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1Author    J.M F G Aerts, A. W. SchramRequires cookie*
 Title    Isolation of Vacuoles from the Upper Epidermis of Petunia Hybrida Petals I. A Comparison of Isolation Procedures  
 Abstract    Vacuoles were isolated from the upper epiderm is o f petals o f Petunia hybrida line R27 using three different procedures. U tilizing the vacuolar localization o f anthocyanin, vacuolar prep­ arations obtained through polybase treatment, osm otic shock and m echanical forces were com ­ pared on purity, yield and stability. The com parison indicated that application o f the polybase procedure results in the best vacuolar preparations. Vacuoles could be obtained which m ain­ tained their acidic pH and retained their anthocyanin content. 
  Reference    Z. Naturforsch. 40c, 189—1 (1985); received June 26/O ctober 26 1984 
  Published    1985 
  Keywords    Petunia hybrida, Vacuoles, Protoplasts, Anthocyanins 
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 TEI-XML for    default:Reihe_C/40/ZNC-1985-40c-0189.pdf 
 Identifier    ZNC-1985-40c-0189 
 Volume    40 
2Author    Natalia Stoicheva, Iana Tsoneva, DimiterS. DimitrovRequires cookie*
 Title    Protoplast Dielectrophoresis in Axisymmetric Fields  
 Abstract    Pea protoplasts dielectrophoretic coefficients were measured in alternating electric fields of frequency 1 M Hz and voltage 5 V applied between two concentric cylindrical metal electrodes of outer and inner radii 0.24 mm and 1 mm, respectively. They do not vary significantly with solution osmolarity, but* show a clear expressed maximum in isotonic conditions; the values in 0.4 m , 0.5 m , 0.6 m and 0.7 m manitol solutions are (6.5, 10.2, 8.8 and 5.6) x lO * 24 A 2s4/kg, respectively. The average cell radii in those conditions are 14.2 (im, 14.6 [im, 13.8 jim and 13.6 [im. The radii of cell-to-electrode contacts follow the same dependence on the osmolarity as the dielectrophoretic coefficients; they do not depend on the applied voltages up to 18V. The times of cell approach near to the electrode were too short to be explained by the action only of the dielectrophoretic force; consequently an attractive force appeared at very close approach. These results may be of use in future studies of membrane adhesion and fusion. 
  Reference    Z. Naturforsch. 40c, 735 (1985); received February 4 1985 
  Published    1985 
  Keywords    Protoplasts, Dielectrophoresis, Axisymmetric Fields 
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 TEI-XML for    default:Reihe_C/40/ZNC-1985-40c-0735.pdf 
 Identifier    ZNC-1985-40c-0735 
 Volume    40 
3Author    GeorgH. Schmid, Pierre ThibaultRequires cookie*
 Title    Characterization of a Light-Induced Oxygen-Uptake in Tobacco Protoplasts  
 Abstract    Protoplasts prepared from the wild type tobacco N. tabacum var. John W illiam 's Broadleaf ex­ hibit photosynthetic oxygen-evolution if the suspension medium is supplemented with bicarbonate. In the absence of bicarbonate no steady state oxygen-evolution is observed with such prepara­ tions. Instead, an appreciable uptake which is mainly insensitive to D C M U and which persists over hours, and therefore is no mduction phenomenon, is seen. Protoplasts of the tobacco aurea mutant Su/su, which is a plant with an exceptionally high photorespiration, show an oxygen con­ 
  Reference    Z. Naturforsch. 34c, 570—575 (1979); received April 2 1979 
  Published    1979 
  Keywords    Protoplasts, Oxygen-Uptake, Photorespiration, Action Spectrum 
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 TEI-XML for    default:Reihe_C/34/ZNC-1979-34c-0570.pdf 
 Identifier    ZNC-1979-34c-0570 
 Volume    34 
4Author    AlanC. Cassells, FrancesM. CockerRequires cookie*
 Title    TMV Inoculation of Tobacco Protoplasts in the Presence of Protoplast Fusion Agents  
 Abstract    Two methods used to induce protoplast fusion, involving polyethylene glycol (PEG) and high calcium/high pH, respectively, have been investigated in procedures for the inoculation of isolated tobacco protoplasts with TMV. It has been shown that inoculation in the presence of fusion agents involves two stages. Firstly, virus precipitation on to the protoplast surface and secondly, entrapment of virus particles between protoplasts in zones (putatively) o f transient protoplast fusion. 
