The Influence of the Proton Gradient on the Activation of Ferredoxin-NADP+-oxidoreductase by Light

Ferredoxin-NADP+-oxidoreductase (FNR, EC 1.18.1.2) has been shown to be activated by light within a few seconds during dark-light transitions and inactivated in the dark. In previous papers this could be pointed out by the correlation of cytochrome f induction kinetics to the rate of NADP-photoreduction and the variable fluorescence. The present study deals with the role of the proton gradient during the activation process. The transition from an inactive to an active form is followed continuously in an in situ system. The steady-state rate of NADP-photoreduction is affected only by ionophores which inhibit a formation of the proton gradient, but not by inhibitors of the electric field. It …

Studies on the mechanism of photosystem II photoinhibition II. The involvement of toxic oxygen species.

In a previous paper it was shown that photoinhibition of reaction centre II of spinach thylakoids was predominantly caused by the degradation of D1-protein. An initial inactivation step at the QB-site was distinguished from its breakdown. The present paper deals with the question as to whether this loss of QB-function is caused by oxygen radical attack. For this purpose the photoinhibition of thylakoids was induced at 20°C in the presence of either superoxide dismutase and catalase or the antioxidants glutathione and ascorbic acid. This resulted in comparable though not total protection of D1-protein, photochemistry and fluorescence from photoinhibition. The combined action of both the enzy…

Die Anpassung des Photosyntheseapparates h�herer Pflanzen an die Lichtbedingungen

Cytochrome F as Indicator for the Interaction of the Two Photosystems in the State 1 and State 2

The transition of plants into a State 1 or a State 2 by an excess of PS I or PS II excitation was described by BONAVENTURA and MYERS [1]. Since the time of their investigations fluorometric methods gained increasing importance in analyzing the distribution of energy between the two photosystems [21. It was possible to correlate the altered energy balance to the phosphorylation of LHCII and it’s migration into non-appressed thylakoid membranes [31. However fluorescence measurements can give only indirect evidences that also changes in the activity of the two photosystems are produced by the changed energy distribution. A direct assessment of the photochemical variations during state transiti…

Chlorophyll b is involved in long-wavelength spectral properties of light-harvesting complexes LHC I and LHC II.

AbstractChlorophyll (Chl) molecules attached to plant light-harvesting complexes (LHC) differ in their spectral behavior. While most Chl a and Chl b molecules give rise to absorption bands between 645 nm and 670 nm, some special Chls absorb at wavelengths longer than 700 nm. Among the Chl a/b-antennae of higher plants these are found exclusively in LHC I. In order to assign this special spectral property to one chlorophyll species we reconstituted LHC of both photosystem I (Lhca4) and photosystem II (Lhcb1) with carotenoids and only Chl a or Chl b and analyzed the effect on pigment binding, absorption and fluorescence properties. In both LHCs the Chl-binding sites of the omitted Chl species…

Measurements of manganese in thylakoids of Sinapis alba grown under high-light and low-light conditions.

The manganese content of thylakoids and tissues was measured in leaves grown under high- and low-light conditions. Especially when grown in a nutrient medium enriched in manganese (20 μM), the thylakoids contained large amounts of manganese, which could be removed by EDTA washing without impairment of the Hill reaction. The unremovable content of manganese was almost the same in thylakoids from plants grown in nutrient media of normal (2 μM) and reduced (0.2 μM) manganese content. Up to this limit of manganese content, Hill activity did not seem to be impaired. 1.2 atoms Mn per 100 molecules chlorophyll were found in low-light thylakoids and 1.6 atoms Mn in high-light thylakoids. This is si…

Investigation on the Photosynthetic Membranes of Spruce Needles in Relation to the Occurrence of Novel Forest Decline I. The Photosynthetic Electron Transport

The investigations described here were carried out in the context of our research project on the physiological, biochemical, and cytomorphological characterization of spruce trees growing in natural habitats and showing damage of varying intensity. Here we report on specific aspects of the photosynthetic apparatus. The aim of the measurements was to analyze whether or not the activity of the photosynthetic electron transport pathway is affected in damaged trees. The investigations were carried out on a 20 to 25-year-old spruce plantation in the Hunsrück mountains and on an 80-year-old spruce plantation in the Westerwald mountains. The photosynthetic electron transport rate was determined by…

The intensification of absorbance changes in leaves by light-dispersion

In dispersive samples, like leaves, the absorbance of pigments is intensified. The intensification is due to a longer optical path through the dispersive sample. However, in chloroplast suspensions the optical path is not much longer than in clear solutions. The factor of intensification β (=the lengthening of the optical path) is calculated by comparing the absorbance of leaves and the absorbance of chloroplast suspensions with equal pigment-content. This method also includes the influence of possible sieve effects which could decrease absorbance. The measurements are carried out with high- and low-light leaves of different thickness and pigment content. The intensification of absorbance w…

Investigation on the Photosynthetic Membranes of Spruce Needles in Relation to the Occurrence of Novel Forest Decline II. The Content of QB-Protein, Cytochrome f, and P-700

In order to obtain an insight into the damage of thylakoid membranes of spruce (Picea abies) trees with damage of varying intensity, investigations were performed on the content of Qв-protein, cytochrome f, and P-700 in chloroplasts of spruce needles from apparently healthy and from damaged trees. Needles from the second and third needle year and the seventh whorl were chosen. The investigations were carried out in 1986 on a 20 to 25-year-old spruce plantation in the Hunsriick mountains and on an 80-year-old spruce plantation in the Westerwald mountains. In damaged trees an unequivocal decrease in the content of Qв-protein, cytochrome f, and P-700 was found, even in needle groups that appea…

Cyclic and noncyclic photophosphorylation during the ontogenesis of high-light and low-light leaves of Sinapis alba.

