A micellar model system for the role of zeaxanthin in the non-photochemical quenching process of photosynthesis--chlorophyll fluorescence quenching by the xanthophylls

To get an insight to the mechanism of the zeaxanthin-dependent non-photochemical quenching in photosystem II of photosynthesis, we probed the interaction of some xanthophylls with excited chlorophyll-a by trapping both pigments in micelles of triton X-100. Optimal distribution of pigments among micelles was obtained by proper control of the micelle concentration, using formamide in the reaction mixture, which varies the micellar aggregation number over three orders of magnitude. The optimal reaction mixture was obtained around 40% (v/v) formamide in 0.2-0.4% (v/v) triton X-100 in water. Zeaxanthin in the micellar solution exhibited initially absorption and circular dichroism spectral features corresponding to a J-type aggregate. The spectrum was transformed over time (half-time values vary-an average characteristic figure is roughly 20 min) to give features representing an H-type aggregate. The isosbestic point in the series of spectral curves favors the supposition of a rather simple reaction between two pure J and H-types dimeric species. Violaxanthin exhibited immediately stable spectral features corresponding to a mixture of J-type and more predominately H-type dimers. Lutein, neoxanthin and beta-carotene did not show any aggregated spectral forms in micelles. The spectral features in micelles were compared to spectra in aqueous acetone, where the assignment to various aggregated types was established previously. The specific tendency of zeaxanthin to form the J-type dimer (or aggregate) could be important for its function in photosynthesis. The abilities of five carotenoids (zeaxanthin, violaxanthin, lutein, neoxanthin and beta-carotene) to quench chlorophyll-a fluorescence were compared. Zeaxanthin, in its two micellar dimeric forms, and beta-carotene were comparable good quenchers of chlorophyll-a fluorescence. Violaxanthin was a much weaker quencher, if at all. Lutein and neoxanthin rather enhanced the fluorescence. The implications to non-photochemical quenching process in photosynthesis are discussed.     

Operation of the xanthophyll cycle in the seagrass Zostera marina in response to variable irradiance

Changes in the photobiology and photosynthetic pigments of the seagrass Zostera marina from Chesapeake Bay (USA) were examined under a range of natural and manipulated irradiance regimes. Photosynthetic activity was assessed using chlorophyll-a fluorescence, and photosynthetic pigments were measured by HPLC. Large changes in the violaxanthin, zeaxanthin, and antheraxanthin content were concomitant with the modulation of non-photochemical quenching (NPQ). Photokinetics (F_v/F_m, rapid light curves (RLC), and non-photochemical quenching) varied as a result of oscillating irradiance and were highly correlated to xanthophyll pigment content. Zeaxanthin and antheraxanthin concentrations increased under elevated light conditions, while violaxanthin increased in darkened conditions. Unusually high concentrations of antheraxanthin were found in Z. marina under a wide range of light conditions, and this was associated with the partial conversion of violaxanthin to zeaxanthin. These results support the idea that xanthophyll intermediate pigments induce a photoprotective response during exposure to high irradiances in this seagrass.     

The xanthophyll cycle pool size controls the kinetics of non-photochemical quenching in Arabidopsis thaliana

Arabidopsis plants overexpressing beta-carotene hydroxylase 1 accumulate over double the amount of zeaxanthin present in wild-type plants. The final amplitude of non-photochemical quenching (NPQ) was found to be the same in these plants, but the kinetics were different. The formation and relaxation of NPQ consistently correlated with the de-epoxidation state of the xanthophyll cycle pool and not the amount of zeaxanthin. These data indicate that zeaxanthin and violaxanthin antagonistically regulate the switch between the light harvesting and photoprotective modes of the light harvesting system and show that control of the xanthophyll cycle pool size is necessary to optimize the kinetics of NPQ.     

