaserine, an inhibitor of glutamate synthase (EC 2.6.1.53), there was no detectable labelling in glutamine although the 15N-labelling of glutamate increased unimpaired. On treating the cephalodia with amino-oxyacetate, an inhibitor of aminotransferase activity, the alanine pool decreased. Evidence was obtained that glutamine synthetase and glutamate synthase were located in the Nostoc, and that glutamate dehydrogenase (EC 1.4.1.4) and various amino-transferases were located in the cephalodial fungus. Possible implications of these findings are discussed.
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  • 4
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    Environmental management 7 (1983), S. 177-187 
    ISSN: 1432-1009
    Keywords: Nitrogen ; Model ; Agriculture ; Mass balance ; Ground-water ; Denitrification ; Immobilization ; Dry deposition ; Nitrogen fixation ; Nitrate ; Florida
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract A detailed nitrogen budget was devised for agricultural activities in the Florida peninsula, based on routine data published by state agricultural agencies. The model demonstrates that important unmonitored fluxes of nitrogen can often be calculated by mass balance on individual model compartments, and that the reasonability of poorly quantified fluxes can be assessed. The results of such models can be very useful in designing and assessing the results of field experiments and in prioritizing environmental monitoring programs.
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  • 5
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Regulation ; Guanosine 5′-diphosphate 3′-diphosphate (ppGpp)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Addition of ammonium to N2 fixing cultures of Azotobacter vinelandii, Klebsiella pneumoniae and Clostridium pasteurianum rapidly reduced the intracellular levels of guanosine 5′-diphosphate 3′-diphosphate (ppGpp) by 70–90%. This change might reflect a regulatory role of ppGpp in nitrogen metabolism.
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  • 6
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    Archives of microbiology 136 (1983), S. 20-25 
    ISSN: 1432-072X
    Keywords: Hydrogen production ; Nitrogen fixation ; Hydrogen recycling ; Hydrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutants of Rhodopseudomonas capsulata lacking uptake hydrogenase activity have been isolated among those unable to grow photoautotrophically. Studies with these mutants showed increases in nitrogenase mediated H2 production from all substrates tested. In addition, photosynthetic synthetic growth on N2 with malate as carbon source was not affeced by the block in H2 uptake even under low light. Under these growth conditions hydrogen was observed to accumulate in mutant but not in wild-type cultures. This finding suggested that H2 was evolved by nitrogenase during N2 fixation by this photosynthetic bacterium and was efficiently recycled in the wild type.
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  • 7
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    Archives of microbiology 135 (1983), S. 287-292 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Respiration ; Nitrogen fixation ; Heterocysts ; K m for O2 ; Anabaena variabilis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Simultaneous measurements of acetylene reduction by Anabaena variabilis and the concentration of dissolved oxygen in the suspension were made using a specially designed vessel which allowed measurements under steady-state conditions. The rate of acetylene reduction in the dark increased with increasing oxygen concentrations until a maximum value was reached at 300 μM O2 (corresponding to 30% O2 in the gas phase at 35°C). This presumably results from a requirement for energy provided by respiration. Measurements of the dependence of respiration rate on dissolved oxygen concentration were made under comparable conditions using an open system to allow conditions close to steady-state to be obtained. The respiration rate of diazotrophically grown Anabaena variabilis had a dependence on oxygen concentration corresponding to the sum of two activities. These had K m values of 1.0 μM and 69 μM and values of V max of similar magnitude. Only the high affinity activity was observed in nitrate-grown cyanobacteria lacking heterocysts, and this presumably represent activity in the vegetative cells. The oxygen concentration dependence of the low affinity activity resembled that for the stimulation of acetylene reduction. We interpret this as the result of oxygen uptake by the heterocysts. The results are consistent with the idea that in intact filaments of cyanobacteria O2 enters heterocysts much more slowly than it enters the vegetative cells.
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  • 8
    ISSN: 1432-072X
    Keywords: Ammonium export ; Ammonium assimilation ; Glutamine synthetase ; Nitrogen fixation ; Rhizobium sp. 32H1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between ammonium assimilation and ammonium export has been studied in free-living, N2-fixing Rhizobium sp. 32H1. After 55 to 67 h of microaerobic growth under a gas phase of 0.2% O2 – 1.0% CO2 – 98.8% Ar high levels of nitrogenase were observed concomitant with a slightly adenylylated glutamine synthetase (GSI) and some glutamine synthetase (GSII) activity. However, after growth of 89 h, or longer, GSI became adenylylated and the level of GSII had decreased. When the gas phase was shifted to 0.2% O2 – 1.0% CO2 – 98.8% N2, a lag was observed before ammonium export could be detected in the 55 to 67 h cultures. No lag in ammonium export was observed in the cultures previously grown for 89 h. The onset of ammonium export in the 55 to 67 h cultures was found to correlate with the adenylylation state of GSI. There appeared to be no correlation between the level of GSII and the export of ammonium. Neither an increase in the adenylylation level of GSI nor ammonium export was observed when the 55 to 67 h cultures were maintained under the Ar gas mixture.
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  • 9
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    Archives of microbiology 129 (1981), S. 238-239 
    ISSN: 1432-072X
    Keywords: Rhizobium ; Disaccharide ; Bacteroid ; Transport ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Slow growing strains of rhizobia appear to lack both uptake systems and catabolic enzymes for disaccharides. In the fast-growing strains of rhizobia there are uptake mechanisms and catabolic enzymes for disaccharide metabolism. In Rhizobium leguminosarum WU 163 and WU235 and R. trifolii WU290, sucrose and maltose uptake appears to be constitutive whereas in R. meliloti WU60 and in cowpea Rhizobium NGR234 uptake of these disaccharides is inducible. There is evidence that there are at least two distinct disaccharide uptake systems in fast-growing rhizobia, one transporting sucrose, maltose and trehalose and the other, lactose. Disaccharide uptake is via an active process since uptake is inhibited by azide, dinitrophenol and carbonyl cyanide m-chlorophenylhydrazone but not by arsenate. Bacteroids of R. leguminosarum WU235 and R. lupini WU8 are unable to accumulate disaccharides.
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  • 10
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    Archives of microbiology 130 (1981), S. 38-43 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Gene expression ; Regulation ; Messenger RNA ; Transcription ; Klebsiella pneumoniae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogenase messenger RNA synthesis in Klebsiella pneumoniae was determined by labelling cells with (3H)uracil and isolating total RnA, which was then hybridized to filterbound recombinant plasmid pSA30 DNA carrying the nitrogenase structural genes nifH, D, and K. Derepression of nitrogenase mRNA starts 1.5 h before the onset of nitrogenase activity (as measured by acetylene reduction). Exposure of nif-derepressed cultures to either NH 4 + , air, or high temperatures (39° C) results in a rapid decrease of the synthesis rates both of nitrogenase mRNA and nitrogenase polypeptides. Nitrogenase mRNA is remarkably stable. After blocking transcription with rifampicin, hybridizable and actively translatable nitrogenase mRNA survives with an average half-life of 18 min. Half-lives are considerably shorter when rifampicin-inhibited cultures are simultaneously shifted to conditions which are non-permissive for nitrogenase synthesis, pointing to some posttranscriptional influence on nitrogenase mRNA stability. In all experiments performed there was no evidence for uncoupling of nitrogenase mRNA synthesis from nitrogenase mRNA translation, indicating that nitrogenase synthesis is regulated solely by transcriptional control.
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  • 11
    ISSN: 1432-072X
    Keywords: Rhizobium trifolii ; Symbiosis ; Nodulation ; Nitrogen fixation ; Symbiotic genes ; Reiterated sequences ; Plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A Rhizobium trifolii symbiotic plasmid specific gene library was constructed and the physical organisation of regions homologous to nifHDK, nifA and nod genes was determined. These symbiotic gene regions were localised to u 25 kb region on the sym-plasmid, pPN1. In addition four copies of a reiterated sequence were identified on this plasmid, with one copy adjacent to nifH. No rearrangement of these reiterated sequences was observed between R. trifolii bacterial and bacteroid DNA. Analysis of a deletion derivative of pPN1 showed that these sequences were spread over a 110 kb region to the left of nifA.
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  • 12
    ISSN: 1432-072X
    Keywords: Ammonia assimilation ; Lichen symbioses ; Nitrogen fixation ; 15N kinetics ; Peltigera canina
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract On following N2-incorporation and subsequent metabolism in the lichen Peltigera canina using 15N as tracer, it was found, over a 30 min period, that greatest initial labelling was into NH 4 + followed by glutamate and the amide-N of glutamine. Labelling of the amino-N of glutamine, aspartate and alanine increased slowly. Pulse-chase experiments using 15N confirmed this pattern. On inhibiting the GS-GOGAT pathway using l-methionine-dl-sulphoximine and azaserine, 15N enrichment of glutamate, alanine and aspartate continued although labelling of glutamine was undetectable. From this and enzymic data, NH 4 + assimilation in the P. canina thallus appears to proceed via GS-GOGAT in the cyanobacterium and via GDH in the fungus; aminotransferases were present in both partners. The cyanobacterium assimilated 44% of the 15N2 fixed; the remainder was liberated almost exclusively as NH 4 + and then assimilated by fungal GDH.
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  • 13
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    Archives of microbiology 135 (1983), S. 103-109 
    ISSN: 1432-072X
    Keywords: RNA polymerase ; Transcription ; Nitrogen fixation ; Symbiosis ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA-dependend RNA polymerase (EC 2.7.7.6) from Rhizobium japonicum was purified. The subunit structure was found to be ββ′α2σ, with the following apparent molecular weights determined by electrophoresis: M r (β and β') 150,000 each, M r (σ) 96,000, M r (α) 40,000, M r (holoenzyme) 490,000, M r (core enzyme) 380,000. The recovery of σ was 28%. RNA polymerase from aerobically grown R. japonicum cells and from nitrogen-fixing cells have the same electrophoretic properties suggesting that no chemical modification of the enzyme occurs when cells undergo this metabolic differentiation. The enzyme is Mg2+-dependent, rifampicin-sensitive, and has optimal activity at alkaline pH (8–10) and at 35–40° C. It binds strongly to bacteriophage T7 promoters, weakly to antibiotic resistance genes, and not at all to cloned R. japonicum nif DNA. Preliminary in vitro transcription experiments, including nif DNA as template, revealed that additional factors may be required for selective transcription from promoters.
