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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Planta 153 (1981), S. 430-435 
    ISSN: 1432-2048
    Keywords: Chloroplast ; Photosynthesis (stress recovery) ; Protoplast ; Spinacia ; Water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reversibility of the inhibition of photosynthetic reactions by water stress was examined with four systems of increasing complexity—stromal enzymes, intact chloroplasts, mesophyll protoplasts, and leaf slices. The inhibition of soluble chloroplast enzymes by high solute concentrations was instantly relieved when solutes were properly diluted. In contrast, photosynthesis was not restored but actually more inhibited when isolated chloroplasts exposed to hypertonic stress were transferred to conditions optimal for photosynthesis of unstressed chloroplasts. Upon transfer, chloroplast volumes increased beyond the volumes of unstressed chloroplasts, and partial envelope rupture occurred. In protoplasts and leaf slices, considerable and rapid, but incomplete restoration of photosynthesis was observed during transfer from hypertonic to isotonic conditions. Chloroplast envelopes did not rupture in situ during water uptake. It is concluded that inhibition of photosynthesis by severe water stress is at the biochemical level brought about in part by reversible inhibition of chloroplast enzymes and in part by membrane damage which requires repair mechanisms for reversibility. Both soluble enzymes and membranes appear to be affected by the increased concentration of internal solutes, which is caused by dehydration.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Key words: Apoplast ; Buffer capacity ; Chloroplast ; Cytosol ; pH regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract.  The regulation of pH in the apoplast, cytosol and chloroplasts of intact leaves was studied by means of fluorescent pH indicators and as a response of photosynthesis to acid stress. The apoplastic pH increased under anaerobiosis. Aeration reversed this effect. Apoplastic responses to CO2, HCl or NH3 differed considerably. Whereas HCl and ammonia caused rapid acidification or alkalinization, the return to initial pH values was slow after cessation of fumigation. Addition of CO2 either did not produce the acidification expected on the basis of known apoplastic buffering or even caused some alkalinization. Removal of CO2 shifted the apoplastic pH into the alkaline range before the pH returned to initial steady-state levels. In the presence of vanadate, the alkaline shift was absent and the apoplastic pH returned slowly to the initial level when CO2 was removed from the atmosphere. In contrast to the response of the apoplast, anaerobiosis acidified the cytosol or, in some species, had little effect on its pH. Acidification was rapidly reversed upon re-admission of oxygen. The CO2-dependent pH changes were very fast in the cytosol. Considerable alkalinization was observed after removal of CO2 under aerobic, but not under anaerobic conditions. Rates of the re-entry of protons into the cytosol during recovery from CO2 stress increased in the presence of oxygen with the length of previous exposure to high CO2. Effective pH regulation in the chloroplasts was indicated by the recovery of photosynthesis after the transient inhibition of photosynthetic electron flow when CO2 was increased from 0.038% to 16% in air. As photosynthesis became inhibited under high CO2, reduction of the electron transport chain increased transiently. The time required for recovery of photosynthesis from inhibition during persistent CO2 stress was similar to the time required for establishing steady-state pH values in the cytosol under acid stress. The high capacity of leaf cells for the rapid re-attainment of pH homeostasis in the apoplast and the cytoplasm under acid or alkaline stress suggested the rapid activation or deactivation of membrane-localised proton-transporting enzymes and corresponding ion channel regulation for co-transport of anions or counter-transport of cations together with proton fluxes. Acidification of the cytoplasm appeared to activate energy-dependent proton export primarily into the vacuoles whereas apoplastic alkalinization resulted in the pumping of protons into the apoplast. Proton export rates from the cytosol into the apoplast after anaerobiosis were about 100 nmol (m2 leaf area)−1 s−1 or less. Proton export under acid stress into the vacuole was about 1200 nmol m−2 s−1. The kinetics of pH responses to the addition or withdrawal of CO2 indicated the presence of carbonic anhydrase in the cytosol, but not in the apoplast.