Carotenoid composition in sun and shade leaves of plants with different life forms

The carotenoid composition of sun leaves of nine species of annual crop plants (some with several varieties) was compared with sun and shade leaves of several other groups of plants, among those sun and shade leaves of several species of perennial shrubs and vines and deep-shade leaves of seven rainforest species. All sun leaves contained considerably greater amounts of the components of the xanthophyll cycle violaxanthin, antheraxanthin and zeaxanthin as well as of β-carotene than the shade leaves, as had previously been reported for a variety of other species by Thayer & Björkman (Photosynthesis Research, 1990, 23, 331–343). Therefore, high light specifically stimulated β,β-carotenoid synthesis. The sun leaves of these crop species did not contain α-carotene which was, however, present in large amounts in all shade leaves and in smaller amounts in sun leaves of three of the four species of perennial shrubs and vines. There was no difference in neoxanthin content on a chlorophyll basis between sun and shade leaves, and there was no consistent general difference in the lutein content between all sun and all shade leaves. The zeaxanthin (and antheraxanthin) content at peak irradiance and the xanthophyll cycle pool size were compared for sun leaves from the different groups of plants with different life forms and different metabolic activities. When growing in full sunlight the annual crop species and a perennial mesophyte had high rates of photosynthesis whereas the perennial shrubs and vines had relatively low photosynthesis rates. More zeaxanthin (and antheraxanthin) were accumulated at noon in full sunlight in those species with the lower photosynthesis rates. However, it was not such that those species also possessed the larger pools of violaxanthin + antheraxanthin + zeaxanthin. Instead, the xanthophyll cycle pools of sun leaves of the annual crop species and the perennial mesophyte were not smaller, and were even possibly larger, than those of sun leaves of the perennial shrubs and vines with low photosynthesis rates. This was so in spite of the fact that the crop species experienced much lesser degrees of excessive light at full sun than the shrubs and vines. Thus, many of the crop species converted only about 30–50% of their xanthophyll cycle pool to zeaxanthin at noon, whereas the shrubs and vines typically converted more than 80% of their pool into zeaxanthin. The crop species also had larger pools of β-carotene than the shrubs and vines but smaller pools of lutein than the majority of the latter species.     

Changes in the quantities of violaxanthin de-epoxidase,xanthophylls and ascorbate in spinach upon shift from low to high light

Zeaxanthin, a carotenoid in the xanthophyll cycle, has been suggested to play a role in the protection against photodestruction. We have studied the importance of the parameters involved in zeaxanthin formation by comparing spinach plants grown in low light (100 to 250 mol m^-2 s^-1) to plants transferred to high light (950 mol m^-2 s^-1). Different parameters were followed for a total of 11 days. Our experiments show that violaxanthin de-epoxidase decreased between 15 and 30%, the quantity of xanthophyll cycle pigments doubled to 100 mmol (mol Chl)^-1, corresponding to 27 mol m^-2, and the rate of violaxanthin to zeaxanthin conversion was doubled. Lutein and neoxanthin increased from 50 to 71 mol m^-2 and from 16 to 23 mol m^-2, respectively. On a leaf area basis, chlorophyll and -carotene levels first decreased and then after 4 days increased. The chlorophyll a/b ratio was unchanged. The quantity of ascorbate was doubled to 2 mmol m^-2, corresponding to an estimated increase in the chloroplasts from 25 to 50 mM. In view of our data, we propose that the increase in xanthophyll cycle pigments and ascorbate only partly explain the increased rate of conversion of violaxanthin to zeaxanthin, but the most probable explanation of the faster conversion is an increased accessibility of violaxanthin in the membrane.     