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  • 14
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    Archives of microbiology 136 (1983), S. 219-221 
    ISSN: 1432-072X
    Keywords: Xanthobacter ; Nitrogen fixation ; Oxygen sensitivity ; Nitrogen metabolism ; Glutamine synthetase ; Glutamate synthase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract N2-fixation was investigated in the chemoautotrophic hydrogen bacterium Xanthobacter H4-14. N2-fixing batch cultures of this organism could only be grown at pO2 values of around 0.02 bar, and in continuous culture dissolved oxygen tensions above 16 μM were found to inhibit N2-fixation. Xanthobacter H4-14 utilized a variety of amino acids, nitrate and ammonia as nitrogen sources. Cell-free extracts from steady-state continuous cultures of ammonia grown, nitrate grown and N2-fixing Xanthobacter were assayed for the presence of ammonia assimilation enzymes. No alanine dehydrogenase or glutamate dehydrogenase activity was detected. Ammonia was assimilated exclusively via the glutamine synthetase/glutamate synthase pathway, irrespective of the extracellular concentration of ammonia.
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  • 15
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrogenase regulation ; Glutamine synthetase ; Methionine suofoximine ; Rhodospirillaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methionine sulfoximine (MSX), an irreversible inhibitor of glutamine synthetase of Rhodopseudomonas palustris restored nitrogenase activity to cells in which nitrogenase had been completely inhibited by ammonia switch-off. After addition of MSX, there was a lag period before nitrogenase activity was fully restored. During this lag, glutamine synthetase activity progressively decreased, and near the time of its complete inhibition, nitrogenase activity resumed. Nitrogenase switch-off by ammonia thus required active glutamine synthetase. Glutamine itself caused nitrogenase inhibition whose reversal by MSX depended on the relative ratio of MSX to glutamine. Unlike ammonia, glutamine inhibited nitrogenase under conditions where glutamine synthetase activity was absent. This indicates that glutamine is the effector molecule in nitrogenase switch-off, for instance by interacting with the enzymatic system for Fe protein inactivation. The effects of glutamine and MSX were also dependent on the culture age. Possible explanation for this and for the competitive effects are a common binding site within the regulatory apparatus for nitrogenase, or, in part, within a common transport system. Some observations with MSX were extended to Rhodopseudomonas capsulata and agreed with those in R. palustris.
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  • 16
    ISSN: 1432-072X
    Keywords: Azotobacter vinelandii ; Continuous culture ; Oxygen control ; Nitrogen fixation ; Respiratory protection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Azotobacter vinelandii strain OP was grown in continuous culture at various dissolved oxygen concentrations of air (100% air saturation of the medium=225 ±14 μM O2). Sucrose was added as carbon source and either dinitrogen or ammonia as nitrogen sources. Irrespective of the nitrogen source steady state cultures showed the following general responses with dissolved oxygen concentrations increasing from about 1% to 30% air saturation: (i) cell protein levels, (ii) the amount of cell protein formed per sucrose consumed as well as (iii) nitrogenase activity decreased by at least a factor of two while (iv) cellular respiration increased. At higher oxygen concentrations the parameters changed only slightly, if at all. Increasing the sucrose concentration in the inflowing medium (s R) from 3 g/l to 15 g/l increased the total level of cellular respiration with nitrogen-fixing cultures but was more pronounced with ammonium-assimilating cultures. With nitrogen-fixing cultures cell protein levels increased five-fold while the ratio of protein formed per sucrose consumed as well as cellular nitrogenase activity remained unaffected. With ammonium-assimilating cultures the cell protein level was only doubled and the level of cell protein formed per sucrose consumed was decreased at the higher s R. Increasing the dilution rate at a constant oxygen concentration of 45% air saturation resulted in an almost parallel increase of both cellular respiratory and nitrogenase activity at low and moderate dilution rates. At high dilution rates nitrogenase activity increased steeply over the respiratory activity. Nitrogen-fixing cultures adapted to various oxygen concentrations were subjected to oxygen stress by increasing the oxygen concentration for 7 min. In all cases, this resulted in a complete inhibition (‘switch-off’) of nitrogenase activity. Upon restoration of the original oxygen concentration nitrogenase activity returned to a decreased level. The discussion arrives at the conclusion that some of the results are incompatible with the concept of respiratory protection of nitrogenase.
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  • 17
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    Archives of microbiology 136 (1983), S. 81-83 
    ISSN: 1432-072X
    Keywords: Ammonia production ; Anabaena ; Cyanobacteria ; Nitrate reductase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the filamentous heterocystous cyanobacterium Anabaena sp. strain ATCC 33047 dinitrogen fixation and nitrate reduction are mutually exclusive processes. Nitrate promotes nitrate reductase synthesis and represses nitrogenase formation. Inhibition of ammonium assimilation by l-methionine-d,l-sulfoximine (MSX) alleviates the repressive effect of nitrate on nitrogenase synthesis, thus indicating that the nitrate effect is indirect through metabolites generated from the ammonium derived from nitrate reduction. In MSX-treated cells both nitrate reduction and dinitrogen fixation take place simultaneously, although at different sites of the filament, without any apparent competition for the required reducing power. The MSX-treated Anabaena cells generate ammonium from both nitrate and dinitrogen, simultaneously.
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  • 18
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    Archives of microbiology 138 (1984), S. 212-216 
    ISSN: 1432-072X
    Keywords: Denitrification ; Nitrate respiration ; Nitrous oxide reduction ; Nitrogen fixation ; Azospirillum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrous oxide reduction can consistently be demonstrated with high activities in cells of Azospirillum brasilense Sp 7 which are grown anaerobically in the presence of low amounts of nitrite. Azospirillum can even grow anaerobically with nitrous oxide in the absence of any other respiratory electron acceptor. Nitrous oxide reduction by Azospirillum is inhibited by acetylene, amytal and weakly by carbon monoxide. Azospirillum converts nitrous oxide to molecular nitrogen without the formation of ammonia. The cells must, therefore, be supplied with ammonia from nitrogen fixation during anaerobic growth with nitrous oxide. When no other nitrogen compound besides nitrous oxide is available in the medium, the bacteria synthesize nitrogenase from protein reserves in about 2 h. Nitrogenase synthesis is blocked by chloramphenicol under these conditions. In contrast, the addition of nitrate or nitrite to the medium represses the synthesis of nitrogenase. Nitrous oxide reduction by Azospirillum and other microorganisms is possibly of ecological significance, because the reaction performed by the bacteria may remove nitrous oxide from soils.
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  • 19
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    Archives of microbiology 130 (1981), S. 96-100 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Nitrate respiration ; Denitrification ; Assimilatory nitrate reduction ; Dissimilatory nitrate reduction ; Acetylene reduction ; Azospirillum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Azospirillum spp. participate in all steps of the nitrogen cycle except nitrification. They can fix molecular nitrogen and perform assimilatory nitrate reduction and nitrate respiration. Culture conditions have been defined under which nitrate is used both as terminal respiratory electron acceptor and as nitrogen source for growth. Nitrate and, possibly to a very limited extent, nitrite, but not sulfate, iron or fumarate support anaerobic respiration. Under anaerobic conditions, nitrate can also supply energy for nitrogen fixation but without supporting growth. Nitrate-dependent nitrogenase activity lasts only for 3–4 h until the enzymes of assimilatory nitrate reduction are synthesized. Nitrite accumulates during this period and inhibits nitrogenase activity at concentrations of about 1 mM.
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  • 20
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    Archives of microbiology 140 (1984), S. 215-217 
    ISSN: 1432-072X
    Keywords: Glutathione reductase ; Cyanobacteria ; Nostoc muscorum ; O2 protection ; Glutathione ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Glutathione reductase activity was detected and characterized in heterocysts and vegetative cells of the cyanobacterium Nostoc muscorum. The activity of the enzyme varied between 50 and 150 nmol reduced glutathione· min-1·mg protein-1, and the apparent Km for NADPH was 0.125 and 0.200 mM for heterocysts and vegetative cells, respectively. The enzyme was found to be sensitive to Zn+2 ions, however, preincubation with oxidized glutathione rendered its resistance to Zn+2 inhibition. Nostoc muscorum filaments were found to contain 0.6–0.7mM glutathione, and it is suggested that glutathione reductase can regenerate reduced glutathione in both cell types. The combined activity of glutathione reductase and isocitrate dehydrogenase in heterocysts was as high as 18 nmol reduced glutathione·min-1·mg protein-1. A relatively high superoxide dismutase activity was found in the two cell types; 34.2 and 64.3 enzyme units·min-1·mg protein-1 in heterocysts and vegetative cells, respectively. We suggest that glutathione reductase plays a role in the protection mechanism which removes oxygen radicals in the N2-fixing cyanobacterium Nostoc muscorum.
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  • 21
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Ultrastructure ; Nitrogen fixation ; Water stress ; Taxonomy ; DNA ; Plasmids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two strains of desiccation-tolerant coccoid cyanobacteria, Chroococcus S24, a marine form, and Chroococcus N41, a cryptoendolith isolated from a hot-desert rock, have been characterized. The mol % DNA base compositions of the strains are 47.1 and 48.9% respectively. Plasmid DNA was not detected in either strain. The pigment contents and nutritional characteristics of the strains are identical. Both lack phycoerythrinoid pigments and, in culture, behave as slow-growing halotolerant marine forms with elevated requirements for Na+, Cl−, Mg2+ and Ca2+. Sucrose was the only carbon source of those tested that supported photoheterotrophic growth. Each strain synthesizes nitrogenase under anaerobic conditions but not in air. Morphologically the two strains are indistinguishable. They are considered to be independent isolates of the same cyanobacterial species. Chroococcus N41 was studied in detail with the electron microscope. When brought to equilibrium at matric water potentials of-168 MPa and lower (to-673 MPa=c0.12a w) the protoplast shrinks, but the cells maintain the same size and diameter as those at-2,156 kPa (MN medium; control); the sheath expands and remains attached to the cell wall outer membrane by fibrils. The cell wall, cell membrane, thylakoid membranes, cyanophycin granules and carboxysomes appeared intact in desiccated cells.
