Strategies providing success in a variable habitat: II. Ecophysiology of photosynthesis of Cladophora glomerata

Cladophora glomerata (L.) Kütz. is the dominant filamentous algae of the river Ilm, Thuringia, Germany. For most of the year it can be found at open as well as at shaded sites. Photosynthetic acclimation of C. glomerata to different light intensities was detected by chlorophyll fluorescence measurements and pigment analysis. Cladophora glomerata from highlight sites showed decreased values of efficiency of open photosystem II (F_v/F_m) as compared with C. glomerata from low‐light sites. Winter populations revealed higher F_v/F_m values than summer populations. A light‐induced decrease in efficiency of the closed photosystem II was observed at increasing irradiance intensities. The decrease was higher in C. glomerata from shaded sites compared with plants from open sites. Differences in the photosynthetic electron transport rate of different populations of C. glomerata were shown by photosynthesis–irradiance curves. Summer populations from high‐light sites yielded higher maximum electron transport rates than plants from low‐light sites, whereas winter populations exhibited significantly decreased values compared with the summer populations. Results of the analysis of photosynthetic pigments corresponded with data from chlorophyll fluorescence measurements. In addition to these long‐term acclimation effects, C. glomerata expressed its ability to cope with rapid changes in the light environment by the de‐epoxidation of violaxanthin during exposure to high light intensities.     

Mechanistic aspects of xanthophyll cycle-dependent photoprotection in higher plant chloroplasts and leaves

Higher plants must dissipate absorbed light energy that exceeds the photosynthetic capacity to avoid molecular damage to the pigments and proteins that comprise the photosynthetic apparatus. Described in this minireview is a current view of the biochemical, biophysical and bioenergetic aspects of the primary photoprotective mechanism responsible for dissipating excess excitation energy as heat from photosystem II (PSII). The photoprotective heat dissipation is measured as nonphotochemical quenching (NPQ) of the PSII chlorophyll a (Chl a) fluorescence. The NPQ mechanism is controlled by the trans-thylakoid membrane pH gradient (ΔpH) and the special xanthophyll cycle pigments. In the NPQ mechanism, the de-epoxidized endgroup moieties and the trans-thylakoid membrane orientations of antheraxanthin (A) and zeaxanthin (Z) strongly affect their interactions with protonated chlorophyll binding proteins (CPs) of the PSII inner antenna. The CP protonation sites and steps are influenced by proton domains sequestered within the proteo-lipid core of the thylakoid membrane. Xanthophyll cycle enrichment around the CPs may explain why changes in the peripheral PSII antenna size do not necessarily affect either the concentration of the xanthophyll cycle pigments on a per PSII unit basis or the NPQ mechanism. Recent time-resolved PSII Chi a fluorescence studies suggest the NPQ mechanism switches PSII units to an increased rate constant of heat dissipation in a series of steps that include xanthophyll de-epoxidation, CP-protonation and binding of the xanthophylls to the protonated CPs; the concerted process can be described with a simple two-step, pH-activation model. The xanthophyll cycle-dependent NPQ mechanism is profoundly influenced by temperatures suboptimal for photosynthesis via their effects on the trans-thylakoid membrane energy coupling system. Further, low temperature effects can be grouped into either short term (minutes to hours) or long term (days to seasonal) series of changes in the content and composition of the PSII pigment-proteins. This minireview concludes by briefly highlighting primary areas of future research interest regarding the NPQ mechanism.     

Arabidopsis plants lacking PsbS protein possess photoprotective energy dissipation

It is commonly accepted that the photosystem II subunit S protein, PsbS, is required for the dissipation of excess light energy in a process termed ‘non‐photochemical quenching’ (NPQ). This process prevents photo‐oxidative damage of photosystem II (PSII) thus avoiding photoinhibition which can decrease plant fitness and productivity. In this study Arabidopsis plants lacking PsbS (the npq4 mutant) were found to possess a competent mechanism of excess energy dissipation that protects against photoinhibitory damage. The process works on a slower timescale, taking about 1 h to reach the same level of NPQ achieved in the wild type in just a few minutes. The NPQ in npq4 was found to display very similar characteristics to the fast NPQ in the wild type. Firstly, it prevented the irreversible light‐induced closure of PSII reaction centres. Secondly, it was uncoupler‐sensitive, and thus triggered by the ΔpH across the thylakoid membrane. Thirdly, it was accompanied by significant quenching of the fluorescence under conditions when all PSII reaction centres were open (F_o state)_. Fourthly, it was accompanied by NPQ‐related absorption changes (ΔA535). Finally, it was modulated by the presence of the xanthophyll cycle carotenoid zeaxanthin. The existence of a mechanism of photoprotective energy dissipation in plants lacking PsbS suggests that this protein plays the role of a kinetic modulator of the energy dissipation process in the PSII light‐harvesting antenna, allowing plants to rapidly track fluctuations of light intensity in the environment, and is not the primary cause of NPQ or a direct carrier of the pigment acting as the non‐photochemical quencher.     

