Multiple Short Term Effects of UV-B Radiation on the Diatom Phaeodactylum Tricornutum

Increases in UV-B irradiance lead to many specific damaging effects upon the plants including damage of the thylakoid membrane, partial inhibition of PS II, decrease of chloroplast ATPase activity, loss of enzyme activities in the calvin cycle and alterations in pigment synthesis (1). Under natural conditions enhanced UV-B light is always accompanied by high intensities of photosynthetic active radiation (PAR). Damaging effects due to photoinhibitory PAR and UV-B light which lead to several oxygen radical species (2) could be reduced by photoprotection mechanisms. One of these protection mechanisms is the xanthophyll cycle. In higher plants and green algae violaxanthin is converted to zeaxa…

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…

Picosecond time-resolved study on the nature of high-energy-state quenching in isolated pea thylakoids different localization of zeaxanthin dependent and independent quenching mechanisms

Abstract The influence of the transthylakoid proton gradient on the kinetics of picosecond fluorescence decay was examined using isolated pea thylakoids having high or low zeaxanthin contents. Fluorescence lifetime measurements were performed with open (Fo) and closed (Fm) PS II reaction centers. Zeaxanthin formation in membrane energized isolated thylakoids led to a marked decrease of the average fluorescence lifetime at both Fm and Fo. In contrast, when zeaxanthin synthesis was blocked by the inhibitor DTT, the fluorescence lifetime decrease was less pronounced in the Fm state and totally missing in the Fo state. Samples containing the uncoupler ammonium chloride did not exhinit any zeaxa…

The Effect of Temperature and Radical Protection on the Photoinhibition of Spinach Thylakoids

Photoinhibition of spinach thylakoids was studied by examination the inactivation of different parts of the electron transport chain, the decline of the variable fluorescence and the loss of atrazine binding sites. The results obtained after photoinhibition at 20° C and 0° C revealed an inactivation at the QB-site of the D1-protein as the first event in the course of photoinhibition. The natural antioxidants glutathione and ascorbate as well as the enzymes SOD and catalase diminished photoinhibition to similar extents. Further protection was achieved through combination of both radical defense systems. In addition to the radical scavenging properties glutathione and ascorbate have reducing …

Changes of the Quantum Yield of Oxygen Evolution and the Electron Transport Capacity of Isolated Spinach Thylakoids during Photoinhibition

Summary Isolated spinach thylakoids can be protected from photoinhibitory loss of electron transport capacity by the radical defense system composed of the enzymes SOD and catalase, as well as the antioxidants glutathione and ascorbate. With these compounds present at saturating concentrations, thylakoids not only retain a high photochemical capacity but also do not suffer D1-protein degradation during photoinhibition. However, a strong decrease in the quantum yield of oxygen evolution, Φ O 2 , occurs with the same thylakoids. These results support the view that the photochemical capacity and the quantum yield of oxygen evolution decline due to different mechanisms during photoinhibition. T…

Light-induced proton slip and proton leak at the thylakoid membrane

A treatment of leaves of Spinacia oleracea L. with light or with the thiol reagent dithiothreitol in the dark led to partly uncoupled thylakoids. After induction in intact leaves, the partial uncoupling was irreversible at the level of isolated thylakoids. We distinguish between uncoupling by proton slip, which means a decrease of the H+/e(-) -ratio due to less efficient proton pumping, and proton leak as defined by enhanced kinetics of proton efflux. Proton slip and proton leak made about equal contributions to the total uncoupling. The enhanced proton efflux kinetics corresponded to reduction of subunit CF1-gamma of the ATP synthase as shown by fluorescence labeling of thylakoid proteins …

Evaluation of positron-emission-tomography for visualisation of migration processes in geomaterials

Abstract Positron-emission-tomography (PET) was applied for direct visualisation of solute transport in order to overcome the limitations of conventional methods for measuring advection and diffusion properties. At intervals from minutes to days the 3D-spatial distribution of the PET-tracer is determined. This spatiotemporal evolution of the tracer concentration can be used as experimental basis for clarification of the relevant transport processes, derivation of transport parameters, and model calibration. Here, 18 F and 124 I in 0.01 M carrier solution of KF and KI, respectively, have been chosen out of the limited number of available PET-tracers, primarily on account of their decay time …

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…

Landslides as Important Disturbance Regimes — Causes and Regeneration

Characterization of the Fast and Slow Reversible Components of Non-Photochemical Quenching in Isolated Pea Thylakoids by Picosecond Time-Resolved Chlorophyll Fluorescence Analysis

The fast and slow reversible components of non-photochemical chlorophyll fluorescence quenching commonly assigned to the qE and the qI mechanism have been studied in isolated pea thylakoids which were prepared from leaves after a moderate photoinhibitory treatment. Chlorophyll fluorescence decays were measured at picosecond resolution and analyzed on the basis of the heterogeneous exciton/radical pair equilibrium model. Our results show that the fast reversible non-photochemical quenching is completely assigned to the PS II antenna and is related to zeaxanthin. The slow reversible qI type quenching is located at the PS II reaction center and involves enhanced nonradiative decay of the prima…

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).

