Three-dimensional model of zeaxanthin binding PsbS protein associated with nonphotochemical quenching of excess quanta of light energy absorbed by the photosynthetic apparatus

A three-dimensional model of the PsbS protein was built with the help of homology-modeling methods. This protein is also known as CP22 and is associated with the protection of photosystem II of thylakoid from excess quanta of light energy absorbed by the photosynthetic apparatus. PsbS is reported to bind two molecules of zeaxanthin at low pH (<5.0) and is believed to be essential for rapid nonphotochemical quenching (qE) of chlorophyll a fluorescence in photosystem II. An attempt was made to explain the pH modulation of the conformation of protein through salt-bridges Glu⁻(122)-Lys⁺(113) and Glu⁻(226)-Lys⁺(217). Binding of two molecules of zeaxanthin in the three-dimensional model of PsbS is postulated. The molecular mechanism of photoprotection by PsbS is explained through the model. 1 Backbone structure of the PsbS protein with two molecules of all trans zeaxanthin (ZEX). Residues Glu 90, 122, 194, 226 and Lys 113, 217 are shown. The figure is drawn with RASMOL (Molecular Visualization Program, RasMol V2.6, Roger Sayle, Glaxo Wellcome Research and Development, Stevenage, Hertfordshire, UK) Electronic Supplementary Material Supplementary material is available for this article at     

The photosystem I trimer of cyanobacteria: molecular organization, excitation dynamics and physiological significance

The photosystem I complex organized in cyanobacterial membranes preferentially in trimeric form participates in electron transport and is also involved in dissipation of excess energy thus protecting the complex against photodamage. A small number of longwave chlorophylls in the core antenna of photosystem I are not located in the close vicinity of P700, but at the periphery, and increase the absorption cross-section substantially. The picosecond fluorescence kinetics of trimers resolved the fastest energy transfer components reflecting the equilibration processes in the core antenna at different redox states of P700. Excitation kinetics in the photosystem I bulk antenna is nearly trap-limited, whereas excitation trapping from longwave chlorophyll pools is diffusion-limited and occurs via the bulk antenna. Charge separation in the photosystem I reaction center is the fastest of all known reaction centers.     

Photoprotection of green plants: a mechanism of ultra-fast thermal energy dissipation in desiccated lichens

In order to survive sunlight in the absence of water, desiccation-tolerant green plants need to be protected against photooxidation. During drying of the chlorolichen Cladonia rangiformis and the cyanolichen Peltigera neckeri, chlorophyll fluorescence decreased and stable light-dependent charge separation in reaction centers of the photosynthetic apparatus was lost. The presence of light during desiccation increased loss of fluorescence in the chlorolichen more than that in the cyanolichen. Heating of desiccated Cladonia thalli, but not of Peltigera thalli, increased fluorescence emission more after the lichen had been dried in the light than after drying in darkness. Activation of zeaxanthin-dependent energy dissipation by protonation of the PsbS protein of thylakoid membranes was not responsible for the increased loss of chlorophyll fluorescence by the chlorolichen during drying in the light. Glutaraldehyde inhibited loss of chlorophyll fluorescence during drying. Desiccation-induced loss of chlorophyll fluorescence and of light-dependent charge separation are interpreted to indicate activation of a highly effective mechanism of photoprotection in the lichens. Activation is based on desiccation-induced conformational changes of a pigment-protein complex. Absorbed light energy is converted into heat within a picosecond or femtosecond time domain. When present during desiccation, light interacts with the structural changes of the protein providing increased photoprotection. Energy dissipation is inactivated and structural changes are reversed when water becomes available again. Reversibility of ultra-fast thermal dissipation of light energy avoids photo-damage in the absence of water and facilitates the use of light for photosynthesis almost as soon as water becomes available.     

Dissipation of excess energy triggered by blue light in cyanobacteria with CP43′ (isiA)

The chlorophyll-protein CP43′ (isiA gene) induced by stress conditions in cyanobacteria is shown to serve as an antenna for Photosystem II (PSII), in addition to its known role as an antenna for Photosystem I (PSI). At high light intensity, this antenna is converted to an efficient trap for chlorophyll excitations that protects system II from photo-inhibition. In contrast to the ‘energy-dependent non-photochemical quenching’ (NPQ) in chloroplasts, this photoprotective energy dissipation in cyanobacteria is triggered by blue light. The induction is proportional to light intensity. Induction and decay of the quenching exhibit the same large temperature-dependence.     

