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  • Nitrogen fixation  (145)
  • Springer  (144)
  • Elsevier  (1)
  • 2015-2019  (1)
  • 2010-2014  (2)
  • 1985-1989  (142)
  • 1
    Publication Date: 2022-05-25
    Description: © The Author(s), 2013. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Ecosystems 16 (2013): 1550-1564, doi:10.1007/s10021-013-9701-0.
    Description: We examined controls of benthic dinitrogen (N2) fixation and primary production in oligotrophic lakes in Arctic Alaska, Toolik Field Station (Arctic Long-Term Ecological Research Site). Primary production in many oligotrophic lakes is limited by nitrogen (N), and benthic processes are important for whole-lake function. Oligotrophic lakes are increasingly susceptible to low-level, non-point source nutrient inputs, yet the effects on benthic processes are not well understood. This study examines the results from a whole-lake fertilization experiment in which N and P were added at a relatively low level (4 times natural loading) in Redfield ratio to a shallow (3 m) and a deep (20 m) oligotrophic lake. The two lakes showed similar responses to fertilization: benthic primary production and respiration (each 50–150 mg C m−2 day−1) remained the same, and benthic N2 fixation declined by a factor of three- to fourfold by the second year of treatment (from ~0.35 to 0.1 mg N m−2 day−1). This showed that the response of benthic N2 fixation was de-coupled from the nutrient limitation status of benthic primary producers and raised questions about the mechanisms, which were examined in separate laboratory experiments. Bioassay experiments in intact cores also showed no response of benthic primary production to added N and P, but contrasted with the whole-lake experiment in that N2 fixation did not respond to added N, either alone or in conjunction with P. This inconsistency was likely a result of nitrogenase activity of existing N2 fixers during the relative short duration (9 days) of the bioassay experiment. N2 fixation showed a positive saturating response when light was increased in the laboratory, but was not statistically related to ambient light level in the field, leading us to conclude that light limitation of the benthos from increasing water-column production was not important. Thus, increased N availability in the sediments through direct uptake likely caused a reduction in N2 fixation. These results show the capacity of the benthos in oligotrophic systems to buffer the whole-system response to nutrient addition by the apparent ability for significant nutrient uptake and the rapid decline in N2 fixation in response to added nutrients. Reduced benthic N2 fixation may be an early indicator of a eutrophication response of lakes which precedes the transition from benthic to water-column-dominated systems.
    Description: This project was supported by NSF-OPP 9732281, NSF-DEB 9810222, NSF-DEB 0423385, and by a Doctoral Dissertation Improvement Grant NSF-DEB 0206173. Additional funding was provided by the Small Grants Program through the NSF-IGERT Program in Biogeochemistry and Environmental Change at Cornell University.
    Keywords: Benthic ; Nitrogen fixation ; Primary production ; Oligotrophic ; Arctic ; Toolik
    Repository Name: Woods Hole Open Access Server
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  • 2
    Publication Date: 2022-05-25
    Description: © The Author(s), 2015. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Deep Sea Research Part I: Oceanographic Research Papers 104 (2015): 72-91, doi:10.1016/j.dsr.2015.06.012.
    Description: Nitrogen fixation is an important yet still incompletely constrained component of the marine nitrogen cycle, particularly in the subsurface. A Video Plankton Recorder (VPR) survey in the subtropical North Atlantic found higher than expected Trichodesmium colony abundances at depth, leading to the hypothesis that deep nitrogen fixation in the North Atlantic may have been previously underestimated. Here, Trichodesmium colony abundances and modeled nitrogen fixation from VPR transects completed on two cruises in the tropical and subtropical North Atlantic in fall 2010 and spring 2011 were used to evaluate that hypothesis. A bio-optical model was developed based on carbon-normalized nitrogen fixation rates measured on those cruises. Estimates of colony abundance and nitrogen fixation were similar in magnitude and vertical and geographical distribution to conventional estimates in a recently compiled climatology. Thus, in the mean, VPR-based estimates of volume-specific nitrogen fixation rates at depth in the tropical North Atlantic were not inconsistent with estimates derived from conventional sampling methods. Based on this analysis, if Trichodesmium nitrogen fixation by colonies is underestimated, it is unlikely that it is due to underestimation of deep abundances by conventional sampling methods.
    Description: We gratefully acknowledge support of this research by NSF and NASA. A NASA Earth and Space Science Fellowship supported E. Olson's graduate studies.
    Keywords: Nitrogen fixation ; Trichodesmium spp. ; North Atlantic ; Video Plankton Recorder
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  • 3
    Publication Date: 2022-05-25
    Description: © The Authors 2009. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License. The definitive version was published in Biogeochemistry 99 (2010): 1-13, doi:10.1007/s10533-009-9392-y.
    Description: Human activities have profoundly altered the global nitrogen (N) cycle. Increases in anthropogenic N have had multiple effects on the atmosphere, on terrestrial, freshwater and marine ecosystems, and even on human health. Unfortunately, methodological limitations challenge our ability to directly measure natural N inputs via biological N fixation (BNF)—the largest natural source of new N to ecosystems. This confounds efforts to quantify the extent of anthropogenic perturbation to the N cycle. To address this gap, we used a pair of indirect methods—analytical modeling and N balance—to generate independent estimates of BNF in a presumed hotspot of N fixation, a tropical rain forest site in central Rondônia in the Brazilian Amazon Basin. Our objectives were to attempt to constrain symbiotic N fixation rates in this site using indirect methods, and to assess strengths and weaknesses of this approach by looking for areas of convergence and disagreement between the estimates. This approach yielded two remarkably similar estimates of N fixation. However, when compared to a previously published bottom-up estimate, our analysis indicated much lower N inputs via symbiotic BNF in the Rondônia site than has been suggested for the tropics as a whole. This discrepancy may reflect errors associated with extrapolating bottom-up fluxes from plot-scale measures, those resulting from the indirect analyses, and/or the relatively low abundance of legumes at the Rondônia site. While indirect methods have some limitations, we suggest that until the technological challenges of directly measuring N fixation are overcome, integrated approaches that employ a combination of model-generated and empirically-derived data offer a promising way of constraining N inputs via BNF in natural ecosystems.
    Description: We acknowledge and are grateful for financial support from the Andrew W. Mellon Foundation (C.C. and B.H.), the National Science Foundation (NSF DEB-0515744 to C.C. and A.T. and DEB-0315656 to C.N.), and the NASA LBA Program (NCC5-285 to C.N.).
    Keywords: Amazon Basin ; Ecosystem modeling ; Mass balance ; Nitrogen fixation ; Nutrient cycling ; Rondonia ; Tropical forest
    Repository Name: Woods Hole Open Access Server
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  • 4
    ISSN: 1432-2048
    Keywords: Bacteroid ; Bradyrhizobium ; Glycine (N2 fixation) ; Nitrate reductase ; Nitrite reductase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Bacteroids of Bradyrhizobium japonicum strain CB1809, unlike CC705, do not have a high level of constitutive nitrate reductase (NR; EC 1.7.99.4) in the soybean (Glycine max. Merr.) nodule. Ex planta both strains have a high activity of NR when cultured on 5 mM nitrate at 2% O2 (v/v). Nitrite reductase (NiR) was active in cultured cells of bradyrhizobia, but activity with succinate as electron donor was not detected in freshly-isolated bacteroids. A low activity was measured with reduced methyl viologen. When bacteroids of CC705 were incubated with nitrate there was a rapid production of nitrite which resulted in repression of NR. Subsequently when NiR was induced, nitrite was utilized and NR activity recovered. Nitrate reductase was induced in bacteroids of strain CB1809 when they were incubated in-vitro with nitrate or nitrite. Increase in NR activity was prevented by rifampicin (10 μg· ml-1) or chloramphenicol (50 μg·ml-1). Nitrite-reductase activity in bacteroids of strain CB1809 was induced in parallel with NR. When nitrate was supplied to soybeans nodulated with strain CC705, nitrite was detected in nodule extracts prepared in aqueous media and it accumulated during storage (1°C) and on further incubation at 25°C. Nitrite was not detected in nodule extracts prepared in ethanol. Thus nitrite accumulation in nodule tissue appears to occur only after maceration and although bacteroids of some strains of B. japonicum have a high level of a constitutive NR, they do not appear to reduce nitrate in the nodule because this anion does not gain access to the bacteroid zone. Soybeans nodulated with strains CC705 and CB1809 were equally sensitive to nitrate inhibition of N2 fixation.
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  • 5
    ISSN: 1432-2048
    Keywords: Glutamate synthase ; Glutamine synthetase ; Nitrogen fixation ; Phaseolus (glutamate synthase) ; Plastid (glutamate synthase) ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The two isoenzymes of NADH-dependent glutamate synthase (NADH-GOGAT; EC 1.4.1.14), previously identified in root nodules of Phaseolus vulgaris L., have both been shown to be located in root-nodule plastids. The nodule specific NADH-GOGAT II accounts for the majority of the activity in root nodules, and is present almost exclusively in the central tissue of the nodule. However about 20% of NADH-GOGAT I activity is present in the nodule cortex, at about the same specific activity as this isoenzyme is found in the central tissue. Glutamine synthetase (GS; EC 6.3.1.2) occurs predominantly as the γ polypeptide in the central tissue, whereas in the cortex, the enzyme is represented mainly by the β polypeptide. Over 90% of both GS and NADH-GOGAT activities are located in the central tissue of the nodule and GS activity exceeds NADH-GOGAT activity by about twofold in this region. Using the above information, a model for the subcellular location and stoichiometry of nitrogen metabolism in the central tissue of P. vulgaris root nodules is presented.
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  • 6
    ISSN: 1432-2048
    Keywords: Nitrogen fixation ; Peroxisome ; Root nodules ; Ureide biogenesis ; Uricase ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cowpea (Vigna unguiculata (L.) Walp.) nodules have been investigated by means of cytochemical and immunocytochemical procedures at the ultrastructural level in order to assess the role of the uninfected cells in ureide biogenesis. Uricase activity in the nodules was shown by cytochemical methods to be localized exclusively in the numberous large peroxisomes confined to the uninfected cells; the small peroxisomes in the infected cells did not stain for uricase. Uricase was also localized in the peroxisomes of uninfected cells by immunogold techniques employing polyclonal antibodies against nodule-specific uricase of soybean. There was no labeling above background of any structures in the infected cells. The results indicate that the uninfected cells are essential for ureide biogenesis in cowpea. Although tubular endoplasmic reticulum, the presumptive site of allantoinase, increases greatly in the uninfected cells during nodule development, it virtually disappears as the nodules mature. The inconsistency between the disappearance of the tubular endoplasmic reticulum from older nodules and the high allantoinase activity reported for older plants remains to be explained.
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  • 7
    ISSN: 1432-2048
    Keywords: Auxin (IAA), production by Rhizobium ; Gibberellin production by Rhizobium ; Mutant (Rhizobium) ; Nitrogen fixation ; Phaseolus (nodulation) ; Rhizobium (mutants) ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Similar ranges of gibberellins (GAs) were detected by high-performance liquid chromatography (HPLC)-immunoassay procedures in ten cultures of wild-type and mutant strains of Rhizobium phaseoli. The major GAs excreted into the culture medium were GA1 and GA4. These identifications were confirmed by combined gas chromatographymass spectrometry. The HPLC-immunoassays also detected smaller amounts of GA9- as well as GA20-like compounds, the latter being present in some but not all cultures. In addition to GAs, all strains excreted indole-3-acetic acid (IAA) but there was no obvious relationship between the amounts of GA and IAA that accumulated. The Rhizobium strains studied included nod − and fix − mutants, making it unlikely that the IAA- and GA-biosynthesis genes are closely linked to the genes for nodulation and nitrogen fixation. The HPLC-immunoassay analyses showed also that nodules and non-nodulated roots of Phaseolus vulgaris L. contained similar spectra of GAs to R. phaseoli culture media. The GA pools in roots and nodules were of similar size, indicating that Rhizobium does not make a major contribution to the GA content of the infected tissue.
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  • 8
    ISSN: 1432-2048
    Keywords: Ammonia/ammonium (assimilation, excretion) ; Anthoceros ; Bryophyta ; Cyanobacteria ; Nitrogen fixation ; Nostoc ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The initial product of fixation of [13N]N2 by pure cultures of the reconstituted symbiotic association between Anthoceros punctatus L. and Nostoc sp. strain ac 7801 was ammonium; it accounted for 75% of the total radioactivity recovered in methanolic extracts after 0.5 min and 14% after 10 min of incubation. Glutamine and glutamate were the primary organic products synthesized from [13N]N2 after incubation times of 0.5–10 min. The kinetics of labeling of these two amino acids were characteristic of a precursor (glutamine) and product (glutamate) relationship. Results of inhibition experiments with methionine sulfoximine (MSX) and diazo-oxonorleucine were also consistent with the assimilation of N2-derived NH 4 + by Anthoceros-Nostoc through the sequential activities of glutamine synthetase (EC 6.3.1.2) and glutamate synthase (EC 1.4.7.1), with little or no assimilation by glutamate dehydrogenase (EC 1.3.1.3). Isolated symbiotic Nostoc assimilated exogenous 13NH 4 + into glutamine and glutamate and their formation was inhibited by MSX, indicating operation of the glutamine synthetase-glutamate synthase (GS-GOGAT) pathway: However, relative to free-living cultures, isolated symbiotic Nostoc assimilated 80% less exogenous ammonium into glutamine and glutamate, implying that symbiotic Nostoc could assimilate only a fraction of N2-derived NH 4 + . This implication was tested by using Anthoceros associations reconstituted with wild-type or MSX-resistant strains of Nostoc incubated with [13N]N2 in the presence of MSX. The results of these experiments indicated that, in situ, symbiotic Nostoc assimilated about 10% of the N2-derived NH 4 + and that NH 4 + was made available to Anthoceros tissue where it was apparently assimilated by the GS-GOGAT pathway. Since less than 1% of the fixed N2 was lost to the suspension medium, it appears that transfer of NH 4 + from symbiont to host tissue was very efficient in this extracellular symbiotic association.
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  • 9
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    Planta 167 (1986), S. 382-386 
    ISSN: 1432-2048
    Keywords: Acetylene/nitrogen molar ratio ; Alnus-Frankia symbiosis ; Nitrogen fixation ; Nitrogenase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Acetylene reduction, 15N2 reduction and H2 evolution were measured in root systems of intact plants of grey alder (Alnus incana (L.) Moench) in symbiosis with Frankia. The ratios of C2H2: 15N2 were compared with C2H2:N2 ratios calculated from C2H2 reduction and H2 evolution, and with C2H2:N2 ratios calculated from accumulated C2H4 production and nitrogen content. It was possible to calculate C2H2:N2 ratios from C2H2 reduction and H2 evolution because this source of Frankia did not show any hydrogenase activity. The ratios obtained using the different methods ranged from 2.72 to 4.42, but these values were not significantly different. It was also shown that enriched 15N could be detected in the shoot after a 1-h incubation of the root-system. It is concluded that the measurement of H2 evolution in combination with C2H2 reduction represents a nondestructive assay for nitrogen fixation in a Frankia symbiosis which shows no detectable hydrogenase activity.
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  • 10
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    Oecologia 79 (1989), S. 566-568 
    ISSN: 1432-1939
    Keywords: Carbon dioxide ; Lichen ; Lobaria ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thalli of Lobaria pulmonaria (L.) Hoffm., a nitrogen-fixing epiphyte common in mesic temperate forests, were collected in a Douglas-fir (Pseudotsuga menziesii Franco) forest near Corvallis, Oregon, and maintained for 20 to 40 days in controlled-environment chambers with atmospheric CO2 concentrations of 374 and 700 μll-1. Nitrogenase activity, which was assayed by the acetylene reduction method, was approximately doubled in the lichen maintained in elevated CO2. Increases in nitrogen fixation by lichens may be an important part of the integrated ecosystem response to rising CO2.
