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  • 1
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The resumption of solute uptake capacity lost after gas-shock of Acer pseudoplatanus L. cell suspension cultures is severely inhibited by low temperatures (1°C) and by inhibitors of transcription and translation of protein synthesis such as 2-mercapto-1 (β-4-pyridethyl) benzimidazole (MPB, 40 μg ml−1), puromycin (around 100 μg ml−1) and actinomycin (100 μg ml−1). Cells that have already attained maximum uptake capacity loose it again after less than 1 h in 40 μg ml−1 MPB. Gel-electrophoresis of the external media of the cells shows that the release of proteins into the solution is affected by shock. The results demonstrate that proteins are involved in the mechanism of solute uptake by the cells, so that these proteins are among the factors altered during shock and recovery, and are important for the understanding of the after-effects of shock.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: At concentrations of 10-−3M, Li+ inhibits the recovery of solute uptake capacity of Acer pseudoplatanus L. cell suspension cultures after gas-shock (i.e. after rapid exchange of the atmosphere in the culture flasks for ambient air). It also reduces solute uptake capacity of cells having already attained high rates of uptake during recovery from gas-shock. The effects of Li+ are much greater in cells which have been cultivated in 7 mM K+ solution than in cells cultivated with higher K+ levels (19 mM). Increasing K+ concentration during recovery reverses the effect of 10–3M Li+ and, with sufficiently high concentrations of K+ (≥ 10-−2M) during recovery, the solute uptake capacity of the fully recovered cells can even become greater than that of the control, at least for the low values of substrate concentration (here sulphate 10-−5M). Since Li+ does not affect the time course of solute uptake measured over 15–20 min, it is thought that it interacts with the synthesis and turnover of the solute uptake machinery of the Acer pseudoplatanus cells. Thermodynamic analysis of the flux data also supports the hypothesis that Li+ inhibits the biosynthesis of specific sites of solute permeation, but it does not rule out the possibility that K+ interferes rather on the forces acting on the transport of the considered solutes than on the catalytic structures of permeation.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 57 (1983), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The electrical resting potential across the plasmalemma of Lemna gibba L. (G 1) cells is −230 to −250 mV and the diffusion potential in the presence of 1 mol m−3 KCN + 1 mol m−3 salicylhydroxamic acid is about −100 mV. A concentration of 0.01 mol m−3 HgCl2 depolarises the transmembrane electrical potential in a largely reversible way. When the cells after 16 min of HgCl2-application are returned to Hg-free solution, the transmembrane electrical potential is only depolarised by 24 × 13 mV (SD, n = 13) compared with the potential prior to HgCl2 treatment. In contrast, a 16 min pretreatment with HgCl2 followed by a wash with mercury-free solution reduces the transient depolarisations of transmembrane potential observed after addition of 5 mol m−3 D-glncose or 1 mol m−3 L-alaoine to about 60% of controls. These transient depolarisations are due to the onset of solute uptake. Accordingly, HgCl2-pretreatment inhibits uptake of 14C-3-O-methyl-d-glucose by more than 50% and uptake of 14C-l-alanine by more than 70%. Washing with 1 mol m−3 1,4-dithiothreitol does not reverse this inhibition. It is, therefore, concluded that Hg2+ irreversibly binds to essential SH-groups of the H+-hexose and the H+-amino-acid cotransport carriers of Lemna gibba and inhibits these carriers without appreciably affecting the electrogenic proton-extrusion pump.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Vacuoles were isolated from leaves of Kalanchoë daigremontiana Hamet et Perrier de la Bathie, and the ionic sensitivity of the vacuolar ATPase was studied in vacuole homogenates desalted on Sephadex G-25. The ATPase activity was dependent on the presence of divalent cations (Mg2+≥ Mn2+≥ Ca2+, Co2+; Zn2+ had no effect). Mg2+-dependent ATPase activity was stimulated by anions (Cl− 〉 malate2+, HCO−3), with maximal stimulation at concentrations above 50 mM. Mg2+-Dependent activity was inhibited by NO−3 above 2 mM, but no saturation was observed up to 100 mM. No stimulation by K+ or Na+ was detected; stimulation by NH+4 was abolished by 0.01% (w/v) Triton X-100, suggesting that the NH+4 effect was due to the permeability of vacuolar membrane vesicles to NH3.Trans-tonoplast electrical potentials (Δψ) and intra-vacuolar pH were measured with glass microelectrodes and antimony covered glass micro-pH-electrodes, respectively. Free vacuofes isolated from Kalanchoë tubiflora (Harv.) Hamet were slightly positive with respect to the suspension medium. This Δψ was insensitive to the protonophore FCCP and depolarized by about 4 mV on addition of 50 mM KCl, still remaining about +5 mV. Upon addition of 7 mM Mg-ATP, vacuoles showed an FCCP-sensitive increase of Δψ from +9.2 ± 2.8 (13) to +17.8 ± 3.7 (12) mV [given as x̄± sd (n)] and an internal acidification from pH 5.4 ± 0.2 (11) to pH 4.3 ± 0.4 (12). Mg-ADP and ATP without Mg2+ had no effect on Δψ.It is concluded that the H4 pumping at the tonoplast is due to the functioning of the anion-sensitive vacuolar ATPase and that this is an essential part of the mechanism of nocturnal acid accumulation in CAM.