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  • 22
    ISSN: 1432-2048
    Keywords: Nitrogen fixation ; Nodule development ; Senescence (nodules) ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cowpea (Vigna unguiculata (L.) Walp cv. Vita 3) seedlings inoculated with Rhizobium strain CB756 were cultured with their root systems maintained in air or in Ar: O2 (80:20, v/v) during early nodule development (up to 24 d after sowing). Compared with those in air, seedlings in Ar:O2 showed progressive N deficiency with inhibited shoot growth, reduced ribulose-1,5-bisphosphate carboxylase and total protein levels and loss of chlorophyll in the leaves. Nodule initiation, differentiation of infected and uninfected nodule tissues and the ultrastructure of bacteriod-containing cells were similar in the air and Ar: O2 treatments up to 16 d after sowing. Thereafter the Ar: O2 treatment caused cessation of growth and development of nodules, reduced protein levels in bacteroids and nodule plant cells, and progressive degeneration of nodule ultrastructure leading to premature senescence of these organs. Provision of NO 3 - (0.1–0.2 mM) to Ar: O2-grown seedlings overcame the abovementioned consequences of N2 deficiency on nodule and plant growth, but merely delayed the degenerative effects of Ar: O2 treatment on nodule structure and senescence. Treatment of Ar: O2-grown seedlings with NO 3 - greatly increased the protein level of nodules but the increase was largely restricted to the plant cell fraction as opposed to the bacteroids. By contrast, NO 3 - treatment of air-grown seedlings increased protein of bacteroid and host nodule fractions to the same relative extents when compared with air-grown plants not supplemented with NO 3 - . These findings, taken together with studies of the distribution of N in nodules of symbiotically effective plants grown from 15N-labeled seed, indicate that direct incorporation of fixation products by bacteroids may be a critical feature in the establishment and continued growth of an effective symbiosis in the cowpea seedling.
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  • 23
    ISSN: 1432-2048
    Keywords: Glutamine synthetase ; Leghaemoglobin ; Nitrogenase ; Nitrogen fixation ; Phaseolus ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The activities of glutamine synthetase (GS), nitrogenase and leghaemoglobin were measured during nodule development in Phaseolus vulgaris infected with wild-type or two non-fixing (Fix-) mutants of Rhizobium phaseoli. The large increase in GS activity which was observed during nodulation with the wild-type rhizobial strain occurred concomitantly with the detection and increase in activity of nitrogenase and the amount of leghaemoglobin. Moreover, this increase in GS was found to be due entirely to the appearance of a novel form of the enzyme (GSn1) in the nodule. The activity of the form (GSn2) similar to the root enzyme (GSr) remained constant throughout the experiment. In nodules produced by infection with the two mutant strains of Rhizobium phaseoli (JL15 and JL19) only trace amounts of GSn1 and leghaemoglobin were detected.
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  • 24
    ISSN: 1573-5036
    Keywords: Clover ; Fertilizer ; Lotus ; Nitrogen fertilizer ; Nitrogen fixation ; Nitrogen source ; Nodulation ; Trifolium repens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Studies under growth cabinet conditions investigated the effect of source and concentration of nitrogen and timing of nitrogen application on the growth and nitrogen fixation byLotus pedunculatus cv. Maku andTrifolium repens cv. S184. KNO3, NaNO3 and NH4NO3 were added at transplanting at the following rates: 3.33, 7.78 and 13.33 mg N/plant. KNO3 was added at 3.33 and 7.78 mg N/plant at 0, 6, 12, 18, 24 or 30 days after transplanting. Lotus shoot weight increased with all increasing nitrogen sources but clover only responded to KNO3 and NaNO3. The root weight of both species increased with increasing KNO3 and NH4NO3. The percentage increase in lotus and clover shoot growth was greater than that of root growth when KNO3 was added within a week of transplanting. Increases in growth by both species resulted from added nitrogen except with lotus when NaNO3 was applied where increased nitrogen fixation also contributed to increased growth. Weight and number of effective nodules on both species were increased with 3.33 mg N per plant as KNO3 but nitrogen fixation was not affected. Addition of 13.33 mg N as NaNO3 reduced weight and number of effective nodules in both species and also nitrogen fixation by lotus. KNO3 increased growth and nodulation of both species when applied within one week after transplanting. Nodulated lotus plants responded to KNO3 by increasing growth but not nodulation. KNO3 appeared to affect infection and development of nodules on lotus and may affect the growth of existing nodules on clover.
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  • 25
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    Plant and soil 59 (1981), S. 201-206 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Coffee plantation ; Inga jinicuil ; Nitrogen fixation ; Nodule biomass ; Woody legume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nodule biomass and yearly C2H2 reduction rates are reported forInga jinicuil, a leguminous tree used for shade in Mexican coffee plantations. Annual fixation by this species approximates 35 kg ha−1; which, when compared to nitrogen additions from fertilizers, represents an important nitrogen input to the coffee ecosystem.
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  • 26
    ISSN: 1573-5036
    Keywords: Azotobacter vinelandii ; Gnotobiotic cultures ; Nitrogen fixation ; Oryza sativa Rhodopseudomonas capsulata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary An all glass tight growth chamber, entirely sterilizable, has been constructed to carry out axenic and gnotobiotic cultures of rice plants (Oryza sativa L.). When grown in liquid medium and in the absence of combined nitrogen but in the presence of the diazotrophsAzotobacter vinelandii andRhodopseudomonas capsulata, rice plants exhibited a complete biological cycle from germination up to ear stage, during a period of time similar to the one encountered in french paddy soil of Camargue. In one experiment, mannitol was given to rice culture medium together withAzotobacter vinelandii andRhodopseudomonas capsulata. In another experiment, mannitol was not given together with Rhodopseudomonas, and still positive nitrogen gain was obtained, although it was less than culture with mannitol. When15N labeled cells of Rhodopseudomonas were added in rice culture medium,15N was partly transferred to rice plant. Among the nitrogen substances excreted from the bacteria in the rhizosphere medium, large organic molecules were shown to be the most abundant in our experimental conditions. Moreover, the concentration of free ammonia or aminoacids present in the rice rhizosphere were always compatible with a bacterial nitrogenase activity.
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  • 27
    ISSN: 1573-5036
    Keywords: Bacteroids ; Cowpea ; Nitrogen fixation ; Peanut ; Rhizobium ; Siratro
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Acetylene reduction activity and nitrogen accumulation in the plant top per unit nodule mass were compared among peanut, cowpea and siratro plants nodulated by six different strains of Rhizobium. Peanut was found to have several fold higher values than cowpea and siratro for both parameters for all strains of Rhizobium which nodulated it effectively, but the bacteroid content of the peanut nodules was similar to those of cowpea and siratro.
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  • 28
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    Plant and soil 62 (1981), S. 331-336 
    ISSN: 1573-5036
    Keywords: Legumes ; Nitrogen fixation ; 15N
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    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The combination of using15N for determining the amount of nitrogen fixed by a legume crop in field experiments and the labelling of only one treatment at a time in each treatment combination is shown to be conceptually and experimentally valid for determining the effect of cultural practices on the amount of nitrogen fixed by a legume crop.
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  • 29
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    Plant and soil 63 (1981), S. 283-289 
    ISSN: 1573-5036
    Keywords: Afzelia quanzensis ; Brachystegia spiciformis ; Ectomycorrhiza ; Miombo woodland ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Ectomycorrhizae were found in root samples of the treesAfzelia quanzensis Welw. andBrachystegia spiciformis Benth. (Caesalpiniaceae), collected in the coastal miombo type woodland 50 km west of Dar-es-Salaam, Tanzania. Root nodules with a structure resembling that of nitrogen-fixing root nodules of other leguminous plants were observed in theA. quanzensis material. The climate of the locality is rather dry, and strongly seasonal. In the tropics, ectomycorrhizae have previously been found only in humid or rain forest climate zones.
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  • 30
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    Plant and soil 72 (1983), S. 321-334 
    ISSN: 1573-5036
    Keywords: Leguminous plants ; Nitrogen fixation ; Protein ; Seed ; Yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Mineral nitrogen did not increase grain yield and seed protein levels ofVicia faba L. andLupinus luteus L. in field trials and pot experiments. Fixed N2 was substituted by mineral nitrogen in these cases because of inhibition of N2 fixation by mineral nitrogen. Contrary to these results mineral nitrogen increased grain yields and seed protein amounts ofLupinus albus L.,Pisum sativum L., andGlycine max. (L.) Merr. The nitrogen effect was caused at an early stage by saving energy due to inhibition of N2 fixation (measurement of gas exchange by means of IRGA). In case of the N application after flowering grain, yields and seed protein levels increased because the mineral N was an additional nitrogen source for plants. At this stage the plants had ceased fixing atmospheric nitrogen. The high sink activity of growing fruits induced a lack of assimilates in nodules (determined by means of14CO2 application). The N effect was therefore the consequence of the lower assimilate pool for supplying root nodules in these plants in comparison withVicia faba L. andLupinus luteus L. Hence it follows that response to mineral nitrogen can be a criterion for discovering more effective Rhizobium-host combinations.
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  • 31
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    Plant and soil 73 (1983), S. 431-434 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; 15N2 diffusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary The kinetic of15N2 diffusion has been measured in a system similar to that for the estimation of N2 fixation in plant microorganism associations cultivated in soil. The15N2 enrichment of the soil atmosphere reached an homogenous value one hour after injection of15N2 and is identical to that obtained by calculation, indicating that no adsorption occurs in the soil particles.
    Notes: Résumé La cinétique de diffusion du15N2 est mesurée sur un système identique à ceux pouvant être utilisés pour la mesure de fixation de l'azote chez les associations plantes-microorganismes cultivées sur sol. L'enrichissement homogène de l'atmosphère du sol est obtenu une heure environ après l'injection de15N2 et correspond à l'enrichissement calculé, ce qui indique qu'aucune adsorption n'a lieu dans les particules du sol.
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  • 32
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    Plant and soil 75 (1983), S. 131-138 
    ISSN: 1573-5036
    Keywords: Hydrogen inhibition ; Nitrogen fixation ; Peas ; Relative efficiency ; Rhizobium leguminosarum ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Experiments were performed to investigate the causes of low relative efficiency, RE, in legume root nodules. Nitrogen fixing activity and RE varied with time of incubation of nodules and with different temperatures and oxygen concentrations. The effects of nitrogen concentration and carbon dioxide concentration were also examined. In each case the RE was inversely related to nitrogen fixing activity; measured by acetylene reduction. Increasing the nitrogen concentration had no effect on either nitrogen fixing activity or RE. Experiments with isolated bacteroids gave higher RE values than the whole nodules from which they were isolated. All the results were consistent with hydrogen inhibition of nitrogenase within the nodule being the cause of low RE.