  Reference    Z. Naturforsch. 35c, 1057—1061 (1980); received June 24/August 22 1980 
  Published    1980 
  Keywords    Polyethylene Glycol, Protoplasts, Tobacco Mosaic Virus 
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 TEI-XML for    default:Reihe_C/35/ZNC-1980-35c-1057.pdf 
 Identifier    ZNC-1980-35c-1057 
 Volume    35 
5Author    Gottfried Küppers, Karl-Josef Diederich, Ulrich ZimRequires cookie*
 Title    Cell Fusion by Simulated Atmospheric Discharges: Further Support for the Hypothesis of Involvement of Electrofusion in Evolution  
 Abstract    Electrofusion of mesophyll cell protoplasts of Avena sativa was induced by simulated atmospheric discharges. It is shown both experimentally and theoretically that the potential differences which occur at quite long distances from the point of lightning stroke are large enough to induce fusion. Besides electromagnetic waves which are emitted during lightning (G. Küppers and U. Zimmermann, FEBS Lett. 164, 323 (1983)) cell fusion may have also occurred directly by means of the voltage built-up on the earth during evolution in response to a lightning stroke. 
  Reference    Z. Naturforsch. 39c, 973—980 (1984); received June 5 1984 
  Published    1984 
  Keywords    Electrofusion, Atmospheric Discharge, Step Voltage, Evolution, Membrane, Protoplasts 
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 TEI-XML for    default:Reihe_C/39/ZNC-1984-39c-0973.pdf 
 Identifier    ZNC-1984-39c-0973 
 Volume    39 
6Author    Ruth Hracky, Jürgen SollRequires cookie*
 Title    Protein Phosphorylation — Dephosphorylation in the Cytosol of Pea Mesophyll Cells  
 Abstract    Soluble protein kinase and protein phosphatase activities were localized in the cytosol of pea mesophyll cells using protoplasts fractionation techniques. The molecular weights of the phos-phorylated cytosolic proteins, as determined by polyacrylamide gel electrophoresis, were 68, 55, 46, 38, 36, 30, 22 and 12 kDa. Histone and, to a much lesser extent, casein but not phosvitin were accepted as exogenous substrates. In every case serine served as acceptor amino acid for the phosphate residue. The protein phosphorylation activity had an alkaline pH optimum, and showed no response to varying Mg2+, Ca2+, Pn cyclo-AMP or calmodulin concentrations. The kinase activity was competitively inhibited by ADP and pyrophosphate with apparent K t values of 0.5 and 0.17 m M , respectively. High ATP concentrations (1 -4 m M) resulted in a strong decrease of radioactivity in the ,2P labeled proteins. It is proposed that the ratio of protein phosphorylation to protein dephosphorylation is regulated by the ATP to ADP ratio in the cytosol. 
  Reference    Z. Naturforsch. 41c, 856 (1986); received July 9/August 5 1986 
  Published    1986 
  Keywords    Protein Kinase, Protein Phosphatase, Cytosol, Protoplasts, Pisum sativum 
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 TEI-XML for    default:Reihe_C/41/ZNC-1986-41c-0856.pdf 
 Identifier    ZNC-1986-41c-0856 
 Volume    41 
7Author    Kirsten Heim, G. Eorg, Kreim Er, M. Ichael, M. Elkonian, Erwin LatzkoRequires cookie*
 Title    Light-Induced Ca2+ Influx into Spinach Protoplasts  
 Abstract    Protoplasts from spinach leaves exhibit a light-induced Ca2+ influx. The half maximum rate of C a2+ influx is achieved at ~ 5 W m"2. The action spectrum o f this influx is similar to that of photosynthesis. Furtherm ore, light-induced Ca2+ influx is abolished by D C M U (2* 0.5 ^ im) and is sensitive to the uncoupler FCCP. Vanadate up to 3 enhances light-induced Ca2+ influx. These data indicate that photosynthetic electron transport is involved in light-induced Ca2+ influx into spinach protoplasts. 