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  • 11
    ISSN: 1432-1939
    Keywords: Lupinus succulentus ; Fabaceae ; Lupine ; Quinolizidine alkaloids ; Nitrogen fixation ; Defoliation ; Plant-herbivore interactions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We examined the effects of nitrogen nutrition and defoliation on the alkaloids, nitrogen levels, and growth of Lupinus succulentus by growing plants under five nitrogen/defoliation treatments: 1) fertilization with a high-nitrate nutrient solution, 2) fertilization with a low-nitrate solution, 3) inoculation with N-fixing bacteria but without available soil nitrogen, 4) high-nitrate solution plus periodic partial defoliation, and 5) low-nitrate plus defoliation. In the absence of defoliation, plants from both the N-fixing and high-N treatments had higher concentrations of alkaloids and nitrogen, and higher growth rates than the low-N plants. Periodic defoliation had little effect on the high-N plants, but defoliated N-fixing plants were severely stunted and had lower alkaloid and nitrogen levels. The experimental treatments also affected the relative concentrations of the alkaloids. Our results indicate that 1) alkaloid composition and concentration in L. succulentus are determined by both nitrogen availability and developmental state, 2) plants relying solely on N-fixation respond quite differently to defoliation than those with adequate soil nitrogen, and 3) the food value of the plant tissue can be affected by an interaction between the effects of defoliation and nitrogen status.
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  • 12
    ISSN: 1432-072X
    Keywords: Alfalfa ; Conjugation ; Cross inoculation ; Host specificity ; Hydrogen uptake ; Nodulation ; Nitrogen fixation ; Rhizobium ; Plasmids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract pIJ1008, a Rhizobium leguminosarum plasmid which determines hydrogen uptake ability and symbiotic functions in pea was transferable to three of seven natural isolates of R. meliloti tested. In these three strains, pIJ1008 was maintained stably with the respective sym megaplasmid indigenous to each R. meliloti strain. These strains carrying both plasmids nodulated alfalfa but not pea. By reisolation and examination of the strains from alfalfa nodule tissue, it was shown that pIJ1008 continued to be maintained but that pea-nodulation ability was suppressed. In one strain of R. meliloti which carries a 200 kb cryptic plasmid (in addition to a megaplasmid), the transfer and selection for pIJ1008 resulted in the loss of the cryptic plasmid. In three separate plant growth experiments, alfalfa nodules induced by each of the R. meliloti strain carrying both sym plasmids were assayed for hydrogen uptake activity. The average activity was 40-, 3.5-and 2-fold higher than with the respective pIJ1008-free strains. However, this higher activity was not accompanied by an increase in plant biomass or nitrogen content of shoots.
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  • 13
    ISSN: 1432-072X
    Keywords: Ammonium assimilation ; Excretion ; Anabaena azollae ; Azolla caroliniana ; Cyanobacteria ; Glutamine ; Glutamate formation ; Nitrogen fixation ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Anabaena azollae was isolated fromAzolla caroliniana by the “gentle roller” method and differential centrifugation. Incubation of suchAnabaena preparations for 10 min with [13N]N2 resulted in the formation of four radioactive compounds; ammonium, glutamine, glutamate and alanine. Ammonium accounted for 66% of the total radioactivity recovered and 58% of the ammonium was in an extracellular fraction. Since essentially no extracellular13N-labeled organic compounds were found, it appears that ammonium is the compound most probably made available toAzolla during dinitrogen-dependent growth of the association. The kinetics of incorporation of exogenous13NH 4 + into glutamine and glutamate were characteristic of a precursor (glutamine)-product (glutamate) relationship and consistent with assimilation by the glutamine synthetase-glutamate synthase pathway. The results of experiments using the glutamine synthetase inhibitor, methionine sulfoximine, the glutamate synthase inhibitor, diazo-oxonorleucine, and increasing the ammonium concentration to greater than 1 mM, provided evidence for assimilation primarily by the glutamine synthetase-glutamate synthase pathway with little or no contribution from biosynthetic glutamate dehydrogenase. While showing that N2 fixation and NH 4 + assimilation were not tightly coupled metabolic processes in symbioticAnabaena, these results reflect a composite picture and do not indicate the extent to which ammonium assimilatory enzymes might be regulated in filaments associated with specific stages in theAzolla-Anabaena developmental profile.
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  • 14
    ISSN: 1432-072X
    Keywords: Oscillatoria ; Cyanobacteria ; Nitrogen fixation ; Oxygen protection of N2-ase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Oscillatoria sp. strain 23 is a filamentous, non-heterocystous cyanobacterium that fixes nitrogen aerobically. Although, in this organism nitrogenase is inactivated by oxygen a high tolerance is observed. Up to a pO2 of 0.15 atm, oxygen does not have any measurable effects on acetylene reduction. Higher concentrations of oxygen inhibited the activity to a relatively high degree. Evidence for two mechanisms of oxygen protection of nitrogenase in this cyanobacterium was obtained. A high rate of synthesis of nitrogenase may allow the organism to maintain a certain amount of active enzyme under aerobic conditions. Secondly, a switch off/on mechanism may reversibly convert the active enzyme into a non-active form which is insensitive to oxygen inactivation after a sudden and short-term exposure to high oxygen concentrations. It is conceived that these mechanisms in addition to a temporal separation of nitrogen fixation from oxygenic photosynthesis sufficiently explain the regulation process of aerobic nitrogen fixation in this organism.
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  • 15
    ISSN: 1432-072X
    Keywords: Oscillatoria ; Cyanobacteria ; Nitrogen fixation ; Light-dark cycles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The non-heterocystous cyanobacterium Oscillatoria sp. strain 23 fixes nitrogen under aerobic conditions. If nitrate-grown cultures were transferred to a medium free of combined nitrogen, nitrogenase was induced within about 1 day. The acetylene reduction showed a diurnal variation under conditions of continuous light. Maximum rates of acetylene reduction steadily increased during 8 successive days. When grown under alternating light-dark cycles, Oscillatoria sp. fixes nitrogen preferably in the dark period. For dark periods longer than 8 h, nitrogenase activity is only present during the dark period. For dark periods of 8 h and less, however, nitrogenase activity appears before the beginning of the dark period. This is most pronounced in cultures grown in a 20 h light – 4 h dark cycle. In that case, nitrogenase activity appears 3–4 h before the beginning of the dark period. According to the light-dark regime applied, nitrogenase activity was observed during 8–11 h. Oscillatoria sp. grown under 16 h light and 8 h dark cycle, also induced nitrogenase at the usual point of time, when suddenly transferred to conditions of continuous light. The activity appeared exactly at the point of time where the dark period used to begin. No nitrogenase activity was observed when chloramphenicol was added to the cultures 3 h before the onset of the dark period. This observation indicated that for each cycle, de novo nitrogenase synthesis is necessary.
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  • 16
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    Archives of microbiology 145 (1986), S. 159-161 
    ISSN: 1432-072X
    Keywords: Clostridium cellobioparum ; Clostridium thermocellum ; Ammonium assimilation ; Nitrogen fixation ; Nitrogen control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Inorganic nitrogen metabolism in two cellulose degrading clostridia, the mesophile Clostridium cellobioparum and the thermophile Clostridium thermocellum was investigated. Both strains show acetylene reduction (i.e. possibly nitrogenase activity), contain glutamine synthetase, glutamate dehydrogenase and glutamate-dependent transaminases. C. cellobioparum additionally contains a NADH-dependent glutamate synthase and a NH 4 + -repressible glycine dehydrogenase (NADPH). Remarkably, acetylene reduction in C. thermocellum is not repressed by ammonium, casting doubt whether this activity is due to nitrogenase. The results are compared with the data from other saccharolytic clostridia.
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  • 17
    ISSN: 1432-072X
    Keywords: Heterocyst isolation ; Osmoregulators ; Cyanobacteria ; Nitrogen fixation ; Anabaena variabilis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method is described for the preparation of cyanobacterial heterocysts with high nitrogen-fixation (acetylene-reduction) activity supported by endogenous reductants. The starting material was Anabaena variabilis ATCC 29413 grown in the light in the presence of fructose. Heterocysts produced from such cyanobacteria were more active than those from photoautotrophically-grown A. variabilis, presumably because higher reserves of carbohydrate were stored within the heterocysts. It proved important to avoid subjecting the cyanobacteria to low temperatures under aerobic conditions, as inhibition of respiration appeared to lead to inactivation of nitrogenase. Low temperatures were not harmful in the absence of O2. A number of potential osmoregulators at various concentrations were tested for use in heterocyst isolation. The optimal concentration (0.2M sucrose) proved to be a compromise between adequate osmotic protection for isolated heterocysts and avoidance of inhibition of nitrogenase by high osmotic strength. Isolated heterocysts without added reductants such as H2 had about half the nitrogen-fixation activity expected on the basis of intact filaments. H2 did not increase the rate of acetylene reduction, suggesting that the supply of reductant from heterocyst metabolism did not limit nitrogen fixation under these conditions. Such heterocysts had linear rates of acetylene reduction for at least 2 h, and retained their full potential for at least 12 h when stored at 0°C under N2.
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  • 18
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    Archives of microbiology 146 (1987), S. 327-331 
    ISSN: 1432-072X
    Keywords: Actinomycetes ; Nitrogen fixation ; Symbiosis ; Immunocytochemistry ; Ultracryotomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immunogoldlabelling on ultrathin cryosections of Frankia sp. Cc1.17 showed specific labelling of nitrogenase in the spherical cells called vesicles. No label was found in the hyphae in any cells grown on a medium with combined nitrogen, nor in those to which no specific antiserum was added. Similar results were obtained with cultures grown under high (20%) and low (2%) oxygen tension in the gas phase.
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  • 19
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    Archives of microbiology 147 (1987), S. 383-388 
    ISSN: 1432-072X
    Keywords: Frankia ; Nitrogen fixation ; Vesicle development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between nitrogen fixation and development of a specialized cell structure, called the vesicle, was studied using four Frankia isolates. Nitrogenase activity was repressed in all four strains during growth with ammonia. Strain CpI1 formed no vesicles during NH4 growth. Strains ACN1 ag , EAN1pec and EUN1f produced low numbers of vesicles in the presence of ammonia. Following transfer to nitrogen-free media, a parallel increase in nitrogenase activity and vesicle numbers occurred with all four isolates. Appearance of nitrogenase activity was more rapid in those strains that possessed some vesicles at the time of shift to N2 as a nitrogen source. The ratio of vesicle numbers to level of nitrogenase activity varied widely among the four strains and in response to different growth conditions and culture age of the individual strains. Optimum conditions of temperature, carbon and energy source, nitrogen source and availability of iron and molybdenum were different for each of the four strains. Those conditions that significantly reduced nitrogenase activity were always associated with decreased numbers of vesicles.
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  • 20
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    Archives of microbiology 150 (1988), S. 326-332 
    ISSN: 1432-072X
    Keywords: Rhizobium leguminosarum ; Plasmids ; Melanin ; Nodulation ; Nitrogen fixation ; Plasmid curing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Rhizobium leguminosarum strain VF39, isolated from nodules of field-grown faba beans in the Federal Republic of Germany, was shown to contain six plasmids ranging in molecular weight from 90 to 400 Md. Hybridisation to nif gene probes, plasmid curing, and mobilisation to other strains of Rhizobium and to Agrobacterium showed that the third largest plasmid, pRleVF39d (220 Md), carried genes for nodulation and nitrogen fixation. This plasmid was incompatible with pRL10JI, the Sym plasmid of R. leguminosarum strain JB300. Of the other plasmids, the two smallest (pRleVF39a and pRleVF39b, 90 and 160 Md respectively) were shown to be self-transmissible at a low frequency. Although melanin production is as yet unreported in strains of R. leguminosarum biovar viceae, strain VF39 produced a dark pigment, which, since it was not produced on minimal media and its production was greatly enhanced by the presence of tyrosine in the media, is probably melanin-like. Derivatives of VF39 cured of pRleVF39a no longer produced this pigment, but regained the ability to produce it when this plasmid was transferred into them. Strains of Agrobacterium tumefaciens, R. meliloti, and some strains of R. leguminosarum carrying pRleVF39a did not produce this pigment, indicating perhaps that some genes elsewhere on the VF39 genome are also involved in pigment production. Plasmid pRleVF39a appeared to be incompatible with the cryptic Rhizobium plasmids pRle336b and pRL8JI (both ca. 100 Md), but was compatible with the R. leguminosarum biovar phaseoli Sym plasmids pRP1JI, pRP2JI and pRph51a, all of which also code for melanin production. The absence of pRleVF39a in cured derivatives of VF39 had no effect on the symbiotic performance or competitive ability of this strain.
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  • 21
    ISSN: 1432-072X
    Keywords: Azorhizobium caulinodans ORS571 ; Hydrogenase ; Nitrogen fixation ; Chemostat cultures ; H2/N2 ratio ; ATP/2e value
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hydrogenase-negative (Hup-) mutants of Azorhizobium caulinodans ORS571 were isolated by means of Tn5 mutagenesis. The colony test used for screening for Hup- strains was based on the absence of reduction of triphenyltetrazolium chloride with hydrogen. Suspensions from cultures of the mutant strains grown under derepressing conditions did not use hydrogen with methylene blue or oxygen as the hydrogen acceptor. The mutants were shown to carry single Tn5 insertions at different locations in the A. caulinodans genome. Molar growth yields (corrected for poly-β-hydroxybutyrate formation) in chemostat cultures of the mutants were similar to those of the wild type. Molar growth yields of the mutants were not increased by passing additional hydrogen through chemostat cultures, which is in agreement with the hydrogenase-negative phenotype of the mutants. H2/N2 ratios (mol H2 formed per mol N2 fixed) were calculated from the hydrogen content of the effluent gas and the N-content of the bacterial dry weight. Low H2/N2 ratios (between 1.2 and 1.9) were found in both energy-limited (oxygen or succinate) cultures and in cultures limited by the supply of an anabolic substrate (Mg2+). ATP/2e values (mol ATP used at the transport of 2e to nitrogen or H+) were calculated from the H2/N2 ratios and the molar growth yields of nitrogen-fixing and ammonia-assimilating cultures. ATP/2e values were between 7 and 11. It was concluded that the calculated ATP/2e values comprise not only 4 mol ATP used at the transport of 2e through nitrogenase but also energy equivalents needed for reversed electron flow from NADH to the low-potential hydrogen donor used by nitrogenase.