Transfer and trapping of excitation energy in Photosystem II as studied by chlorophyll a2 fluorescence quenching by dinitrobenzene and carotenoid triplet. The matrix model

1. The curves representing the reciprocal fluorescence yield of chlorophyll a of Photosystem II (PS II) in Chlorella vulgaris as a function of the concentration of m-dinitrobenzene in the states P Q and P Q^-, are found to be straight parallel lines; P is the primary donor and Q the primary acceptor of PS II. In the weakly trapping state P Q^- the half-quenching of dinitrobenzene is about 0.2 mM, in vitro it is of the order of 10 mM. The fluorescence yield as a function of the concentration of a quencher is described for three models for the structure of pigment systems: the model of separate units, the model of limited energy transfer between the units, and the matrix model. If it is assumed that the rate constant of quenching by dinitrobenzene is high and thus the number of dinitrobenzene molecules per reaction center low, it can be concluded that the pigment system of PS II in C. vulgaris is a matrix of chlorophyll molecules in which the reaction centers are embedded. Theoretical and experimental evidence is consistent with such an assumption.

For Cyanidium caldarium the zero fluorescence yield Ф₀ and its quenching by dinitrobenzene were found to be much smaller than the corresponding quantities for C. vulgaris. Nevertheless, our measurements on C. caldarium could be interpreted by the assumption that the essential properties (rate constants, dinitrobenzene quenching) of PS II are the same for these two species belonging to such widely different groups.

2. The measured dinitrobenzene concentrations required for half-quenching in vivo and other observations are explained by (non-rate-limiting) energy transfer between the chlorophyll a molecules of PS II and by the assumptions that dinitrobenzene is approximately distributed at random in the membrane and does not diffuse during excitation.

3. The fluorescence kinetics of C. vulgaris during a 350 ns laser flash of variable intensity could be simulated on a computer using the matrix model. From the observed fluorescence quenching by the carotenoid triplet (C^T) and the measurement of the number of C^T per reaction center via difference absorption spectroscopy, the rate constant for quenching of C^T is calculated to be k_T = 3.3 · 10¹¹ s⁻¹ which is almost equal to the rate constant of trapping by an open reaction center (Duysens, L.N.M. (1979) CIBA Foundation Symposium 61 (New Series), pp. 323–340).

4. The fluorescence quenching by C^T in non-treated spinach chloroplasts after a 500 ns laser flash (Breton, J., Geacintov, N.E. and Swenberg, C.E. (1979) Biochim. Biophys. Acta 548, 616–635) could be explained within the framework of the matrix model when the value for k_T is used as given in point 3.

5. The observations mentioned under point 1 indicate that the fluorescence yield Ф₀ for centers in trapping state P Q is probably for a fraction exceeding 0.8 emitted by PS II.     

Organization of the photosynthetic apparatus of the chlorina-f2 mutant of barley using chlorophyll fluorescence decay kinetics

The time-resolved chlorophyll fluorescence emission of higher plant chloroplasts monitors the primary processes of photosynthesis and reflects photosynthetic membrane organization. In the present study we compare measurements of the chlorophyll fluorescence decay kinetics of the chlorophyll-b-less chlorina-f2 barley mutant and wild-type barley to investigate the effect of alterations in thylakoid membrane composition on chlorophyll fluorescence. Our analysis characterizes the fluorescence decay of chlorina-f2 barley chloroplasts by three exponential components with lifetimes of approx. 100 ps, 400 ps and 2 ns. The majority of the chlorophyll fluorescence originates in the two faster decay components. Although photo-induced and cation-induced effects on fluorescence yields are evident, the fluorescence lifetimes are independent of the state of the Photosystem-II reaction centers and the degree of grana stacking. Wild-type barley chloroplasts also exhibit three kinetic fluorescence components, but they are distinguished from those of the chlorina-f2 chloroplasts by a slow decay component which displays cation- and photo-induced yield and lifetime changes. A comparison is presented of the kinetic analysis of the chlorina-f2 barley fluorescence to the decay kinetics previously measured for intermittent-light-grown peas (Karukstis, K. and Sauer, K. (1983) Biochim. Biophys. Acta 725, 384–393). We propose that similarities in the fluorescence decay kinetics of both species are a consequence of analogous rearrangements of the thylakoid membrane organization due to the deficiencies present in the light-harvesting chlorophyll $\text{a}\text{b}$ complex.     

