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  • 1
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 113 (2001), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The effect of copper on photoinhibition of photosystem II (PSII) in vitro was studied in bean (Phaseolus vulgaris L. cv. Dufrix) and pumpkin (Cucurbita pepo L.) thylakoids. The thylakoids were illuminated at 200–2 000 μmol photons m−2 s−1 in the presence of 70–1 830 added Cu2+ ions per PSII. Three lines of evidence show that the irreversible damage of PSII caused by illumination of thylakoids in the presence of Cu2+ was mainly due to donor-side photoinhibition resulting from inhibition of the PSII donor side by Cu2+. First, addition of an artificial electron donor partially restored PSII activity of thylakoids that had been illuminated in the presence of Cu2+. Second, already moderate light was enough to cause rapid inhibition of PSII, and the inhibition could be saturated by light. Third, the extrinsic polypeptides of the oxygen-evolving complex were found to become oxidized by the combined effect of Cu2+ and light. The presence of oxygen was not necessary for the copper-induced enhancement of photoinhibition of PSII. When the illumination was prolonged, copper caused a gradual collapse of the thylakoid structure by increasing degradation of thylakoid proteins.
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We studied photoinhibition in two cultivars of tobacco (Nicotiana tabacum L.) expressing the bacterial gor gene in the cytosol and in four lines of poplar (Populus tremula×P. alba) expressing the FeSOD gene of Arabidopsis thaliana in the chloroplast. The respective total activities of glutathione reductase (EC 1.6.4.2) in leaves of gor tobaccos and superoxide dismutase (EC 1.15.1.1) in the FeSOD poplars were 5–8 times higher than in the respective untransformed control plants. Leaves of control and transformed plants were subjected to high-light stress at 20°C, and photoinhibition of photosystem II (PSII) was measured by oxygen evolution and chlorophyll fluorescence. The leaves were illuminated both in the presence and absence of lincomycin, which inhibits chloroplast protein synthesis. In both cases, the time course of loss of PSII activity was identical in plants overproducing superoxide dismutase (SOD) and in the untransformed controls, suggesting that the ability to convert superoxide to hydrogen peroxide is not a limiting factor in protection against photoinhibition, or in the repair of photoinhibitory damage or that the site of O2− production is not accessible to the transgene product. The rate constant of photoinhibition, measured in lincomycin-treated leaves, was smaller in glutathione reductase (GR) overproducing tobacco cv. Samsun than in the respective wild-type, but this difference was not seen in cv. Bel W3. The steady-state level of PSII activity measured when the PSII repair cycle was allowed to equilibrate with photoinhibitory damage under high light was not higher in the GR overproducing cv. Samsun, suggesting that the repair of photoinhibitory damage was not enhanced in plants overproducing GR in the cytosol.
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  • 3
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The ‘stay-green’ mutation cytG in soybean (Glycine max) partially inhibits the degradation of the light-harvesting complex II (LHCII) and the associated chlorophyll during monocarpic senescence. cytG did not alter the breakdown of the cytochrome b6/f complex, thylakoid ATP synthase or components of Photosystem I. In contrast, cytG accelerated the loss of oxygen evolution activity and PSII reaction-centre proteins. These data suggest that LHCII and other thylakoid components are degraded by separate pathways. In leaves induced to senesce by darkness, cytG inhibited the breakdown of LHCII and chlorophyll, but it did not enhance the loss of PSII-core components, indicating that the accelerated degradation of PSII reaction centre proteins in cytG was light dependent. Illumination of mature and senescent leaves of wild-type soybean in the presence of an inhibitor (lincomycin) of chloroplast protein synthesis revealed that senescence per se did not affect the rate of photoinhibition in leaves. Likewise, mature leaves of the cytG mutant did not show more photoinhibition than wild-type leaves. However, in senescent cytG leaves, photoinhibition proceeded more rapidly than in the wild-type. We conclude that the cytG mutation enhances photoinhibition in senescing leaves, and photoinhibition causes the rapid loss of PSII reaction-centre proteins during senescence in cytG.