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  • 22
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    Archives of microbiology 151 (1989), S. 445-453 
    ISSN: 1432-072X
    Keywords: Denitrification ; Growth yield measurements ; Nitrate respiration ; Nitrogen fixation ; Proton translocations in respirations ; Azospirillum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract For Azospirillum brasilense Sp7, the energy transformation efficiencies were measured in anaerobic respirations with either nitrate, nitrite or nitrous oxide as respiratory electron acceptors by determining the maximal molar growth yields and the H+-translocations using the oxidant pulse method. In continuous cultures grown with malate limiting, the maximal molar growth yields (Y s max -values) were essentially the same with O2 or N2O but were 1/3 and 2/3 lower with NO 2 - or NO 3 - , respectively, as respiratory electron acceptors. Both the maximal molar growth yields and the maintenance energy coefficients were surprisingly high when Azospirillum was grown with nitrite as the sole electron acceptor and source for N-assimilation. Growth under N2-fixing conditions drastically reduced the Y s max -values in the N2O and O2-respiring cells. In the H+-translocation measurements, the $$\vec H^ + $$ /oxidant ratios were 5.6 for O2→H2O, 2.5–2.8 for NO 3 - →NO 2 - , 2.2 for NO 2 - →N2O and 3.1 for N2O→N2 respirations when the cells were preincubated with valinomycin and K+. All the values were enhanced when the experiments were performed with valinomycin plus methyltriphenylphosphonium (=TPMP+) cation. The uncoupler carbonyl cyanide-m-chlorophenyl-hydrazone diminished the H+-excretion indicating that this translocation was due to vectorial flow across the membrane. In the absence of any ionophore, nitrate and nitrite respirations were accompanied by a H+-uptake $$(NO_3^ - \to N_2 = - 2.9 \vec H^ + /NO_3^ - and NO_2^ - \to N_2 = - 2.5 \vec H^ + /NO_2^ - )$$ . Any significant H+-translocation could not be detected in N2O- and O2-respirations under these conditions. It is concluded that nitrate reduction proceeds inside the cytoplasmic membrane, whereas nitrite is reduced extramembraneously. The data are not conclusive for the location of nitrous oxide reductase. The maximal molar growth yield determinations and the absence of any H+-uptake in untreated cells indicate a cytoplasmic orientation of the enzyme similar to the terminal cytochrome oxidase of respiration. The low H+-extrusion values for N2O-respiration compared to O2-respiration in cells treated with valinomycin plus TPMP+ are, however, not in accord with such an interpretation.
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  • 23
    ISSN: 1432-072X
    Keywords: Bradyrhizobium ; Gene cloning ; Heme ; Marker exchange mutagenesis ; Nitrogen fixation ; Respiration ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Random and site-directed Tn5-induced mutagenesis of Bradyrhizobium japonicum yielded two mutations, one in strain 2960 and the other in strain 2606::Tn5-20, which mapped close to each other but in separate genes. The corresponding wild-type genes were cloned, and their approximate location on the cloned DNA was determined. Mutant 2960 was Fix- and formed green nodules on soybean, whereas strain 2606::Tn5-20 had ca. 4% of wild-type Fix activity and formed white nodules. Cytochrome oxidase assays (Nadi tests) showed a negative reaction with both mutants, indicating a functional deficiency of cytochrome c or its terminal oxidase or both. However, the mutants grew well under aerobic conditions on minimal media with different carbon sources. Furthermore, mutant 2960 had a reduced activity in hydrogen uptake, was unable to grow anaerobically with nitrate as the terminal electron acceptor and 2960-infected soybean nodules contained little, if any, functional leghemoglobin. Southern blot analysis showed that a B. japonicum heme biosynthesis mutant [strain LO505: O'Brian MR, Kirshbom PM, Maier RJ (1987) Proc Natl Acad Sci USA 84: 8390–8393] had its mutation close to the Tn5 insertion site of our mutant 2606::Tn5-20. This finding, combined with the observed phenotypes, suggested that the genes affected in mutants 2960 and 2606::Tn5-20 were involved in some steps of heme biosynthesis thus explaining the pleiotropic respiratory deficiencies of the mutants. Similar to strain LO505, the mutant 2606::Tn5-20 (but not 2960) was defective in the activity of protoporphyrinogen IX oxidase which catalyzes the penultimate step in the heme biosynthesis pathway. This suggests that one of the two cloned genes may code for this enzyme.
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  • 24
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    Archives of microbiology 141 (1985), S. 364-370 
    ISSN: 1432-072X
    Keywords: Nitrogen fixation ; Denitrification ; Associative symbiosis ; Acetylene reduction ; Nitrous oxide formation ; Azospirillum
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract A model system is described where Azospirillum and germinated wheat seeds were grown in association for a week and then assayed for nitrogen fixation (C2H2-reduction) and denitrification (N2O-formation) activities. The association performed C2H2-reduction and N2O-formation under microaerobic conditions. Both activities were measurable after already 3–5 h of incubation with substantial rates and were strictly dependent on the presence of both plants and bacteria. During the week of the growth of the association, the bacteria had lived exclusively from the carbon compounds supplied by the roots of the plants. C2H2-reduction activity by the association was more or less the same with all the Azospirillum brasilense strains, but lower with A. lipoferum and with the A. amazonense strains tested. Two nitrogenase negative mutants of Azospirillum brasilense showed virtually no activity in the association. C2H2-reduction activity was strongly dependent on the growth temperature of the association. Denitrification (N2O-formation) was high also at higher temperatures and at pH-values in the medium around 7.8 but not at neutrality and was strictly dependent on nitrate. The Azospirillum strain used strongly determined the rate of the N2O-formation in the association. It is suggested that Azospirillum may be beneficial to crops particularly under tropical conditions.
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  • 25
    ISSN: 1432-072X
    Keywords: Phototrophic bacteria ; Rhodospirillaceae ; Glutamine synthetase ; Nitrogen metabolism ; Nitrogen fixation
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    Notes: Abstract The phylogenetically related phototrophic bacteria Rhodospirillum tenue and Rhodocyclus purpureus modulate activity of their glutamine synthetases by adenylylation/deadenylylation. Evidence for covalent modification includes the inhibitory effect of Mg2+ on the activity of glutamine synthetase extracted from cells of either species grown on excess ammonia, and the lack of Mg2+ inhibition of activity of the enzyme isolated from N2-(R. tenue) or glutamine (R. purpureus)-grown cells. In addition, snake venom phosphodiesterase treatment of glutamine synthetase from either species grown on excess ammonia relieved Mg2+ inhibition of the enzyme (as measured via the γ-glutamyl transferase assay), and changed the cation specificity from Mn2+ to Mg2+ (in the biosynthetic assay).
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  • 26
    ISSN: 1432-072X
    Keywords: Evolution ; Nif genes ; Nitrogen fixation ; Nitrogenase ; Nucleotide sequence ; Phylogeny ; Rhizobium ; 16S rRNA cataloguing
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    Notes: Abstract It was known that nitrogenase genes and proteins are well conserved even though they are present in a large variety of phylogenetically diverse nitrogen fixing bacteria. This has lead to the speculation, among others, that nitrogen fixation (nif) genes were spread by lateral gene transfer relatively late in evolution. Here we report an attempt to test this hypothesis. We had previously established the complete nucleotide sequences of the three nitrogenase genes from Bradyrhizobium japonicum, and have now analyzed their homologies (or the amino acid sequence homologies of their gene products) with corresponding genes (and proteins) from other nitrogen fixing bacteria. There was a considerable sequence conservation which certainly reflects the strict structural requirements of the nitrogenase iron-sulfur proteins for catalytic functioning. Despite this, the sequences were divergent enough to classify them into an evolutionary scheme that was conceptually not different from the phylogenetic positions, based on 16S rRNA homology, of the species or genera harboring these genes. Only the relation of nif genes of slow-growing rhizobia (to which B. japonicum belongs) and fast-growing rhizobia was unexpectedly distant. We have, therefore, performed oligonucleotide cataloguing of their 16S rRNA, and found that there was indeed only a similarity of S AB=0.53 between fast- and slowgrowing rhizobia. In conclusion, the results suggest that nif genes may have evolved to a large degree in a similar fashion as the bacteria which carry them. This interpretation would speak against the idea of a recent lateral distribution of nif genes among microorganisms.
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  • 27
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    Archives of microbiology 143 (1985), S. 185-191 
    ISSN: 1432-072X
    Keywords: Heterocyst ; Pyruvate: ferredoxin oxidoreductase ; Nitrogen fixation ; Electron transport to nitrogenase ; Ferredoxin ; Cyanobacteria ; Anabaena cylindrica ; Anabaena variabilis
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    Notes: Abstract Various electron donors were found to stimulate C2H2 reduction (N2 fixation) by isolated heterocysts from Anabaena variabilis and Anabaena cylindrica. Intermediates of glycolysis and the tricarboxylic acid cycle as well as unphosphorylated sugars like glucose, fructose and erythrose were among these electron donors. The transfer of electrons from donors like H2, NADH, glyoxylate and glycollate was strictly light-dependent, whereas others like NADPH or pyruvate plus coenzyme A supported C2H2 reduction also in the dark. In all cases, the overall activity was enhanced by light. The stimulation by light was more distinct with heterocysts from A. variabilis than with heterocysts from A. cylindrica. The present communication establishes that pyruvate supports C2H2 reduction by heterocysts from either A. variabilis or A. cylindrica with rates comparable to those with other electron donors. Pyruvate could, however, support C2H2 reduction only in the presence of coenzyme A, and the concentrations of both coenzyme A and pyruvate were crucial. A pyruvate-dependent reduction of ferredoxin by extracts from heterocysts was recorded spectrophotometrically. Glyoxylate, which is an inhibitor of thiamine pyrophosphate-dependent decarboxylations, inhibited pyruvate-dependent C2H2 reduction. This result supports the conclusion that pyruvate is metabolised by pyruvate: ferredoxin oxidoreductase in heterocysts. High concentrations of pyruvate and other electron donors inhibited C2H2 reduction which suggests that nitrogenase activity in heterocysts may be controlled by the availability of electron donors.
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  • 28
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    Archives of microbiology 143 (1986), S. 330-336 
    ISSN: 1432-072X
    Keywords: Phototrophic bacteria ; Green sulfur bacteria ; Chtorobium ; Nitrogen fixation ; Nitrogenase
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    Topics: Biology
    Notes: Abstract Four strains of the green sulfur bacterium Chlorobium were studied in respect to nitrogen nutrition and nitrogen fixation. All strains grew on ammonia, N2, or glutamine as sole nitrogen sources; certain strains also grew on other amino acids. Acetylene-reducing activity was detectable in all strains grown on N2 or on amino acids (except for glutamine). In N2 grown Chlorobium thiosulfatophilum strain 8327 1 mM ammonia served to “switch-off” nitrogenase activity, but the effect of ammonia was much less dramatic in glutamate or limiting ammonia grown cells. The glutamine synthetase inhibitor methionine sulfoximine inhibited ammonia “switch-off” in all but one strain. Cell extracts of glutamate grown strain 8327 reduced acetylene and required Mg2+ and dithionite, but not Mn2+, for activity. Partially purified preparations of Rhodospirillum rubrum nitrogenase reductase (iron protein) activating enzyme slightly stimulated acetylene reduction in extracts of strain 8327, but no evidence for an indigenous Chlorobium activating enzyme was obtained. The results suggest that certain Chlorobium strains are fairly versatile in their nitrogen nutrition and that at least in vivo, nitrogenase activity in green bacteria is controlled by ammonia in a fashion similar to that described in nonsulfur purple bacteria and in Chromatium.
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  • 29
    ISSN: 1432-072X
    Keywords: Bradyrhizobium ; Electron microscopy ; Mutants ; Nitrogen fixation ; Nodulation ; Soybean ; Symbiosis ; Transposon Tn5
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genome of the slow-growing Bradyrhizobium japonicum (strain 110) was mutagenized with transposon Tn5. A total of 1623 kanamycin/streptomycin resistant derivatives were screened in soybean infection tests for nodulation (Nod) and symbiotic nitrogen fixation (Fix). In this report we describe 14 strains possessing a stable, reproducible Nod+Fix- phenotype. These strains were also grown under microaerobic culture conditions to test them for free-living nitrogen fixation activity (Nif). In addition to strains having reduced Fix and Nif activities, there were also strains that had reduced symbiotic Fix activity but were Nif+ ex planta. Analysis of the genomic structure revealed that the majority of the strains had a single Tn5 insertion without any further apparent physical alteration. A few strains had additional insertions (by Tn5 or IS50), or a deletion, or had cointegrated part of the vector used for Tn5 mutagenesis. One of the insertions was found in a known nif gene (nifD) whereas all other mutations seem to affect different, hitherto unknown genes or operons. Several mutant strains had an altered nodulation phenotype, inducing numerous, small, widely distributed nodules. Light and electron microscopy revealed that most of these mutants were defective in different stages of bacteroid development and/or bacteroid persistence. The protein patterns of the mutants were inspected by two-dimensional gel electrophoresis after labelling microaerobic cultures with l-(35S)methionine. Of particular interest were mutants lacking a group of proteins the synthesis of which was known to be under oxygen control. Such strains can be regarded as potential regulatory mutants.
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  • 30
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    Archives of microbiology 146 (1986), S. 12-18 
    ISSN: 1432-072X
    Keywords: Arctic rhizobia ; Arctic legumes ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Forty-eight strains of Rhizobium isolated from the root nodules of three species of legumes indigenous to the high tundra (Astragalus alpinus, Oxytropis maydelliana andOxytropis arctobia) are phenotypically heterogenous with respect to intrinsic antibiotic resistance, expression of nitrogenase activityex planta and plasmid content. All of the strains possess a 250–300 kb plasmid and are homologous to each other on the genomic DNA level but have little DNA homology with selected reference strains of well characterized species of rhizobia. The arctic rhizobia have an optimum growth temperature of 23°C and can grow slowly at 5°C. The DNA from four of the isolates, which were selected for detailed investigation, have sequences homologous tonif andnod genes fromRhizobium trifolii.
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  • 31
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    Archives of microbiology 146 (1986), S. 239-244 
    ISSN: 1432-072X
    Keywords: Cicer arietinum ; nif Genes ; Plasmids ; Rhizobia ; DNA/DNA hybridization ; Nitrogen fixation ; Chickpeas
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    Notes: Abstract We examined 27 strains of chickpea rhizobia from different geographic origins for indigenous plasmids, location and organization of nitrogen fixation (nif) genes, and cultural properties currently used to separate fast- and slow-growing groups of rhizobia. By using an in-well lysis and electrophoresis procedure one to three plasmids of molecular weights ranging from 35 to higher than 380 Mdal were demonstrated in each of 19 strains, whereas no plasmids were detected in the eight remaining strains. Nitrogenase structural genes homologous to Rhizobium meliloti nifHD, were not detected in plasmids of 26 out of the 27 strains tested. Hybridization of EcoRI digested total DNA from these 26 strains to the nif probe from R. meliloti indicated that the organization of nifHD genes was highly conserved in chickpea rhizobia. The only exception was strain IC-72 M which harboured a plasmid of 140 Mdal with homology to the R. meliloti nif DNA and exhibited also a unique organization of nifHD genes. The chickpea rhizobia strains showed a wide variation of growth rates (generation times ranged from 4.0 to 14.5 h) in yeast extract-mannitol medium but appear to be relatively homogeneous in terms of acid production in this medium and acid reaction in litmus milk. Although strains with fast and slow growth rates were identified, DNA/DNA hybridization experiments using a nifHD-specific probe, and the cultural properties examined so far do not support the separation of chickpea rhizobia into two distinct groups of the classical fast- and slow-growing types of rhizobia.
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  • 32
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    Archives of microbiology 151 (1988), S. 44-48 
    ISSN: 1432-072X
    Keywords: Vanadium ; Molybdenum ; Methanogenesis ; Nitrogen fixation ; Archaebacterium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation of the Methanosarcina barkeri strains “Fusaro” (DSM 804) and “227” (DSM 1538) was found to be dependent on the presence of vanadium or molybdenum whereby molybdenum (added as Na2-molybdate) was preferred to vanadium (added as VCl3). Strain “227” showed less pronounced effects on diazotrophic growth with respect to vanadium and molybdenum. Rhenium (ReCl3) or tungsten (Na2-tungstate) could not replace vanadium or molybdenum. The optimum concentrations were found to be 2μM for vanadium and 5μM for molybdenum (strain “Fusaro”). This Mo optimum of methanogenesis was 10-fold higher with N2 than with NH4Cl as nitrogen source. A vanadium requirement with NH4Cl could not be detected. No interferences were observed if molybdenum and vanadium were added simultaneously under diazotrophic conditions. Growth yields were smallest for strain “227” grown diazotrophically ( $$Y_{CH_3 OH}$$ =0.6g dw/mol in the presence of vanadium and $$Y_{CH_3 OH}$$ =0.9g dw/mol in the presence of molybdenum), obviously higher for strain “Fusaro” grown diazotrophically ( $$Y_{CH_3 OH}$$ =1.15g dw/mol in the presence of V and $$Y_{CH_3 OH}$$ =1.4g dw/mol with Mo) and highest if M. barkeri was grown on NH4Cl as N-source ( $$Y_{CH_3 OH}$$ =3.4g dw/mol with Mo, strain “Fusaro”).