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  • 33
    ISSN: 1573-5036
    Keywords: Actinorhizae ; Alnus nitrida ; Frankia ; Host-specificity ; Nitrogen fixation ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Two different strains, An 1 and An 2, were obtained from root nodules ofAlnus nitida Endl., collected from one locality in the area of its natural habitat near Bahrin, District Swat, Pakistan. The light and electron microscopy of the isolates revealed the occurrence of septate and branched hyphae bearing sporangia and vesicles. The strains differed in their growth requirements, nitrogen-fixing ability and production of extracellular pigments, thus indicating the existence of more than oneFrankia strain in the same locality. In the absence of combined nitrogen in the medium strain An 1 formed vesicles and fixed N2 (up to 200 nmol C2H4. mg protein−1.h−1), while strain An 2 under the experimental conditions formed only few vesicles and fixed N2 at a very low rate (ca 10 nmol C2H4. mg protein−1 .h−1). The nitrogenase activity of strain An 1 was strongly affected by the O2 concentration.Frankia An 1 and An 2 were infective and effective onA. nitida andA. glutinosa but not onDatisca cannabina andElaeagnus umbellata. Both An 1 and An 2 strains were more infective and effective onA. glutinosa thanFrankia strains AvcIl and CpI1.
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  • 34
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    Plant and soil 77 (1984), S. 253-261 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Beijerinckia ; Nitrogen fixation ; Paddy ; Phyllosphere ; Silicification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary An investigation was conducted to study the levels of nitrogen fixation on the leaf or sheath surfaces of four cultivars of paddy plants by using acetylene reduction technique. Varying levels of positive nitrogenase activity were observed on all the leaf surfaces. Sheath of IET 1991 cultivar showed a higher rate of fixation than the leaf surface. All the nitrogen-fixing organisms on the leaf or sheath surfaces belonged to the genus Beijerinckia. There was no correlation between the bacterial density and the level of fixation. Scanning electron microscopic data revealed that the upper surface of IET 1991 leaf was highly silicified and the microflora was either scanty or nil while the lower surface appeared quite different and harboured more micro-organisms. Similarly, the inner surface of sheath was devoid of silicification and showed the presence of micro-organisms.
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  • 35
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    Plant and soil 78 (1984), S. 29-43 
    ISSN: 1573-5036
    Keywords: Alnus glutinosa ; Endophyte ; Frankia ; Infectivity ; Nitrogen fixation ; Sporulation ; Variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Frankia sp. LDAgpl, an isolate from spore positive nodules ofAlnus glutinosa, only slowly infects its host plant. Reisolates obtained from occasional nodules caused by infection with LDAgpl, are capable of infecting the alder much more rapidly. A variability analysis of LDAgpl has been performed to obtain more insight into the question whether these reisolates constitute a different genotype within LDAgpl and if the plant is exerting an influence during plant passage. High dilutions of mildly sonicatedFrankia suspensions were plated to obtain genetically homogeneous colonies. Clones thus generated showed differences in growth pattern, sporulation and C2H2-reduction on media containing propionic acid as sole C-source (P-medium). Differences in sporulation on P-medium indicate that LDAgpl was a highly heterogeneous strain. Comparisons of sporulation on several different media gave evidence that the differences in sporulation between LDAgpl clones are the result of differences in efficiency of propionic acid utilization. The differences observed between the reisolates and LDAgpl clones indicate that the reisolates constitute a different genotype, which could be selected for by the plant during the infection process. Comparison with similar changes in phenotype occuring in a spore negative type strain fromA. glutinosa is discussed.
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  • 36
    ISSN: 1573-5036
    Keywords: Actinorhizas ; Alnus nitida ; Amino acid composition ; Datisca cannabina ; Hydrogen uptake ; Inoculation ; Nitrogen fixation ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The nodulation and the morphology and physiology of the nodules were studied onDatisca cannabina, a perennial herb from northern Pakistan andAlnus nitida, a nodulated tree in the same locality. Both species bear coralloid clusters of actinorhizal nodules. The main free amino acid inD. cannabina nodules was arginine while the predominant free amino acid inA. nitida nodules was citrulline. The infectivity of crushed nodules of both types of plants on their respective host was about 106 infective particles per gram of nodule fresh wt. In cross-inoculation experiments crushed nodule inoculum fromA. nitida failed to induce nodulation onD. cannabina seedlings but the crushed nodule inoculum fromD. cannabina caused low nodulation on seedlings ofA. nitida (103 infective particles. g. nodule fresh wt.). The activity of nitrogenase, hydrogenase and respiration (O2 uptake) were measured in detached nodules, nodule homogenates and the 20 μm residue and 20 μm filtrate preparations from the nodules of both species. Both species showed similar patterns of activities except that only the nodule homogenate and 20 μm residue preparations fromD. cannabina showed pronounced enhancement of the O2 uptake by succinate which was further stimulated by ADP. This has in part been explained by the presence of mitochondria in close connection with the endophyte.
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  • 37
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    Plant and soil 79 (1984), S. 227-234 
    ISSN: 1573-5036
    Keywords: Bacteria ; Inoculation ; Microorganisms ; Nitrogen fixation ; Phosphate solubilization ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Inoculation effect ofA. chroococcum, P. striata andA. awamorii on yield and nutrients uptake in rice was studied under green house conditions. The organisms appreciably increased the yield and uptake of nutrients with or without chemical fertilizers. Phosphorussolubilizing microorganisms and a mixture of the three showed better response than the rest of the treatments among single and mixed culture inoculations respectively. Chemical fertilizers further improved the yield and nutrients uptake. The yield response remained unaffected by replacing superphosphate with rock phosphate and microbial inoculations.
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  • 38
    ISSN: 1573-5036
    Keywords: Mesquite ; Nitrogen fixation ; 15N natural abundance ; Nodule morphology ; Rhizobium bacteria ; Woody legumes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The relative nitrogen fixation efficiencies (RE 1-[H2 evolved÷C2H2 reduced]·100) of four mesquite (Prosopis glandulosa var.torreyana) rhizobia (Strains WR 1001, WR 1002, L5, L9) and a cowpea rhizobia (Strain 176A32) on mesquite were evaluated in a glasshouse experiment. Plant yield, shoot N accumulation, and the natural15N abundance (δ15N) of nodule tissue were determined. Strain WR 1002 failed to nodulate mesquite and strain L5 produced ineffective nodules. Among the three effective strains (WR 1001, L9, 176A32) the cowpea strain (176A32) and strain L9 had significantly higher RE than strain WR 1001. Differences in RE, however, were not accompanied by significantly higher plant yield and shoot N accumulation. The difference in15N abundance between foliar tissue and nodules (nodules minus leaves) was 0.47 δ15N for the ineffective L5 nodules, while for the effective WR 1001, L9, and 176A32 nodules, respectively, this difference was 8.35, 7.81, and 8.35 δ15N. This indicates a similar relationship between N2-fixing effectiveness and natural15N enrichment of nodules that was previously observed in soybeans (Glycine max, L. Merr.). Strains WR 1001 and L9 produced elongate, indeterminate nodules typical for mesquite. The ineffective L5 nodules had few infected cells and an abundance of cortical amyloplasts. Mesquite nodules produced by the cowpea strain were spherical and were somewhat more similar in internal morphology to determinate nodules typical of cowpea than indeterminate nodules normally associated with mesquite.
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  • 39
    ISSN: 1573-5036
    Keywords: Actinorhizae ; Datisca cannabina ; Frankia ; Nitrogen fixation ; Root nodules ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The fine structures of the microsymbiont inside the root nodules ofDatisca cannabina have been studied by light, by transmission- and by scanning-electron microscopy. The endophyte is prokaryotic and actinomycetal in nature. The hyphae are septate and branched, diameter 0.3–0.5 μm. The tips of hyphae are swollen to form electron-dense, clubshaped to filamentous vesicles, ranging in diameter: 0.4–1.4 μm. The endophyte penetrates through walls of the cortial cells. The infected zone is kidney shaped and confined to one side of the acentric stele. The orientation of infection is reversed from other actinorhizae exceptCoriaria. The hyphae are near the host cell wall and vesicles are directed towards the central vacuole. Vesicles are aseptate and no collapsing of the vesicle cell wall (void area) has been observed. Vesicle clusters structures are globular with an opening at one side of the cluster. The host cell is multinucleate or contains a lobed nucleus. Groups of mitochondria are located in between the hyphae, suggesting a strong association between the host and the endophyte for energy supply and amino acid production. The consequences of the inability to separate the mitochondria from the vesicle clusters in nodule homogenates in physiological studies have been discussed. Isolated vesicles clusters showed dehydrogenase activity, indicated by the presence of formazan crystals, after incubation with NADH and NBT. Strongest reducing activity was found within the vesicles. The possible role of filamentous vesicles in nitrogen fixation has been discussed.
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  • 40
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Actinomycete-nodulated ; Legume ; Lime ; Mine spoil ; Nitrogen fixation ; Reclamation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary An acid mine spoil in Southern Indiana was amended with lime (CaCO3) (0.0, 12.5, 25 and 39t/ha) and planted withElaegnus umbellata Thunb.,Alnus glutinosa Gaertn.,Robinia pseudoacacia L.,Robinia fertilis Ashe, ‘Arnot’,Myrica pensylvania Lois,Caragana arborescens L. andShepherdia argentea Nutt. Survival and soil data were collected periodically and plants were harvested 15 months after planting. Nodule and top dry weights were determined and acetylene reduction assays performed on the nodules. Addition of lime caused significant increases in pH, and 39 t/ha of lime were required to maintain a pH above 5.5. Survival of plant material was greatest at the highest lime addition, although response of individual species varied.Elaeagnus umbellata, R. pseudoacacia, R. fertilis ‘Arnot’, andA. glutinosa appeared more tolerant of the harsh conditions. OnlyC. arborescens showed a linear increase in top dry weight due to lime addition.Alnus glutinosa andS. argentea achieved statistically the same growth regardless of pH, andR. fertilis ‘Arnot’ andE. umbellata did not increase in top dry weight above an addition of 25 t/ha.Robinia pseudoacacia achieved maximum top dry weight at 25 t/ha, whereasM. pensylvanica growth declined with increasing pH. Nodule dry weights increased with increasing pH; however,S. argentea showed greater nodule dry weights at lower lime levels. Acetylene reduction rates increased with lime addition.Elaegnus umbellata did not respond above 25 t/ha lime, whereasA. glutinosa did not show an increase until this point.