Iron deficiency-induced changes in the photosynthetic pigment composition of field-grown pear (Pyrus communis L) leaves

In this work we characterize the changes induced by iron deficiency in the pigment composition of pear (Pyrus communis L.) leaves grown under high light intensities in field conditions in Spain. Iron deficiency induced decreases in neoxanthin and β-carotene concomitantly with decreases in chlorophyll a, whereas lutein and carotenoids within the xanthophyll cycle were less affected. Iron deficiency caused major increases in the lutein/chlorophyll a and xanthophyll cycle pigments/chlorophyll a molar ratios. The chlorophyll a/chlorophyll b ratio increased in response to iron deficiency. The carotenoids within the xanthophyll cycle in iron-deficient and in iron-sufficient (control) leaves underwent epoxidations and de-epoxidations in response to ambient light conditions. In control leaves dark-adapted for several hours, most of the xanthophyll cycle pigment pool was in the epoxidated form vio-laxanthin, whereas iron-deficient leaves had significant amounts of zeaxanthin. Iron-deficient leaves also exhibited an increased non-photochemical quenching, supporting the possibility of a role for pigments within the xanthophyll cycle in photoprotection.     

Non-photochemical quenching kinetics during the dark to light transition in relation to the formation of antheraxanthin and zeaxanthin

Nonlinear regression analysis (NLR) is applied to quantify the dynamic response of non-photochemical fluorescence quenching (NPQ) of Trifolium repens cv. Regal upon dark to light transition. Commonly, only steady-state levels of NPQ are evaluated, ignoring transient kinetics. Experimental NPQ kinetics are fitted best with a sum of two functions: a sigmoidal Hill function plus a transient logarithmic normal function. It is shown that not only steady-state level of NPQ, but also the speed at which steady state is reached, increased with light intensity. The question is raised which biological processes cause the induction of the components of NPQ kinetics. The NPQ kinetics are found to resemble the kinetics of antheraxanthin and zeaxanthin formation during a dark to light transition. Furthermore, both molecules are known to induce NPQ. The hypothesis is put forward that a transient phase of NPQ (0-2 min after transition) is dependent upon concentrations of antheraxanthin, whereas the saturating phase corresponds with the production of zeaxanthin. A mathematical model, based on the presented hypothesis, predicts the effect of increasing light intensity on concentrations of antheraxanthin and zeaxanthin which correspond with experimental results. Implications of the hypothesis are discussed as well as the role of NLR in evaluating chlorophyll a fluorescence kinetics.     

Cation radicals of xanthophylls

Carotenes and xanthophylls are well known to act as electron donors in redox processes. This ability is thought to be associated with the inhibition of oxidative reactions in reaction centers and light-harvesting pigment–protein complexes of photosystem II (PSII). In this work, cation radicals of neoxanthin, violaxanthin, lutein, zeaxanthin, β-cryptoxanthin, β-carotene, and lycopene were generated in solution using ferric chloride as an oxidant and then studied by absorption spectroscopy. The investigation provides a view toward understanding the molecular features that determine the spectral properties of cation radicals of carotenoids. The absorption spectral data reveal a shift to longer wavelength with increasing π-chain length. However, zeaxanthin and β-cryptoxanthin exhibit cation radical spectra blue-shifted compared to that of β-carotene, despite all of these molecules having 11 conjugated carbon–carbon double bonds. CIS molecular orbital theory quantum computations interpret this effect as due to the hydroxyl groups in the terminal rings selectively stabilizing the highest occupied molecular orbitals of preferentially populated s-trans-isomers. The data are expected to be useful in the analysis of spectral results from PSII pigment–protein complexes seeking to understand the role of carotene and xanthophyll cation radicals in regulating excited state energy flow, in protecting PSII reaction centers against photoinhibition, and in dissipating excess light energy absorbed by photosynthetic organisms but not used for photosynthesis.     