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  • 5
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Transcellular electrical profiles ofKalanchoë leaf cells were obtained by pushing a glass micro-saltbridge through cells with the tip consecutively in the cell wall, cytoplasm, and vacuole. The electrical resistance of the cell wall was too small to be detectable, that of the plasmalemma and tonoplast was about 0.18–0.21 and 0.16–0.18 Ωm2, respectively. The electrical potential difference between the cytoplasm and the external medium,ψ co , was ≈−180 mV, the potential difference between the vacuole and the medium,ψ vo , was ≈−155mV, and thus the mean potential difference at the tonoplast,ψ vc , was about +25 mV. Potential difference,ψ vo , was independent of proton concentration in the external medium between pH 9 and 5.5, and behaved like an H+-electrode between pH 5 and 3. Depolarizations and hyperpolarizations ofψ vo obtained by increasing and decreasing, respectively, the Na+-concentrations in the medium were smaller than with changing K+-concentrations, suggesting that permeabilities areP Na +/P K +≈-0.23. Assessment of K+-compartmentation by flux analysis gave K+-concentrations in the cytoplasm including chloroplasts (c c) and vacuole (c v) asc c between 200 and 400 mmol kg−1 FrWt andc v ≈-15 mmol kg−1 FrWt. The Nernst criterion suggests that metabolically regulated K+ transport out of the vacuoles concentrates K+ in the cytoplasm. Fusicoccin (10−5 m) hyperpolarizedψ co by about 100 mV and depolarized the positiveψ vc by about 10 mV, the latter presumably being an insignificant effect. The evidence for the existence of proton pumps exchanging H+ and K+ at the plasmalemma and at the tonoplast is discussed.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 81 (1984), S. 149-158 
    ISSN: 1432-1424
    Keywords: crassulacean acid metabolism ; Kalanchoë ; malic acid ; tonoplast ; membrane permeability ; lipid-solution mechanism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary An analysis was carried out of the mechanism of malic-acid efflux from vacuoles of mesophyll cells of the crassulacean acid metabolism (CAM) plantKalanchoë daigremontiana. Following its accumulation in the vacuole as a result of nocturnal CO2 fixation, the malic acid is passively transported back across the tonoplast in the subsequent light period and is decarboxylated in the cytoplasm. Malic-acid efflux was studied using leaf slices in solution or by following malic-acid utilization (deacidification) in leaves of intact plants. Samples of leaf-cell sap were taken at different times during the day-night rhythm to establish the relation between cell-sap pH and malate content. From the empirically determined pK values for malic acid in the cell sap, it was then possible to calculate the proportion of malate existing as the undissociated acid (H2mal0) and in the anionic forms (Hmal1− and mal2−) for all times during the CAM rhythm. In leaf-slice experiments it has been found that the rate of malic-acid efflux increases exponentially with the malic-acid content of the tissue. This is shown to be related to the increasing amounts of H2mal0 present at high malic-acid contents. At low malic-acid contents (〈65 mol m−3), when H2mal0 is not present in significant amounts, efflux must be in the form of Hmal−1 and/or mal2−. At high malic-acid contents it is suggested that efflux occurs predominantly in the form of passive, noncatalyzed diffusion of H2mal0 across the tonoplast by a ‘lipid-solution’ mechanism. This is supported by the fact that the slope of the curve relating efflux to H2mal0 concentration, when corrected for the presumed contributions from Hmal1− and mal2− transport and plotted on a log-log basis, approaches 1.0 at the highest malic-acid contents. Moreover, the permeability coefficient required to be consistent with such a mechanism $$(P_{H_2 mal^0 } = 1.0to2.0 \times 10^{ - 8} m\sec ^{ - 1} )$$ is similar to that estimated from a Collander plot, using the partition coefficient of malic acid between ether and water. We suggest that $$P_{H_2 mal^0 } $$ may be important in determining the maximum amounts of malic acid that can be accumulated during the CAM rhythm.
