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  • 1
    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.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 68 (1986), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The reasons for CO2 release by some CAM plants during the light period were investigated by comparative gas-exchange studies with three leaf-succulent species, Kalanchoë tubiflora Hamet, Sedum morganianum E. Walth. and Sempervivum tectorum L. Sempervivum showed net CO2 output during the day at constant day-night temperatures of both 15 and 25°C, whereas Kalanchoë and Sedum showed no CO2 release at this time at either temperature. Net nocturnal CO2 uptake was markedly reduced at 25°C compared with 15°C in Kalanchoë and Sempervivum, while CO2 was released at night at higher temperature by Sedum if leaf/air vapour pressure difference was also increased.Values for intercellular partial pressure of CO2 (piCO2) were calculated from leaf conductance and were characteristically lowest in the dark and highest in the light period. Maximum values of piCO2 ranged from 264 Pa in Sedum to 87 Pa in Kalanchoë, but were somewhat lower than those reported from direct measurements of the gas phase in other CAM plants. Leaf conductance declined with increasing piCO2 in all three species. Minimum values of approximately 0.2 mmol m−2 s−1 were observed at piCO2= 33 Pa in Kalanchoë and Sedum, with no further decrease at higher piCO2, implying that this largely represented the conductance of the cuticle. In Sempervivum, however, leaf conductance declined more gradually, and was as much as 0.8 mmol m−2 s−1 at the highest observed values of piCO2. The cause of net CO2 output in the light by Sempervivum thus appeared to be lower stomatal sensitivity to CO2, resulting in relatively high leaf conductance even at times when piCO2 greatly exceeded that of ambient air.
    Type of Medium: Electronic Resource
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  • 3
    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.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Approximately 400 species of metal-hyperaccumulating plants are currently known, of which about 75% are endemic to ultra-mafic soils and hyperaccumulate nickel (defined as nickel concentrations exceeding 0.1% of above-ground dry biomass in field samples)1"3. To compare closely related ...
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Planta 148 (1980), S. 35-41 
    ISSN: 1432-2048
    Keywords: Phloem transport ; Ricinus ; Sap exudation ; Solute flux ; Water culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The osmotic characteristics of phloem-sap exudation were examined in soil-grown and watercultured plants of Ricinus communis L. Prolonged exudation occurred from bark incisions in water-cultured plants. Fresh incisions caused large alterations in solute flux, but phloem-sap solute potential Ψs changed by less than ±8% over a period of 7 h. This was associated with a constancy in the levels of sucrose and K+, the principal solutes in the sap. Studies with foliar-applied tracers and leaf-excision experiments suggested that exudation was maintained by solute loading from mature leaves. A wide range of mass transfer values through the phloem was found, these being a function of exudation rate. We consider that the exudation process possesses essentially similar characteristics to phloem transport in the intact plant. The way in which bark incisions bring about large changes in solute flux is discussed in terms of the physical properties of the sieve-tube system.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2048
    Keywords: ATP ; Citrate transport ; Hevea ; Proton pump ; Vacuole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between the electrochemical proton gradient, Δ μH+ − , and citrate transport has been studied in tonoplast vesicles from Hevea brasiliensis (the rubber tree). Vesicles were generated from lyophilized samples of fresh vacuoles obtained from the latex sap. Methylamine was used to measure intravesicular pH and lipophilic ions to determine the electrical potential difference (ΔΨ) across the tonoplast. When incubated at pH 7.5 in the absence of ATP, the tonoplast vesicles showed a ΔpH of 0.6 units (interior acid) and a ΔΨ of about-100 mV (interior negative). This potential is thought to be made up of contributions from an H+ diffusion potential, diffusion potentials from other cations and a Donnan potential arising from the presence of internal citrate. In the presence of 5 mol m-3 MgATP the ΔpH was increased to about 1.0 unit and the ΔΨ to about-10 mV. Under these conditions the proton-motive force (Δ p Δ μH+ − /F) became positive and reached +50 mV. These effects were specific to MgATP (ADP and Mg2+ having no significant effect) and were prevented by the protonophore p-trifluoromethoxycarbonylcyanidephenylhydrazone (FCCP). Citrate uptake by the vesicles was markedly stimulated by MgATP; ADP and Mg2+ again had no effect. Nigericin greatly increased ΔpH and this was associated with a large increase in citrate accumulation. The results indicate that the vesicle membrane possesses a functional H+-translocating ATPase. The Δ μH+ − generated by this ATPase can be used to drive citrate uptake into the vesicles. The properties of the tonoplast vesicles are compared with those of the fresh latex vacuoles.