The peculiar NPQ regulation in the stramenopile Phaeomonas sp. challenges the xanthophyll cycle dogma

In changing light conditions, photosynthetic organisms develop different strategies to maintain a fine balance between light harvesting, photochemistry, and photoprotection. One of the most widespread photoprotective mechanisms consists in the dissipation of excess light energy in the form of heat in the photosystem II antenna, which participates to the Non Photochemical Quenching (NPQ) of chlorophyll fluorescence. It is tightly related to the reversible epoxidation of xanthophyll pigments, catalyzed by the two enzymes, the violaxanthin deepoxidase and the zeaxanthin epoxidase. In Phaeomonas sp. (Pinguiophyte, Stramenopiles), we show that the regulation of the heat dissipation process is different from that of the green lineage: the NPQ is strictly proportional to the amount of the xanthophyll pigment zeaxanthin and the xanthophyll cycle enzymes are differently regulated. The violaxanthin deepoxidase is already active in the dark, because of a low luminal pH, and the zeaxanthin epoxidase shows a maximal activity under moderate light conditions, being almost inactive in the dark and under high light. This light-dependency mirrors the one of NPQ: Phaeomonas sp. displays a large NPQ in the dark as well as under high light, which recovers under moderate light. Our results pinpoint zeaxanthin epoxidase activity as the prime regulator of NPQ in Phaeomonas sp. and therefore challenge the deepoxidase-regulated xanthophyll cycle dogma.     

Photoadaptation of two members of the Chlorophyta (Scenedesmus and Chlorella) in laboratory and outdoor cultures: changes in chlorophyll fluorescence quenching and the xanthophyll cycle

The role of the xanthophyll cycle in the adaptation of two chlorococcal algae Scenedesmus quadricauda and Chlorella sorokiniana to high irradiance was studied under laboratory and outdoor conditions. We wished to elucidate whether the xanthophyll cycle plays a key role in dissipating the excesses of absorbed light, as in higher plants, and to characterise the relationship between chlorophyll fluorescence parameters and the content of xanthophyll-cycle pigments. The xanthophyll cycle was found to be operative in both species; however, its contribution to overall non-photochemical quenching (NPQ) could only be distinguished in Scenedesmus (15–20% of total NPQ). The Scenedesmus cultures showed a larger pool of xanthophyll-cycle pigments than Chlorella, and lower sensitivity to photoinhibition as judged from the reduction of maximum quantum yield of photosystem II. In general, both algae had a larger xanthophyll-cycle pool when grown outdoors than in laboratory cultures. Comparing the two species, Scenedesmus exhibited a higher capacity to adapt to high irradiance, due to an effective quenching mechanism and high photosynthetic capacity; in contrast, Chlorella represents a species with a larger antennae system, less-efficient quenching and lower photosynthetic performance. Non-photochemical quenching (NPQ) induced through the xanthophyll cycle can, to a limited extent, represent a regulatory factor in diluted algal cultures grown in outdoor solar photobioreactors, as well as in natural algal phytoplankton populations exposed transiently to high irradiance. However, it does not play an appreciable role in dense, well-mixed microalgal suspensions. Received: 6 August 1998 / Accepted: 12 February 1999     

Seasonal differences in xanthophyll cycle characteristics and antioxidants in Mahonia repens growing in different light environments