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  • 41
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    Plant and soil 74 (1983), S. 395-406 
    ISSN: 1573-5036
    Keywords: Fix− mutants ; Fix+ revertants ; Macroptilium lathyroides ; Nitrogen fixation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Eight ineffective mutant strains were isolated from N-methyl-N'-nitro-N-nitrosoguanidine mutagenized cultures of cowpea Rhizobium strain 32H1. Strains CR1, CR2, CR3, CR4, CR5 and CR6 induced more, but smaller, nodules than the wild type. With the exception of strain CR2, these mutant strains reduced less than 1% of the amount of acetylene reduced by the wild type, in both the free-living and symbiotic assays. Strain CR2 reduced acetylene in the free-living assay but not in the symbiotic assay. Strains CR7 and CR8 responded variably (5–20% of the wild type) in free-living and symbiotic acetylene reduction assays. Nodules also varied from small white to normal-sized pink nodules. The phenotypic characteristics of the mutant strains were consistant with all leguminous plants tested and were stable upon reisolation from nodules. Fully effective revertants were selected from 4 of the ineffective mutant strains by the use of the leguminous plant,Macroptilium lathyroides. Serology, patterns of resistance to anti-bacterial agents, phage-typing, and antibiotic resistance markers were used to confirm strain identification.
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  • 42
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    Plant and soil 72 (1983), S. 95-105 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Inga jinicuil ; Nitrogen fixation ; Nodulation ; Nodule biomass ; Nutrients ; Shading ; Tropics ; Woody legume
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A pot experiment was conducted to assess the effects of different fertilizers and soil shading on nodulation and acetylene reduction ofInga jinicuil seedlings. Initially, seedlings produced maximum nodule biomass when grown with high levels of phosphorus but reduced the most acetylene under intermediate phosphorus fertilization. These response differences, however, gradually diminished with age, being negligible when the seedlings were a year old. Nitrogen fertilization inhibited nodulation and acetylene reduction throughout the experiment. Potassium did not significantly affect nodulation, but low levels of potassium stimulated, and high levels inhibited acetylene reduction activity relative to unfertilized control plants. Neither magnesium nor molybdenum affected nodulation or acetylene reduction. Soil shading resulted in decreased nodule biomass and less nitrogen-fixing activity during summer months. However, the data suggest that shading may favour nitrogen fixation in colder periods by moderating soil temperatures. These results confirm findings from an earlier field study and show that nodulation and nitrogen-fixing activity by leguminous trees is influenced by the types and amounts of nutrients supplied. This suggests that the quantity of nitrogen fixed by leguminous shade trees in coffee plantations may be amenable to manipulation through simple management techniques.
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  • 43
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    Plant and soil 75 (1983), S. 309-342 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Amino-acid composition ; Chromosome number ; Cross-inoculation ; Growth various substrates ; Medium composition ; Nitrogen fixation ; Parasponia parviflora ; Root nodule ; Temperate Rhizobium species
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Cross-inoculation experiments withParasponia parviflora plants and a large number of Rhizobium strains isolated from legumes, mainly of temperate origin, showed that strains ofRhizobium trifolii andR. lupini could produce root nodulation in Parasponia. Root nodulation was also obtained with some tropical Rhizobium isolates such as those from Arachis and Albizzia, but not withRhizobium japonicum strains. In addition, strains ofR. leguminosarum, R. phaseoli andR. meliloti produced abnormal root nodules or pseudo-root nodules in Parasponia seedlings. All root nodules induced in Parasponia by foreign Rhizobium species lacked, however, nitrogenase activity. Testing the reverse combination Parasponia-Rhizobium on legumes yielded with regard to root nodulation negative results, except withVigna sinensis and twoMacroptilium species (M. atropurpureum andM. lathyroides). The root nodules produced in the latter legumes showed nitrogenase activity.Trema cannabina seedlings inoculated with Parasponia-Rhizobium invariably did not produce root nodules. The feasibility of Parasponia to accept foreign rhizobia bacteria or to produce pseudo-nodulation is probably genetically determined. In a karyological study it was shown thatParasponia parviflora had the chromosome number 2n=20. Growth experiments revealed thatP. parviflora can be cultivated in water culture, perlite and in soil (pot) culture. It can also be grown in agar tubes, although growth under these conditions is sometimes rather variable. Nitrogenase (C2H2) tests showed that water-culture root nodules were 5–10 times less active than those from perlite culture, soil or agar tubes. On the whole, nitrogenase activity of root nodules grown on the various substrates and receiving different treatments, was rather variable.Vigna sinensis plants inoculated with Parasponia-Rhizobium showed on basis of fresh nodule weight a 3–4 times higher nitrogenase activity as compared with the same strain on Parasponia. However, as shown by acetylene reduction tests of nodule slices, the main activity of mature Parasponia root nodules is situated in the apical part of the root nodule. On fresh weight basis the nitrogenase activity of this nodule section is of the same order as that of leguminous root nodules. Intact Parasponia plants showed higher nitrogenase activities than excised root nodules. The maximal nitrogen fixation rate ofParasponia parviflora plants was 20–25 μmol. C2H4·g−1 fresh weight nodule tissue.h−1, which value is comparable to that of legumes and higher than estimates obtained by Parasponia in the field. No decrease of the nitrogen-fixation rate was observed in Parasponia plants tested in a diurnal cycle during the 12-hours dark period. Amino-acid analyses of Parasponia root nodules showed large quantities of aspartic acid, asparagine, glutamic acid and glutamine suggesting a role of these amino acids in the nitrogen fixation or transport processes. In contrast to actinorhizal root nodules Parasponia root nodules do not contain citrulline. The latter amino acid was, however, found—although in rather low concentration—inVigna sinensis andMacroptilium atropurpureum nodules obtained withParasponia-Rhizobium.
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  • 44
    ISSN: 1573-5036
    Keywords: Genetic control ; Nitrogen fixation ; Pisum sativum L. ; Rhizobium Maternal effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary In pea cv. Afghanistan a recessive gene sym 6 prevents the full expression of nitrogenase activity in root nodules, induced byRhizobium leguminosarum strain F 13. In contrast, nitrogenase activity is fully expressed in pea cv. Iran. A comparison of the reciprocal hybrids of these two plants showed that the size of the plant was determined by the mother plant (maternal effect). Therefore the shoot weight and the total amount of nitrogen fixed are not suitable as parameters for a genetic analysis. The % nitrogen of the shoot and the specific activity of the nodules per gram of nodules are more reliable, but for practical purposes the specific activity of the nodules expressed per gram of shoot tissue can be used.
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  • 45
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    Plant and soil 77 (1984), S. 3-14 
    ISSN: 1573-5036
    Keywords: Asian-type soybean ; Fast-growingRhizobium japonicum ; Glycine max ; Nitrogen fixation ; Rhizobium japonicum ; Rhizobium physiology ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Physiological and symbiotic characteristics were identified in fast-growing (FG)Rhizobium japonicum. Carbon nutritional patterns linked these rhizobia to other FG rhizobia. They were able to use hexoses, pentoses, disaccharides, trioses, and organic acids for growth, but they were unable to use dulcitol or citrate. These rhizobia produced acid with all carbon sources except intermediates of the Krebs cycle. FGR. japonicum showed no vitamin requirements and were tolerant to 1% NaCl but not to 2%. They nodulated cowpea, pigeon pea, and mung bean but not peanut. Effective, nitrogen-fixing symbioses were observed only with cowpea and pigeon pea. In addition, FGR. japonicum formed effective symbioses with Asian-type soybeans. We concluded that although the physiological characteristics of FGR. japonicum were similar to other FG rhizobia, their symbiotic properties were similar to slow-growing rhizobia of the cowpea miscellany.
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  • 46
    ISSN: 1573-5036
    Keywords: Actinorhizal plants ; Bacterial isolation techniques ; Frankia ; Nitrogen fixation ; Root nodules ; Soil ; Sucrose density fractionation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The isolation and pure culture of the symbiotic nitrogen-fixing frankiae has always been difficult. In the past the isolation of these actinomycetes directly from soil samples has proven impossible and isolations from root nodules of many genera has been only poorly successful. We report here a modified sucrose fractionation procedure which increased the success of isolations from root nodules and which permitted the isolation ofFrankia directly from soil samples. Crushed nodule suspensions or soil suspensions were incubated briefly in 0.7% phenol (carbolic acid) just before application to a sucrose density gradient. This phenol incubation decreased the number of contaminating eubacteria and fungi but more importantly increased the number ofFrankia developing on the isolation plates. If the phenol incubation was used solely without sucrose fractionation noFrankia were isolated, suggesting the death of the organisms due to phenol toxicity. The use of selective nitrogen-deficient media proved important for the isolation of frankiae from soils.
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  • 47
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    Plant and soil 78 (1984), S. 105-128 
    ISSN: 1573-5036
    Keywords: Cytology of root nodules ; Dryas drummondii ; Frankia ; Geographic distribution ; Nitrogen fixation ; Rubus ellipticus ; Ultrastructure of endophyte
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Root nodules ofDryas drummondii are of the coralloid type (Alnus type). The endophyte is present in the middle cortical cells of the root-nodule tissue. Transmission electron micrographs revealed an actinorhizal endophyte with septate hyphae and non-septate spherical or ovoid vesicles. Vesicles always possess at the base a septum; septa formation in the endophyte is always associated with the presence of mesosomes. Branching of the endophyte is not necessarily correlated with septum formation. Hyphal structures are more prominent in the apical part of the root nodule and vesicles are more numerous in a broad zone below this. In the middle and towards the base of the root nodule the endophytic structures appear in a stage of disintegration. Vesicles appear in a broad region near the periphery of the host cell and regularly show no strict orientation towards the host-cell wall. In the center of the host cells only hyphae occur. In the intercellular spaces between the host cells theFrankia endophyte produces spore-like structures although the outline of the sporangia is often faint. The coralloid root ofRubus ellipticus shows characteristically a basal rootlet initiated below the dichotomous branching of the nodular lobes, but extending beyond the root nodule. The endophyte is only present in the outer cortex of the root nodule in a 1–2 cell wide layer. This endophytic layer is bounded, internally as well as externally, with a 4–5 cell wide layer of tannin-filled host cells. The implications of this situation are discussed. Tannin-filled cells occur regularly inRubus species and their arrangement has been used for taxonomic purposes within the genus. TheRubus endophyte is aFrankia species with septate hyphae and distinctly septate spherical vesicles. The ultrastructure of the vesicles of theRubus endophyte is very similar to that of theAlnus endophyte.