Inhibition of photosynthesis of sunflower leaves by an endogenous solute and interdependence of different photosynthetic reactions

The relationship between components of non-photochemical quenching of chlorophyll fluorescence yield (q_NP) and dissipation of excessive excitation energy was determined in cotton leaves using concurrent measurements of fluorescence and gas-exchange at 2% and 20% O₂ under a range of photon flux densities and CO₂ pressures. A nearly stoichiometric relationship was obtained between dissipation of energy not used in photosynthetic CO₂ fixation or photorespiration and q_NP provided that a component, probably associated with state transitions, was not included in q_NP. Although two distinct components of q_NP were resolved on the basis of their relaxation kinetics, both components appear effective in energy dissipation. The photon yield of open photosystem-II reaction centers decreased linearly with increases in q_NP, indicating that much of the energy dissipation occurs in the pigment bed. However, increases in q_NP appear dependent on the redox state of these centers. The results are discussed in relation to current hypotheses of the molecular basis of non-radiative energy dissipation. It is concluded that determinations of q_NP can provide a quantitative measure of the dissipation of excessive excitation energy if precautions are taken to ensure that the maximum fluorescence yield is measured under conditions that provide complete closure of the photosystem-II reaction centers. It is also concluded that such dissipation can prevent photoinhibitory damage in cotton leaves even under extreme conditions where as much as 80% of the excitation energy is excessive.Abbreviations and symbols F _M, F _O, F _V, F _S fluorescence yield when all PSII centers are closed, when all centers are open, F_M-F_O, at steady state in the light - PFD photon flux density (photon fluence rate) - P(CO₂) sum of rates of CO₂ uptake and dark respiration - P(ET) sum of P(CO₂) and rate of oxygenation - PSI, PSII photosystem I, II - q_NP, q_P non-photochemical, photochemical fluorescence quenching - Q the acceptor for PSII - Q _r/Q _t the fraction of reduced Q or closed PSII centers - _r/ _t intrinsic photon yield of CO₂ fixation in the absence of photorespiration of O₂ evolution - _a P(ET)/PFD (absorbed light) C.I.W. Publication No. 1016     

Growth of cotton under continuous salinity stress: influence on allocation pattern,stomatal and non-stomatal components of photosynthesis and dissipation of excess light energy

Cotton (Gossypium hirsutum L.) plants were grown in flowing-culture solutions containing 0%, 26% and 55% natural seawater under controlled and otherwise identical conditions. Leaf Na⁺ content rose to 360 mM in 55% seawater, yet the K⁺ content was maintained above 100 mM. The K⁺/Na⁺ selectivity ratio was much greater in the saline plants than in the control plants. All plants were healthy and able to complete the life cycle but relative growth rate fell by 46% in 26% seawater and by 83% in 55% seawater. Much of this reduction in growth was caused by a decreased allocation of carbon to leaf growth versus root growth. The ratio of leaf area/plant dry weight fell by 32% in 26% seawater and by 50% in 55 % seawater while the rate of carbon gain per unit leaf area fell by only 20% in 26% seawater and by as much as 66% in 55% seawater. Partial stomatal closure accounted for nearly all of the fall in the photosynthesis rate in 26% seawater but in 55% seawater much of the fall also can be attributed to non-stomatal factors. As a result of the greater effect of salinity on stomatal conductance than on CO₂-uptake rate, photosynthetic water-use efficiency was markedly improved by salinity. This was also confirmed by stablecarbon-isotope analyses of leaf sugar and of leaf cellulose and starch. — Although non-stomatal photosynthetic capacity at the growth light was reduced by as much as 42% in 55% seawater, no effects were detected on the intrinsic photon yield of photosynthesis nor on the efficiency of photosystem II photochemistry, chlorophyll a/b ratio, carotenoid composition or the operation of the xanthophyll cycle. Whereas salinity caused in increase in mesophyll thickness and content of chloroplast pigments it caused a decrease in total leaf nitrogen content. The results indicate that the salinity-induced reduction in non-stomatal photosynthetic capacity was not caused by any detrimental effect on the photosynthetic apparatus but reflects a decreased allocation to enzymes of carbon fixation. — Rates of energy dissipation via CO₂ fixation and photorespiration, calculated from gas-exchange measurements, were insufficient to balance the rate of light-energy absorption at the growth light. Salinity therefore would have been expected to cause the excess excitation energy to rise, leading to an increased nonradiative dissipation in the pigment bed and resulting increases in non-photochemical fluorescence quenching and zeaxanthin formation. However, no such changes could be detected, implying that salinity may have increased energy dissipation via a yet unidentified energy-consuming process. This lack of a response to salinity stress is in contrast to the responses elicited by short-term water stress which caused strong non-photochemical quenching and massive zeaxanthin formation.Abbreviations and Symbols A net rate of CO₂ uptake - A_c calculated rate of CO₂ uptake at constant p_i - Chl chlorophyll - E rate of transpiration - EPS epoxidation state of xanthophyll cycle components - F, F_m fluorescence emission at the actual, full reduction of PSII reaction centers - F_v variable fluorescence - g_s stomatal conductance to water vapor - g_w conductance to CO₂transfer from intercellular spaces to chloroplasts - NPQ non-photochemical fluorescence quenching - p_a, p_i, p_c atmospheric, intercellular and chloroplastic partial pressures of CO₂ - PCRO photosynthetic carbon reduction and oxygenation cycle sum of the rate of carboxylation and oxygenation - PFD photon flux density - PSII photosystem II - V+A+Z pool size of xanthophyll cycle components - ¹³C carbon-isotope composition - (_PSII) photon yield of PSII photochemistry at the actual reduction state in the light * C.I.W.-D.P.B. Publication No. 1115, CNR-RAISA paper No. XXXWe thank Connie Shih for skilful assistance in growing plants and for conducting HPLC analyses and Barbara Mortimer for conducting the nitrogen analyses. Thanks are also due to C. Barry Osmond (now, Australian National University, Research School of Biological Sciences, Canberra) and Larry Giles of the Department of Botany, Duke University, Durham, N.C., for conducting carbonisotope analysis. E.B. was partially supported by the National Research Council of Italy, Special Project RAISA, Sub-Project No. 2. A Carnegie Institution Fellowship to E.B. is also gratefully acknowledged. This work was supported by grant No. 89-37-280-4902 of the Competitive Grant Program of the U.S. Department of Agriculture to O.B.     