Carotenoids of dragonflies,from the perspective of comparative biochemical and chemical ecological studies

Carotenoids of 20 species of dragonflies (including 14 species of Anisoptera and six species of Zygoptera) were investigated from the viewpoints of comparative biochemistry and chemical ecology. In larvae, β-carotene, β-cryptoxanthin, lutein, and fucoxanthin were found to be major carotenoids in both Anisoptera and Zygoptera. These carotenoids were assumed to have originated from aquatic insects, water fleas, tadpoles, and small fish, which dragonfly larvae feed on. Furthermore, β-caroten-2-ol and echinenone were also found in all species of larvae investigated. In adult dragonflies, β-carotene was found to be a major carotenoid along with lutein, zeaxanthin, β-caroten-2-ol, and echinenone in both Anisoptera and Zygoptera. On the other hand, unique carotenoids, β-zeacarotene, β,ψ-carotene (γ-carotene), torulene, β,γ-carotene, and γ,γ-carotene, were present in both Anisoptera and Zygoptera dragonflies. These carotenoids were not found in larvae. Food chain studies of dragonflies suggested that these carotenoids originated from aphids, and/or possibly from aphidophagous ladybird beetles and spiders, which dragonflies feed on. Lutein and zeaxanthin in adult dragonflies were also assumed to have originated from flying insects they feed on, such as flies, mosquitoes, butterflies, moths, and planthoppers, as well as spiders. β-Caroten-2-ol and echinenone were found in both dragonfly adults and larvae. They were assumed to be metabolites of β-carotene in dragonflies themselves. Carotenoids of dragonflies well reflect the food chain during their lifecycle.     

Antisense inhibition of the beta-carotene hydroxylase enzyme in Arabidopsis and the implications for carotenoid accumulation,photoprotection and antenna assembly

The xanthophylls are oxygenated carotenoids and are important structural components of the photosynthetic apparatus. Xanthophylls contribute to the assembly and stability of light-harvesting complex apoproteins (LHC) and contribute to photoprotection via non-photochemical quenching of chlorophyll fluorescence (NPQ) in oxygenic photosynthetic organisms. Previously, mutations have been described that disrupt many steps in the xanthophyll biosynthetic pathway. However, there are no definitive reports of a lesion that effects the -hydroxylase enzyme, which catalyzes hydroxylation of the -rings of -carotene and -carotene, and is thus necessary for synthesis of essentially all xanthophylls of higher plant chloroplasts. We have utilized an antisense approach to effectively reduce levels of -hydroxylase in Arabidopsis thaliana in order to examine how a reduction in this enzyme impacts carotenoid biosynthesis and plant viability. Expression of the antisense -hydroxylase transgene resulted in a maximal reduction in violaxanthin of 64% and a maximal reduction in neoxanthin of 41%. This reduction was reflected in a 22% increase in -carotene and a reduction in the total carotenoid pool, whereas lutein levels were relatively unaltered. Despite the reduction in violaxanthin and neoxanthin, the antisense -hydroxylase plants had a wild-type complement of chlorophylls and LHCs on a fresh weight basis. Under high light stress, the unconverted pool of violaxanthin was the same size as in wild type and thus there was an even greater proportional reduction in zeaxanthin of 75%. Despite this marked decrease in zeaxanthin, NPQ only declined by 16%.This revised version was published online in October 2005 with corrections to the Cover Date.     

Photoinhibition, xanthophyll cycle and in vivo chlorophyll fluorescence quenching of chilling-tolerant Oxyria digyna and chilling-sensitive Zea mays

The relationship between susceptibility to photoinhibition, zeaxanthin formation and chlorophyll fluorescence quenching at suboptimal temperatures was studied in chilling-sensitive maize and in non-acclimated and cold-acclimated Oxyria digyna , a chilling-tolerant plant of arctic and alpine habitats. In maize, zeaxanthin formation was strongly suppressed by chilling. Zeaxanthin formed during preillumination at 20°C did not protect maize leaves from photoinhibition during a subsequent high-light, low-temperature treatment, as judged from the ratios of variable to maximal fluorescence, F_v/F_m. However, such preillumination significantly increased non-photochemical quenching (q_N) at low temperatures, mainly due to an enhancement of the fast-relaxing q_N component (i.e., of energy-dependent quenching. q_E). In O. digyna , cold-acclimation resulted in an increased zeaxanthin formation in the temperature range of 2.5–20°C. Cold-acclimation substantially decreased the susceptibility towards photoinhibition at 4°C, but q_N remained nearly unchanged between 2 and 38°C, as compared to control plants. Effects of cold acclimation on photosynthesis, photochemical quenching and quantum efficiency of photosystem II were small and indicated a slight amelioration only of the function of the photosynthetic apparatus at suboptimal temperatures (2–20°Ct. I) is concluded, that the xanthophyll cycle is strongly influenced by cold acclimation, while effects on the photosynthetic carbon assimilation only play a minor role in O. digyna.     