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  • 48
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Glycine max L. ; Nitrogen fixation ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A system for employing open-ended root chambers to measurein situ acetylene reduction rates under field conditions is described. Gas mixtures containing about 2 mbar acetylene were continuously flowed through the chambers providing a continuous record of acetylene reduction. These chambers have been used to measure acetylene reduction rates of soybeans during three growing seasons. The system has proved to be reliable with a high degree of precision. The large amount of plant-to-plant variability observed in N2 fixation research has been confirmed by the data collected with this system. However, such variability in physiological studies can be reduced by using a non-destructive system to compare the response of an individual plant with its rates before treatment.
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  • 49
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    Plant and soil 73 (1983), S. 117-128 
    ISSN: 1573-5036
    Keywords: Light quality ; Nitrogen fixation ; Trifolium repens ; White clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The possibilities of using light quality treatments to gain an understanding of the mechanisms controlling the allocation of photosynthate for symbiotic nitrogen fixation were studied. White clover (Trifolium repens) plants were grown at the same photon irradiance in red, blue and green light treatments. Growth, nodulation and the carbon/nitrogen economies of the plants were measured. Both photosynthetic rates per unit leaf area and shoot-root ratios were affected by the treatments. However, the carbon/nitrogen economies of the plants and the fraction of the total plant weight allocated to nodule growth were unaffected.
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  • 50
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    Plant and soil 73 (1983), S. 151-153 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Beijerinckia ; C3 and C4 plants ; Nitrogen fixation ; Phyllosphere ; Wettability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nitrogen fixation on leaf surfaces of sugarcane, sorghum, ragi, bamboo and mulberry plants was determined by using acetylene reduction assay. The data revealed varying levels of positive nitrogenase activity on the surfaces. Beijerinckia was observed to be the predominant diazotroph on the leaves. No correlation between fixation rates and C3 or C4 plant species was discerned. The possiblity of improving phyllosphere nitrogen fixation has been discussed in light of the above observations.
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  • 51
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    Plant and soil 73 (1983), S. 299-305 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; Rhizobium trifolii populations
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effectiveness ofRhizobium trifolii isolates from five locations in southern Britain representing contrasting soil types has been examined with five white clover varieties. The average effectiveness of Rhizobium isolates varied considerably as did the average productivity of plant varieties. The largest differences were, however, associated with Rhizobium population × plant variety interactions. These were often large enough to reverse relative yield differences between white clover varieties. The implications of these results for improving clover productivity in nitrogen fixation are discussed.
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  • 52
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    Plant and soil 76 (1984), S. 77-91 
    ISSN: 1573-5036
    Keywords: Microorganisms ; Mycorrhizas ; Nitrogen fixation ; Phosphate uptake ; Rhizosphere
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary No root systems in nature are without a microbial population. These may be freeliving or symbiotic. The incidence and nutrition of the freeliving microorganisms is discussed. Shortage of substrate makes it unlikely that the N-fixers in the population can fix useful amounts of N. There is a possibility that P supply is improved, but an analysis of possible processes shows them to be rather unlikely, and evidence for them to be poor. Manganese and iron uptake can be altered by microbial activity. Growth of plants can be affected by non-nutritional bacterial effects. The ecology of Rhizobium in the soil is briefly discussed, and the varying needs of different identified strains is stressed. Mycorrhizal infection of plants leads to large growth increases in appropriate conditions. This is almost always linked to increased P uptake, but zinc and copper nutrition can also be improved. The processes involved are briefly discussed. Rapid and extensive infection is important; it is very sensitive to temperature. New modelling methods are now becoming available to measure the behaviour of the fungal infections. The microorganisms require C compounds from the plant, and new measurements of this cost are discussed. The possibility of practical use of mycorrhizal fungi seem to be improving.
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  • 53
    ISSN: 1573-5036
    Keywords: Hill soils ; Lime ; Mycorrhiza ; Nitrogen fixation ; Nodulation ; Phosphorus ; Rhizobium ; Symbioses ; White clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The responses of white clover (cv NZ Grasslands Huia grown in four UK hill soil types) to additions of lime and P, to inoculation with Rhizobium and mycorrhizal fungi, and to differences in soil water status were assessed in pot and field experiments. With a deep peat soil in pots, shoot production, nodulation and N fixation by clover were increased by 160, 130 and 85% respectively following inoculation with mycorrhiza, but in the field, despite a doubling of root infection, there was no response in growth. On a brown earth soil in the field inoculation with one endophyte (Glomus mosseae L1) out of four tested depressed production of white clover shoots by 42% but enhanced that of leeks (Allium porrum) by 50%; the others were without effect. With dry peaty podzol and brown earth soils in pots, clover shoot production was highest with added P when a water holding capacity of 80% was maintained, but roots from the latter had only 2.6 compared to 68 nodules per plant from the former. Further work is required to explain poor nodulation in the brown earth soils.
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  • 54
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Field method ; Medicago sativa L. ; Mucilaginous substances ; Nitrogen fixation ; Trigonella foenum-graecum L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary A non-destructive method using a special device for measuringin situ acetylene reducting activity by nitrogen fixing plants is described. Plant roots are isolated from external atmosphere with a mucilaginous material laid on the soil around the plants. Acetylene or ethylene is directly injected into or taken from the soil around the nodules. Using this device the values of acetylene reducing activity ofTrigonella foenum graecum L. andMedicago sativa L. are much higher than those obtained with the same plants placed under polyethylene bags. This method is not expensive and allows the detection of low enzyme activities. It doesn't perturb plant physiology and can be used for young plants as well as for plants with ramified stems at their base.
    Notes: Résumé Les auteurs décrivent un dispositif qui permet de mesurer l'activité nitrogénasein situ par réduction de l'acétylène chez les plantes fixatrices d'azote, sans perturber leur physiologie et sans les détruire. Le système racinaire est isolé de l'atmosphère extérieure par une substance mucilagineuse déposée sur le sol autour des plantes testées. Les gaz acétylène ou éthylène sont injectés ou prélevés directement dans le sol au niveau des nodosités. Avec ce nouveau dispositif les valeurs de l'activité réductrice d'acétylène obtenues surTrigonella foenum-graecum L. etMedicago sativa L. cultivés en plein champ, sont très supérieures à celles mesurées sur ces mêmes végétaux enfermés dans une enceinte en polyéthylène. Cette méthode peu onéreuse, permet de détecter des activités réductrices faibles; elle est utilisable sur des jeunes plantes et également sur des individus ramifiés à la base.
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  • 55
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    Plant and soil 79 (1984), S. 37-49 
    ISSN: 1573-5036
    Keywords: Azotobacter ; Nitrogen fixation ; N2-fixing efficiency ; Respiration ; Soil ; Water potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Respiration and N2-fixation (acetylene reduction) ofAzotobacter vinelandii have been studied at a variety of soil water potentials. Both processes were strictly linked and strongly reduced at water potentials between −0.6 and −1.3 MPa. Complete inhibition occurred below −2.1MPa. Osmotic potentials in soil compared to matric potentials of the same value were less inhibitory to respiration and acetylene reduction by Azotobacter. The N2-fixing efficiency (mg N/g glucose) was not influenced by water potentials ranging from −0.1 to −2.1 MPa.
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  • 56
    ISSN: 1573-5036
    Keywords: C2H2 reduction ; Intact-plant assay ; Millet ; Nitrogen fixation ; Nitrogenase activity ; Sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A non destructive intact-plant assay for estimating nitrogenase activity (C2H2 reduction) of pot-grown sorghum and millet plants is described. Plants with intact shoots sustained more activity than plants whose tops were removed prior to the assay. With this technique individual plants can be assayed several times during their life cycle. The C2H2 reduction was linear up to 16h incubation in this assay procedure. More rapid diffusion of C2H2 was achieved by injection through a Suba seal in the bottom of the pot. The equlibration of injected C2H2 in the gas phase of the pots filled with sand and sand:FYM media was completed within 1 h. Significantly higher nitrogenase activity and better growth of sorghum and millet plants occurred when plants were grown in a mixture of sand and farmyard manure (FYM) than when plants were grown in vermiculite, soil, or sand + soil medium. Nitrogenase activity and plant growth were greater in a mixture of sand with 2 and 3% FYM than with 0.5 and 1% FYM. Activity was higher when the plants were incubated at 33°C and 40°C than at 27°C. Activity also increased with increasing soil moisture. There were significant differences amongst 15 sorghum cultivars screened for associated nitrogenase activity. This new technique has good prospects for screening cultivars of millet, sorghum and other grain crops for their nitrogen-fixing ability.
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  • 57
    ISSN: 1573-5036
    Keywords: Azospirillum brasilense ; Azotobacter chroococcum ; Effect on yield ; Inoculation ; Nitrogen fixation ; Setaria italica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Under controlled conditions in pots filled with sand, vermiculite and field soil, inoculation withA. brasilense-Cd ATCC 29729 or withAzotobacter chroococcum caused increases above controls in the weight and N content of panicles ofSetaria italica. In no case, however, did N increases in test plants exceed the initial total N content. High acetylene reduction activities (1,000–2,000 nmole/h/pot) could be found only in plants inoculated withAzospirillum. Inoculation withAzospirillum (strain-Cd) in the field caused a significant increase above noninoculated controls of 18.5% in shoot dry weight, ofSetaria italica. Azotobacter caused a non significant increase of 8%. No significant differences were found between yields ofSetaria italica grown in soil inoculated withA. chroococcum, and those of plants grown in the presence ofA. vinelandii. A. brasilense-Cd was more effective in the field thanA. brasilense Sp-7 ATCC 29145. The results suggest that Azospirillum may increase yields ofS. italica more efficiently than Azotobacter under local field conditions.
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  • 58
    ISSN: 1573-5036
    Keywords: Clover ; Nitrogen fixation ; Rhizobium ; Root nodule ; Selection and plant breeding ; Symbiosis ; Trifolium pratense ; T. subterraneum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary This paper reviews (i) basic studies on the genetics of symbiosis in red clover (a self-sterile species) and subterranean clover (cleistogamous) and (ii) work on selection and plant breeding to increase nitrogen fixation in these hosts. Symbiotic effectiveness in red clover is influenced by many major and minor genes. The highly effective phenotype is inherited in a complex manner associated with early nodulation and the formation of large amounts of persistent bacteroid-containing tissue. Lines bred to fix more nitrogen with one strain ofRhizobium trifolii do so with most but not all other strains examined. They also show slightly increased vigour when grown on nitrate. The highly effective response is correlated with abundant nodulation and an early flowering habit, the evidence from breeding studies indicating that this correlation is not absolute. Normally effective and highly effective nodules have the same specific nitrogenase activities. The expression of the highly effective response is relatively little affected by environmental factors (temperature, light intensity, day length, supplementary carbon-di-oxide). Inbreeding substantially degrades the symbiotic response. Heterosis is shown in crosses between cultivars of subterranean clover but otherwise selection to increase effectiveness in this host was unsuccessful. The relevance of these results (and their physiological aspects) for the improvement of grain legumes is discussed.