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  • 33
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    Archives of microbiology 151 (1989), S. 180-182 
    ISSN: 1432-072X
    Keywords: Klebsiella pneumoniae ; Nitrogen fixation ; nifL ; Regulation ; Oxygen control ; Nitrogen control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A number of in-frame deletions have been constructed in the Klebsiella pneumoniae regulatory gene nifL. The effects of each nifL mutation on NifA-mediated expression from the nifH promoter of K. pneumoniae have then been assessed with respect to both nitrogen and oxygen control. These experiments indicate that, in contrast to the situation with the homologous regulatory proteins NtrB and NtrC, NifA activity is not impaired in the absence of NifL. We conclude that the only function of NifL is to inactivate NifA in response to an increase in the nitrogen or oxygen status of the cell.
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  • 34
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    Archives of microbiology 148 (1987), S. 286-291 
    ISSN: 1432-072X
    Keywords: Sulfate-reducing bacteria ; Desulfobacter species ; Acetate ; Hydrogen ; Autotrophic growth ; Nitrogen fixation
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    Notes: Abstract Sulfate-reducing bacteria with oval to rod-shaped cells (strains AcRS1, AcRS2) and vibrio-shaped cells (strains AcRM3, AcRM4, AcRM5) differing by size were isolated from anaerobic marine sediment with acetate as the only electron donor. A vibrio-shaped type (strain AcKo) was also isolated from freshwater sediment. Two strains (AcRS1, AcRM3) used ethanol and pyruvate in addition to acetate, and one strain (AcRS1) grew autotrophically with H2, sulfate and CO2. Higher fatty acids or lactate were never utilized. All isolates were able to grow in ammonia-free medium in the presence of N2. Nitrogenase activity under such conditions was demonstrated by the acetylene reduction test. The facultatively lithoautotrophic strain (AcRS1), a strain (AcRS2) with unusually large cells (2×5 μm), and a vibrio-shaped strain (AcRM3) are described as new Desulfobacter species, D. hydrogenophilus, D. latus, and D. curvatus, respectively.
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  • 35
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    Biology and fertility of soils 5 (1987), S. 83-87 
    ISSN: 1432-0789
    Keywords: Inoculation ; Inoculum dose ; Nitrogen fixation ; Chickpea ; Rhizobium spp. ; Cicer arietinum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The influence of three inoculum rates on the performance of three chickpea (Cicer arietinum L.) Rhizobium strains was examined in the field on a Mollisol soil. Increasing amounts of inoculum improved the performance of the strains. A normal dose (104 cells per seed) applied at different intervals gave non-significant increases in nodulation, nitrogenase activity (acetylene reduction assay), nitrogen uptake and grain yield. A ten-fold increase in inoculum increased nodule number, shoot dry weight, nitrogenase activity (ARA) and grain yield, but increases over the control were significant only for nodule dry weight and nitrogen uptake by shoot and grain. The highest level of inoculum (100 × normal) significantly increased nodule dry weight, grain yield, total nitrogenase activity (ARA) and nitrogen uptake by shoot and grain. Strain TAL 620 was more effective than the other two. Combined nitrogen (60 kg N ha−1) suppressed nodulation and nitrogenase activity (ARA).
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  • 36
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    Biology and fertility of soils 6 (1988), S. 279-281 
    ISSN: 1432-0789
    Keywords: Sesbania rostrata ; Green manure ; Biofertilizer ; Nitrogen fixation ; Stem nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Ratooning and stem cutting were compared with seeding in order to reduce the amount of seeds of Sesbania rostrata for green-manure growth. Both methods increased the biofertilizer yield highly significantly within a 6-week growth period.
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  • 37
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    Archives of microbiology 149 (1987), S. 24-29 
    ISSN: 1432-072X
    Keywords: Frankia ; Nitrogen fixation ; Calcium ; Vesicle development
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    Topics: Biology
    Notes: Abstract A calcium requirement was shown for both vesicle development and nitrogenase activity by Frankia strains EAN1pec and CpI1. Washing cells with EGTA or EDTA inhibited both vesicle development and nitrogenase activity. The inhibition of both was reversed by the addition of calcium. A variety of agents known to affect calcium-dependent biological processes, such as a Ca-ATPase inhibitor, Ca-channel blockers, Ca-ionophores, calmodulin antagonists and the local anaesthetics, tetracaine and dibucaine, inhibited nitrogenase activity. Respiratory studies showed that a CN-insensitive respiration process occurred only under nitrogen derepressing conditions. Respiration by NH4Cl-grown cells was completely inhibited by KCN while N2-grown cells were inhibited by only 70%. Removal of calcium ions by EGTA or by the addition of dibucaine or tetracaine blocked the CN-insensitive respiration. This CN-insensitive respiration may be involved in protecting nitrogenase inside the vesicles from oxygen.
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  • 38
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    Archives of microbiology 141 (1985), S. 40-43 
    ISSN: 1432-072X
    Keywords: Nitrogenase ; Nitrogen fixation ; Regulation ; Photosynthetic bacteria ; Chromatium ; Ammonia switch off
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Nitrogenase in Chromatium vinosum was rapidly, but reversibly inhibited by NH 4 + . Activity of the Fe protin component of nitrogenase required both Mn2+ and activating enzyme. Activating enzyme from Rhodospirillum rubrum could replace Chromatium chromatophores in activating the Chromatium Fe protein, and conversely, a protein fraction prepared from Chromatium chromatophores was effective in activating R. rubrum Fe protein. Inactive Chromatium Fe protein contained a peptide covalently modified by a phosphate-containing molecule, which migrated the same in SDS-polyacrylamide gels as the modified subunit of R. rubrum Fe protein. In sum, these observations suggest that Chromatium nitrogenase activity is regulated by a covalent modification of the Fe protein in a manner similar to that of R. rubrum.
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  • 39
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    Archives of microbiology 141 (1985), S. 105-111 
    ISSN: 1432-072X
    Keywords: Ammonia ; Anabaena ; Cyanobacteria ; Nitrogen fixation ; Nitrogenase ; Nostoc
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Incubation in the dark of photoautotrophically grown N2-fixing heterocystous cyanobacteria leads to a loss of nitrogenase activity. Original levels of nitrogenase activity are rapidly regained upon re-illumination of the filaments, in a process dependent on de novo protein synthesis. Ammonia, acting indirectly through some of its metabolic derivatives, inhibits the light-promoted development of nitrogenase activity in filaments of Anabaena sp. ATCC 33047 and several other cyanobacteria containing mature heterocysts. The ammonia-mediated control system is also operative in N2-fixing filaments in the absence of any added source of combined nitrogen, with the ammonia resulting from N2-fixation already partially inhibiting full expression of nitrogenase. High nitrogenase levels, about two-fold higher than those in normal N2-fixing Anabaena sp. ATCC 33047, are found in cell suspensions which have been treated with the glutamine synthetase inhibitor l-methionine-d,l-sulfoximine or subjected to nitrogen starvation. Filaments treated in either way are insensitive to the ammonia-promoted inhibition of nitrogenase development, although this insensitivity is only transitory for the nitrogen-starved filaments, which become ammonia-sensitive once they regain their normal nitrogen status.
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    Archives of microbiology 141 (1985), S. 244-248 
    ISSN: 1432-072X
    Keywords: Ammonia analogues ; Anabaena variabilis ; Cyanobacteria ; Ethylenediamine ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cyanobacterium Anabaena variabilis showed a pH dependent uptake of ethylenediamine. No uptake of ethylenediamine was detected at pH 7.0. At higher pH values (e.g. pH 8.0 and pH 9.0) accumulation did occur and was attributed to diffusion of uncharged ethylenediamine in response to a pH gradient. A biphasic pattern of uptake was observed at these higher pH values. Treatment with l-methionine-d,l-sulphoximine (MSX) to inactivate glutamine synthetase (GS) inhibited the second slower phase of uptake without any significant alteration of the initial uptake. Therefore for sustained uptake, metabolism of ethylenediamine via GS was required. NH 4 + did not alter the uptake of ethylenediamine. Ethylenediamine was converted in the second phase of uptake to an analogue of glutamine which could not be detected in uptake experiments at pH 7.0 or in uptake experiments at pH 9.0 following pretreatment of cells with MSX. Ethylenediamine treatment inhibited nitrogenase activity and this inhibition was greatest at high pH values.
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  • 41
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    Archives of microbiology 142 (1985), S. 289-294 
    ISSN: 1432-072X
    Keywords: Proteus mirabilis ; Nitrogen fixation ; nif genes ; nif plasmids ; Klebsiella pneumoniae
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    Topics: Biology
    Notes: Abstract Self-transmissible plasmids carryinghis andnif genes fromKlebsiella pneumoniae have been introduced into threehis mutants ofProteus mirabilis: strains 5006-1, WR19 and WR20. Expression ofhis by the transconjugants was unequivocal, if slightly temperature-sensitive, but none was Nif+ when tested for acetylene reduction in anaerobic glucose medium using inocula from rich or glucose-minimal aerobic agar cultures. Succinate or pyruvate in place of glucose, low glucose, lower temperature or elevated Na2MoO4 did not allownif expression and no nitrogenase MoFe-protein peptide was detected immunologically after exposure to conditions in which diazotrophic enterobacteria, normal or genetically constructed, derepressnif. One strain,P. mirabilis WR19, carrying thehis nif Kmr plasmid pMF250 was examined in detail. Thenif activator genenifA was introduced on the plasmid pCK1. Such derivatives remained Nif- when tested, after aerobic growth on rich agar media, with normal or low glucose, with succinate or with elevated Mo. However, pre-conditioning by aerobic growth on glucose-minimal agar led to subsequent anaerobic expression ofnif in glucose medium from pMF250 in WR19 carrying pCK1. NH 4 + or proline could serve as N-source in the glucose-minimal agar. Maximum activity was about 5% of that ofK. pneumoniae in our assay conditions. Material cross-reacting with anti-serum to the nitrogenase MoFe protein was formed. Nitrogenase activity was not ‘switched off’ by NH 4 + .P. mirabilis WR19 (pCK1) showed NH 4 + -constitutive temperature-sensitive kanamycin resistance (anif-related phenotype of this plasmid) in aerobic glucose minimal medium. Expression ofnif inP. mirabilis WR19 (pCK1, pMF250) was NH 4 + -repressible despite the constitutivenifA character of pCK1 and introduction of thentrA + plasmid pMM17 did not alter this phenotype. However, pCK1 did not give rise to NH 4 + -constitutive diazotrophy in the wild-typeK. pneumoniae M5al. A construct of WR19 carrying pMF250 and constitutiventrC plasmid (pMD45) remained Nif- even after pre-growth on glucose-minimal media. We conclude (a) thatP. mirabilis forms a gene product functionally equivalent to that ofntrA inK. pneumoniae, (b) that it forms no functional equivalent of thentrC product in our growth conditions. The need for pre-conditioning on aerobic glucose media remains perplexing.
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  • 42
    ISSN: 1432-072X
    Keywords: Transposon mutagenesis ; Soybean ; Nitrogen fixation ; Root nodules ; Auxotrophy ; Bradyrhizobium japonicum ; Glycine ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Four histidine auxotrophs of Bradyrhizobium japonicum strain USDA 122 were isolated by random transposon Tn5 mutagenesis. These mutants arose from different, single transposition events as shown by the comparison of EcoRI and XhoI-generated Tn5 flanking sequences of genomic DNA. The mutants grew on minimal medium supplemented with l-histidine or l-histidinol but failed to grow with l-histidinol phosphate. While two of the muants were symbiotically defective and did not form nodules on Glycine max cvs. Lee and Peking and on Glycine soja, the other two mutants were symbiotically competent. Reversion to prototrophy occurred at a frequency of about 10-7 on growth medium without added antibiotics, but prototrophs could not be isolated from growth medium containing 200 μg/ml kanamycin and streptomycin. The prototrophic revertants formed nodules on all the soybean cultivars examined. When histidine was supplied to the plant growth medium, both nodulation deficient mutants formed effective symbioses. On histidine unamended plants, nodules were observed infrequently. Three classes of bacterial colonies were isolated from such infrequent nodules: class 1 were kanamycin resistant-auxotrophs; class 2 were kanamycin sensitive-prototrophs; and class 3 were kanamycin-sensitive auxotrophs. Our results suggest that two Tn5 insertion mutations in B. japonicum leading to histidine auxotrophy, affect nodulation in some way. These mutations are in regions that show no homology to the Rhizobium meliloti common nodulation genes.
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    Archives of microbiology 145 (1986), S. 403-407 
    ISSN: 1432-072X
    Keywords: Propene ; 1-Butene ; Xanthobacter ; Mono-oxygenase ; Nitrogen fixation
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    Topics: Biology
    Notes: Abstract Yellow-pigmented bacteria showing typical characteristics of Xanthobacter spp. were isolated from enrichments with propene and 1-butene, using classical techniques. The generation time for growth on propene and 1-butene of these bacteria ranged from 5 to 7h. A NADH-dependent mono-oxygenase was identified in cell-free extract of Xanthobacter Py2. This mono-oxygenase was not influenced by potential inhibitors tested indicating that propene mono-oxygenase is different from other hydrocarbon mono-oxygenases described until now. Nitrogenase activity could be measured using the acetylene reduction assay with propene as energy source, because acetylene did not inhibit the mono-oxygenase activity.
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  • 44
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    Archives of microbiology 146 (1986), S. 74-79 
    ISSN: 1432-072X
    Keywords: Azotobacter chroococcum ; Nitrogen fixation ; Nitrogenase genes ; Nif gene reiteration ; Deletion mutagenesis
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    Notes: Abstract Strains of the obligately aerobic nitrogen fixing organismAzotobacter chroococcum were constructed which contained defined chromosomal deletions in which the nitrogenase structural genenifHDK cluster (nifH for the polypeptide of the Fe-protein component of nitrogenase andnifD andnifK for the alpha and beta subunits respectively of the MoFe-protein component of the enzyme) was replaced by a kanamycin resistance gene. N2 fixation was nevertheless observed in deletion strains though only in a molybdenum-deficient medium or in spontaneously arising tungstate-resistant derivatives. In comparison with the parent strain growing in molybdenum-sufficient medium, diazotrophic growth was slow and the nitrogenase activity in vivo was characterised by disproportionately low rates of C2H2-reduction compared to H2-evolution and relative insensitivity of H2-evolution to inhibition by C2H2. The findings show reiteration of functional structural genes for nitrogenase inA. chroococcum consistent with our previous observation of twonifH genes in this organism and detection in this work of a secondnifK-like sequence in the genomes of both parent and deletion strains whenA. chroococcum nifK DNA was used as a probe.