Noncyclic electron transport to ferricyanide and photophosphorylation as well as the methylviologen mediated aerobic and anaerobic photophosphorylation with dichlorophenolindophenol-ascorbate as the electron donor of photosystem I were measured during the development of high-light and low-light adapted leaves of Sinapis alba. Anaerobic methylviologen-catalyzed phosphorylation is more than twice as high as aerobic phosphorylation. The difference between the rates of aerobic and anaerobic phosphorylation is sensitive to dibromothymoquinone. Thus, under anaerobic conditions, methylviologen mediates a cyclic phosphorylation including plastoquinone. All photochemical activities of high-light chl…

Studies on the mechanism of photosystem II photoinhibition I. A two-step degradation of D1-protein

The role of D1-protein in photoinhibition was examined. Photoinhibition of spinach thylakoids at 20°C caused considerable degradation of D1-protein and a parallel loss of variable fluorescence, QB-independent electron flow and QB-dependent electron flow. The breakdown of D1-protein as well as the loss of variable fluorescence and QB-independent electron flow were largely prevented when thylakoids were photoinhibited at 0°C. The QB-dependent electron flow markedly decreased under the same conditions. This inactivation may represent the primary event in photoinhibition and could be the result of some modification at the QB-site of D1-protein. Evidence for this comes from fluorescence relaxati…

The Effect of Different Growth Light Intensities On Photosystem II Components

Light is essential not only as the driving force of photosynthesis but also as a trigger and a modulator of morphogenic processes. Physiological and morphological factors are modified when plants are grown at different light intensities and light qualities. Many plants are able to adapt the photosynthetic rate to the environmental factor light in a wide range. Low-light (LL) and high- light(HL) plants differ in a number of component processes of photosynthesis (1, 2). The adaptation process consists in a complex well coordinated change of many structural and biochemical components of the series of photosynthetic subprocesses (3).

Structure and function of ferredoxin-NADP+-oxidoreductase

The redox-enzyme ferredoxin-NADP-oxidoreductase has been shown to be activated by light and inactivated in the dark. This review will summarize recent data concerning the biochemical characterization of the enzyme compared to its in-vivo activation. Further-more the mechanism of this activation process is discussed as a conformational change caused by the light-driven proton gradient.

The Mechanism of Photoinhibition of Spinach Thylakoids

There is conflicting evidence as to wether D1-protein is the primary target of photoinhibition [1] or P680, the reaction centre of photo-system II [2]. The present paper desribes photoinhibition within a two step process consisting of an oxygen radical induced inactivation at the QB-site followed by damage to reaction centre II through the degradation of the D1-protein.

Preparation of Native and Recombinant Light-Harvesting Chlorophyll-<I>a/b </I>Complex

Procedures to isolate native light-harvesting chlorophyll-a/b complex (LHCIIb) and to reconstitute recombinant LHCIIb are described. Separation of trimeric from monomeric forms and free pigment by sucrose density-gradient ultracentrifugation can be applied to both native and reconstituted complexes. The preparations are characterized by their pigment composition, protein pattern, and spectral properties.

Measurements of cytochrome f and P-700 in intact leaves of Sinapis alba grown under high-light and low-light conditions

The oxidation and reduction of cytochrome f and P-700 is measured spectrophotometrically in leaves of low-light and high-light plants. After illumination with red light, an induction phenomenon for cytochrome f oxidation is observed which indicates a regulation of photosystem I activity through energy distribution between the pigment systems by the energy state of the membrane. After far-red excitation the reduction of cytochrome f in the dark is much slower in low-light leaves. This shows that cyclic electron transport is not improved in low-light plants under these conditions. P-700 is oxidized on excitation with far-red light. However, with high intensities of far-red light, P-700 is par…

The Effect of Phosphinothricin (Glufosinate) on Photosynthesis II. The Causes of Inhibition of Photosynthesis

It was shown in the previous study that phosphinothricin (glufosinate) causes an accumulation of ammonia and inhibition of photosynthesis. The extent to which there is a connection between these two processes is now investigated in the present study. First of all. the role of NH3 per se in the impairment of photosynthesis was to be clarified. For this purpose, the inhibition of photosyn­thesis was investigated in relation to exogenously applied ammonia in chloroplasts, protoplasts and entire leaves. The comparison with the experimental results in leaves in which the ammonia was formed endogenously (by action of phosphinothricin) shows that the ammonia toxicity at least cannot be solely resp…

Chlorophyll-protein-complexes of thylakoids of wild type and chlorophyll b mutants of Arabidopsis thaliana

Pigment-protein-complexes of two chlorophyll b deficient mutants of Arabidopsis and from the wild type were separated electrophoretically. Light-harvesting proteins were absent in the chlorophyll b free mutant ch(1) and their amount was reduced in the mutant ch(2) which has a reduced content of chlorophyll b. The ratio of CPa:CP I increased with decreasing chlorophyll b content which indicated that the stoichiometry of photosystem II to photosystem I is not constant.