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  • 1
    Online Resource
    Online Resource
    Hannover : Univ., Inst. für Zierpflanzenbau, Baumschule u. Pflanzenzüchtung, Abt. Spezielle Ertragsphysiologie
    Keywords: Forschungsbericht
    Type of Medium: Online Resource
    Pages: Online-Ressource (30 S., 5,41 MB) , Ill., graph. Darst
    Language: German
    Note: Förderkennzeichen BMBF 50 WB 0010/ 50 BW 0010. - Literaturangaben , Unterschiede zwischen dem gedruckten Dokument und der elektronischen Ressource können nicht ausgeschlossen werden , Auch als gedr. Ausg. vorh , Systemvoraussetzungen: Acrobat reader.
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  • 2
    ISSN: 1432-2048
    Keywords: Cucurbita ; ATPase (protonated) ; Plasma membrane (H+-ATPase) ; Vacuole (H+-ATPase)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A new method of preparing sealed vesicles from membrane fractions of pumpkin hypocotyls in ethanolamine-containing buffers was used to investigate the subcellular localization of H+-ATPase measured as nigericin-stimulated ATPase. In a fluorescence-quench assay, the H+ pump was directly demonstrated. The H+ pump was substrate-specific for Mg·ATP and 0.1 mM diethylstilbestrol completely prevented the development of a Δ pH. The presence of unsupecific phosphatase hampered the detection of nigericin-stimulated ATPase. Unspecific phosphatases could be demonstrated by comparing the broad substrate specificity of the hydrolytic activities of the fractions with the clear preference for Mg·ATP as the substrate for the proton pump. Inhibitor studies showed that neither orthovanadate nor molybdate are absolutely specific for ATPase or acid phosphatase, respectively. Diethylstilbestrol seemed to be a specific inhibitor of ATPase activity in fractions containing nigericin-stimulated ATPase, but it stimulated acid phosphatase which tended to obscure its effect on ATPase activity. Nigericin-stimulated ATPase had its optimum at pH 6.0 and the nigericin effect was K+-dependent. The combination of valinomycin and carbonylcyanide m-chlorophenylhydrazone had a similar effect to nigericin, but singly these ionophores were much less stimulatory. After prolonged centrifugation on linear sucrose gradients, nigericin-stimulated ATPase correlated in dense fractions with plasma membrane markers but a part of it remained at the interphase. This lessdense part of the nigericin-stimulated ATPase could be derived from tonoplast vesicles because α-mannosidase, an enzyme of the vacuolar sap, remained in the upper part of the gradient. Nigericinstimulated ATPase did not correlate with the mitochondrial marker, cytochrome c oxidase, whereas azide inhibition of ATPase activity did.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Glycine ; Vacuolar H−+-ATP-ase ; Tonoplast (negative staining)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Highly purified tonoplast fractions isolated by preparative free-flow electrophoresis from hypocotyls of etiolated soybean (Glycine max L. (Merr.)) were examined by negative-staining electron microscopy, and many but not all vesicles were found to exhibit head and stalk structures resembling the 9-nm stalked F1 ATPase particles reported previously for Neurospora (Bowman et al., 1989, J. Biol. Chem. 264, 15606–15612). The structures show distinguishing characteristics similar to those for Neurospora. These include a cleft in the particle not exhibited by mitochondrial F1 ATPase and a tendency to disappear from the membrane when treated with nitrate plus Mg−2+-ATP-containing solutions. The position of the stalked ATPase structures, indicates that some of the tonoplast vesicles were oriented cytoplasmic side out whereas others were oriented cytoplasmic side in.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Auxin receptor ; Cucurbita (auxin action) ; Growth ; Phospholipase A in auxin action ; Signal transduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microsomal vesicles were prepared from zucchini (Cucurbita pepo L.) hypocotyls containing radioactive phosphatidylethanolamine or phosphatidylcholine, and these lipids were used as substrates by phospholipase A which is activated by auxins. Phospholipase D and phospholipase C hydrolysed the same substrates but were not influenced by auxin. Phospholipase A was activated by the auxins indolyl-3-acetic acid, 2,4-dichlorophenoxyacetic acid and, to a lesser extent, by α-naphthaleneacetic acid whereas the weak auxins 2,3-dichlorophenoxyacetic acid and β-naphthaleneacetic acid were almost inactive. This hormone specificity was also found in growth tests with etiolated zucchini hypocotyls. Phospholipase A activation by auxin was blocked by a polyclonal antibody against the maize auxin-binding protein. We propose that phospholipase A activation is a primary reaction in the signal transduction leading from hormone-binding to the growth response.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Addition of the animal ether phospholipid platelet-activating factor, 1-0-alkyl-2-acetyl-sn-glycero-3-phosphocholine, (PAF) stimulates medium acidification in cultured soybean (Glycine max L.) cells. The pH of the medium after 8–10 hours is on the average one pH unit lower than in controls. With fusicoccin an average pH difference of 1.7 units is reached. Phospholipids, glycerol, 1-oleyl-2-acetyl-sn-glycerol, 1-0-hexadecyl-sn-glycerol, and triolein at the same concentrations as PAF had no stimulatory effect on medium acidification. The detergents CHAPS and deoxycholate lead to alkalinization of the medium whereas lysophosphatidylcholine (LPC), a detergent with structural similarity to PAF, shows no effect.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Planta 161 (1984), S. 394-397 
    ISSN: 1432-2048
    Keywords: Acid growth theory ; ATPase ; Auxin (action mechanism) ; Cucurbita
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of auxin on membrane-bound ATPase activity was studied in a plasma membrane-enriched fraction from zucchini hypocotyls. The apparent KM of ATPase activity for ATP was decreased in the presence of 10-6 M auxin so that at very low ATP concentrations the stimulation of ATPase activity was most obvious. The weak auxin analogue, 2,3-dichlorophenoxyacetic acid, stimulated much less than the active auxin 2,4-dichlorophenoxyacetic acid.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Planta 151 (1981), S. 434-438 