    Type of Medium: Electronic Resource
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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
    ISSN: 1432-1939
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An investigation was carried out into the photosynthetic pathways of the complete bromeliad flora of Trinidad (West Indies). Carbon-isotope ratios (δ13C values) were used to distinguish obligate C3 and crassulacean acid metabolism (CAM) species. Measurements were also carried out on some species in the field to test for day-night changes in leaf titratable acidity. A wide range of δ13C values was found. The obligate CAM species had values of -10 to -20‰ and the obligate C3 species of -23 to -35‰ CAM was found (a) in the majority of Tillandsia spp. (Tillandsioideae) and (b) in all species of Bromelioideae. The other genera of the Tillandsioideae appeared to be at least predominantly C3. One species, Guzmania monostachia var. monostachia, was identified as a C3-CAM intermediate, and others may well exist in the Trinidad flora. The influence of factors such as source CO2, photosynthetic photon flux density and ambient humidity in determining the δ13C values is discussed. The taxonomic distribution of C3 and CAM species within the Bromeliaceae is analyzed in terms of the life-forms and ecological types recognized by Pittendrigh (1948). The most xerophytic species (the light-demanding “atmospherics”) all show CAM and are restricted to the drier parts of the island. Most of the species with waterstoring “tanks” have a wide geographic distribution: these include light-demanding C3 plants and less light-demanding CAM plants. The shade-tolerant bromeliads, which show a requirement for high ambient humidity, are all C3 plants. We discuss the phylogenetic origins of CAM and the epiphytic habit in the Bromeliaceae.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-2048
    Keywords: Anion transport ; ATPase (tonoplast) ; Crassulacean acid metabolism ; Kalanchoë ; Malate ; Proton transport ; Pyrophosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tonoplast vesicles were prepared from leaf mesophyll homogenates of the crassulacean-acid-metabolism plant Kalanchoë daigremontiana Hamet et Perrier de la Bâthie to study the effects of anions on ATP- and inorganic-pyrophosphate (PPi)-dependent H+ transport. In the presence of gramicidin, substrate hydrolysis by the tonoplast ATPase was characteristically stimulated by chloride and inhibited by nitrate, but was unaffected by malate and a wide range of other organic-acid anions; the PPiase was anion-insensitive. Malate was more effective than chloride both in stimulating ATP- and PPi-dependent vesicle acidification (measured as quinacrine-fluorescence quenching) and in dissipating a pre-existing inside-positive membrane potential (measured as oxonol-V-fluorescence quenching), indicating that malate was more readily transported across the tonoplast. Certain other four-carbon dicarboxylates also supported high rates of vesicle acidification, their order of effectiveness being fumarate ≫ malate ∼-succinate 〉 oxalacetate ∼- tartrate; the five-carbon dicarboxylates 2-oxoglutarate and glutarate were also transported, although at lower rates. Experiments with non-naturally occurring anions indicated that the malate transporter was not stereospecific, but that it required the trans-carboxyl configuration for transport. Shorter-chain or longer-chain dicarboxylates were not transported, and neither were monocarboxylates, the amino-acid anions aspartate and glutamate, nor the tricarboxylate isocitrate. The non-permeant anions maleate and tartronate appeared to be competitive inhibitors of malate transport but did not affect chloride transport, indicating that malate and chloride influx at the tonoplast might be mediated by separate transporters.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2048
    Keywords: Kalanchoë ; Reflection coefficient ; Osmotic pressure ; Turgor pressure ; Xylem sap ; Xylem tension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a study with the crassulacean acid metabolism (CAM) plant Kalanchoë daigremontiana Hamet et Perr. using the pressure probe, Rygol et al. (1987, Planta 172, 487–4493) calculated a value for the reflection coefficient (σ) for malate of 0.6. This value was derived from the relationship between measured changes in cell turgor pressure and malic-acid concentration, and would imply that malate was a relatively ineffective osmoticum. Here we show that the calculation of Rygol et al. (1987) involved the implicit assumption that xylem tension was constant with changing cell turgor pressure and osmotic pressure. This has been shown not to be the case using the pressure-chamber technique. We present an alternative method of deriving a weighted-mean value of a for K. daigremontiana and show that it is not significantly different from 1.0.
    Type of Medium: Electronic Resource
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