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  • Nitrate reductase  (7)
  • Microbial biomass  (5)
  • Springer  (12)
  • American Institute of Physics
  • Molecular Diversity Preservation International
  • Nature Publishing Group
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Publisher
  • Springer  (12)
  • American Institute of Physics
  • Molecular Diversity Preservation International
  • Nature Publishing Group
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Biology and fertility of soils 22 (1996), S. 299-304 
    ISSN: 1432-0789
    Keywords: Microbial biomass ; Fungal biomass ; Ergosterol ; Fumigation extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Ergosterol and microbial biomass C were measured in 26 arable, 16 grassland and 30 forest soils. The ergosterol content ranged from 0.75 to 12.94 μg g-1 soil. The geometric mean ergosterol content of grassland and forest soils was around 5.5 μg g-1, that of the arable soils 2.14 μg g-1. The ergosterol was significantly correlated with biomass C in the entire group of soils, but not in the subgroups of grassland and forest soils. The geometric mean of the ergosterol: microbial biomass C ratio was 6.0 mg g-1, increasing in the order grassland (5.1), arable land (5.4) and woodland (7.2). The ergosterol:microbial biomass C ratio had a strong negative relationship with the decreasing cation exchange capacity and soil pH, indicating that the fungal part of the total microbial biomass in soils increased when the buffer capacity decreased. The average ergosterol concentration calculated from literature data was 5.1 mg g-1 fungal dry weight. Assuming that fungi contain 46% C, the conversion factor from micrograms ergosterol to micrograms fungal biomass C is 90. For soil samples, neither saponification of the extract nor the more effective direct saponification during extraction seems to be really necessary.
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  • 2
    ISSN: 1432-0789
    Keywords: Analytical variations ; Root intenference ; Root pre-extraction ; Fumigation-extraction ; Microbial biomass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A control soil stored at 4°C was analyzed 38 times by fumigation-extraction during a period of 11 months to correct for variations caused by the analytical procedure. The difference in extractable C between fumigated and unfumigated samples oscillated around the average without a positive or negative trend. When data from contemporaneously extracted field samples were corrected with control soil data the variations were lowered. The deviations between corrected and uncorrected biomass C values had maxima of ±12%. Data obtained for seven dates using pre-extraction, wet-sieving, and centrifuging were compared with data obtained by the conventional procedure without any pretreatment. A negative difference from data obtained without pretreatment was found when the soil water content was decreased to 6%. The largest positive difference (+38%) was found in May during the period of highest root growth.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Biology and fertility of soils 19 (1995), S. 215-219 
    ISSN: 1432-0789
    Keywords: Microbial biomass ; Acidification ; Beech forest ; Soil organic C ; Total P ; Fagus sylvatica
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Thirty-eight soils from forest sites in central Germany dominated by beech trees (Fagus sylvatica L.) were sampled to a depth of about 10 cm after careful removal of the overlying organic layers. Microbial biomass P was estimated by the fumigation — extraction method, measuring the increase in NaHCO3-extractable phosphate. The size of the microbial P pool varied between 17.7 and 174.3 μg P g-1 soil and was on average more than seven times larger than NaHCO3-extractable phosphate. Microbial P was positively correlated with soil organic C and total P, reflecting the importance of soil organic matter as a P source. The mean microbial P concentration was 13.1% of total P, varying in most soils between 6 and 18. Microbial P and microbial C were significantly correlated with each other and had a mean ratio of 14.3. A wide (5.1–26.3) microbial C: P ratio indicates that there is no simple relatinship between these two parameters. The microbial C: P ratio showed strong and positive correlations with soil pH and cation exchange capacity.