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  • 45
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    Archives of microbiology 150 (1988), S. 224-229 
    ISSN: 1432-072X
    Keywords: Vibrio ; V. diazotrophicus ; V. natriegens ; V. pelagius ; V. cincinnatiensis ; Nitrogenase ; Nitrogen fixation ; Oxygen sensitivity
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract Virtually all Vibrio spp. known and available in culture collections and several newly isolated Vibrio sp. were tested for their ability to fix molecular nitrogen, using the acetylene reduction technique, the fixation of the heavy isotope 15N, and by growth on media devoid of combined nitrogen. Among the 27 species tested, four, including V. diazotrophicus, proved to be nitrogenase-positive. The potential of nitrogen fixation was now also discovered in V. natriegens, V. pelagius and V. cincinnatiensis. Among the 9 newly isolated strains, 4 were nitrogenase-positive. These strains were classified as V. diazotrophicus on the basis of DNA homology studies. Nitrogenase was only induced during growth under anaerobic conditions. Dissolved oxygen as low as 1 μM inhibited nitrogenase completely. This inhibition at low oxygen concentration, however, was reversible. 50–100 μM dissolved oxygen inhibited nitrogenase irreversibly.
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  • 46
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    Theoretical and applied genetics 71 (1986), S. 724-729 
    ISSN: 1432-2242
    Keywords: Nitrogen fixation ; Alfalfa cultivars ; Rhizobium strains ; Acetylene reduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two experiments were conducted in the greenhouse to study the interaction between alfalfa cultivars (Medicago sativa L. and M. falcata L.) and strains of Rhizobium meliloti Dang. for acetylene reduction rate, plant height and dry weights of shoot, root and whole plant. Fifteen alfalfa cultivars were inoculated with 10 strains of Rhizobium in Experiment I. Variance component analysis revealed that more than 30% of the total variance was due to alfalfa cultivars for acetylene reduction rate and 26% was accounted for by Rhizobium strains. More than 36% of the total variation was attributed to the interaction between alfalfa cultivars and Rhizobium strains for this character. Twenty-five host cultivars and 11 Rhizobium strains were included in Experiment II. The results also showed that the interaction of alfalfa cultivars and Rhizobium strains contributed the largest portion of the total variation for dry weights of shoot, root and whole plant and acetylene reduction rate. The results clearly demonstrated that the non-additive effects were the major component of variation for these characters associated with nitrogen fixation in alfalfa. Therefore, an effective way of improving nitrogen fixation in alfalfa is to select for a favourable combination of specific Rhizobium strains and alfalfa cultivars.
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    Theoretical and applied genetics 78 (1989), S. 433-435 
    ISSN: 1432-2242
    Keywords: Rhizobium ; Irradiation ; Nitrogen fixation ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Sumary The objective of this work was to know the behaviour and variability of Rhizobium leguminosarum after irradiation. The induced variation was tested under greenhouse conditions on the variety JV 3 of broad beans (Vicia faba) in six replications. Induced genetic variabilty was observed for strain, parent and mutant versus parent. Out of 24 irradiated strains, strain 93-32 performed better with a greater number of nodules and higher dry weight of nodules per plant and biological yield. Environment played an important role in the expression of characters observed. High heritability and genetic advance of these traits indicated that the nitrogen fixation ability of Rhizobium can easily be improved by selection.
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  • 48
    ISSN: 1432-2048
    Keywords: Carbon dioxide fixation ; Citrulline ; Coralloid roots ; Cycads (nitrogen fixation) ; Nitrogen fixation ; Nitrogen transport ; Nostoc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Freshly detached coralloid roots of several cycad species were found to bleed spontaneously from xylem, permitting identification of products of nitrogen transfer from symbiotic organ to host. Structural features relevant to the export of fixed N were described for Macrozamia riedlei (Fisch. ex Gaud.) Gardn. the principal species studied. Citrulline (Cit), glutamine (Gln) and glutamic acid (Glu), the latter usually in a lesser amount, were the principal translocated solutes in Macrozamia (5 spp.), Encephalartos (4 spp.) and Lepidozamia (1 sp.), while Gln and a smaller amount of Glu, but no Cit were present in xylem sap of Bowenia (1 sp.),and Cycas (2 spp.). Time-course studies of 15N enrichment of the different tissue zones and the xylem sap of 15N2-pulse-fed coralloid roots of M. riedlei showed earlier 15N incorporation into Gln than into Cit, and a subsequent net decline in the 15N of Gln of the coralloid-root tissues, whereas Cit labeling continued to increase in inner cortex and stele and in the xylem sap. Hydrolysis of the 15N-labeled Cit and Gln consistently demonstrated much more intense labeling of the respective carbamyl and amide groups than of the other N-atoms. Coralloid roots of M. riedlei pulse-fed 14CO2 in darkness showed 14C labeling of aspartic acid (Asp) and Cit in all tissue zones and of Cit of xylem bleeding sap. Lateral roots and uninfected apogeotropic roots of M. riedlei and M. moorei also incorporated 14CO2 into Cit. The 14C of Cit was restricted to the carbamyl-C. Comparable 15N2 and CO2-feeding studies on corallid roots of Cycas revoluta showed Gln to be the dominant product of N2 fixation, with Asp and alanine as other major 14C-labeled amino compounds, but a total absence of Cit in labeled or unlabeled form.
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  • 49
    ISSN: 1432-0789
    Keywords: Nodule damage ; Rivellia angulata ; Nitrogen fixation ; Cajanus cajan ; Pigeonpea ; Vertisol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Damage caused by Rivellia angulata larvae to pigeonpea root nodules at the ICRISAT center in India was greater in the crop grown on Vertisols (up to 86%) compared to that on Alfisols (20%). Attempts to quantify the field effects of nodule damage on growth and yield of pigeonpea in a Vertisol, involving many heavy applications of soil insecticides (aldrin and hexachlorocyclohexane) failed because the insecticides did not control the pest and adversely affected the growth of the pigeonpea and the subsequent crop of sorghum (Sorgorum bicolor L. Moench). The impact of nodule damage on pigeonpea growth, yield and nutrient uptake was successfully studied in greenhouse-grown plants at three N levels. In this pot study, artificial inoculation with Rivellia sp. led to substantial nodule damage (70%). The results of this damage were a significant overall reduction in nodule dry weight (46%), acetylene reduction activity (31%), total leaf area (36%), chlorophyll content of leaves (39%) and shoot dry weight (23%) 68 days after sowing. At maturity, Rivellia sp. infestation caused significant reductions in top dry weight (22%), root and nodule dry weight (27%), seed dry weight (14%), and total N (29%) and P uptake (19%). The problems and prospects of manipulating nodule damage so as to reduce N losses in pigeonpea are discussed.
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    Biology and fertility of soils 4 (1987), S. 61-66 
    ISSN: 1432-0789
    Keywords: Stem nodulation ; Aeschynomene afraspera ; Legume ; Nitrogen fixation ; Acetylene reduction assay (ARA)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Aeschynomene afraspera is a wild annual legume growing in periodically waterlogged soils in western Africa. This legume is characterized by a profuse stem nodulation. Nodules are formed on the stem at the emergence of lateral root primordia, called nodulation sites. These sites are irregularly distributed on vertical rows all along the stem and branches. Stem nodules are hemispherically shaped. Their outside is dark green and they contain a red-pigmented central zone. Stem nodules exhibit a high nitrogen-fixing potential. Acetylene reduction assays result in stem nodule activity of 309 μmol C2H4 g−1 dry nodule h−1. Field-grown stem nodulated Aeschynomene accumulated more N (51 g N m−2 in 10 weeks) than the root nodulated one. Because of this nitrogenfixing potential and its ability to grow in waterlogged conditions, A. afraspera could probably be introduced into tropical rice cropping systems.
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  • 51
    ISSN: 1432-0789
    Keywords: Triticum aestivum ; T. turgidum ; Nitrogen fixation ; Field inoculation ; Acetylene reduction assay (ARA)
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    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Eight commercial Israeli spring wheat cultivars (six Triticum aestivum and two T. turgidum) grown with 40 and 120 kg N/ha were tested for responses to inoculation with Azospirillum brasilense. At the low level of N fertilization (40 kg/ha), five cultivars showed significant increases in plant dry weight measured at the milky ripe stage; however, by maturation only the cultivar “Miriam” showed a significant increase in grain yield. Two cultivars, which had shown a positive inoculation effect at the earlier stages, had a significant decrease in grain yield. No significant effect of inoculation was found at the high N level. To confirm those results, four wheat (T. aestivum) cultivars were tested separately over 4 years in 4 different locations under varying N levels. Only Miriam showed a consistently positive effect of Azospirillum inoculation on grain yield. Inoculation increased the number of roots per plant on Miriam compared with uninoculated plants. This effect was found at all N levels. Nutrient (N, P and K) accumulation and number of fertile tillers per unit area were also enhanced by Azospirillum, but these parameters were greatly affected by the level of applied N. It is suggested that the positive response of the spring wheat cultivar “Miriam” to Azospirillum inoculation is due to its capacity to escape water stresses at the end of the growth season.
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  • 52
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    Biology and fertility of soils 4 (1987), S. 15-19 
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; N-balance studies ; Azolla ; Blue-green algae ; Chemical N fertilization ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A nitrogen balance study conducted in ceramic pots under net house conditions for four seasons showed that flooded rice soil leaves a positive nitrogen balance (N increase) in soil after rice cropping in both fertilized and unfertilized soil. Recovery of nitrogen from rice soil was more than its input in unfertilized soil, but it was reverse in fertilized soil. Incorporation of Azolla or BGA twice as basal and 20 days after transplanting (DAT) alone or in combination showed higher nitrogen balance and N2-fixation (N gain) in soil than in that where it was applied once either as basal or 20 DAT. Planted soil showed more N2-fixation than that of fallow rice, and flooded soil fixed more nitrogen in comparison to non-flooded soil in light but less in dark. Soil exposed to light fixed more nitrogen than that of unexposed soil in both flooded and non-flooded conditions.
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  • 53
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    Biology and fertility of soils 4 (1987), S. 9-14 
    ISSN: 1432-0789
    Keywords: Rhizosphere ; Nitrogen fixation ; Root exudates ; Soil bacteria ; Carbon budget ; Rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The association of rice seedlings (cv. Delta) with different strains of Azospirillum was studied under monoxenic conditions in the dark. Axenic 3-day-old seedlings were obtained on a C- and N-free medium and inoculated with 6 · 107 bacteria per plant in a closed vial. Seven days later, different components of a carbon budget were evaluated on them and on sterile controls: respired CO2, carbon of shoot and roots, bacterial and soluble carbon in the medium. Two strains (A. lipoferum 4B and A. brasilense A95) isolated from the rhizosphere of rice caused an increase in exudation, + 36% and + 17% respectively compared with sterile control. Shoot carbon incorporation and respiration were reduced by inoculation. A third strain (A. brasilense R07) caused no significant change in exudation. A. lipoferum B7C isolated from maize did not stimulate rice exudation either. We further investigated a possible effect of nitrogen fixation on this phenomenon: inhibition of nitrogen fixation by 10% C2H2 did not modify the extent of C exudation by rice associated with A. lipoferum 4B or with the non-motile A. lipoferum 4T.
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  • 54
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    Biology and fertility of soils 7 (1989), S. 269-274 
    ISSN: 1432-0789
    Keywords: Nitrogen fixation ; Frankia-Ceanothus spp. association ; Acetylene reduction assay (ARA) ; Microsymbiont population ; Nodules ; Actinomycetes
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    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Wildland shrub improvement is needed for sound range and disturbed land revegetation practice. The possibility of selecting superior N2-fixingFrankia-Ceanothus spp. actinorhizal associations was examined. Greenhouse tests were used to expose various soil-borne microsymbiont andCeanothus sp. population accessions in reciprocal combination. The acetylene reduction rate was used as a measure of N2-fixation capacity. There was no significant interaction between host and microsymbiont regardless of source for all variables measured. The acetylene reduction rate, nodule number and mass, plant biomass, and root: shoot ratio were significantly different among soil sources. The acetylene reduction rate was not significantly different amongCeanothus sp. accessions. Neither was it strongly correlated with other variables. It was concluded that the N2-fixation rate is more a function ofFrankia sp. than the hostCeanothus sp. in actinorhizal associations. It appears possible to select soil sources with superior N2-fixing microsymbiont populations.
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  • 55
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    Biology and fertility of soils 6 (1988), S. 39-44 
    ISSN: 1432-0789
    Keywords: Alnus ; Energy forestry ; Frankia ; Meadow soil ; Nitrogen fixation ; Nodulation ; Peat soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Use of the N2-fixing grey alder, Alnus incana (L.) Moench, as a short-rotation crop for energy production is currently being explored. To evaluate the need for inoculation of alders, the distribution of infective propagules of Frankia in the soil at potential sites for alder plantations was examined. Uninoculated grey alder seedlings were grown in three types of soil. Frequent nodulation was found in a meadow soil which had been free from actinorhizal plants for nearly 60 years, but the alder seedlings failed to nodulate in peat soil from two different bog sites. One of these bogs had been exploited for peat and the surface layer of the peat had been removed, so that the soil samples were taken from deep layers of the peat. At the other site, an area of cultivated peat, there were no infective propagules of Frankia in plots without alders; the infective Frankia was present in plots only where it had been introduced by inoculated alders. There was no detectable air-borne dispersal of Frankia. Instead, water movement might account for the dispersal of Frankia in peat. Although the apparent absence of Frankia in these peat soils necessitates inoculation of alder seedlings before planting out, this makes it possible to introduce and maintain Frankia strains with selected beneficial characteristics, since there is no competition from an indigenous Frankia flora.
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  • 56
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    Biology and fertility of soils 8 (1989), S. 356-368 
    ISSN: 1432-0789
    Keywords: Plant-root associations ; Azospirillum spp ; Rhizosphere ; Nitrogen fixation ; Acetylene reduction assay (ARA) ; Phytohormones
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Bacteria of the genus Azospirillum are extensively studied for their plant-growth promoting effect following inoculation. Physiological and biochemical studies of these diazotrophic bacteria are now benefiting from recent breakthroughs in the development of genetic tools for Azospirilum. Moreover, the identification and cloning of Azospirillum genes involved in N2 fixation, plant interaction, and phytohormone production have given new life to many research projects on Azospirillum. The finding that Azospirillum genes can complement specific mutations in other intensively studied rhizosphere bacteria like Rhizobia will certainly trigger the exploration of new areas in rhizosphere biology. Therefore a review of the Azospirillum-plant interactions is particularly timely.
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  • 57
    ISSN: 1476-5535
    Keywords: Sulfate-reducing bacteria ; Hydrogen metabolism ; Nitrogen fixation ; Deuterium-proton exchange
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Hydrogenase and nitrogenase activities of sulfate-reducing bacteria allow their adaptation to different nutritional habits even under adverse conditions. These exceptional capabilities of adaptation are important factors in the understanding of their predominant role in problems related to anaerobic metal corrosion. Although the D2−H+ exchange reaction indicated thatDesulfovibrio desulfuricans strain Berre-Sol andDesulfovibrio gigas hydrogenases were reversible, the predominant activity in vivo was hydrogen uptake. Hydrogen production was restricted to some particular conditions such as sulfate or nitrogen starvation. Under diazotrophic conditions, a transient hydrogen evolution was followed by uptake when dinitrogen was effectively fixed. In contrast, hydrogen evolution proceeded when acetylene was substituted as the nitrogenase substrate. Hydrogen can thus serve as an electron donor in sulfate reduction and nitrogen metabolism.