Photosynthetic Characteristics and the Ratios of Chlorophyll, β-Carotene,and the Components of the Xanthophyll Cycle Upon a Sudden Increase in Growth Light Regime in Several Plant Species*

Among three species, Gossypium hirsutum, Rhizophora mangle, and Monstera deliciosa, which were transferred from low to high growth PFD, only small decreases in the efficiency of photochemical energy conversion were observed in those plants which exhibited an increase in photosynthetic capacity. Leaves of plants which showed no increase in photosynthetic capacity experienced a continuous decrease in photochemical efficiency, accompanied by a more pronounced loss of chlorophyll than that observed in the former group. In all species marked increases in the xanthophyll/β-carotene ratio resulted from small increases in lutein, and several-fold increases in the sum of the three components of the xanthophyll cycle, zeaxanthin, antheraxanthin, and violaxanthin. A strong increase in the level of zeaxanthin was only partially matched by a decrease of violaxanthin to zero, and was further paralleled by a decrease in β-carotene. Antiparallel changes in the sum of zeaxanthin + antheraxanthin + violaxanthin and β-carotene between morning and evening were observed in all species. These diel changes were overlaid on a net increase in β-carotene as well as total carotenoid content in those plants in which photosynthetic capacity increased. In those, however, which exhibited no photosynthetic acclimation upon transfer to high light, a decrease in both β-carotene and total carotenoid content was observed. Rhizophora mangle grown at 100 % seawater exhibited a particularly high capacity for increasing the level of zeaxanthin in response to high light.     

HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY OF PHYTOPLANKTON PIGMENTS USING A POLYMERIC REVERSED-PHASE C18 COLUMN1

A high-performance liquid chromatographic (HPLC) method is described that allows improved resolution of several chemotaxonomically significant phytoplankton pigments. The protocol, which employs two pumps and a modified Mantoura and Llewellyn (1983) solvent system, can be easily adapted for many HPLC systems currently in use. The most unique aspect of the method is the use of a polymeric C₁₈ reversed phase HPLC column (VYDAC 201TP). In comparison to the monomeric C₁₈ columns typically used in the characterization of phytoplankton pigments, polymeric C₁₈ columns offer superior selectivity for structurally similar compounds. The protocol was evaluated for the ability to resolve most of the phytoplankton pigments of diagnostic importance using algal cultures from nine classes. Pigment pairs that were resolved by the method include a) lutein and zeaxanthin, b) neoxanthin and 19′-hexanoyloxyfucoxanthin, and c) α-carotene and β-carotene, and partial resolution of chlorophyll c₁ and chlorophyll c₂.     

Differential effects of carotenoids on lipid peroxidation due to membrane interactions: X-ray diffraction analysis

The biological benefits of certain carotenoids may be due to their potent antioxidant properties attributed to specific physico-chemical interactions with membranes. To test this hypothesis, we measured the effects of various carotenoids on rates of lipid peroxidation and correlated these findings with their membrane interactions, as determined by small angle X-ray diffraction approaches. The effects of the homochiral carotenoids (astaxanthin, zeaxanthin, lutein, β-carotene, lycopene) on lipid hydroperoxide (LOOH) generation were evaluated in membranes enriched with polyunsaturated fatty acids. Apolar carotenoids, such as lycopene and β-carotene, disordered the membrane bilayer and showed a potent pro-oxidant effect (> 85% increase in LOOH levels) while astaxanthin preserved membrane structure and exhibited significant antioxidant activity (40% decrease in LOOH levels). These findings indicate distinct effects of carotenoids on lipid peroxidation due to membrane structure changes. These contrasting effects of carotenoids on lipid peroxidation may explain differences in their biological activity.     

