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  • Guard cell protoplasts  (2)
  • Vicia (guard cells)  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Planta 161 (1984), S. 27-31 
    ISSN: 1432-2048
    Keywords: Guard cell vacuoles ; Malic acid ; Vacuole (swelling process) ; Vicia (guard cells)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method for the preparation of vacuoles from guard cells ofVicia faba L. is described. Vacuoles were released from guard-cell protoplasts by osmotic shock and purified on a Ficoll gradient. Contamination of the vacuoles was examined by assaying marker enzymes, such as fumarase, glucose-6-phosphate dehydrogenase, phosphofructokinase, acid phosphatase and mannosidase. Potassium ions in the incubation medium caused increases in the volume of the vacuoles by a factor of about 2.6, while the malate level remained unchanged. In contrast, malate synthesis was stimulated during the swelling phase when complete guard-cell protoplasts were exposed to K+. The possible role of K+ as an efficient osmotic effector is discussed.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Planta 149 (1980), S. 52-58 
    ISSN: 1432-2048
    Keywords: Allium ; CO2 Fixation ; Gluconeogenesis ; Guard cell protoplasts ; Malate ; Protoplasts (Guard cells) ; Starch ; Stomates ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Isolated, purified mesophyll and guard-cell protoplasts of Vicia faba L. and Allium cepa L. were exposed to 14CO2 in the light and in the dark. The guard-cell protoplasts of Vicia and Allium did not show any labeling in phosphorylated products of the Calvin cycle, thus appearing to lack the ability to reduce CO2 photosynthetically. In Vicia, high amounts of radioactivity (35%) appeared in starch after 60-s pulses of 14CO2 both in the light and in the dark. Presumably, the 14CO2 is fixed into the malate via PEP carboxylase and then metabolized into starch as the final product of gluconeogenesis. This is supported by the fact that guard-cell protoplasts exposed to malic acid uniformly labeled with 14CO2 showed high amounts of labeled starch after the incubation, whereas cells labeled with [4-14C]malate had minimal amounts of labeled starch (1/120). In contrast, the starch-deficient Allium, guard-cell protoplasts did not show any significant 14CO2 fixation. However, adding PEP to an homogenate stimulated 14CO2 uptake, thus supporting the interpretation that the presence of starch as a source of PEP is necessary for incorporating CO2 and delivering malate. With starch-containing Vicia guard-cell protoplasts, the correlation between changes in volume and the interconversion of malate and starch was demonstrated. It was shown that the rapid gluconeogenic conversion of malate into starch prevents an increase of the volume of the protoplasts, whereas the degradation of starch to malate is accompanied by a swelling of the protoplasts.
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  • 3
    ISSN: 1432-2048
    Keywords: Guard cell protoplast ; Malate sensitivity ; Phosphoenolpyruvate carboxylase ; Protoplast (guard cell) ; Vicia (guard cells)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Properties of phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31) obtained from isolated guard-cell protoplasts of Vicia faba L. were determined following rapidly desalting of the extract on a Sephadex G 25 column. The activity of PEP carboxylase was measured as a function of PEP and malate concentration, pH and K+ concentration within 2–3 min after homogenization of the guard-cell protoplasts. The activity of this enzyme was stimulated by PEP concentrations of 0.1 to 0.75 mM and by K+ ions (12 mM), but inhibited by PEP concentrations above 1 mM and by malate. Changes in the Km(PEP) and Vmax values with increasing malate concentrations (2.5 and 5 mM) indicate that the malate level, varying in relation to the physiological state of guard cells, plays an important role in regulating the properties of phosphoenolpyruvate carboxylase.
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  • 4
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Allium ; Fusicoccin ; Guard cell protoplasts ; Vicia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Guard cell protoplasts from starch-containing Vicia faba and starch-deficient Allium cepa stomata were isolated, stabilized and recovered with an efficiency — in relation to the potential yield — of approx. 62% and 77%, respectively. In vitro, guard cell protoplasts (GCP) respond to abscisic acid and fusicoccin by respectively contracting and swelling, that is, decreasing or increasing in diameter by about 15% and more in comparison to the control. This in vitro response correlates with, but is more than 4 times as rapid as, the in vivo response of the stomata. Among the advantages presented by working with isolated GCPs are: greater sensitivity in response; freedom from influences of cuticular ridges, cell walls, subsidiary cells, and epidermal cells; and direct and parallel comparisons of starch-containing and starch-deficient GCP systems.
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