    ISSN: 1432-2048
    Keywords: ATPase ; Auxin ; Cucurbita ; Hypocotyl ; Membrane fractionation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Membrane fractions from Cucurbita maxima hypocotyls were isolated in a medium which inhibits the action of endogenous phospholipases. After removal of soluble phosphatases by Sepharose 2B-CL column chromatography, an auxin-stimulated ATPase activity was found in membrane fractions from linear sucrose gradients. In the presence of 10-4 M phenylacetic acid (PAA), the stimulation by indol-3-acetic acid (IAA) exhibited a bimodal concentration dependence with maximal stimulation of about 50% at 10-6 M IAA. Without PAA, only a high concentration of 10-4 M IAA was stimulatory, whereas 10-6 M IAA had no apparent effect and 10-8 M IAA exhibited weak inhibition. PAA alone had only weak or no effects. The effects of IAA must be considered as hormone-specific. The ATPase activity in the presence of 10-4 M PAA was activated only by 2,4-dichlorophenoxyacetic acid (2,4-D), an active auxin analogue, but not by the inactive stereoisomers, 2,3-D and 3,5-D. Comparison with marker enzyme profiles suggested that part of the auxin-stimulated ATPase was localized on plasma membranes as well as other compartments. Thus, the auxin-stimulated ATPase may become a useful tool in the investigation of the mechanism of action of auxin.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2048
    Keywords: Cucurbita (phospholipid) ; Ether phospholipid ; H+ transport ; Platelet activating factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A plant lipid was isolated from zucchini (Cucurbita pepo L.) membranes and from soybean (Glycine max [L.] Merr) phospholipids by thinlayer chromatography and further purified by high-performance liquid chromatography. This plant lipid was chromatographically very similar to the platelet-activating factor, an ether phospho-lipid with hormone-like properties found in mammals. Both the plant lipid and the platelet-activating factor stimulated ATP-dependent H+ transport in isolated membrane vesicles from zucchini hypocotyls.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-2048
    Keywords: H+-ATPase ; Cucurbita ; Plasma membrane ; Platelet-activating factor ; Phospholipid ; Protein kinase, phospholipid-activated ; Tonoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine, an ether phospholipid from mammals known as platelet-activating factor (PAF), specifically stimulates proton transport in zucchini (Cucurbita pepo L.) microsomes (G.F.E. Scherer, 1985, Biochem. Biophys. Res. Commm. 133, 1160–1167). When plant lipids were analyzed by two-dimensional thin-layer chromatography a lipid was found with chromatographic properties very similar to the PAF (G.F.E. Scherer and B. Stoffel, 1987, Planta, 172, 127–130). This lipid was isolated from zucchini hypocotyls, red beet root, lupin root, maize seedlings and crude soybean phospholipids. It had biological activity similar to that of the PAF, based on phosphorus content, and stimulated the steady-state ΔpH in zucchini hypocotyl microsomes about twofold. Other phospholipids, monoglyceride, diglyceride, triglyceride, oleic acid, phorbol ester, and 1-O-alkylglycerol did not stimulate proton transport. When microsomes were washed the PAF was ineffective but when soluble protein was added the PAF stimulation of H+ transport was reconstituted. The soluble protein responsible for the PAF-dependent stimulation of transport activity could be partially purified by diethylaminoethyl Sephacel column chromatography. In the same fractions where the PAF-dependent transport-stimulatory protien was found, a protein kinase was active. This protein kinase was stimulated twofold either by the PAF or by Ca2+. When Ca2+ was present the PAF did not stimulate protein-kinase activity. When either the PAF, protein kinase, or both were added to membranes isolated on a linear sucrose gradient, ATPase activity was stimulated up to 30%. Comparison with marker enzymes indicated the possibility that tonoplast and plasma-membrane H+-ATPase might be stimulated by the PAF and protein kinase. We speculate that a PAF-dependent protein kinase is involved in the regulation of proton transport in plants in vitro and in vivo.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2048
    Keywords: Cucurbita ; Filipin ; H+-ATPase ; Lepidium ; Plasma membrane ; Tonoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Large-scale preparations of highly purified tonoplast and plasma-membrane vesicles were obtained from roots (garden cress, Lepidium sativum L.) and shoots (etiolated zucchini hypocotyl, Cucurbita pepo L.) of representative dicotyledonous seedlings. When tonoplast-enriched fractions of cress roots were prepared by centrifugation and then subjected to free-flow electrophoresis a highly purified tonoplast fraction was obtained. This fraction from cress roots was characterized by morphometry of filipin-treated freeze-fractured preparations and by enzymology to be about 90% homogeneous. Using latency of nitrate-inhibited ATPase and H+-pumping as criteria we found that the majority of the tonoplast vesicles from both sources were oriented right(cytoplasmic)-side-out. Plasma-membrane vesicles were first purified by two-phase partitioning and then subjected to free-flow electrophoresis for further purification. From cress roots, the fraction of highest purity contained 89% plasma-membrane vesicles as judged by morphometry of filipin-treated, freeze-fractured preparations and by enzymology. From both sources, the major plasma-membrane subfraction in the upper phase after two-phase partitioning was shown to have the least electrophoretic mobility in free-flow electrophoresis and to be oriented right(extracytoplasmic)-side-out a slightly more mobile plasma-membrane subfraction was oriented inside-out and originated after freezing thawing from outside-out plasma-membrane vesicles.
    Type of Medium: Electronic Resource
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