The induction of CP43′ by iron-stress in Synechococcus sp. PCC 7942 is associated with carotenoid accumulation and enhanced fatty acid unsaturation

Comparative lipid analysis demonstrated reduced amount of PG (50%) and lower ratio of MGDG/DGDG in iron-stressed Synechococcus sp. PCC 7942 cells compared to cells grown under iron sufficient conditions. In parallel, the monoenoic (C:1) fatty acids in MGDG, DGDG and PG increased from 46.8%, 43.7% and 45.6%, respectively in control cells to 51.6%, 48.8% and 48.7%, respectively in iron-stressed cells. This suggests increased membrane dynamics, which may facilitate the diffusion of PQ and keep the PQ pool in relatively more oxidized state in iron-stressed compared to control cells. This was confirmed by chlorophyll fluorescence and thermoluminescence measurements. Analysis of carotenoid composition demonstrated that the induction of isiA (CP43′) protein in response to iron stress is accompanied by significant increase of the relative abundance of all carotenoids. The quantity of carotenoids calculated on a Chl basis increased differentially with nostoxanthin, cryptoxanthin, zeaxanthin and β-carotene showing 2.6-, 3.1-, 1.9- and 1.9-fold increases, respectively, while the relative amount of caloxanthin was increased only by 30%. HPLC analyses of the pigment composition of Chl-protein complexes separated by non-denaturating SDS-PAGE demonstrated even higher relative carotenoids content, especially of cryptoxanthin, in trimer and monomer PSI Chl-protein complexes co-migrating with CP43′ from iron-stressed cells than in PSI complexes from control cells where CP43′ is not present. This implies a carotenoid-binding role for the CP43′ protein which supports our previous suggestion for effective energy quenching and photoprotective role of CP43′ protein in cyanobacteria under iron stress.     

Mechanisms of photoprotection and nonphotochemical quenching in pea light-harvesting complex at 2.5 A resolution

The plant light-harvesting complex of photosystem II (LHC-II) collects and transmits solar energy for photosynthesis in chloroplast membranes and has essential roles in regulation of photosynthesis and in photoprotection. The 2.5 A structure of pea LHC-II determined by X-ray crystallography of stacked two-dimensional crystals shows how membranes interact to form chloroplast grana, and reveals the mutual arrangement of 42 chlorophylls a and b, 12 carotenoids and six lipids in the LHC-II trimer. Spectral assignment of individual chlorophylls indicates the flow of energy in the complex and the mechanism of photoprotection in two close chlorophyll a-lutein pairs. We propose a simple mechanism for the xanthophyll-related, slow component of nonphotochemical quenching in LHC-II, by which excess energy is transferred to a zeaxanthin replacing violaxanthin in its binding site, and dissipated as heat. Our structure shows the complex in a quenched state, which may be relevant for the rapid, pH-induced component of nonphotochemical quenching.     

The synthesis of NPQ-effective zeaxanthin depends on the presence of a transmembrane proton gradient and a slightly basic stromal side of the thylakoid membrane