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Chloroplast metabolism (metabolism regulation) ; Photosystem I (P700 oxidation) ; Electron transport ; Ferredoxin-NADP reductase ; Glyceraldehydephosphate dehydrogenase ; Photosynthesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract P700 is rapidly, but only transiently photooxidized upon illuminating dark-adapted leaves. Initial oxidation is followed by a reductive phase even under far-red illumination which excites predominantly photosystem (PS) I. In this phase, oxidized P700 is reduced by electrons coming from PSII. Charge separation in the reaction center of PSI is prevented by the unavailability of electron acceptors on the reducing side of PSI. It is subsequently made possible by the opening of an electron gate which is situated between PSI and the electron acceptor phosphoglycerate. Electron acceptors immediately available for reduction while the gate is closed corresponded to 10 nmol · (mg chlorophyll)−1 electrons in geranium leaves, 16 nmol · (mg chlorophyll)−1 in sunflower and 22 nmol · (mg chlorophyll)−1 in oleander. Reduction of NADP during the initial phase of P700 oxidation showed that the electron gate was not represented by ferredoxin-NADP reductase. Availability of ATP indicated that electron flow was not hindered by deactivation of the thylakoid ATP synthetase. It is concluded that NADP-dependent glyceraldehydephosphate dehydrogenase is completely deactivated in the dark and activated in the light. The rate of activation depends on the length of the preceding dark period. As chloroplasts contain both NAD- and NADP-dependent glyceraldehydephosphate dehydrogenases, deactivation of the NADP-dependent enzyme disconnects chloroplast NAD and NADP systems and prevents phosphoglycerate reduction in the dark at the expense of NADPH and ATP which are generated by glucose-6-phosphate oxidation and glycolytic starch breakdown, respectively.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Planta 182 (1990), S. 253-261 
    ISSN: 1432-2048
    Keywords: Cytosol ; Metabolite ; Photosynthesis ; pH regulation ; Transport ; Vacuoles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Illumination of leaves of C3 plants caused cytosolic alkalization and vacuolar acidification in the mesophyll cells. Both phenomena were particularly pronounced when CO2 was absent, were suppressed by CO2, and were related to the activation state of the photosynthetic apparatus. The cytosolic alkalization reaction has at least two major components. Trivalent cytosolic phosphoglycerate must be protonated before it can be transferred into the chloroplasts for reduction. Pumping of protons from the cytosol into the vacuole also contributes to cytosolic alkalization. The dependence of light scattering by chloroplast thylakoids on the energy fluence rate was closely related to that of vacuolar acidification under different conditions for chloroplast energization. This indicates (i) transport of energy from the chloroplasts to the cytosol in the light and (ii) use of this energy for the transport of protons into the vacuoles. The light-dependent vacuolar acidification is interpreted to be caused by the increase in the activity of a proton-translocating enzyme of the tonoplast. The decrease of vacuolar acidification during photosynthetic carbon reduction or photorespiration is indicative of decreased cytosolic energization. In low light, the light-dependent vacuolar acidification was stimulated in the absence of CO2 when photorespiration was inhibited. The data do not support the view that photorespiration is capable of increasing the cytosolic energy state in the light.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Planta 182 (1990), S. 244-252 
    ISSN: 1432-2048
    Keywords: Chloroplast ; Cytosol ; Photosynthesis ; pH ; Vacuole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chloroplasts, mesophyll protoplasts, cytoplasts, vacuoplasts, vacuoles and leaves were stained with pH-indicating fluorescent dyes of differing pK values. Fluorescence microscopy was used to obtain information on the intracellular and intercellular distribution of the probes. The kinetics of blue or green fluorescence emitted from chloroplasts, protoplasts, cytoplasts and leaves was measured during illumination with red light. The intensity of light used for fluorescence excitation was so low that it had little effect on photosynthesis. In leaves, fluorescence signals emitted from chloroplasts were small and usually insignificant compared to signals originating from the cytosol. Both indicated light-dependent alkalization and reversal of alkalization on darkening. Vacuolar signals were opposite in sign to cytosolic signals. They indicated acidification of the vacuole in the light-dark transient and reversal of this effect on darkening.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-5079