  Reference    Z. Naturforsch. 42c, 283 (1987); received O ctober 24 1986 
  Published    1987 
  Keywords    Calcium-Influx, Protoplasts, Spinach, Light-Induced 
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 TEI-XML for    default:Reihe_C/42/ZNC-1987-42c-0283.pdf 
 Identifier    ZNC-1987-42c-0283 
 Volume    42 
8Author    Ulrike Mackenbrock, Ralph Vogelsang, Wolfgang BarzRequires cookie*
 Title    Isoflavone and Pterocarpan Malonylglucosides and ß -l ,3 -G l u c a n -and Chitin-Hydrolases are Vacuolar Constituents in Chickpea (Cicer arietinum L.)  
 Abstract    Cell suspension cultures of chickpea (Cicer arietinum L.) were used to prepare protoplasts and vacuoles. The vacuolar preparation revealed only slight contaminations of cytoplasmic marker enzymes. H PLC analysis of the vacuolar extract showed that the malonylglucosides of isoflavones, isoflavanones and pterocarpans are exclusively located in the vacuole. Experi­ ments designed to determine the subcellular localization of the isoflavone malonylglucoside: malonylesterase suggest an association of this enzyme with the vacuolar membrane. Finally, a ß-l,3-glucanase and a chitinase with basic isoelectric points were also found to be localized in the chickpea vacuoles. 
  Reference    Z. Naturforsch. 47c, 815—822 (1992); received August 17 1992 
  Published    1992 
  Keywords    Chickpea, Cicer arietinum, Cell Suspension Cultures, Protoplast, Vacuole 
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 TEI-XML for    default:Reihe_C/47/ZNC-1992-47c-0815.pdf 
 Identifier    ZNC-1992-47c-0815 
 Volume    47 
9Author    Arkadiusz Kozubeka, Jacek SkałabRequires cookie*
 Title    Fusiogenic Activity of Natural Amphiphiles, 5-fl-Alkylresorcinols in a Yeast Protoplast System  
 Abstract    Two hom ologues of cereal grain resorcinolic lipids, 5-/7-heptadecylresorcinol and 5-/7-hepta-decenylresorcinol studied in the system employing yeast cell protoplasts showed marked fusiogenic activity. The frequency of hybrid formation induced by studied amphiphiles was significantly higher than that obtained with the use of 40% (w/v) polyethylene glycol 4000. The resorcinolic lipids as fusion-inducing agents did no affect regeneration of the cellular wall. The fusiogenic activity of resorcinolic lipids lost when calcium ions were absent in the medium. Fusiogenic activity of studied amphiphiles is related to their ability to induce non­ bilayer structures within the cellular membranes. 
  Reference    Z. Naturforsch. 50c, 656 (1995); received May 17. June 8. 1995 
  Published    1995 
  Keywords    Phenolic Lipids, 5-Alkylresorcinols, Fusion Yeast, Protoplast 
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 TEI-XML for    default:Reihe_C/50/ZNC-1995-50c-0656.pdf 
 Identifier    ZNC-1995-50c-0656 
 Volume    50 
10Author    Florenz Sasse, Dietlinde Backs-Hüsemann, Wolfgang BarzRequires cookie*
 Title    Isolation and Characterization of Vacuoles from Cell Suspension Cultures of Daucus carota  
 Abstract    A reliable procedure for the isolation o f vacuoles from anthocyanin-containing cells o f Daucus carota cell suspension cultures has been developed. From cells o f the late linear growth phase, protoplasts were prepared and purified in a sucrose/sorbitol gradient. Vacuoles were liberated from these protoplasts by osm otic shock and purified in a M etrizamide step gradient. The vacuole-containing fractions were analysed for their anthocyanin content as a measure for the yield o f vac­ uoles. The purity o f vacuoles was examined by assaying various marker enzym es in both proto­ plasts and vacuoles. The purest vacuolar fraction had 8% o f the total activity o f glucose-6-phos-phate dehydrogenase (marker for cytosol), 8% o f cytochrom e oxidase (m itochondria) and 10% o f N A DPH -cytochrom e c reductase (ER). 55% o f the acid phosphatase activity o f the protoplasts and 35% o f the total malate were recovered in the vacuoles. The vacuolar pool o f am ino acids is quite large. D ata for 15 am ino acids show that 44 to 73% are being located in the vacuole. 
