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  • 1
    ISSN: 1432-2048
    Keywords: Acetate uptake ; Beta-Glutamine transport ; Plasma membrane ; Ricinus ; Sucrose transport ; Tetraphenylphosphonium uptake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plasma membrane vesicles, purified by aqueous two-phase partitioning, were used to investigate the presence of sugar and amino acid carriers in cotyledons and roots of Ricinus communis L. and in roots of red beet (Beta vulgaris L.). Artificial pH and electrical gradients were generated across the plasma membrane, and [14C]acetate and [14C]tetraphenylphosphonium were used to demonstrate the presence of an internal alkaline pH gradient and an internal negative membrane potential, respectively. In Ricinus cotyledons, uptake of sucrose was more strongly inhibited than that of glutamine by p-chloromercuribenzenesulphonic acid, phlorizin and phenylglyoxal. The sucrose transport system showed a high degree of substrate specificity with only the presence of maltose and phenyl-α-glucoside significantly affecting sucrose uptake; in contrast, the glutamine transport system was inhibited by a number of other amino acids. ΔpH+gDψ-driven glutamine uptake showed saturation kinetics with a K m of 0.35 mol · m−3. Sucrose and glutamine Δψ-driven uptake was pH dependent with an optimum in the acidic range (pH 6.25) and a decrease at higher pH values. Vesicles obtained from cotyledons and roots of Ricinus showed different transport properties. In the cotyledons, gDH+gDψ-driven transport for both sucrose and glutamine were observed at similar levels; however, in the root tissue, δpH-Δψ-driven glutamine transport was the dominant uptake process. Uptake rates for glucose and fructose were low in the cotyledons whereas, in the roots, glucose and sucrose transport were slightly higher than that of fructose. In vesicles from red beet tissue there was a different uptake profile, with evidence of proton-coupled cotransport systems for sucrose and glucose, but lower uptake of glutamine and fructose. The results are discussed in relation to the reported different pathways for loading and unloading of solutes in these tissues.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Auxin ; Cell elongation ; Cytochalasin B ; Dictyosomes ; Secretion ; Triticum ; Zea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cytochalasin B (CB) inhibits the elongation growth of maize roots, and that of wheat coleoptile segments incubated in indolyl-3-acetic acid, by over 30% after a lag period of about 60 min. This long lag is not due to poor tissue penetration by the inhibitor, but seems to reflect a property of the process inhibited by CB. The only visible ultrastructural change accompanying growth inhibition is the accumulation of secretory vesicles in the vicinity of dictyosomes, which occurs between 90 and 300 min. However, a massive accumulation of vesicles is seen after 120 min in root cap cells which possess very active dictyosomes. The results indicate that CB does not inhibit elongation growth by interfering with cytoplasmic streaming. Instead, they indicate that the drug acts to inhibit the secretion of cell wall components at some stage after vesicle production, but prior to their transport.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Glutamine transport ; Plasma membrane ; Proton cotransport ; Ricinus (solute transport) ; Sucrose transport ; Valinomycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Evidence is presented for the proton-coupled transport of sucrose and glutamine in purified plasma membrane vesicles isolated from cotyledons ofRicinus communis. Imposition of a pH gradient (internal alkaline) across the plasma membrane resulted in a rapid uptake of sucrose and glutamine which was inhibited in the presence of carbonyl cyanide-m-chlorophenyl hydrazone. Imposition of a pH gradient plus an internal negative membrane potential stimulated uptake further. Glucose and fructose uptakes were negligible under these conditions. Sucrose uptake into the vesicles demonstrated saturation kinetics with a Km of 0.87 mol·m-3, indicating carrier-mediated transport. In support of this, uptake was very sensitive to the protein-modifying reagentp-chloromercuribenzenesulphonic acid. N-Ethylmaleimide, another sulphydryl reagent, was only slightly inhibitory. However, both reagents strongly inhibited sucrose uptake into intact cotyledons; the possible reasons for the difference between the intact and isolated systems are assessed. The value of this system for the study of sucrose and amino acid carriers is discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: ATPase ; Plasma membrane ; Pyrophosphatase ; Ricinus (solute transport) ; Sucrose transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A highly enriched plasma membrane fraction has been isolated from dark-grown cotyledons ofRicinus communis by phase partitioning. This is demonstrated by the properties of the associated ATPase: high vanadate sensitivity, azide and nitrate insensitivity, sharp pH optimum around 6.5, and high specificity for ATP as substrate. The upper plasma membrane fraction also contained a pyrophosphatase activity, normally considered to be located on the tonoplast or Golgi membranes, which showed a specific activity higher than that in the lower phase. Sucrose gradient centrifugation of both microsomal and upper phase fractions showed a comigration of some pyrophosphatase activity with the plasma membrane fraction. Sucrose uptake changes with development inRicinus cotyledons. The ATPase activity in the upper (plasma membrane) phase also varied in a similar way with development, whereas activity in the lower phase showed little change. Pyrophosphatase activity in the upper phase also increased with development but did not show a peak and fall as seen for sucrose uptake and ATPase. The possibility that changes in plasma membrane ATPase may contribute to changes in sucrose uptake capacity and the possible cellular origin and physiological significance of the pyrophosphatase activity are discussed.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2048
    Keywords: Auxin ; Cell elongation ; Epidermis peeling ; Fusicoccin ; Pisum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effects of peeling and wounding on the indole-3-acetic acid (IAA) and fusicoccin (FC) growth response of etiolated Pisum sativum L. cv. Alaska stem tissue were examined. Over a 5 h growth period, peeling was found to virtually eliminate the IAA response, but about 30% of the FC response remained. In contrast, unpeeled segments wounded with six vertical slits exhibited significant responses to both IAA and FC, indicating that peeling does not act by damaging the tissue. Microscopy showed that the epidermis was removed intact and that the underlying tissue was essentially undamaged. Neither the addition of 2% sucrose to the incubation medium nor the use of a range of IAA concentrations down to 10-8 M restored IAA-induced growth in peeled segments, suggesting that lack of osmotic solutes and supra-optimal uptake of IAA were not important factors over this time period. It is concluded that, although the possibility remains that peeling merely allows leakage of hydrogen ions into the medium, it seems more likely that peeling off the epidermis removes the auxin responsive tissue.
    Type of Medium: Electronic Resource
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