    Keywords: chlorophyll fluorescence ; energy dissipation ; light scattering ; photosynthesis ; state transition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The light-induced induction of components of non-photochemical quenching of chlorophyll fluorescence which are distinguished by different rates of dark relaxation (qNf, rapidly relaxing and qNs, slowly relaxing or not relaxing at all in the presence brief saturating light pulses which interrupt darkness at low frequencies) was studied in leaves of spinach. After dark adaptation of the leaves, a fast relaxing component developed in low light only after a lag phase. Quenching increased towards a maximum with increasing photon flux density. This ‘fast’ component of quenching was identified as energy-dependent quenching qE. It required formation of an appreciable transthylakoid ΔpH and was insignificant when darkened spinach leaves received 1 s pulses of light every 30 s even though zeaxanthin was formed from violaxanthin under these conditions. Another quenching component termed qNs developed in low light without a lag phase. It was not dependent on a transthylakoid pH gradient, decayed exponentially with a long half time of relaxation and was about 20% of total quenching irrespective of light intensity. When darkened leaves were flashed at frequencies higher than 0.004 Hz with 1 s light pulses, this quenching also appeared. Its extent was very considerable, and it did not require formation of zeaxanthin. Relaxation was accelerated by far-red light, and this acceleration was abolished by NaF. We suggest that qNs is the result of a so-called state transition, in which LHC II moves after its phosphorylation from fluorescent PS II to nonfluorescent PS I. This state transition was capable of decreasing in darkened leaves the potential maximum quantum efficiency of electron flow through Photosystem II by about 20%.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-5079
    Keywords: chlorophyll fluorescence ; cyclic electron transport ; photorespiration ; photosynthesis ; Photosystem II ; proton gradient
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chlorophyll fluorescence, light scattering, the electrochromic shift P515 and levels of some photosynthetic intermediates were measured in illuminated leaves. Oxygen and CO2 concentrations in the gas phase were varied in order to obtain information on control of Photosystem II activity under conditions such as produced by water stress, when stomatal closure restricts access of CO2 to the photosynthetic apparatus. Light scattering and energy-dependent fluorescence quenching indicated a high level of chloroplast energization under high intensity illumination even when linear electron transport was curtailed in CO2-free air or in 1% oxygen with 35 μll-1 CO2. Calculations of the phosphorylation potential based on measurements of phosphoglycerate, dihydroxyacetone phosphate and NADP revealed ratios of intrathylakoid to extrathylakoid proton concentrations, which were only somewhat higher in air containing 35 μl l-1 CO2 than in CO2-free air or 1% oxygen/35 μl l-1 CO2. Anaerobic conditions prevented appreciable chloroplast energization. Acceptor-limitation of electron flow resulted in a high reduction level of the electron transport chain, which is characterized by decreased oxidation of P700, not only under anaerobic conditions, but also in air, when CO2 was absent, and in 1% oxygen, when the CO2 concentration was reduced to 35 μll-1. Efficient control of electron transport was indicated by the photoaccumulation of P700 + at or close to the CO2 compensation point in air. It is proposed to require the interplay between photorespiratory and photosynthetic electron flows, electron flow to oxygen and cyclic electron flow. The field-indicating electrochromic shift (P515) measured as a rapid absorption decrease on switching the light off followed closely the extent of photoaccumulation of P700 + in the light.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1573-5079
    Keywords: 9-aminoacridine fluorescence ; chlorophyll fluorescence ; cyclic electron transport ; light scattering ; photosynthesis ; transthylakoid proton gradient
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transthylakoid proton transport based on Photosystem I-dependent cyclic electron transport has been demonstrated in isolated intact spinach chloroplasts already at very low photon flux densities when the acceptor side of Photosystem I (PS I) was largely closed. It was under strict redox control. In spinach leaves, high intensity flashes given every 50 s on top of far-red, but not on top of red background light decreased the activity of Photosystem II (PS II) in the absence of appreciable linear electron transport even when excitation of PS II by the background light was extremely weak. Downregulation of PS II was a consequence of cyclic electron transport as shown by differences in the redox state of P700 in the absence and the presence of CO2 which drained electrons from the cyclic pathway eliminating control of PS II. In the presence of CO2, cyclic electron transport comes into play only at higher photon flux densities. At H+/e=3 in linear electron transport, it does not appear to contribute much ATP for carbon reduction in C3 plants. Rather, its function is to control the activity of PS II. Control is necessary to prevent excessive reduction of the electron transport chain. This helps to protect the photosynthetic apparatus of leaves against photoinactivation under light stress.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1573-5079