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  • 4
    ISSN: 1432-0789
    Keywords: Key words Analytical variations ; Root interference ; Root pre-extraction ; Fumigation-extraction ; Microbial biomass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A control soil stored at 4°C was analyzed 38 times by fumigation-extraction during a period of 11 months to correct for variations caused by the analytical procedure. The difference in extractable C between fumigated and unfumigated samples oscillated around the average without a positive or negative trend. When data from contemporaneously extracted field samples were corrected with control soil data the variations were lowered. The deviations between corrected and uncorrected biomass C values had maxima of ±12%. Data obtained for seven dates using pre-extraction, wet-sieving, and centrifuging were compared with data obtained by the conventional procedure without any pretreatment. A negative difference from data obtained without pretreatment was found when the soil water content was decreased to 6%. The largest positive difference (+38%) was found in May during the period of highest root growth.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Biology and fertility of soils 23 (1996), S. 38-42 
    ISSN: 1432-0789
    Keywords: Microbial biomass ; Depth profile ; Fumigation-extraction method ; Soil organic matter ; Dormant population
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We measured microbial biomass C and soil organic C in soils from one grassland and two arable sites at depths of between 0 and 90 cm. The microbial biomass C content decreased from a maximum of 1147 (0–10 cm layer) to 24 μg g-1 soil (70–90 cm layer) at the grassland site, from 178 (acidic site) and 264 μg g-1 soil (neutral site) at 10–20 cm to values of between 13 and 12 μg g-1 soil (70–90 cm layer) at the two arable sites. No significant depth gradient was observed within the plough layer (0–30 cm depth) for biomass C and soil organic C contents. In general, the microbial biomass C to soil organic C ratio decreased with depth from a maximum of between 1.4 and 2.6% to a minimum of between 0.5 and 0.7% at 70–90 cm in the three soils. Over a 24-week incubation period at 25°C, we examined the survival of microbial biomass in our three soils at depths of between 0 and 90 cm without external substrate. At the end of the incubation experiment, the contents of microbial biomass C at 0–30 cm were significantly lower than the initial values. At depths of between 30 and 90 cm, the microbial biomass C content showed no significant decline in any of the four soils and remained constant up to the end of the experiment. On average, 5.8% of soil organic C was mineralized at 0–30 cm in the three soils and 4.8% at 30–90 cm. Generally, the metabolic quotient qCO2 values increased with depth and were especially large at 70–90 cm in depth.
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  • 6
    ISSN: 1432-072X
    Keywords: Rhodobacter capsulatus ; Periplasmic enzymes ; Nitrate reductase ; Trimethylamine-N-oxide/dimethylsulphoxide/chlorate reductase ; Molybdenum cofactor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The periplasmic dissimilatory nitrate reductase from Rhodobacter capsulatus N22DNAR+ has been purified. It comprises a single type of polypeptide chain with subunit molecular weight 90,000 and does not contain heme. Chlorate is not an alternative substrate. A molybdenum cofactor, of the pterin type found in both nitrate reductases and molybdoenzymes from various sources, is present in nitrate reductase from R. capsulatus at an approximate stoichiometry of 1 molecule per polypeptide chain. This is the first report of the occurrence of the cofactor in a periplasmic enzyme. Trimethylamine-N-oxide reductase activity was fractionated by ion exchange chromatography of periplasmic proteins. The fractionated material was active towards dimethylsulphoxide, chlorate and methionine sulphoxide, but not nitrate. A catalytic polypeptide of molecular weight 46,000 was identified by staining for trimethylamine-N-oxide reductase activity after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. The same polypeptide also stained for dimethylsulphoxide reductase activity which indicates that trimethylamine-N-oxide and dimethylsulphoxide share a common reductase.
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  • 7
    ISSN: 1432-072X
    Keywords: Nitrobacter hamburgensis ; Nitrite oxidoreductase ; Nitrate reductase ; Molybdenum iron-sulfur protein ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrite oxidoreductase, the essential enzyme complex of nitrite oxidizing membranes, was isolated from cells of the nitrifying bacterium Nitrobacter hamburgensis. The enzyme system was solubilized and purified in the presence of 0.25% sodium deoxycholate. Nitrite oxidoreductase oxidized nitrite to nitrate in the presence of ferricyanide. The pH optimum was 8.0, and the apparent K m value for nitrite amounted to 3.6 mM. With reduced methyl-and benzylviologen nitrite oxidoreductase exhibited nitrate reductase activity with an apparent K m value of 0.9 mM for nitrate. NADH was also a suitable electron donor for nitrate reduction. The pH optimum was 7.0. Treatment with SDS resulted in the dissociation into 3 subunits of 116,000, 65,000 and 32,000. The enzyme complex contained iron, molydbenum, sulfur and copper. A c-type cytochrome was present. Isolated nitrite oxidoreductase is a particle of 95±30 Å in diameter.