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  • 58
    ISSN: 1440-1703
    Keywords: Acetylene reduction ; Antarctica ; Cyanobacteria ; Moss community ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation by cyanobacteria in a moss community on East Ongul Island (69°00'S 39°35'E), Antarctica was investigated using the acetylene reduction method. The mean acetylene reduction rate at 10°C and 200 μE·m−2·s−1 photosynthetically active radiation was 7.12 nmol C2H4 per square centimeter of moss community per hour. The effects of temperature, radiation, desiccation and rehydration on the acetylene reduction rates were examined. A simple predictive model was constructed in order to estimate the amount of nitrogen fixed in the field. Using this model, the daily amount of nitrogen fixation was calculated from microclimatic data (temperature and radiation) measured in the experimental field at Syowa Station on East Ongul Island between 1983 and 1984. The cumulative amount of nitrogen fixation in the growing season during this period was estimated to be 329 mg N per square meter of moss community. It is suggested that nitrogen fixation by cyanobacteria in the moss community is important as a nitrogen source for the community growth on East Ongul Island.
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  • 59
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    Plant and soil 86 (1985), S. 57-67 
    ISSN: 1573-5036
    Keywords: Azolla ; N2-fixing efficiency ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The productivity of three species of Azolla (A. pinnata, A. filiculoides andA. caroliniana) in outdoor culture has been evaluated at different planting densities. The highest yields were obtained with biomass concentration ranging from 40 to 70g d.w. m−2. The mean productivity over a 90 days period (from May 10th to August 10th) ranged from 10g d.w. m−2 day−1 forA. filiculoides up to 11.5 g d.w. m−2 day−1 forA. caroliniana. The nitrogen content of the dried biomasses was 48.3 mg (g d.w.)−1 forA. pinnata, 51.5mg (g d.w.)−1 forA. filiculoides and 52.3 mg (g d.w.)−1 forA. caroliniana. Very little variations of the nitrogen content of the ferns during the experimental period were observed. The nitrogen-fixing efficiency of the Azolla-Anabaena azollae symbiosis grown in outdoor conditions was evaluated both by direct measurement of the amount of N2 fixed by the culture and by the C2H2-reduction and H2-evolution tests in an air atmosphere. These tests were performed outdoor under the same environmental conditions as the growing cultures. For all the species the ratios of C2H2-reduced to N2-fixed were unexpectedly low, ranging from 2.04 (A. pinnata) to 1.50 (A. caroliniana). The results suggest that the reliability of the C2H2-reduction assay, particularly when applied to complex biological N2-fixing systems, must be re-examined.
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  • 60
    ISSN: 1573-5036
    Keywords: Glycine max ; Isotope dilution ; 15N techniques ; Nitrogen availability ; Nitrogen fixation ; Plant breeding ; Varietal comparison
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Soybeans (Glycine max (L.) Merr.) have a high N requirement which is fulfilled by soil N uptake and N2-fixation. This study was concerned with the effects of past yield selection on N2-fixation in soybeans. The soybean cultivars, ‘Lincoln’, ‘Shelby’, and ‘Williams’, which represent successive improvements in the ‘Lincoln’ germplasm, and a non-nodulating control were planted in a soil containing15N labelled organic matter. Two replications occurred on soil previously cropped to alfalfa and two on soil previously cropped to soybeans. Plants were harvested at five growth stages and leaf area, plant weight, total N, and atom percent15N were determined. Mature grain was harvested and yield components were also determined, as well as the total N and15N content. Cultivar differences in total dry matter were only evident at physiological maturity, when Williams contained the greatest dry matter. Williams exhibited the longest period of seed formation and seed fill and also had the highest grain yield which resulted from a larger weight per seed. The N content of the cultivars did not vary until physiological maturity when Williams contained the highest percent N. The quantity of N fixed at physiological maturity was highest for Williams and lowest for Lincoln. Fixed N contained in the harvested grain was greater for Williams than for the other two cultivars. The fraction of the total plant N derived from fixation was not greatly affected by cultivar and all cultivars acquired an average of 50% of their total N through N2-fixation. Previous cropping history greatly affected the quantity of N fixed and the fraction of the total plant N derived from fixation. Soybeans following soybeans were more dependent upon N2-fixation than soybeans following alfalfa with the former deriving 65% of the total plant N from fixation and the latter only 32%. These soybean cultivars apparently utilized soil N first and then used N2-fixation to satisfy their N requirement. The past selection for higher yield has resulted in soybean cultivars with improved capacities to fix atmospheric N2 and an improved ability to take up available soil N.
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  • 61
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    Plant and soil 87 (1985), S. 49-60 
    ISSN: 1573-5036
    Keywords: Actinorhizae ; Alnus ; Frankia ; Nitrogen fixation ; Plasmids ; Restriction analysis ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary After the initial screening of more than 200Frankia strains, the plasmid DNA observed in eight Frankiae was analyzed.In situ lysis was performed to obtain an estimate of their copy number and molecular weight. Four plasmid classes were distinguished, 7–9, 18–20, 30–35 and 50–55 kb. Twelve plasmids were thus analysed with restriction enzymes to determine their plasmid restriction patterns. While someFrankia plasmids with comparable molecular weights were found to be heterologous in their restriction enzyme pattern, an 8 kb plasmid found in bothFrankia sp. ArI3, isolated fromAlnus rubra andFrankia sp. CpI1 isolated fromComptonia peregrina showed undistinguishable ‘fingerprints’. Furthermore, an 18 kb plasmid found in the same two strains, also showed homologous restriction enzyme patterns. However, the copy numbers of the two ArI3 plasmids were higher than those of the CpI1 plasmids. Similarly, strains ACN1AG, $$TX41_{b^--- }^{AC} and TX38_{b^--- }^{AC} $$ , isolated fromAlnus crispa all contained a 50 kb plasmid, and the three plasmids were found upon restriction analysis to be undistinguishable. In one strain, ARgX17c isolated fromAlnus rugosa, it was found through restriction enzyme analysis that two plasmids of a similar molecular weight were in fact heterologous. The possible origin of the homologous plasmids and their potential as specificFrankia markers to be used in ecological studies are discussed.
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  • 62
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    Plant and soil 87 (1985), S. 1-16 
    ISSN: 1573-5036
    Keywords: Allocasuarina ; Casuarina ; Frankia ; Nitrogen fixation ; Nodulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Seedlings ofCasuarina spp. andAllocasuarina spp. were grown from seed in the greenhouse and inoculated with a nodule suspension fromC. equisetifolia. Plants ofCasuarina spp. nodulated regularly and were effective in nitrogen-fixation. Only one species ofAllocasuariona, A. lehmanniana formed root nodules. Using these plants as source of inoculum, the isolation of a newFrankia sp. HFPA11I1 (HFP022 801) was made and the strain was grown in pure culture.Frankia sp. HFPA11I1 grows well in a defined medium and shows typical morphological characteristics. In media lacking combined nitrogen, the filamentours bacterium forms terminal vesicles in abundance and differentiaties large intrahyphal or terminal sporangia containing numerous spores. This strain, used as inoculum, nodulates effectively seedlings ofC. equisietifolia andC. cunninghamiana, forming nodules with verically-growing nodule roots. Although effective in acetylene reduction, the endophyte within the nodules is filamentous and lacks veiscles. When used to inoculated seedlings ofA llocasuarina lehmanniana, Frankia sp. HFPA11I1 induces root nodules which are coralloid and lacking nodule roots. The nodules are effective in acetylene reduction and the filamentous hyphae ofFrankia within the nodule lobes lack vesicles. Effective nodulation inA. Lehmanniana depends upon environmental conditions of the seedlings and proceeds much more slowly than in Casuariana.
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  • 63
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    Plant and soil 87 (1985), S. 143-152 
    ISSN: 1573-5036
    Keywords: Actinorhizae ; Elaeagnus angustifolia ; Frankia ; In vitro propagation ; Nitrogen fixation ; Nodulation ; Russian Olive ; Tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Following the evaluation of the nutritional requirements for thein vitro propagation ofElaeagnus angustifolia, this actinorhizal species was routinely multiplied on MS, supplemented with 100 mM sucrose and 5 μM kinetin. On this medium, at a 3 week-interval, a multiplication rate of 5–10 was observed. A morphological variant occurred in culture (wet type) but it was converted into the normal type (pubescent type) by a passage on 1/2 macro MS and 1.5% agar. One hundred percent rooting was achieved in liquid medium containing 1/2 MS without growth regulators. The plantlets were transferred aseptically to a nitrogen-free artificial soil substrate and inoculated with pure cultures of differentFrankia strains which had been isolated from Elaeagnus, Shepherdia and Hippophae host plants. We thus ascertained that afterin vitro propagation, the plants retained their capacity to nodulate and sustain nitrogen fixation.
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  • 64
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    Plant and soil 87 (1985), S. 223-231 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; Rhizobia ; Vigna unguiculata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Biological nitrogen fixation is considered an important trait of cowpeas (Vigna unguiculata (L.) Walp. var. ‘California Blackeye’ No. 5) for economical production yet the process does not alone provide the quantity of nitrogen required by the plant for maximum productivity. Two experiments were undertaken to determine the potential of an increase in nodule mass and number of bacteroids resulting in increased nitrogen fixation. Cowpeas were grown in a glasshouse for 7 weeks under conditions forcing near total dependence on biological nitrogen fixation for growth. Nodule mass on the roots was varied by inoculating seeds with various ratios of effective and ineffective rhizobia that could be identified serologically and by the color of nodule formed. The results of both experiments demonstrated a linear relationship between total nodule mass formed by the effective rhizobia and quantity of nitrogen fixed. The regression coefficients were high in both experiments (r=0.99** and 0.91**). The relationship between total nitrogen fixed and total number of bacteroids of the effective strain was not consistent. In one experiment the regression coefficient was 0.93** but in the other experiment it was 0.65**. From these results it appears that there is good potential for increasing nitrogen fixation in cowpeas by increasing nodule mass. An increase in nodule mass would also result in an increase in the number of bacteroids.
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  • 65
    ISSN: 1573-5036
    Keywords: Azospirillum ; Mustard ; Nitrogen fixation ; Rhizosphere soil ; Root association ; Seed inoculation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A microplot field experiment was conducted in the presence or absence of P and N application to evaluate the influence of the seed inoculation of mustard (cv. Baruna T59) withAzospirillum lipoferum on N2-fixation in rhizosphere, association of the bacteria with the roots and grain yield and N uptake. Inoculation significantly increased the N content in rhizosphere soil particularly at early stage (40 days) of plant growth, which was accompanied by the increased association of the bacteria (A. lipoferum) in rhizosphere soil, root surface washing and surface-sterilized macerated root. A significant increase in grain yield and N uptake was also observed due to inoculation. Application of P particularly at the 20 kg. ha−1 level further enhanced the beneficial effect ofAzospirillum lipoferum inoculation, while N addition markedly reduced such an effect.
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  • 66
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    Plant and soil 87 (1985), S. 445-447 
    ISSN: 1573-5036
    Keywords: Glomus fasciculatum ; Lentil ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Responses of lentil in unsterile soils at low, medium and high levels of plant available soil P toGlomus fasciculatum inoculation were evaluated. It was observed that growth, dry matter accumulation, nodulation, and nitrogen fixation were considerably improved in VAM inoculated plants over uninoculated control at low and medium levels of plant available soil P.
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  • 67
    ISSN: 1573-5036
    Keywords: Breeding ; Nitrate nitrogen ; Nitrogen fixation ; Nodulation ; Trifolium repens ; White clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sodium nitrate applications ranging from 0.36 to 22.84 mM N were shown to depress rates of nodule formation and reduce total nitrogen fixation (acetylene reduction) in white clover plants grown in aseptic test tube culture. Low nitrate levels gave an initial depression in symbiotic activity but the reduction was of short duration and these treatments were subsequently associated with enhanced rates of nodule formation and nitrogen fixation. As a result, phenotypic variation appeared to be strongly differentially affected by the amount of nitrate present. A subsequent experiment suggested that much of the variation was a consequence of early enhancement of plant growth rates by low levels of nitrate followed by rapid depletion thus giving a transitory inhibitory effect. This was confirmed in a third experiment in which the range of nitrate concentration was held constant. Differential effects on variability in nodule formation and nitrogen fixation were then greatly reduced but there was still a residual level of plant-to-plant variation. The results have clear implications for selecting genetic variants capable of fixing di-nitrogen in the presence of combined N. The provision of a single limiting dose of combined nitrogen to a population containing individuals with inherently different growth rates can bring about variations in the phenotypic expression of symbiotic characters. These variations are unlikely to be based on genetic factors which have a direct and stable effect on nodule development and nitrogenase activity. The implications of the results for plant breeding are discussed.
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  • 68
    ISSN: 1573-5036
    Keywords: Nitrate nitrogen ; Nitrate tolerance ; Nitrogen fixation ; Nodulation ; Trifolium repens ; Symbiotic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The variability in nodulation and nitrogen fixation traits ofTrifolium repens (cv. S. 100) supplied with abundant nitrate nitrogen was examined using an aseptic tube culture technique. Nodule numbers varied from 0 to 20 per plant and acetylene reduction activity from 0 to 49 nmol C2H4/plant per h. Plants were selected from this variable population for high, low and zero nodule numbers. Plants within groups were intercrossed and progeny used to establish high, low and zero nodule families. These were subsequently tested for nodulation and acetylene reduction activity in the presence and absence of combined nitrogen. When grown with nitrate the control populations (unselected S. 100 and Pronitro) averaged 3.05 nodules per plant, the high family averaged 4 per plant while the low line averaged 1 nodule per plant (P〈0.05). Nodule numbers were strongly correlated with acetylene reduction activity (r= +0.92) but C2H4 production was generally low, averaging respectively 4.9 and 3.1 nmol/plant per h in the high and low lines. In the absence of nitrate nodule numbers increased to an average of 50 per plant and acetylene reduction activity rose to 175.5 nmol/plant per h. There was no correlation between ability of selection lines to nodulate in the presence and absence of nitrate. The results demonstrate the existence of heritable genetic factors controlling nodulation and nitrogenase activity in the presence of nitrate, but the phenotypic expression of these characters is so strongly inhibited by nitrate that increases in nitrogen fixation in the presence of combined nitrogen which are likely to be of agricultural significance would require much greater genetic improvements than those demonstrated here. The possibilities for such improvements are discussed.
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  • 69
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Bean cultivars ; Harvest index ; 15N dilution ; Nitrogen fixation ; Nodulation ; Phaseolus vulgaris ; Rhizobium inoculation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A field experiment was performed to assess the effects of Rhizobium inoculation and nitrogen fertilizer (100 kg N ha−1) on four cultivars of Phaseolus beans; Carioca, Negro Argel, Venezuela 350 and Rio Tibagi. In the inoculated treatment 2.5 kg N ha−1 of15N labelled fertilizer was added in order to apply the isotope dilution technique to quantify the contribution of N2 fixation to the nutrition of these cultivars. Nodulation of all cultivars in the uninoculated treatments was poor, but the cultivars Carioca and Negro Argel were well nodulated when inoculated. Even when inoculated, nodulation of the cultivars Venezuela 350 and Rio Tibagi was poor and these cultivars showed little response to inoculation in terms of nitrogen accumulation or grain yield. The estimates of the contribution of N2 fixation estimated using the isotope dilution technique, for the Carioca and Negro Argel cultivars, amounted to 31.7 and 18.4 kg N ha−1 respectively. These two cultivars produced 991 and 883 kg ha−1 of grain, respectively, when inoculated and 663 and 620 kg ha−1 with the addition of 100 kg N ha−1 of N fertilizer. The response to nitrogen was particularly poor due to high leaching losses in the very sandy soil at the experimental site. The Venezuela 350 and Rio Tibagi cultivars only responded to N fertilizer and not to inoculation with Rhizobium which stresses the great importance of selecting plant cultivars for nitrogen fixation in the field.