    Keywords: chlorophyll fluorescence ; cyclic electron flow ; high temperature ; light scattering ; photosynthesis ; Photosystems II and I
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In illuminated intact spinach chloroplasts, warming to and beyond 40 °C increased the proton permeability of thylakoids before linear electron transport through Photosystem II was inhibited. Simultaneously, antimycin A-sensitive cyclic electron transport around Photosystem II was activated with oxygen or CO2, but not with nitrite as electron acceptors. Between 40 to 42 °C, activation of cyclic electron transport balanced the loss of protons so that a sizeable transthylakoid proton gradient was maintained. When the temperature of darkened spinach leaves was slowly increased to 40°C, reduction of the quinone acceptor of Photosystem II, QA, increased particularly when respiratory CO2 production and autoxidation of plastoquinones was inhibited by decreasing the oxygen content of the atmosphere from 21 to 1%. Simultaneously, Photosystem II activity was partially lost. The enhanced dark QA reduction disappeared after the leaf temperature was decreased to 20 °C. No membrane energization was detected by light-scattering measurements during heating the leaf in the dark. In illuminated spinach leaves, light scattering and nonphotochemical quenching of chlorophyll fluorescence increased during warming to about 40 °C while Photosystem II activity was lost, suggesting extra energization of thylakoid membranes that is unrelated to Photosystem II functioning. After P700 was oxidized by far-red light, its reduction in the dark was biphasic. It was accelerated by factors of up to 10 (fast component) or even 25 (slow component) after short heat exposure of the leaves. Similar acceleration was observed at 20 °C when anaerobiosis or KCN were used to inhibit respiratory oxidation of reductants. Methyl viologen, which accepts electrons from reducing side of Photosystem II, completely abolished heat-induced acceleration of P700+ reduction after far-red light. The data show that increasing the temperature of isolated chloroplasts or intact spinach leaves to about 40 °C not only inhibits linear electron flow through Photosystem II but also activates Photosystem I-driven cyclic electron transport pathways capable of contributing to the transthylakoid proton gradient. Heterogeneity of the kinetics of P700+ reduction after far-red oxidation is discussed in terms of Photosystem I-dependent cyclic electron transport in stroma lamellae and grana margins.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1573-5079
    Keywords: light scattering ; photoinactivation ; proton gradient ; P700 photooxidation ; quenching of chlorophyll fluorescence ; redox poising
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Leaves of the C3 plant Brassica oleracea were illuminated with red and/or far-red light of different photon flux densities, with or without additional short pulses of high intensity red light, in air or in an atmosphere containing reduced levels of CO2 and/or oxygen. In the absence of CO2, far-red light increased light scattering, an indicator of the transthylakoid proton gradient, more than red light, although the red and far-red beams were balanced so as to excite Photosystem II to a comparable extent. On red background light, far-red supported a transthylakoid electrical field as indicated by the electrochromic P515 signal. Reducing the oxygen content of the gas phase increased far-red induced light scattering and caused a secondary decrease in the small light scattering signal induced by red light. CO2 inhibited the light-induced scattering responses irrespective of the mode of excitation. Short pulses of high intensity red light given to a background to red and/or far-red light induced appreciable additional light scattering after the flashes only, when CO2 levels were decreased to or below the CO2 compensation point, and when far-red background light was present. While pulse-induced light scattering increased, non-photochemical fluorescence quenching increased and F0 fluorescence decreased indicating increased radiationless dissipation of excitation energy even when the quinone acceptor QA in the reaction center of Photosystem II was largely oxidized. The observations indicate that in the presence of proper redox poising of the chloroplast electron transport chain cyclic electron transport supports a transthylakoid proton gradient which is capable of controlling Photosystem II activity. The data are discussed in relation to protection of the photosynthetic apparatus against photoinactivation.
    Type of Medium: Electronic Resource
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