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  • 8
    ISSN: 1617-4623
    Keywords: Nitrate reductase ; cDNA expression cloning ; Tobacco ; Sequence ; Cytochrome b5
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Messenger RNAs encoding the nitrate reductase apoenzyme from tobacco can be translated in a cell-free system. Poly(A)+ mRNA fractions from the 23-32 S area of a sucrose gradient were used to build a cDNA library in the expression vector λgt11 with an efficiency of cloning of approximately 104 recombinants/ng mRNA. Recombinant clones were screened with a rabbit polyclonal antibody directed against the corn nitrate reductase, which cross reacts specifically with the nitrate reductases from dicotyledons. Among 240000 recombinant plaques, eight clones were isolated containing inserts of sizes ranging from 1.6 kb to 2.1 kb and sharing sequence homologies. Seven of these clones contained a common internal 1.6 kb EcoRI fragment. The identity of these clones was confirmed as follows. A fusion protein of 170 kDa inducible by IPTG and recognized by the rabbit nitrate reductase antibody was expressed by a lysogen derived from one of the recombinants. The antibodies binding the fused protein were eluted and shown to be inhibitory to the catalytic activity of tobacco nitrate reductase. Two monoclonal antibodies directed against nitrate reductase were also able to bind the hybrid protein. The 1.6 kb EcoRI fragment was sequenced by the method of Sanger. The open reading frame corresponding to a translational fusion with the β-galactosidase coding sequence of the vector shared strong homology at the amino acid level with the heme-binding domain of proteins of the cytochrome b5 superfamily and with human erythrocyte cytochrome b5 reductase. When the 1.6 kb EcoRI fragment was used as a probe for Northern blot experiments a signal corresponding to a 3.5 kb RNA was detected in tobacco and in Nicotiana plumbaginifolia mRNA preparations but no cross-hybridization with corn mRNAs was detected. The probe hybridized with low copy number sequences in genomic blots of tobacco DNA.
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  • 9
    ISSN: 1617-4623
    Keywords: Nicotiana tabacum ; Nitrate reductase ; Retrotransposon ; Transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effects of Tntl retrotransposon insertion on nitrate reductase (NR) gene transcription have been analyzed in three NR-deficient insertional, mutants of Nicotiana tabacum. In the three mutants, named h9-Nia4, h9-Nia5 and h9-Nia6, Tnt1 was inserted into exon 3, exon 2 and exon 1 of the nia2 NR alloallelle, respectively. The mutants h9-Nia4 and h9-Nia6, which contained Tnt1 insertions that were oriented opposite to the direction of nia2 gene transcription, expressed chimaeric nia2-Tnt1 RNAs, respectively 12 kb and 10 kb long. The size observed in h9-Nia6 was close to the expected size for a full-length hybrid transcript starting and ending under the control of nia2 signals (about 9 kb). The larger transcript found in h9-Nia4 was shown to be due to a failure to splice the nia2 intron 2. The mutant h9-Nia5, which contained a Tntl insertion oriented in parallel with the direction of nia2 transcription expressed two truncated nia2-Tnt1 RNAs, 2 kb and 6.7 kb long. These transcripts arose from termination in the long terminal repeats (LTRs) of Tull. Since no full-length hybrid RNA was detected, we suggest that Tnt1 carries efficient termination signals, which are more efficiently recognized in the 3′ LTR than in the 5′ LTR.
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  • 10
    ISSN: 1617-4623
    Keywords: Nitrate reductase ; Reporter gene ; Nicotiana tabacum ; Nicotiana plumbaginifolia ; Transgenic plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Reporter gene techniques and mutant analysis were used to identify the molecular basis of the regulation of the expression of nitrate reductase (NR) by nitrate and nitrate-, or ammonium-derived metabolites (N-metabolites), in the true diploïd species Nicotiana plumbaginifolia and in the amphidiploïd species Nicotiana tabacum. The N. plumbaginifolia mutant E23 results from the insertion of a Tnt1-like retrotransposon (Tnp2) in the first exon of the single-copy nia gene, which encodes nitrate reductase. One of the resulting transcripts ends in the 5′ LTR (long terminal repeat) sequence of this retrotransposon, and another one in the 3′ LTR. Nitrate and N-metabolites modulate the expression of these truncated transcripts, indicating that intron splicing and termination processes are not essential to these regulatory events. A GUS reporter sequence was transcriptionally linked to the promoter of the nia-1 gene of N. tabacum. This fusion was functional in transient expression assays done with protoplasts derived from mesophyll cells of N. tabacum. However none of the regulatory mechanisms known to affect steady-state levels of the nia-1 transcript were operative under these experimental conditions. Transgenic plants carrying either this fusion or translational fusions of GUS linked to the promoter of either the nia-1 or nia-2 gene of N. tabacum were obtained by Agrobacterium-mediated transfer. A low proportion of the transgenic plants (22 out of 105 independent transformants) expressed GUS activity although at a low level. Only 4 plants exhibited a detectable level of GUS mRNA. The concentration of this mRNA increased significantly in an NR-deficient background, indicating regulation by N-metabolites. Only 2 plants, however, showed regulation (induction) by nitrate. Attempts to use aux2 or nptII reporter sequences linked to either the nia-1 or nia-2 promoter as marker genes for the selection of regulatory mutants of the nitrate assimilation pathway were unsuccessful because of our inability to isolate transgenic plants in which these reporter genes were properly regulated by nitrate. The implications of these results are discussed.
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