Role of ΔpH in the mechanism of zeaxanthin-dependent amplification of qE

Abstract The influence of zeaxanthin on the high-energy-state fluorescence quenching (qE) and the pH dependence of the maximum chlorophyll fluorescence yield (Fm) was examined in spinach thylakoids. High contents of zeaxanthin were achieved using different pretreatments. A pronounced, zeaxanthin-dependent amplification of non-photochemical quenching (NPQ) was exclusively found in thylakoids containing zeaxanthin, synthesized in the dark via the buildup of an artificial ΔpH. These thylakoids also showed a significant quenching of chlorophyll fluorescence in the range pH 5.5–6.3, where no or only slight quenching was visible in zeaxanthin-free thylakoids. Thylakoids containing high amounts of…

The importance of a highly active and DeltapH-regulated diatoxanthin epoxidase for the regulation of the PS II antenna function in diadinoxanthin cycle containing algae.

The present study focuses on the regulation of diatoxanthin (Dtx) epoxidation in the diadinoxanthin (Ddx) cycle containing algae Phaeodactylum tricornutum, Thalassiosira pseudonana, Cyclotella meneghiniana and Prymnesium parvum and its significance for the control of the photosystem II (PS II) antenna function. Our data show that Dtx epoxidase can exhibit extremely high activities when algal cells are transferred from high light (HL) to low light (LL). Under HL conditions, Dtx epoxidation is strongly inhibited by the light-driven proton gradient. Uncoupling of the cells during HL illumination restores the high epoxidation rates observed during LL. In Ddx cycle containing algae, non-photoche…

Zeaxanthin Dependent and Zeaxanthin Independent Changes in Nonphotochemical Energy Dissipation

Summary The influence of zeaxanthin on high-energy-state quenching (qE) of room temperature chlorophyll fluorescence and on the quantum yield of oxygen evolution(ФO2) has been studied with isolated spinach thylakoids. A set of three different pretreatments was tested which all led to thylakoids with high contents of zeaxanthin. Depending on the presence of light the pretreatments differed strongly with respect to their effect on nonphotochemical fluorescence quenching. Independently from the extent of changes in nonphotochemical energy dissipation in the membrane energized state as reflected by nonphotochemical quenching the light pretreatments caused also changes in nonphotochemical energy…

Supplementary Ultraviolet-B Radiation Induces a Rapid Reversal of the Diadinoxanthin Cycle in the Strong Light-Exposed DiatomPhaeodactylum tricornutum 

AbstractA treatment of the diatom Phaeodactylum tricornutum with high light (HL) in the visible range led to the conversion of diadinoxanthin (Dd) to diatoxanthin (Dt). In a following treatment with HL plus supplementary ultraviolet (UV)-B, the Dt was rapidly epoxidized to Dd. Photosynthesis of the cells was inhibited under HL + UV-B. This is accounted for by direct damage by UV-B and damage because of the UV-B-induced reversal of the Dd cycle and the associated loss of photoprotection. The reversal of the Dd cycle by UV-B was faster in the presence of dithiothreitol, an inhibitor of the Dd de-epoxidase. Our results imply that the reversal of the Dd cycle by HL + UV-B was caused by an incre…

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.

Pigment composition of PS II pigment protein complexes purified by anion exchange chromatography. identification of xanthophyll cycle pigment binding proteins

Summary The pigment composition of the chlorophyll binding proteins of Photosystem II (PS II) of spinach ( Spinacea oleracea L.) has been determined using sucrose gradient ultracentrifugation, anion exchange chromatography and HPLC based pigment analysis. The xanthophyll cycle pigments violaxanthin, antheraxanthin and zeaxanthin were exclusively found in the proteins of the outer PS II antenna, with the highest amounts being present in the minor chlorophyll alb binding proteins CP 29 and CP 26. PS II core particles containing the reaction centre proteins D1, D2, cytochrome b 559 and the proteins of the inner antenna CP 47 and CP 43 bind β-carotene as the only carotenoid. The presence of the…

Picosecond Time Resolved Analysis of the Fast and Slow Reversible Non-Photochemical Chlorophyll Fluorescence Quenching

Photosystem II, which is a potential target of adverse effects of supersaturating light, is strongly dependent on a mechanism, which allows to switch over between efficient photochemical energy conversion at limiting light intensity and efficient photothermal energy conversion under strong light. The mechanisms for the thermal dissipation of light absorbed in excess are reflected by the socalled non-photochemical quenching of chlorophyll fluorescence (NPQ). Under excessive illumination two major components contribute to the overall NPQ which can be distinguished by their different kinetics of dark relaxation. The fast reversible component is supposed to be linked to the light-induced format…