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  • 4
    Electronic Resource
    Electronic Resource
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 93 (1995), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Reversible phosphorylation of the D1 reaction centre protein of photosystem II (PSII) occurs in thylakoid membranes of higher plants. The significance of D1 protein phosphorylation in the function of PSII is not yet clear. This paper summarizes the data implying that phosphorylation of D1 protein in higher plants is involved in the regulation of the repair cycle of photoinhibited PSII centres. Photoinhibition of PSII, D1 protein phosphorylation and degradation have been studied in vivo in higher plant leaves acclimated to different growth irradiances. It is shown that photoinhibitory illumination induces maximal phosphorylation of the D1 protein. Under these conditions D1 turnover is also saturated. We postulate that phosphorylation retards the degradation of damaged D1 protein under conditions where rapid replacement by a new D1 copy is not possible. This would protect PSII from total disassembly and degradation of all PSII subunits. We conclude that the phosphorylation of D1 protein and the regulation of D1 protein degradation may have evolved together. Furthermore, these characteristics seem to be related to the highly organized structure of higher-plant type thylakoid membranes, since the capability to phosphorylate D1 protein is restricted to seed plants.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 79 (1990), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Attached leaves of pumpkin (Cucurbita pepo L. cv. Jattiläismeloni) were exposed to high light intensity at room temperature (ca 23°C) and at 1°C. Fluorescence parameters and electron transport activities measured from isolated thylakoids indicated faster photoinhibition of PSII at low temperature. Separation of the α and β components of the complementary area above the fluorescence induction curve of dichlorophenyl-dimethylurea-poisoned thylakoids revealed that at low temperature only the α-centers declined during exposure to high light intensity while the content of functional β-centers remained constant. Freeze-fracture electron microscopy showed no decrease in the density of particles on the appressed exoplasmic fracture face, indicating that the photoinhibited α-centers remained in the appressed membranes at 1°C. Because of the function of the repair and protective mechanisms of PSII, strong light induced less photoinhibition at room temperature, but more complicated changes occurred in the α/β-heterogeneity of PSII. During the first 30 min at high light intensity the decrease in α-centers was almost as large as at 1°C, but in contrast to the situation at low temperature the decrease in α-centers was compensated for by a significant increase in PSIIβ-centers. Changes in the density and size of freeze-fracture particles suggest that this increase in β-centers was due to migration of phosphorylated light-harvesting complex from appressed to non-appressed thylakoid membranes while the PSII core remained in the appressed membranes. This situation, however, was only transient and was followed by a rapid decrease in the functionalβ-centers.
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Photosynthetica 37 (1999), S. 225-237 
    ISSN: 1573-9058
    Keywords: CP43 chlorophyll a binding protein ; D1 protein mutant ; 77 K fluorescence emission ; Synechocystis 6803
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Excitation-emission maps were constructed by measuring emission spectra from tobacco thylakoids and from thylakoids and intact cells of the cyanobacterium Synechocystis 6803. The measurement of such maps is greatly facilitated by the current diode-array detector technology. We show that excitation-emission maps are valuable tools for studies of the structure and energy transfer pathways in photosynthetic systems.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Photosynthesis research 56 (1998), S. 223-227 
    ISSN: 1573-5079
    Keywords: luminescence quenching ; oxygen electrode ; oxygen optode ; photosynthesis ; pressure sensitive paint
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We measured the light response curve of photosynthetic oxygen evolution by illuminating a leaf disc in an air-tight windowed chamber. Oxygen production was measured by monitoring the quenching of luminescence of an organometallic ruthenium compound. A photodiode based chlorophyll a fluorometer was used to measure the luminescence intensity. Oxygen evolution measurements with a traditional oxygen electrode gave the same numerical values at different light intensities when the same leaf disk was tested. The quality of the measurement signal of the new method was found to be similar to that obtained with the oxygen electrode method. The new luminescence based system is more stable against electrical disturbances than an oxygen electrode, its response to oxygen pressure changes is very rapid, and the new method allows the same basic equipment to be used for chlorophyll fluorescence and oxygen measurements.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Photosynthesis research 62 (1999), S. 55-66 
    ISSN: 1573-5079
    Keywords: bicarbonate ; chlorophyll fluorescence ; electron transfer ; heat ; QA ; redox potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High-temperature-induced inhibition of the acceptor side of Photosystem II (PS II) was studied in tobacco thylakoids using oxygen evolution, chlorophyll a (Chl a) fluorescence and redox potential measurements. When thylakoids were heated at 2 °C/min from 25 to 50 °C, the oxygen evolving complex became inhibited between 32 and 45 °C, whereas the acceptor side of PS II tolerated higher temperatures. Variable Chl a fluorescence decreased more slowly than oxygen evolution, suggesting that transitions between some S-states occurred even after heat-induced inhibition of the oxygen evolving activity. 77 K emission spectroscopy reveals that heating does not cause detachment of the light-harvesting complex II from PS II, and thus the heat-induced increase in the initial F0 fluorescence is due to loss of exciton trapping in the heated PS II centers. Redox titrations showed a heat-induced increase in the midpoint potential of the QA/QA -) couple from the control value of –80 mV to +40 mV at 50 °C, indicating a loss of the reducing power of QA -). When its driving force thus decreased, electron transfer from QA -) to QB in the PS II centers that still could reduce QA became gradually inhibited, as shown by measurements of the decay of Chl a fluorescence yield after a single turnover flash. Interestingly, the heat-induced loss of variable fluorescence and inhibition of electron transfer from QA -) to QB could be partially prevented by the presence of 5 mM bicarbonate during heating, suggesting that high temperatures cause release of the bicarbonate bound to PS II. We speculate that both the upshift in the redox potential of the QA/QA -) couple and the release of bicarbonate may be caused by a heat-induced structural change in the transmembrane D1 or D2 proteins. This structural change may, in turn, be caused by the inhibition of the oxygen evolving complex during heating.