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  • 59
    ISSN: 1573-5036
    Keywords: Assimilates distribution ; 14CO2 and15N2 labelling ; Glycine max L. Merrill ; Nitrogen fixation ; Respiration costs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Summary Distribution and use of photoassimilated carbon by nodulated soybeans (Glycine max L. Merr.) grown in natural conditions have been studied during two consecutive years by successive and simultaneous exposures of the plants to14CO2 and15N2 during one day. This method, together with detailed analysis of CO2 efflux by root systems made it possible to follow the fate of carbon in relation to dinitrogen fixation. The results showed that the stage of development at which the plants were labelled exerted a high influence on the initial distribution of carbon. The growing organs always attracted more of the recently assimilated carbon. So, the reproductive parts accumulated increasing proportions and by day 100 about 70% of the current net production was recovered in pods and seeds. Nodules stored from 3 to 5% of the daily production, from day 50 to day 100, when their fixation activity already dropped by day 85. During the period of optimal fixation the amount of carbon lost as CO2 due to the only process of N2 reduction ranged between 2.5 and 7 mg per mg of N2 fixed. This was equivalent to the amount of carbon directed towards roots and nodules structures during the same time. Secondary transfers by remobilization occurred from leaves and pods to the grains but mainly from the carbon assimilated during the podfilling stage (after day 80). However, these amounts were small indicating that grain development is mainly supported by current assimilates.
    Notes: Resumé La distribution et l'utilisation du carbone photoassimilé par des sojas (Glycine max L. Merr.) nodulés, cultivés en conditions naturelles ont été suivies durant deux années consécutives par expositions répétées et simultanées de plantes à du14CO2 et15N2 pendant une journée. Cette méthode couplée à une analyse détaillée des flux respiratoires au niveau des systèmes racinaires a permis de suivre le devenir du carbone en relation avec l'activité fixatrice d'azote. Les résultats montrent que le stade de développement des plantes au moment du marquage exerce une forte influence sur la distribution initiale du carbone. Ce sont les organes en forte croissance qui attirent le plus d'assimilats récents. Ainsi les organes reproducteurs, accumulent des quantités croissantes à partir de leur formation si bien qu'aux environs du jour 100, ils attirent près de 70% de la production nette. Les nodules eux stockent da 3 à 5% de cette production journalière entre les jours 50 et 100 alors que leur activité fixatrice chute déjà fortement dès le jour 85. Pendant la phase d'activité fixatrice optimale, les quantités de carbone perdues sous forme de CO2, pour le seul processus de réduction du N2, varient entre 2,5 et 7 mg par mg d'azote fixé. Ceci équivaut à la quantité de carbone destinée aux structures “racines et nodules”. Les transferts secondaires d'organes à organes se font à partir des organes végétatifs et puis des gousses vers les graines, mais essentiellement à partir de carbone assimilé durant la phase de remplissage des gousses (après le jour 80). Cependant, les quantités remobilisées sont faibles, ce qui indique que la croissance des graines dépend surtout d'une alimentation directe à partir de la photosynthèse.
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  • 60
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    Plant and soil 82 (1984), S. 273-284 
    ISSN: 1573-5036
    Keywords: Legume ; Nitrogen fixation ; Rhizobium ; C and N economy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Studies of the C and N economy of a range of temperate and tropical legume/Rhizobium symbioses indicate considerable variation (up to three-fold) in the cost of N2 fixation. Comparisons between and within symbioses indicate that the proportion of net photosynthate utilized in nodule functioning varies almost ten-fold from as low as 3% to as high as 25%. Factors possibly responsible for variation in efficiency of C use in nodules and in the proportioning of translocated photosynthetic products to nodules are discussed.
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  • 61
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    Plant and soil 81 (1984), S. 133-141 
    ISSN: 1573-5036
    Keywords: Benomyl ; Pesticides ; Nitrogen fixation ; Soybeans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Benomyl applied to the seeds of soybean (Glycine max L. Merrill) inoculated with a benomyl resistant strain ofRhizobium japonicum increased the relative abundance of nodules formed by the inoculum strain and the numbers of the added rhizobium on the roots, the total N content, the percentage N, the yield at one plant density and, in one of four soils, the pod weight of soybeans grown in the greenhouse. Oxamyl applied to the seeds, foliage or both of soybeans inoculated with an oxamyl resistant strain ofR. japonicum increased the yield, N content, percentage N, and weight of nodules, pods and grain and enhanced the relative frequency of nodules formed by the inoculum strain. It is suggested that pesticides or other antimicrobial agents and rhizobia resistant to these inhibitors may provide a new means for increasing nitrogen fixation by soybeans.
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  • 62
    ISSN: 1573-5036
    Keywords: Azospirillum lipoferum ; Grain yield ; N-content ; Nitrogen fixation ; Triticum aestivum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Spring wheat (Triticum aestivum var. Arkas) was associated withAzospirillum lipoferum under greenhouse and field conditions of a temperate region. Controls were treated with autoclaved bacteria. The soils used were: sand, sandy loam, and a peat-clay mixture. In experiments run over a period of three years, there were increases in grain yield, N-yield of the grains, and 1000 grain weight. Depending from environmental conditions, increase changed from year to year, and within one given year. There was, however, no experiment without positive response to the inoculation. Highest grain yield increase (70%) was found on sand supplemented with P and K only, but up to 32% were also obtained on peat-clay soil containing 0.28% total N. Under greenhouse conditions, one third of technical N-fertilizer could be saved by bacterial activities. With high probability the effects observed have been at least partly due to bacterial N2-fixation, because the N-yield of the grains was increased (up to 33%), and the most pronounced response was found on sand without any N-fertilizer added.
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  • 63
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    Plant and soil 82 (1984), S. 329-335 
    ISSN: 1573-5036
    Keywords: Legume breeding ; Medicago sativa ; Nitrogen fixation ; Rhizobium ; Trifolium repens ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary This paper examines evidence which quantifies the relative importance of legume and Rhizobium genotypes as determinants of phenotypic variation in symbiotic nitrogen fixation. It demonstrates potentially large and unpredictable effects of the Rhizobium genotype. The likely importance of such effects on crop yield is considered. The information is then used to assess ways in which legume breeding programmes may be altered to encompass the effects of genetic variation in Rhizobium.
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  • 64
    ISSN: 1573-5036
    Keywords: Co-evolution ; Domestication ; Gene centre ; Genetic variability ; Nitrogen fixation ; Pisum sativum L ; Rhizobium leguminosarum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Soil samples from several European countries; Sweden, the Netherlands, Spain, Italy and Greece, contained rhizobial populations capable of forming an effective symbiosis with the cultivated pea cv. Rondo from the Netherlands. The range of variation among the European Rhizobium strains, as expressed on pea cv. Rondo, was not so large and almost the same variation could be found within the rhizobial population within each country. Superior Rhizobium strains for the Dutch pea were not restricted to soils from the Netherlands but were also found in those from Sweden and Italy. Soils from Turkey and Israel also contained Rhizobium strains capable of nodulating pea cv. Rondo. However, the genetic variation among these Middle East Rhizobium strains was much larger than that of the European strains. When tested on pea cv. Rondo the majority of the Middle East strains belonged to the medium or low effective classes and only a few strains were comparable with European Rhizobium strains. Dutch Rhizobium strains induced effective nodules on both the Dutch pea cv. Rondo and the Swedish cv. L 110. However, in association with a Turkish Rhizobium strain effective nodules were formed on pea cv. Rondo and ineffective nodules on cv. L 110. We suggest that the genetic uniformity of EuropeanR. leguminosarum strains is the result of selection and domestication of Rhizobium strains originally derived from the gene centres of the pea plant.
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  • 65
    ISSN: 1573-5036
    Keywords: Alfalfa ; Correlation coefficient ; Factor analysis ; Leaf area ratio ; Nitrogen fixation ; Net assimilation rate ; Path-coefficient analysis ; Relative growth rate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Twelve alfalfa cultivars inoculated with an indigenous strain (RM9) ofRhizobium meliloti, were compared for their seedling morphological characters, and growth characters, including net assimilation rate (NAR), relative growth rate (RGR), leaf area ratio (LAR) and relative nitrogen assimilation rate (RN). Highly significant differences were obtained between cultivars for most characters. Simple correlation showed that NAR influenced RGR (r=0.91) more than leaf area ratio (LAR) (r=−0.44), and that most characters measured were highly correlated with seedling dry weight. Factor analysis showed that NAR, RGR and RN contributed 25% of the total variation in the dependence structure. The grouping indicated that the higher the NAR and RN the greater was the RGR. Path-coefficient analysis showed that NAR had more important direct and indirect effects than RN in dry matter accumulation. The relationship implied that selection for plants with high NAR, or high efficiency in converting light energy to dry matter production could contribute greater N2 fixation in alfalfa.
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  • 66
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    Plant and soil 59 (1981), S. 473-477 
    ISSN: 1573-5036
    Keywords: Hexachloroxyclohexane ; Nitrogenase activity ; Nitrogen fixation ; Redox potential ; Rhizosphere ; Rice plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The influence of application of hexachlorocyclohexane (HCH=gamma BHC), to a submerged tropical field soil at rates equivalent to recommended field rates (1–2.5kg a.i./ha) and twice this level, upon the rhizosphere soil nitrogenase, nitrogen fixers, and soil redox potential (Eh) was investigated. The rhizosphere soil from HCH-treated field exhibited significantly higher nitrogenase activity than that from untreated fields. HCH retarded the drop in redox potential of the field soil upto 80 days after transplantation under submerged conditions. Populations of nitrogen-fixingAzospirillum sp. and Azotobacter, to a greater extent, and anaerobic organisms, to a lesser extent, were stimulated in HCH-treated soils. Results indicate the stimulation of heterotrophic nitrogen-fixing bacteria by HCH in submerged paddy soils.