Regulation of excitation energy distribution in Photosystem-II fragments by magnesium ions

Fluorescence characteristics of Photosystem-II subchloroplasts (TSF-II and TSF-IIa) fractionated by Triton X-100 treatment were studied in relation to cation-induced regulation of excitation-energy distribution within subchloroplast fragments. Absorption spectra and fluorescence-emission spectra at 77 K showed that TSF-II contains the light-harvesting chlorophyll-protein complex in addition to the reaction-center complex, which is present alone in TSF-IIa.Mg²⁺ increased the ratio of F_695nm to F_685nm in the fluorescence-emission spectrum of TSF-II particles at 77 K, but had no effect on TSF-IIa particles. Mg²⁺ also induced a quenching of chlorophyll fluorescence at room temperature in TSF-II, an effect that was insensitive to the presence of DCMU. The DCMU-insensitive fluorescence quenching was not observed in the TSF-IIa preparation. These results suggest an existence of cation-induced regulation of excitation-energy transfer in TSF-II preparations. Presence of antenna chlorophyll molecules alone does not seem to be sufficient for observing energytransfer regulation by cations in Photosystem-II preparations.     

Modulation of photosynthetic electron transport in the absence of terminal electron acceptors: Characterization of the rbcL deletion mutant of tobacco

Tobacco rbcL deletion mutant, which lacks the key enzyme Rubisco for photosynthetic carbon assimilation, was characterized with respect to thylakoid functional properties and protein composition. The ΔrbcL plants showed an enhanced capacity for dissipation of light energy by non-photochemical quenching which was accompanied by low photochemical quenching and low overall photosynthetic electron transport rate. Flash-induced fluorescence relaxation and thermoluminescence measurements revealed a slow electron transfer and decreased redox gap between Q_A and Q_B, whereas the donor side function of the Photosystem II (PSII) complex was not affected. The 77 K fluorescence emission spectrum of ΔrbcL plant thylakoids implied a presence of free light harvesting complexes. Mutant plants also had a low amount of photooxidisible P700 and an increased ratio of PSII to Photosystem I (PSI). On the other hand, an elevated level of plastid terminal oxidase and the lack of F₀ ‘dark rise’ in fluorescence measurements suggest an enhanced plastid terminal oxidase-mediated electron flow to O₂ in ΔrbcL thylakoids. Modified electron transfer routes together with flexible dissipation of excitation energy through PSII probably have a crucial role in protection of PSI from irreversible protein damage in the ΔrbcL mutant under growth conditions. This protective capacity was rapidly exceeded in ΔrbcL mutant when the light level was elevated resulting in severe degradation of PSI complexes.