  Reference    Z. Naturforsch. 34c, 848—853 (1979); received M ay 15 1979 
  Published    1979 
  Keywords    Daucus carota, Protoplasts, Vacuoles, Lysosomes, A m ino Acids, Anthocyanin 
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 TEI-XML for    default:Reihe_C/34/ZNC-1979-34c-0848.pdf 
 Identifier    ZNC-1979-34c-0848 
 Volume    34 
11Author    Bdel-B AssetRequires cookie*
 Title    Calcium/Calmodulin Regulated Cell Wall Regeneration in Zea mays Mesophyll Protoplasts  
 Abstract    The composition of newly synthesized cell walls starting from enzymatically isolated Zea mays mesophyll protoplasts was studied. The contents of pectin, cellulose and hemicellulose in addition to the wall-associated proteins were followed in the presence of increasing con­ centrations of calcium ions with or without the calcium channel blockers like lanthanum, lithium ions, verapamil, nifedipine and the calmodulin antagonist trifluperazine. Pectin accu­ mulation was inhibited by all antagonists except lanthanum. Cellulose formation, however, was increased by organic antagonists trifluperazine, verapamil and maximally by nifedipine while it was not affected by the inorganic ions lanthanum or lithium. Hemicellulose accumu­ lated with nifedipine present but significantly decreased by all other antagonists. A dded Ca2+ (5 -1 0 m M) reversed most of the blockers-induced inhibition on pectin and hemicellulose. In the case of cellulose, however, calcium concentration which reversed the inhibitory action of these antagonists was dependent on the antagonist. Starch exhibited little alterations indicat­ ing its minor role in deposition of wall components. Cell wall-associated proteins were nega­ tively affected by lanthanum and verapamil and positively by lithium and nifedipine. G en­ erally, this fraction was found in a negative correlation with pectin levels. Chlorophyll contents were lowered after growth for 48 h; this might be due to repeated cell division. These results are discussed in relation to the mechanism of antagonists and effects of calcium. 
  Reference    Z. Naturforsch. 53c, 33—3 (1998); received November ll/D ecem ber 8 1997 
  Published    1998 
  Keywords    Calcium, Calcium Antagonists, Cell Wall Polysaccharides, Protoplasts, Vicia Faba 
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 TEI-XML for    default:Reihe_C/53/ZNC-1998-53c-0033.pdf 
 Identifier    ZNC-1998-53c-0033 
 Volume    53 
12Author    J.R 0 Dawson, P. E. Dickerson, JanetM. King, F. Sakai, A.R H Trim, J. W. WattsRequires cookie*
 Title    Improved Methods for Infection of Plant Protoplasts with Viral Ribonucleic Acid  
 Abstract    Infection of tobacco protoplasts with R N A of tobacco mosaic virus and the bromoviruses has been studied. The quality of the R N A was very important. Ribonuclease activity was controlled by incorporating 5 mM ZnCl2 in the inoculum. Up to 60% infection has been observed with bromovirus R N A but infection with T M V R N A was much less efficient. A procedure for inoculating in 40% polyethylene glycol was developed. High levels of infection were obtained with all the viruses and viral R N A examined. 