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  • 67
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    Plant and soil 60 (1981), S. 139-142 
    ISSN: 1573-5036
    Keywords: Legumes ; Nitrogen fixation ; Nodulation ; Winged beans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Only legumes of the cowpea cross-inoculation group, including the winged bean (Psophocarpus tetragonolobus) were found to form nodules in a temperate zone soil with no previous history of legume cropping. Isolates from root nodules from uninoculated winged beans grown in the field only nodulated legumes in the cowpea cross-inoculation group.Rhizobium japonicum formed ineffective nodules with the winged bean.
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  • 68
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    Plant and soil 60 (1981), S. 309-315 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Aeschynomene americana ; Flooding ; Leaf water potential ; Nitrogen fixation ; Soil moisture deficits
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Studies on the tolerance ofAeschynomene americana L. to periods of flooding or soil moisture deficit were conducted in an attempt to elucidate nitrogen fixation as affected by soil moisture. Nitrogenase activity was not reduced significantly in pot-grown Aeschynomene plants subjected to flooding in greenhouse conditions. After 20 days of withholding water from the soil, nitrogenase activities of the drought-stressed plants were much lower than those of either the well-watered or flooded plants. Leaf water potentials were similar in flooded and control plants; however, the droughted plants had leaf water potentials that were 4 bars lower than those of the control plants. Aeschynomene plants were tolerant to long-term periods of flooding, but exhibited a reduction in nitrogenase activity and leaf water status when subjected to soil moisture deficits.
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  • 69
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    Plant and soil 61 (1981), S. 93-111 
    ISSN: 1573-5036
    Keywords: Ineffective nodules ; Medicago laciniata ; Nitrogen fixation ; Nodulation ; Root temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Medicago laciniata, an annual leguminous plant of Saharo-Sindian origin, is particularly refractory to root nodulation by most strains ofRhizobium meliloti. Using a series of such bacterial strains belonging to the 8 groups of Brockwell and Hely, and a variety of environmental conditions, it was noted that several normally non-nodulating strains (at 20°C) produced ineffective nodules at root temperatures of 24°C to 28°C. Nodulation at 20°C failed to occur in the presence of a wide variety of test compounds and physical conditions. No phytoalexins or anti-Rhizobium growth inhibitors were isolated from inoculated root tissue at any temperature. Temperature shift experiments indicated no infection of the root hairs at 20°C, and infection threads produced at the permissive root temperature failed to elongate after transfer to 20°C. However, if meristematic activity had been initiated in the inner root-cortical cells as a result of infection thread penetration at 28°C, no blockage of nodule maturation occurred upon subsequent transfer to 20°C root temperature. Nodules produced at 28°C were completely devoid of nitrogenase activity, although the apical (but not the distal) regions contained normal-appearing bacteriods, surrounded by enclosing membranes, and possessed a fully functional leghaemoglobin. A shortage of metabolic energy did not appear to be involved in the ineffective response. A hypothesis to explain the nodulation phenomenon observed was based on the observation in the roots of 2 factors present at 20°C but not at 28°C.
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  • 70
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    Plant and soil 61 (1981), S. 135-143 
    ISSN: 1573-5036
    Keywords: Hydrogen ; Nitrogen fixation ; Nodulated roots ; Pea ; Respiration ; Soya bean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Oxygen uptake, carbon dioxide evolution and nitrogenase activity, measured either as hydrogen evolution (under argon 80%, oxygen 20%) or as the reduction of acetylene to ethylene, were assayed over the same time period by a direct mass-spectrometric method. When carbon dioxide evolution was used to estimate carbohydrate consumption, the results agreed with other work on whole plants. The RQ values obtained in these experiments were always less than 1.0 and thus the carbohydrate consumption calculated from oxygen uptake suggests that previous estimates, using carbon dioxide evolution as a measure of the cost of nitrogen fixation may be underestimates. Lag periods observed in the reduction of acetylene to ethylene suggest that there is a resistance to diffusion of gases in the root nodules.
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  • 71
    ISSN: 1573-5036
    Keywords: Biofuel ; Leguminous trees ; Nitrogen fixation ; Semi-arid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sand culture pot experiments were carried out with Proposis seedlings in the greenhouse on a nitrogen free nutrient solution with increasing levels of sodium chloride. All species tolerated a 6,000 mg/l salinity with no reduction in growth.P. velutina was the only species that poorly tolerated the 12,000 mg/l salinity level.P. articulata, P. pallida, andP. tamarugo tolerated 18,000 mg/l NaCl with little reduction in growth and grew slightly in a salinity (36,000 mg/l NaCl) greater than seawater. This is the first legume known to grown in salinities equivalent to seawater.
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  • 72
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    Plant and soil 61 (1981), S. 53-63 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Inoculation ; Klebsiella oxytoca ; Nitrogen fixation ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Of 45 fermentative gram negative bacterial isolates examined from wheat roots, three were capable of fixing atmospheric nitrogen as determined by the acetylene reduction technique and by protein contents of cells. A gram negative non-motile facultatively anaerobic bacterial strain capable of N2 fixation was identified asKlebsiella oxytoca ZMK-2. Optimal growth and N2 fixation occurred at pH 6.5. The optimum temperatures for growth under anaerobic conditions ranged between 30°–37°C. Acetylene reduction by intact cells was strikingly inhibited by 0.1 atm. or greater partial pressure of O2. Furthermore, the accumulation of H2 in the gas phase over cultures ofKlebsiella oxytoca ZMK-2 at partial pressures greater than 0.02 atm. resulted in a striking inhibition in the rate of C2H2 reduction. The addition of suspensions of eitherKlebsiella oxytoca ZMK-2 orAzotobacter vinelandii or a mixed culture of these two organisms to axenic cultures of wheat plants produced no significant increase in plant growth as measured by plant dry weight or nitrogen content of plants.
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  • 73
    ISSN: 1573-5036
    Keywords: Azospirillum brasilense ; Cereals ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The potential of the nitrogen fixing bacteriumAzospirillum brasilense to enhance development and increase growth of several gramineae was investigated. In both sterilized and non-sterilized systems heading and flowering occurred earlier in the inoculated plants as compared to the noninoculated ones. Total shoot and root weights, total-N content, plant height and leaf length were significantly increased by inoculation.
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  • 74
    ISSN: 1573-5036
    Keywords: Host selection ; Nitrogen fixation ; Nodule structure ; Rhizobium trifolii ; TemperatureTrifolium pratense
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary 1. Nodule structure was examined in the red clover cultivar S123 and in lines (designated H2) separately bred for high yield withRhizobium trifolii strains RCR 0403 and RCR 5. Plants were grown at 16/11, 22/17 and 27/22°C in a 16 h day at 25,000 lux. 2. The larger yields of bred lines compared with S123 were directly correlated with larger aggregate nodule size, larger infected zones and with more bacteroid tissue. Over all treatments the correlation of plant dry weight with aggregate areas of bacteroid tissue, assessed in median longitudinal section, accounted for 79% of the variance. 3. Yields were unrelated to the proportion of uninfected cells and vacuoles within the bacteroid zone. 4. Effects of host type and strain on nodule structure and yield were greatest at the moderate temperature and least and most irregular at high temperature. 5. Bacteriod tissue degenerated more rapidly in S123 than in the bred lines, more rapidly with RCR 0403 than RCR 5 and more rapidly at the high temperature than at moderate or low temperatures. 6. Aggregate nodule size irrespective of treatment was correlated with the aggregate sizes of nodule meristem and differentiating tissues. Nodule comtex comprised a larger proportion of the nodule section at 17 days than subsequently. Other than as specified above, the size relationships of the different tissues of the nodule were unaffected by host selection, temperature, bacterial strain or plant age. 7. At 22/17°C over a 17–25 day period the increment in dry matter per mm2 active nodule tissue was similar for H2 and S123 plants and was less for RCR 0403 (3.33 mg) than for RCR 5 (6.87 mg). At 27/22°C the effeciency of bacteroid tissue was much less,viz. 0.67 and 1.23 mg per mm2 respectively. 8. Nodule tissue areas and volumes were closely related so that unit volumes of active bacteriod tissue in general promoted similar dry matter increment in S123 and bred lines,viz. about 50.5 mg mm−3 at 22/17° for RCR 0403 and 80.8 mg mm−3 for RCR 5. The latter was associated with a fixation rate of 0.157 mg N mm−3 active tissue day−1.
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  • 75
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    Plant and soil 61 (1981), S. 189-202 
    ISSN: 1573-5036
    Keywords: Actinomycete symbiosis ; Alder ; Alnus glutinosa ; Endophyte ; Farmyard manure ; Forestry ; Frankia ; Inoculation ; Nitrogen fixation ; Nodulation ; Root nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The occurrence and the infectivity of Frankia, the root-nodule endophyte ofAlnus glutinosa, were studied in different kinds of soil in the Netherlands. Both field and pot experiments indicated that many soils, on which alders have not been grown before, had low numbers of endogenous Frankia or none at all. Inoculation of these soils usually enhanced growth and nodulation of alders. The effect of fertilizer treatments on growth and nodulation ofA. glutinosa were studied in experimental plots. Alders grown in sandy soils, dressed with farmyard manure had the highest yield and the most nodules. The influence of inoculation with homogenates of Sp(+) and Sp(−) nodules and with a pure culture of Frankia AvcIl were studied in pot experiments. The quantity of different kinds of inoculum needed to obtain good growth and nodulation of alder was estimated. The results indicated that addition of a nodule homogenate of 90 g fresh AvcIl Sp(+) nodules is sufficient to inoculate one hectare of nursery soil to produce 10 nodules per plant, while a thousand times larger amount of inoculum is necessary when Sp(−) nodules are used. The limitations and the potentials of using nodule homogenates and pure cultures of Frankia for inoculation in forestry are discussed.
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  • 76
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    Plant and soil 82 (1984), S. 315-327 
    ISSN: 1573-5036
    Keywords: Breeding ; Competition ; Glycine max ; Nitrogen fixation ; Nodulation ; Phaseolus vulgaris
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary While symbiotic nitrogen (N2) fixation byG. max andP. vulgaris reduces their need for combined N, N2 fixation under field conditions is rarely maximized. This paper reviews constraints to N2 fixation in these species, then examines the genetic variability recorded for traits affecting N2 fixation and the further work needed in this area. It considers emerging programs for the improvement of N2 fixation inG. max andP. vulgaris and pays particular attention to methodological considerations.
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