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  • 70
    ISSN: 1573-5036
    Keywords: Adaptation ; Carbon balance ; Carbon dioxide ; Environmental stress ; Heat stress ; Maintenance respiration ; Nitrogen fixation ; Salinity stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The fitter of two species that use different strategies to overcome the same stress may be the one that expends the least resources to cope with this stress. However, this concept has proven difficult to quantify. It is proposed here that the increase in maintenance respiration in response to stress factors such as high temperature, salinity or a high-oxygen atmosphere (one indirect effect of which is nitrogen deficiency) may provide a measure of the cost of adaptation, in terms of expenditure of assimilated carbon. A corrolary to this is that, where it can be shown that an adaptive strategy results in the expenditure of assimilates, adaptation may be enhanced by increasing carbon assimilation. Results are presented supporting the hypothesis and its corrolary.
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  • 71
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    Plant and soil 90 (1986), S. 141-150 
    ISSN: 1573-5036
    Keywords: Aromatic degradation ; Bacteria ; Nitrogen fixation ; Phenolics ; Physiology ; Soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A microaerobic diazotrophic bacterium tentatively identified as aPseudomonas species was isolated from a forest soil. Its nitrogenase (C2H2 reduction) activity in liquid medium was significantly supported by phenolic compounds when compared with glucose-, mannitol- or malate-supported activity. The utilization of phenolics was dependent on substrate induction and the appropriate oxygen concentration. At a pO2 of 0.05 protocatechuate was a better carbon source for N2 fixation than glucose. In the case ofLignobacter protocatechuate was a better carbon source for N2 fixation than glucose at pO2 0.2 but not at pO2 0.05. It is suggested that certain monomeric phenols can support nitrogenase activities in many carbon-limited soil environments.
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  • 72
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; Nucleotide pools ; Rhodospirillum rubrum ; Switch off
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary When ammonium ions are added to a nitrogen fixing culture ofRhodospirillum rubrum, nitrogenase activity decreases due to inactivation of the Fe-protein. We have studied the adenylate and pyridine nucleotide pools during switch-off using the sensitive bioluminescence method. Immediately after the addition of ammonium ions there is a decrease in the ATP pool which is quickly reversed and no change is seen during the switch-off period. The pyridine nucleotide pools also do not change significantly during the switch-off. Consequently we conclude that changes in the pools studied were not the signal promoting inactivation of the Fe-protein.
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  • 73
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    Plant and soil 90 (1986), S. 193-202 
    ISSN: 1573-5036
    Keywords: azospirillum ; Denitrification ; Nitrate respiration ; Nitrite reductase ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Model experiments were performed to investigate the nitrogen fixation (C2H2 reduction) and denitrification (N2O formation) capabilities ofAzospirillum spp. in association with wheat. Plants and bacteria were grown together for a week and then assayed for activities. This association performed C2H2 reduction or N2O formation, depending on the concentrations of nitrate and oxygen in the vessels. Both activities depended on theAzospirillum strains used. The newly isolatedAzospirillum amazonense strains Y1 and Y6 showed significant C2H2 reduction and low N2O formation in association with wheat under the conditions employed and are possibly useful in practice. A cell-free preparation fromAzospirillum brasilense Sp 7 possessed a cytochrome cd type dissimilatory nitrite reductase.
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  • 74
    ISSN: 1573-5036
    Keywords: Blue grass ; Klebsiella pneumoniae ; Poa pratensis ; Triticum aestivum ; Nitrogen fixation ; Nitrogen nutrition ; 15N isotope dilution ; Spring wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The technique of15N isotope dilution was used to verify that nitrogen was fixed and transferred to the plant byKlebsiella pneumoniae strain Pp in association withPoa pratensis orTriticum aestivum. Surface sterilized, sprouting seeds were inoculated withK. pneumoniae and grown in sand in modified Leonard jars. Potassium nitrate enriched with15N was used to provide N concentrations ranging from 10–40 mg Nl−1 nutrient solution. After 10–18 weeks the shoots and roots were analyzed separately for dry matter, N content, total N, and atom %15N excess. The acetylene reduction technique was used to test for the presence of N2-fixing organisms on the roots. The data from15N isotope dilution demonstrated that up to 33.8% of N in the shoots ofP. pratensis and 15.9% in those ofT. aestivum were derived from associative N2 fixation byK. pneumoniae. In most experiments the dry matter yield, N content, and total N yield of the shoots ofP. pratensis were increased byK. pneumoniae inoculation, whereas inoculation had no significant effect on the dry matter yield, N content or total N of the shoots ofT. aestivum.
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  • 75
    ISSN: 1573-5036
    Keywords: Brachiaria spp ; Nitrogen fixation ; 15N techniques ; Paspalum notatum ; Pasture grasses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Six pasture grasses,Paspalum notatum cv batatais,P. notatum cv pensacola,Brachiaria radicans, B. ruziziensis, B. decumbens andB. humidicola, were grown in concrete cylinders (60 cm diameter) in the field for 31 months. The soil was amended with either a single addition of15N labelled organic matter or frequent small (2 kg N. ha−1) additions of15N enriched (NH4)2SO4. In the labelled fertilizer treatment soil analysis revealed that there was a very drastic change in15N enrichment in plant-available nitrogen (NO 3 − +NH 4 + ) with depth. The different grass cultivars recovered different quantities of applied labelled N, and evidence was obtained to suggest that the roots exploited the soil to different depths thus obtaining different15N enrichments in soil derived N. This invalidated the application of the isotope dilution technique to estimate the contribution of nitrogen fixation to the grass cultivars in this treatment. In the labelled organic matter treatment the15N label in the plant-available N declined at a decreasing rate during the experiment until in the last 12 months the decrease was only from 0.274 to 0.222 atom % excess. There was little change in15N enrichment of available N with depth, hence it was concluded that although the grasses recovered different quantities of labelled N, they all obtained virtually the same15N enrichment in soil derived N. Data from the final harvests of this treatment indicated thatB. humidicola andB. decumbens obtained 30 and 40% respectively of their nitrogen from N2 fixation amounting to an input of 30 and 45 kg N.ha−1 year−1 respectively.
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  • 76
    ISSN: 1573-5036
    Keywords: Azolla caroliniana (Willd.) ; Isotope dilution ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The symbiotic association of the water fernAzolla with the blue-green algaAnabaena azollae can fix 30–60 kg N ha−1 per rice cropping season. The value of this fixed N for rice production, however, is only realized once the N is released from theAzolla biomass and taken up by the rice plants. The availability of N applied asAzolla or as urea was measured in field experiments by two15N methods. In the first,Azolla caroliniana (Willd.) was labelled with15N in nutrient solution and incorporated into the soil at a rate of 144 kg N ha−1. The recovery ofAzolla-N in the above ground parts of rice [Oryza sativa (L) cv. Nucleoryza] was found to be 32% vs. 26% for urea applied at a rate of 100 kg N/ha; there was no significant difference in recovery. In the second, 100 kg N/ha of15N-urea was applied separately or in combination with either 250 or 330 kg N ha−1 of unlabelledAzolla. At the higher rate, the recovery ofAzolla-N was significantly greater than that of urea. There was a significant interaction when both N sources were applied together, which resulted in a greater recovery of N from each source in comparison to that source applied separately. Increasing the combined urea andAzolla application rate from 350 kg N ha−1 to 430 kg N ha−1 increased the N yield but had no effect on the dry matter yield of rice plants. The additional N taken up at the higher level of N application accumulated to a greater extent in the straw compared to the panicles. Since no assumptions need to be made about the contribution of soil N in the method using15N-labelledAzolla, this method is preferable to the15N dilution technique for assessing the availability ofAzolla-N to rice. Pot trials usingAzolla stored at −20°C or following oven-drying showed that both treatments decreased the recovery of N by one third in comparison to freshAzolla.
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  • 77
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    Plant and soil 90 (1986), S. 335-342 
    ISSN: 1573-5036
    Keywords: Azospirillum brasilense ; Effect on yield ; Inoculation ; Legumes ; Nitrogen fixation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Inoculation of naturally nodulatedPisum sativum L. (garden pea) withAzospirillum in the greenhouse caused a significant increase in nodule numbers above controls. Field inoculation of garden peas in the winter 1981–1982 andCicer arietinum L. (chick pea), in winter 1982–1983, withAzospirillum one week after plant emergence, produced a significant increase in seed yield, but did not affect plant dry matter yield. ForVicia sativa L. (vetch) grown in soil in the greenhouse and in the field for forage, winter 1980–1981, inoculation significantly increased dry matter yield, %N, N-content, and acetylene reduction (nitrogen fixation) activity. InHedysarum coronarium L. (sulla clover), winter 1981–1982, inoculated with both its specificRhizobium (by the slurry method) andAzospirillum, 7 days after emergence, there was an increase in acetylene reduction above controls inoculated withRhizobium alone. These results suggest that it is possible, under conditions tested in this work, to increase nodulation, nitrogen fixation, and crop yields of winter legumes by inoculation withAzospirillum.
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  • 78
    ISSN: 1573-5036
    Keywords: Alnus rubra ; Alnus glutinosa ; Fatty acids ; Frankia ; Nitrogen fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Alnus species are used widely in Britain for land reclamation, forestry and other purposes. Rapid juvenile growth of the AmericanAlnus rubra makes it an attractive species for planting on N-deficient soils, particularly those of low organic content. In small plot trials, this species is nodulated by indigenous soil frankiae as effectively asAlnus glutinosa. Over a three year period both species return similar amounts of N to the ecosystem, estimated at up to 10–12 kg N ha−1. Several strains ofFrankia have been isolated from local (Lennox Forest)A. rubra nodules. These differ morphologically and in their growth on different culture media, both from each other and fromA. glutinosa nodule isolates. AllAlnus isolates, however, have a total cellular fatty acid composition qualitatively similar to some other Group B frankiae. Glasshouse tests in N free culture suggest thatA. rubra nodules formed after inoculation of seedlings with American spore (−) isolates are three times more effective in N fixation than those inoculated with LennoxA. rubra spore (+) nodule homogenates. By contrast, the early growth of seedlings inoculated with spore (−)Frankia strains suggests at best a 35% improvement in N fixing activity over seedlings inoculated with LennoxA. rubra nodule isolates. Nevertheless, this improvement in activity, together with the better performance of seedlings inoculated with isolates compared with those treated with crushed nodule preparations, suggest that it would be worthwhile commercially to inoculate nursery stock with a spore (−)Frankia strain.
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  • 79
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    Plant and soil 90 (1986), S. 429-453 
    ISSN: 1573-5036
    Keywords: Actinorhizae ; Frankia ; Genetics ; Nitrogen fixation ; Nodulation ; Symbiosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
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  • 80
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    Plant and soil 91 (1986), S. 43-49 
    ISSN: 1573-5036
    Keywords: Alnus incana ; Frankia ; Leaf litter ; Nitrogen fertilization ; Nitrogen fixation ; Root exudation ; Root litter ; Shoot litter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A pot experiment withAlnus incana (L.) Moench growing in sand was set up to compare the amounts of nitrogen released from plants shoot litter with that released below ground as root litter and/or root exudation. No nitrogen fixation by free-living microorganisms was found in the sand and the increased nitrogen content of the plant + soil system was therefore due to nitrogen fixation byFrankia in the alder root-nodules. Most of the nitrogen released from the plants was in the nitrogen-rich leaf and other shoot litter. Only small amounts of nitrogen were found in the drainage water from the pots and were recorded as increased nitrogen content of the sand.
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  • 81
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    Plant and soil 91 (1986), S. 147-160 
    ISSN: 1573-5036
    Keywords: Cowania ; Frankia ; Microbial ecology ; Nitrogen fixation ; Purshia ; Revegetation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nitrogen fixing trees and shrubs may be useful in revegetation efforts. The possibility that soil and environmental factors may influence a soil's capability to produce nodulated seedlings was explored.Purshia tridentata andCowania mexicana var. Stansburiana seedlings were grown in greenhouse trials using ten soils from native sites for each of the two genera. Treatments included a control and a six mmole nitrogen amendment as NH4NO3 for both surface and subsurface samples. Nodulation was often sparse for seedlings grown in surface collected samples. Although nodulation was usually better in subsoil samples, even some subsoils produced few or no nodules. Nitrogen additions inhibit nodulation and although soil nitrogen may be inhibitory in some unamended surface soils it is probably not a general cause of sparse nodulation. Nodule masses showed the same trends as nodule number but varied less with treatment and depth of soil source. Seedlings compensated for sparse nodulation with an increase in mass per nodule. Incidence of nodulation was related to some soil and environmental factors. Multiple regression analysis explained a substantial portion of nodulation variability. Soils from lower elevations with less precipitation did not produce well nodulated seedlings even in well watered greenhouse trials. Micronutrient cations, potassium, and phosphorus are positively correlated with nodulation incidence. The two genera were generally similar in nodulation responses to soil and environmental factors.
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  • 82
    ISSN: 1573-5036
    Keywords: A-value ; Barley ; Field bean ; Isotope dilution ; Nitrogen fixation ; 15N ; Non-fixing reference crop ; Pea ; Pisum sativum ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The total amount of nitrogen derived from symbiotic nitrogen fixation in two pea and one field bean cultivar, supplied with 50 kg N ha−1 at sowing (‘starter’-N), was estimated to 165, 136, and 186 kg N ha−1, respectively (three-year means). However, estimates varied considerably between the three years. At the full bloom/flat pod growth stage from 30 to 59 per cent of total N2 fixation had taken place. The proportion of total N derived from N2 fixation at maturity was higher in seeds than in vegetative plant parts and amounted to 59.5, 51.3 and 66.3 per cent of total above-ground plant N in the two pea cultivars and field bean, respectively (three-year means). The recovery of fertilizer N was 62.2, 70.2, 52.1, and 69.5 per cent in the two pea cultivars, field bean and barley, respectively. Growth analysis indicated that barley did not meet the claims for an ideal reference crop in the15N fertilizer dilution technique for estimating N2 fixation in pea and field bean. ‘Starter’-N neither increased the seed yield nor the N content of the grain legumes.
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  • 83
    ISSN: 1573-5036
    Keywords: Mycorrhiza ; Nitrogen fixation ; Nodulation ; Soybean ; Symbioses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Glycine max (L. Merr. cv. Amsoy 71) plants were grown in a greenhouse in a sand/perlite medium low in plant-available N and P. Plants were either inoculated with a vesicular-arbuscular mycorrhizal (VAM) fungus alone, a strain ofRhizobium japonicum alone, both endophytes together or were left non-inoculated to serve as a control. All combinations received a N-and P-free nutrient solution. Nodulated plants contained 4 to 5 times the phytomass of non-inoculated controls, and plants colonized with both the VAM fungus and Rhizobium were 18% greater in dry weight than nodulated, non-VAM plants due to a positive VAM times Rhizobium interaction. Nitrogen fixation, calculated from C2H4 and H2 data, was significantly higher in the tripartite symbiosis, with 80% of the increase attributable to increased nodule mass and 20% due to increases in specific nodule activity. Colonization by the VAM fungus and the development of vesicles increased significantly following nodulation. The synergistic interactions between the microsymbionts suggests that the response of the host to dual colonization is complex and depends on a balance between the three members of the symbiosis.
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  • 84
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    Plant and soil 92 (1986), S. 55-62 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Environmental factors ; Low temperature ; Nitrogen fixation ; Rhizobium ; Trifolium pratense
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Red clover Rhizobium strains, isolated from different locations between latitudes 60° and 63°30′ N in Finland, were tested for their adaptation to low temperatures. 31 strains were tested for growth at 5°C, 10°C, 15°C and 18°C in pure culture. No strain grew at 5°C. At the other temperatures there were differences between the strains, but the same strains grew fast at all temperatures. Ten strains were investigated for nodulation and acetylene reduction in phytotrons in two different climates, one simulating the growing season in southern and the other in northern Finland. There were differences between the strains in their ability to nodulate their host plant, and northern strains showed higher nitrogenase activity than southern strains in the cold climate.