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  • 7
    ISSN: 1432-2048
    Keywords: ATPase ; Crassulacean acid metabolism ; Kalanchoë ; Protoplast lysis (polybaseinduced) ; Vacuole (ATPase)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A technique is described that allows a relatively rapid and controlled isolation of vacuoles from leaves of the crassulacean acid metabolism (CAM) plant Kalanchoë daigremontiana. The method involves polybase-induced lysis of mesophyllcell protoplasts and isolation of vacuoles on a discontinuous density gradient. ATPase activity is associated with the isolated vacuoles and is not attributable to contamination by cytoplasmic constituents. It is suggested that this ATPase is responsible for the energization of malic-acid accumulation in the vacuole in CAM plants.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Planta 59 (1962), S. 108-114 
    ISSN: 1432-2048
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Zusammenfassung 1. Der Gehalt einiger floralen und extrafloralen Nektare an K+, Na+, Ca++ und Mg++ wurde geprüft. Zum Vergleich werden Ergebnisse von Phosphatbestimmungen und von Analysen der ninhydrinpositiven Substanzen angegeben. 2. Die molaren Verhältnisse lösl. Mg/lösl. Ca und Gesamt-Mg/Gesamt-Ca wurden in verschiedenen Nektargeweben und vergleichsweise in anderen, den Nektarien möglichst benachbarten Pflanzenteilen untersucht. Das Verhältnis lösl. Mg/lösl. Ca ist in den Nektarien desrs hoch. Daraus werden Rückschlüsse aus die Bedeutung des Mg++, bei der Nektarsekretion gezogen.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Planta 63 (1964), S. 103-117 
    ISSN: 1432-2048
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Zusammenfassung A.Nepenthes. 1. Casein und Ovalbumin wurden durch das Sekret offener Kannen mit Fang und geschlossener Kannen verdaut. Mit Casein als Substrat ergaben sich zwei pH-Optima und zwar bei pH 2,1 und bei pH 6,0–6,5, mit Ovalbumin war nur das saurere Optimum nachzuweisen. Das Temperaturoptimum der Caseinverdauung bei pH 6,0 liegt bei 50°C, die Proteinaseaktivität bei pH 2,1 erreicht ihr Optimum bei noch höheren Temperaturen. Es bleibt ungeklärt, ob es sich um zwei verschiedenen Enzyme handelt. 2. Leucinaminopeptidase fand sich nur im Saft offener Kannen mit Fang. Ihr Vorhandensein ist auf die Tätigkeit von Mikroorganismen zurückzuführen. Man könnte demnach von einer Symbiose zwischenNepenthes und den Mikroben sprechen. Triaminopeptidase und Dipeptidasen wurden nicht gefunden. 3. Eine Anregung zur Sekretion von Enzymen durch Substrate wie Casein und Ovalbumin konnte beiNepenthes nicht beobachtet werden. B.Dionaea. Im Saft mit Fleisch gefütterter Klappfallen vonDionaea konnten die Aktivität einer Proteinase (bei Casein als Substrat!), ferner Leucinaminopeptidase, Glycyl-glycin-dipeptidase, Glycyl-l-leucin-dipeptidase und (mit Einschrändkung) Prolidase nachgewiesen werden.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2048
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Zusammenfassung 1. Die SO 4 −− -Aufnahme durch Maiswurzeln mit einer gut ausgebildeten primären Endodermis mit Casparyschem Streifen wurde mikroautoradiographisch untersucht. Die wichtigsten Ergebnisse sind in Abb. 4 zusammengefaßt. a) Bei normalen Bedingungen war die Sulfatkonzentration bei Abbruch der Versuche in der Epidermis sehr groß, in der Rinde und in der Endodermis sehr viel geringer, aber noch deutlich über dem Blindwert, und im Perizykel und Xylem wieder sehr hoch. Im Phloem war die Konzentration etwa halb so groß wie im Xylem. b) Bei Vergiftung der Atmung mit Azid fand sich in der Epidermis ebenfalls eine hohe SO 4 −− -Kozentration. Auch in der Rinde war die Konzentration hier verhältnismäßig groß und sank in der Endodermis und im Zentralzylinder auf ein niedrigeres Niveau ab. c) Es ist daraus zu folgern, daß der Sulfattransport durch die Endodermis unter unseren Versuchsbedingungen (mäßige Sulfatkonzentration; starke Verminderung der Transpiration) überwiegend metabolisch gesteuert wird. Ein Restbetrag an inaktiver Sulfatbewegung durch die Endodermis bleibt bei 98% Hemmung der Atmung erhalten. Der Sulfattransport in der Rinde und durch die Epidermis erfolgt unabhängig von der Atmung. Ein „Siebeffekt” der Epidermis wird erörtert. 2. Durch azidvergiftete Keimwurzeln wurde das Sulfat im ersten Millimeter hinter der Calyptra in das Periblem bis zu dessen Grenze zum Plerom aufgenommen, wobei die Konzentration bis zu dieser Grenze hin anstieg. 3. Junge Seitenwurzeln nahmen mehr Sulfat in das Xylem auf als die Hauptwurzeln an der Einmündungsstelle der Seitenwurzeln.
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