In the present study we address the question which factors during the synthesis of zeaxanthin determine its capacity to act as a non-photochemical quencher of chlorophyll fluorescence. Our results show that zeaxanthin has to be synthesized in the presence of a transmembrane proton gradient. However, it is not essential that the proton gradient is generated by the light-driven electron transport. NPQ-effective zeaxanthin can also be formed by an artificial proton gradient in the dark due to ATP hydrolysis. Zeaxanthin that is synthesized in the dark in the absence of a proton gradient by the low pH-dependent activation of violaxanthin de-epoxidase is not able to induce NPQ. The second important factor during the synthesis of zeaxanthin is the pH-value of the stromal side of the thylakoid membrane. Here we show that the stromal side has to be neutral or slightly basic in order to generate zeaxanthin which is able to induce NPQ. Thylakoid membranes in reaction medium pH 5.2, which experience low pH-values on both sides of the membrane, are unable to generate NPQ-effective zeaxanthin, even in the presence of an additional light-driven proton gradient. Analysing the pigment contents of purified photosystem II light-harvesting complexes we are further able to show that the NPQ ineffectiveness of zeaxanthin formed in the absence of a proton gradient is not caused by changes in its rebinding to the light-harvesting proteins. Purified monomeric and trimeric light-harvesting complexes contain comparable amounts of zeaxanthin when they are isolated from thylakoid membranes enriched in either NPQ-effective or ineffective zeaxanthin.     

In vitro and in vivo biosynthesis of xanthophylls by the cyanobacterium Aphanocapsa

Of the six carotenoids identified in the cyanobacterium Aphanocapsa, β-carotene, zeaxanthin, echinenone and myxoxanthophyll are the major pigments, whilst β-cryptoxanthin and 3-hydroxy-4-keto-β-carotene are present only in trace amounts. With the exception of zeaxanthin, the other xanthophylls could be formed in vitro from [¹⁴C]phytoene in high yields, especially β-cryptoxanthin and 3-hydroxy-4-keto-β-carotene. In a time course experiment of xanthopyll biosynthesis the flow of radioactivity from [¹⁴C]phytoene was followed through the pools of phytofluene, lycopene, and β-carotene. The reaction sequence from phytoene to xanthophylls is sensitive in vitro to both difunone, an inhibitor of carotene desaturation, and CPTA, an inhibitor of cyclization.     

The Lhcb protein and xanthophyll composition of the light harvesting antenna controls the DeltapH-dependency of non-photochemical quenching in Arabidopsis thaliana

Nonphotochemical quenching (NPQ) is the photoprotective dissipation of energy in photosynthetic membranes. The hypothesis that the DeltapH-dependent component of NPQ (qE) component of non-photochemical quenching is controlled allosterically by the xanthophyll cycle has been tested using Arabidopsis mutants with different xanthophyll content and composition of Lhcb proteins. The titration curves of qE against DeltapH were different in chloroplasts containing zeaxanthin or violaxanthin, proving their roles as allosteric activator and inhibitor, respectively. The curves differed in mutants deficient in lutein and specific Lhcb proteins. The results show that qE is determined by xanthophyll occupancy and the structural interactions within the antenna that govern allostericity.     

Photosynthesis and chlorophyllafluorescence during flag leaf senescence of field-grown wheat plants

The characteristics of photosynthetic gas exchange, chlorophyll a fluorescence, and xanthophyll cycle pigments during flag leaf senescence of field-grown wheat plants were investigated. With senescence progressing, the light-saturated net CO₂ assimilation rate expressed either on a basis of leaf area or chlorophyll decreased significantly. The apparent quantum yield of net photosynthesis decreased when expressed on a leaf area basis but increased when expressed on a chlorophyll basis. The maximal efficiency of PSII photochemistry decreased very little while actual PSII efficiency, photochemical quenching, and the efficiency of excitation capture by open PSII centers decreased considerably. At the same time, non-photochemical quenching increased significantly. A substantial decrease in the contents of violaxanthin and zeaxanthin, but a slight decrease in the content of antheraxanthin were observed. However, the de-epoxidation status of the xanthophyll cycle was positively correlated with progressive senescence. This increase was due mainly to a smaller decrease in zeaxanthin than in violaxanthin. Our results suggest that PSII apparatus remained functional, but a down-regulation of PSII occurred under the steady state of photosynthesis in senescent flag leaves. Such a down-regulation was associated with the closure of PSII centers and an enhanced xanthophyll cycle-related thermal dissipation in the PSII antennae.