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  • 85
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    Plant and soil 92 (1986), S. 171-180 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Associative nitrogen fixation ; Azospirillum ; Nitrogen fixation ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Spring-wheat plant seedlings were inoculated with various isolates of nitrogen-fixing rhizosphere bacteria, includingAzospirillum brasilense, in gnotobiotic sand cultures. Bacteria which had lost their acetylene reduction activity (ARA) during purification did not regain it in the presence of the plant. Bacteria with stable ARA were stimulated to low ARA (maximum 5.6 nmol C2H4 plant−1 h−1) by young (22–32-day) wheat seedlings.
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  • 86
    ISSN: 1573-5036
    Keywords: Mutualism ; Nitrogen fixation ; Sorghum bicolor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Sorghum plants were inoculated withAzospirillum brasilense or received an N-amended nutrient solution. Azospirillum inoculation increased plant dry weight and nitrogen assimilation by 25%. Most plant growth responses to Azospirillum were comparable to application of 2.0 mM N. Increased scavenging of nutrients, altered root permeability or nitrogen fixation are possible explanations for these effects.
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  • 87
    ISSN: 1573-5036
    Keywords: Cowpea ; Nitrogen fixation ; Nodulation ; Pea ; Pesticide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Six carbamate pesticides namely 1-naphthol, sevin, dimetilan, trematan, NaDDC and dymid were studied to see their effect on nodulation and nitrogen fixation inPisum sativum andVigna sinensis. Low concentrations of the pesticides have little effect on nodulation and nitrogen fixation, whereas higher concentrations adversely effect these processes. The results also indicate that then sensitivity depends upon the species of the Rhizobium and also the type of the pesticide. Pesticides belonging to the carbamate group differ in their capacity to affect nodulation and nitroge fixation.
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  • 88
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    Plant and soil 95 (1986), S. 301-313 
    ISSN: 1573-5036
    Keywords: Alnus glutinosa ; Ammonium ; Nitrate ; Nitrate reductase activity ; Nitrogen fixation ; Phyllosphere
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Thein vivo nitrate reductase activity (NRA) was determined inAlnus glutinosa plants grown nonsymbiotically on ammonium, nitrate, a combination of both, or symbiotically with atmospheric nitrogen as the only nitrogen source. Root NRA was absent when ammonium or atmospheric nitrogen was the nitrogen source. With nitrate in the culture solution the roots showed a high NRA. However, the leaf NRA behaved quite differently: with negligible activities on all nitrogen sources except atmospheric nitrogen. The foliar NRA measured, however, is likely not due to the activity of the plant but of microbial origin. Methods commonly used to facilitate produced nitrite to leak out of the tissue, such as addition of propanol and cutting the plant material, did not increase the nitrite release from the leaves. A turbidity developed when testing the samples for nitrite which was positively correlated with the NRA. Populations of microorganisms in the phyllosphere did not differ between the nutritional treatments. Bacteria, able to grow on a low-nitrogen medium, were present on the leaves. Nitrifiers could not be detected. The bacteria on the leaves appear to produce nitrite when incubated with leaf material.
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  • 89
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    Plant and soil 96 (1986), S. 205-213 
    ISSN: 1573-5036
    Keywords: Actinorhizal plants ; Alnus glutinosa ; Alnus incana ; Frankia ; Nitrogen fixation ; Nodules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The distribution of spore positive (Sp+) and spore negative (Sp−) nodules on the two native alder species (A. incana andA. glutinosa) in Finland was investigated. Nodules were collected throughout the country from different ecosystems (forests, swamps, lake- sea- and riversides, old pastures and fields as well as from alder plantations). OnA. incana Sp+ nodules predominated, whereas onA. glutinosa the vast majority of the nodules were of the Sp− type. Sp+ nodules onA. glutinosa were found only at sites where the two alder species grew close together. This distribution pattern indicates an association of nodule type with alder species, the reasons for which are discussed. Indications of saprophytic growth in the Sp− strain were also found.
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  • 90
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    Plant and soil 96 (1986), S. 215-224 
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Daviesia mimosoides ; Eucalypt forest ; Nitrogen fixation ; Nodulation ; Understorey
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Daviesia mimosoides is a common understorey legume in Eucalyptus forests of the Brindabella Range in southeastern Australia, capable of fixing atmospheric nitrogen. Rates of N fixation were measured by the acetylene-reduction technique over a growing season in the field. Pot trials under controlled conditions were also carried out to elucidate effects of soil moisture, temperature, and light. Average rates in the field varied from about 1–5 μ mol C2H4/g/h (wet weight of nodule), but rates up to 14 μ mol C2H4/g/h were measured in optimum controlled conditions. Annual N-fixation rates approximate 4.5–7.0 kg/ha. In pot trials, rate of acetylene reduction decreased with soil moisture to about−10 MPa tension, with a marked depression at about−6 MPa, but within the normal field range of soil moisture there was little correlation of moisture with average acetylene reduction rate. Rates were similar in the temperature range of 20–30°C, but were depressed by either low or high temperature (〈10 or 〉30°C). Diurnal fluctuations in acetylene reduction rates were not correlated with solar radiation, but rates were limited by high mid-day temperatures.
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  • 91
    ISSN: 1573-5036
    Keywords: Acetylene reduction ; Minesoil ; Nitrogen fixation ; Nitrogen source ; Reclamation ; Sericea lespedeza ; Surface mining
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Two growth chamber experiments were conducted to determine the response ofLespedeza cuneata (Dumont) G. Don. (sericea lespedeza) to delayed inoculation and low levels of nitrogen fertilization. Nitrogen was supplied either as NH 4 + or as NO 3 − in solution. At 0.5 and 5.0 ppm nitrogen early growth and N2(C2H2) fixation was inhibited by NH 4 + and promoted by NO 3 − . Inoculation at seeding did not negatively affect growth prior to the onset of N2(C2H2) fixation. Delayed inoculation until the trifoliate stage thus did not increase growth or N2 fixation during the first 40 days of growth. After 40 days, specific nitrogenase activity was highest for plants inoculated at the first trifoliate stage of growth. In contrast, growth and total shoot nitrogen accumulation were higher in plants inoculated at planting. The experimental results suggest that delaying inoculation is not a useful technique for improving early growth ofL. cuneata for surface mine reclamation.
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  • 92
    ISSN: 1573-5036
    Keywords: Fertilizer ; Lupinus ; Nitrogen ; Nitrogen fixation ; Nutrient availability ; Pinus ; Water potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effects ofLupinus angustifolius, L. arboreus. and nitrogen fertilizer plus weedicide on soil inorganic nitrogen, and the growth, nutrition and water status ofPinus radiata seedlings were compared on a sandy podzol in Victoria Australia. Both lupins increased the concentration of soil inorganic nitrogen from 6 μg g−1 to at least 11 μg g−1 one year after treatment.Lupinus arboreus (a perennial) led to high mortality ofP. radiata seedlings and no growth response in survivors because it competed for water during summer. In contrast,L. angustifolius (an annual) senesced prior to summer and resulted in increased fascicle size and stem growth rates ofP. radiata. Growth ofP. radiata withL. angustifolius was similar to that with weedicide only. The concentration. of phosphorus in foliage ofP. radiata seedlings was highest when growth withL. angustifolius. However, the highest growth rate ofP. radiata occurred in the nitrogen fertilizer plus weedicide treatment.
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  • 93
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    Plant and soil 92 (1986), S. 249-254 
    ISSN: 1573-5036
    Keywords: Ecology ; Nitrogen fixation ; Rhizobium leguminosarum populations ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The size and symbiotic effectiveness, withVicia faba, ofRhizobium leguminosarum populations from five locations in southern Britain has been estimated. Population numbers varied from 4.54×103 to 1.69×105. Nitrogen fixing potential differed by up to 30%. The implications of the results for improving the productivity of field beans are discussed.
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  • 94
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    Plant and soil 94 (1986), S. 147-151 
    ISSN: 1573-5036
    Keywords: Nitrogen fixation ; 15N Rhizobia ; Vigna radiata ; Nitrogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Remobilization of15N from vegetative tissue of mungbean (Vigna radiata (L.) Wilczek) into pods was measured during the reproductive phase of growth. Plant tissue was labelled with15N during vegetative development. Experiments were conducted in the field at two sites. At one site the soil provided cowpeas with most of their N but at the other site N fixation provided most of the N. Remobilized N from vegetative tissue to pods occurred soon after they began to develop. The quantity of the labelled N ultimately remobilized to the pods amounted to 50% for one cultivar (Tx33) at the high soil N site and 70% at the low N site. For the other cultivar (Tx13) the values were 25% and 30%, respectively. The two cultivars performed very differently with respect to partitioning of N into pods and the rate of N fixation. Even though more N was accumulated in the shoots of the high N fixing cultivar (Tx13) less total N was contained in the pods.
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  • 95
    ISSN: 1573-5036
    Keywords: Combined nitrogen ; Grain yield ; Inoculation ; Nitrogen fixation ; Rhizobium leguminosarum ; Vicia faba
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nitrogen limitations to the yield of a field crop ofVicia faba have been examined. Application of nitrogen totalling 560 kg/ha increased dry matter yield at flowering by 674 kg/ha (32%) and grain yield at final harvest by 1.6 tonnes/ha (24%). Attempts to reduce nitrogen limitations by replacing the native rhizobia with strains ofRhizobium leguminosarum selected for high rates of nitrogen fixation were unsuccessful but the introduction of poor rhizobia reduced grain yield. The reasons for this and the implications of the results for crop improvement are discussed.
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  • 96
    ISSN: 1573-5036
    Keywords: Alfalfa ; Barley ; Corn ; Management ; Nitrogen ; Nitrogen fixation ; Plow down ; Red clover ; Sweetclover ; Temperate climate ; Trefoil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Under some conditions the plow down of forage legumes increases the yield of subsequent crops, which is usually caused by improved soil N. However, better soil structure is also a contributing factor. Three experiments were conducted to measure the effect of legume plow down on the yield of subsequent corn crops grown at the Ottawa Research Station (ORS), Ottawa, Canada. In all experiments, corn yields were not affected by legume species, legume cultivars, and/or planting methods. Corn yields from barley plots receiving 0, 60, or 120 kg N ha−1 did not differ until two years after establishment in one experiment and three years in another. The data from these experiments indicated that soil N was high at the ORS, which may inhibit N2-fixation by forage legumès in the establishment year. Therefore, legume plow down was not beneficial to subsequent crops under these conditions. Two other experiments were conducted to measure the effect of legume plow down on the yield of subsequent barley crops. In both experiments, barley yields in the field were not affected by legume type or legume seeding density. Greenhouse and field data indicated that the two cuts with removal strategy benefitted the most to succeeding crops. Data from the greenhouse test indicated that soil N levels were not low in the establishment year, and that some cultivars improved soil fertility more than others.
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  • 97
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    Plant and soil 96 (1986), S. 17-29 
    ISSN: 1573-5036
    Keywords: Drought stress ; Growth ; Harvest ; Legume ; Medicago sativa L. ; Nitrogen fixation ; Nitrogen assimilation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Symbiotic N2 fixation, NO 3 − assimilation and protein accumulation in the shoots were measured simultaneously in alfalfa (Medicago sativa L.) grown in the field or in pots, in order to study how the balance between the two modes of nitrogen nutrition could be influenced by agronomic factors, such as harvest, mineral nitrogen supply and drought stress. During periods of rapid growth, fixation and assimilation may function simultaneously; they are antagonistic at the beginning and at the end of the growth cycle, when the nitrogen requirement of the plant is lower. When nitrogen nutrition does not limit growth, mineral nitrogen supply favours assimilation at the expense of fixation, but does not modify the amount of nitrogen accumulated, which is adjusted to the growth capacity of the plant. After cutting, nitrate assimilation compensated for the decrease in fixation and supplied the plant with the nitrogen required by the regrowth, the proliferation of which determined the fixation recovery. Drought stress decreased N2 fixation much more than NO 3 − assimilation. The latter made growth recovery possible when water supply conditions became normal again. These results suggested the existence of an optimum level of nitrate assimilation, which differed depending on the age of the plants and allowed both maximum growth and fixing activity.
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  • 98
    ISSN: 1573-5036
    Keywords: Foliar fertilization ; Fertilizer uptake efficiency ; Late fertilization ; Nitrogen-15 ; Nitrogen fixation ; Nitrogen translocation ; Soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Field experiments were conducted to determine the effects of the amount, time and method of fertilizer N application on the efficiency of N uptake, N2 fixatio and yield of soybean. Soil and foliar fertilizer N, applied during the pod-filling stage were absorbed by plants with equal and high efficiency, compared to an appreciably lower utilization efficiency for N applied before seedling emergence. These results reveal that the soybean roots were active in N uptake during these late stages of growth. Nitrogen fertilization during pod-filling resulted in significant yield increases over the control treatment which received an early application of 20 Kg N/ha. Seed yield increases were, however, more pronounced than total dry matter yield, and virtually all of the late-applied N was translocated into the pods. Nitrogen fixation in soybean was not influenced by the application of 40 kg N/ha to plants as soil or foliar N during the pod-filling stage. However, 80 kg N/ha supplied during pod-filling as 40 kg soil plus 40 kg foliar N/ha significantly reduced the amount of N2 fixed. The results obtained in these studies suggest that inadequate N supply during pod-filling limited soybean yields, and that by the judicious application of fertilizer N during the late stages of growth, it was possible to enhance soybean yields without necessarily inhibiting N2 fixation.
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  • 99
    ISSN: 1573-5036
    Keywords: Azolla ; Blue green alga ; Evapo-transpiratio chamber ; Humidity ; Light condition ; Nitrogen fixation ; Plant environment ; Symbiotic system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The construction and application of a new type of growth chamber, in which different growth conditionsi.e.: temperature, humidity, pH, light intensity, light colour, change in nutrient composition and gas exchange can easily be controlled, are presented. The method has previously been applied to twoAzolla speciesviz. Azolla filiculoides, which is cold tolerant andAzolla pinnata (distinguished in Vietnam as the form Xanh), which is heat tolerant. In the growth chamber natural growth conditions of the Azolla —Anabaena azollae symbiotic association were imitated as much as possible. For testing the system, methods discussed earlier8,14 and some previously presented data, concerning photosynthetic activities, such as oxygen evolution and nitrogen fixation (acetylene reduction) of twoAzolla species39, were partially used. Biomass ofA. filiculoides was measured and reactions to its environment at conditions when grown in the field and in the growth chamber, were studied. Growth and photosynthesis measurements were performed under special light conditions and with whole plants grown under laboratory conditions. Anthocyanin synthesis was studied in relation with humidity. Anthocyanin spectra were analyzed by means of a spectrum-deconvolution method.
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  • 100
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    Plant and soil 100 (1987), S. 157-169 
    ISSN: 1573-5036
    Keywords: Legume ; Nitrogen fixation ; Nodule ; Translocation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Nitrogen (N2) fixed by Rhizobium bacteroids in the legume nodule is excreted as ammonia to the surrounding host cell where it is efficiently assimilated into the amide group of glutamine. Generally glutamine is a minor exported solute of nitrogen, being further metabolised to asparagine in temperate species and to the ureides, allantoin and allantoic acid in tropical species. These solutes serve as the principal translocated forms of nitrogen in xylem. Compartmentalisation of the pathways of nitrogen metabolism and the role of ammonia in regulation of their activity is examined in nodules of both asparagine-forming (Lupinus albus L.) and ureide-forming (Vigna unguiculata L. Walp) symbioses.
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