  Reference    Z. Naturforsch. 33c, 548—551 (1978); received May 10 1978 
  Published    1978 
  Keywords    Protoplasts, R N A, Plant Viruses, Tobacco M osaic Virus, Polyethylene Glycol 
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 TEI-XML for    default:Reihe_C/33/ZNC-1978-33c-0548.pdf 
 Identifier    ZNC-1978-33c-0548 
 Volume    33 
13Author    Margareta Proksch, Dieter Strack, Gottfried WeissenböckRequires cookie*
 Title    Incorporation of [I4C]Phenylalanine and [14C|Cinnamic Acid into Leaf Pieces and Mesophyll Protoplasts from Oat Primary Leaves for Studies on Flavonoid Metabolism at the Tissue and Cell Level  
 Abstract    When the abaxial epidermis was peeled from 5 to 6 day old oat primary leaves, and 3 cm segments were floated on radioactive phenylalanine or cinnamic acid solutions, more than 90 per cent of the radioactivity was incorporated within 3 to 7 h depending on the developmental stage of the leaf. C-glycosylflavones were labelled within 15 min and radioactivity in these compounds increased for several hours. Pulse labelling and pulse chase experiments with either phenylalanine or cinnamic acid, unequivocally demonstrate that oat flavones are stable end products o f metabolism. However, this procedure does not distinguish between sequential biosynthesis of various flavones and their interconversion. Cinnamic acid was more efficiently (ca. 20 x) converted into oat leaf flavones than was phenylalanine, when the precursor was fed to leaf pieces, and flavones recovered from mesophyll protoplasts. Different labelling patterns were obtained with whole leaf segments and protoplasts which apparently reflect differences in tissue specific flavone biosynthesis o f mesophyll and epidermis. Isolated mesophyll protoplasts incubated with [14C]cinnamic acid synthesize 14C-labelled flavones characteristic o f the mesophyll, as well as several unidentified phenylpropanoid derivatives not found in the intact tissue. Data suggest that photosynthetically active mesophyll cells are a main site o f tissue specific flavone biosynthesis. 
  Reference    Z. Naturforsch. 36c, 222 (1981); received January 141981 
  Published    1981 
  Keywords    Avena sativa, Primary Leaf Tissues, Protoplasts, [14C]Phenylpropanoid Incorporation, C-Glycosyl-flavones 
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 TEI-XML for    default:Reihe_C/36/ZNC-1981-36c-0222.pdf 
 Identifier    ZNC-1981-36c-0222 
 Volume    36 
14Author    Jörg-Peter Schnitzler, HannsUlrich SeitzRequires cookie*
 Title    Rapid Responses of Cultured Carrot Cells and Protoplasts to an Elicitor from the Cell Wall of Pythium aphanidermatum (Edson) Fitzp  
 Abstract    , A llgem eine Botanik und P flanzenphysiologie, A u f der M orgenstelle Suspension cultured cells of an anthocyanin-containing cell line (D C b) from Daucus carota L. ssp. sativus respond very rapidly to treatment with a soluble carbohydrate elicitor from the wall of the oom ycete Pythium aphanidermatum by synthesizing 4-hydroxybenzoic acid which is incorpo­ rated into the plant cell wall. Enzym es of phenol m etabolism , phenylalanine am m onia-lyase (P A L) and chalcone synthase (C H S), respond to elicitor treatment in different ways. Both the catalytic activity o f PAL and its subunit concentration, measured by means o f im m unoblotting, show a transient increase upon elicitation, whereas CHS the initial enzym e o f the flavonoid pathway, is inhibited after administration of the elicitor. and consequently anthocyanin accum ula­ tion ceases. In protoplasts derived from the cultured cells a very similar elicitor-induced response has been observed. Lacking a cell wall, the protoplasts secrete the 4-hydroxybenzoic acid into the culture fluid. The carrot protoplasts isolated by wall degrading enzym es retain their responsive­ ness to the fungal carbohydrate elicitor. The value o f the protoplast system for studying the im m ediate events following elicitor treatment is discussed. 
  Reference    Z. Naturforsch. 44c, 1020—1028 (1989); received Septem ber 13 1989 
  Published    1989 
  Keywords    Carrot Cell Cultures, Elicitor, 4-Hydroxybenzoic A cid, Protoplasts, Pythium aphanidermatum 
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 TEI-XML for    default:Reihe_C/44/ZNC-1989-44c-1020.pdf 
 Identifier    ZNC-1989-44c-1020 
 Volume    44