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  • 9
    ISSN: 1573-5028
    Keywords: D-E loop ; D1 protein ; photosystem II ; psbA-2 gene expression ; Synechocystis sp. PCC 6803
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence connecting α-helices D and E of the D1 protein in photosystem II (PSII) is longer than that found in the corresponding loop of the L subunit in the rhodobacterial reaction centre. This sequence was mutated in order to determine its role in oxygenic photosynthesis. Site-specific mutants, including point mutations and deletions of different size, of the PEST-like region and the putative cleavage area in the D-E loop of the D1 protein were constructed in Synechocystis sp. PCC 6803. The effects of mutations on the functional and structural properties of PSII and turnover of the D1 protein were examined. Our results demonstrate that deletion of either the PEST-like sequence ( Δ R225-F239) or the putative cleavage region ( Δ G240-V249, Δ R225-V249) of the D1 protein resulted in severe perturbations on the function of the QB electron acceptor of PSII. However, PSII centres of the mutant with deleted PEST region remained functional enough to support autotrophic growth whereas deletions of the putative cleavage region prevented autotrophic growth. Although enhanced degradation rates of the mutant D1 proteins under low-light growth conditions demonstrate that neither the PEST-like sequence nor the putative cleavage region are required for D1 proteolysis, it became clear that the extension in the D-E loop of the D1 protein is essential for proper PSII assembly and photoautotrophic growth. Moreover, modifications of the D-E loop resulted in transcriptional activation of the psbA gene, indicating that neither light intensity, as such, nor the activity of the electron transfer chain are the only determinants in regulation of psbA gene transcription.
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  • 10
    ISSN: 1573-5079
    Keywords: chlorophyll fluorescence ; non-photochemical quenching ; flash-induced oxygen evolution ; D1 protein ; lincomycin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pumpkin (Cucurbita pepo L.) leaves in which chloroplast protein synthesis was inhibited with lincomycin were exposed to strong photoinhibitory light, and changes in FO, FM, FV/FM and in the amount of functional Photosystem II (O2 evolution induced by saturating single-turnover flashes) were monitored during the high-light exposure and subsequent dark or low-light incubation. In the course of the photoinhibitory illumination, FM, FV/FM and the amount of functional PS II declined continuously whereas FO dropped rapidly to some extent and then slowly increased. If the experiments were done at room temperature, termination of the photoinhibitory illumination resulted in partial relaxation of the FV/FM ratio and in an increase in FO and FM. The relaxation was completed in 10–15 min after short-term (15 min) photoinhibitory treatment but continued 30–40 min if the exposure to high light was longer than 1 h. No changes in the amount of functional PS II accompanied the relaxation of FV/FM in darkness or in low light, in the presence of lincomycin. Transferring the leaves to low temperature (+4°C) after the room-temperature illumination (2 h) completely inhibited the relaxation of FV/FM. Low temperature did not suppress the relaxation if the photoinhibitory illumination had also been done at low temperature. The results indicate that illumination of lincomycin-poisoned pumpkin leaves at room temperature does not lead to accumulation of a reversibly photoinactivated intermediate.
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