We investigated differences between summer and winter in photosynthesis, xanthophyll cycle-dependent energy dissipation, and antioxidant systems in populations of Mahonia repens (Lindley) Don growing in the eastern foothills of the Colorado Rocky Mountains in deep shade, full exposure, and under a single-layered canopy of Pinus ponderosa (partially shaded). In summer, increasing growth irradiance (from deep shade to partial shade to full exposure) was associated with increased xanthophyll cycle-dependent energy dissipation in PSII and an increased capacity to detoxify reactive reduced oxygen species, as measured by increases in the activities of ascorbate peroxidase, superoxide scavenging, glutathione reductase, and monodehydroascorbate reductase, as well as increases in leaf ascorbate and glutathione content. Leaves of exposed and partially shaded plants exhibited decreased capacities for photosynthetic O₂ evolution in winter compared to summer, while in the deeply shaded plants this parameter did not differ seasonally. Seasonal differences in the levels of antioxidants generally exhibited an inverse response to photosynthesis, being higher in winter compared to summer in the exposed and partially shaded populations, but remaining unchanged in the deeply shaded population. In addition, total pool size and conversion state of the xanthophyll cycle were higher in winter than in summer in all populations. These trends suggest that both xanthophyll cycle-dependent energy dissipation in PSII and the capacity to detoxify reactive reduced oxygen species responded to the level of excess light absorption. Received: 23 October 1997 / Accepted: 23 March 1998     

Photoprotective role of rhodoxanthin during cold acclimation in Cryptomeria japonica

To examine the role of rhodoxanthin in long‐term acclimation to low temperatures, we monitored seasonal changes in pigment composition, photosynthesis, chlorophyll fluorescence and the level of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) in needles of wild‐type and mutant forms of Cryptomeria japonica. In winter, rhodoxanthin accumulated in sun‐exposed needles of wild‐type plants, but not in those of the mutant. The level of chlorophyll decreased in both types of plant in winter. In contrast, the level of the xanthophyll cycle pool increased in both cases. The level of the pool in the mutant was twice that in the wild type in winter, on a Chl basis, even though the levels in both were similar in summer. The synthesis of rhodoxanthin might be triggered by photo‐inhibitory conditions, as suggested by the sustained elevated levels of zeaxanthin (Z) and antheraxanthin (A). In the wild type and the mutant, the quantum yield of CO₂ fixation (φ), the photosynthetic capacity, the photochemical efficiency of photosystem II (PSII), the photochemical quenching and the level of Rubisco in summer were similar. However, all these values for the wild type were higher than those for the mutant in winter. The non‐photochemical quenching (NPQ) in the mutant in winter increased rapidly even under low light conditions due to the high sustained levels of Z and A. In contrast, in the wild type, the conversion of Z via A to rhodoxanthin prevented the rapid increase in NPQ to maintain the relatively high level of φ. These findings suggest that rhodoxanthin might play an important photoprotective role in long‐term acclimation to cold. The dynamic regulation of the amount of rhodoxanthin relative to the level of the xanthophyll cycle pool might act to maintain an appropriate balance between light absorption, photosynthesis and the thermal dissipation of energy due to excess absorbed light in winter.     

Diversity in Xanthophyll Cycle Pigments Content and Related Nonphotochemical Quenching (NPQ) Among Microalgae: Implications for Growth Strategy and Ecology

Xanthophyll cycle-related nonphotochemical quenching (NPQ), which is present in most photoautotrophs, allows dissipation of excess light energy. Xanthophyll cycle-related NPQ depends principally on xanthophyll cycle pigments composition and their effective involvement in NPQ. Xanthophyll cycle-related NPQ is tightly controlled by environmental conditions in a species-/strain-specific manner. These features are especially relevant in microalgae living in a complex and highly variable environment. The goal of this study was to perform a comparative assessment of NPQ ecophysiologies across microalgal taxa in order to underline the specific involvement of NPQ in growth adaptations and strategies. We used both published results and data acquired in our laboratory to understand the relationships between growth conditions (irradiance, temperature, and nutrient availability), xanthophyll cycle composition, and xanthophyll cycle pigments quenching efficiency in microalgae from various taxa. We found that in diadinoxanthin-containing species, the xanthophyll cycle pigment pool is controlled by energy pressure in all species. At any given energy pressure, however, the diatoxanthin content is higher in diatoms than in other diadinoxanthin-containing species. XC pigments quenching efficiency is species-specific and decreases with acclimation to higher irradiances. We found a clear link between the natural light environment of species/ecotypes and quenching efficiency amplitude. The presence of diatoxanthin or zeaxanthin at steady state in all species examined at moderate and high irradiances suggests that cells maintain a light-harvesting capacity in excess to cope with potential decrease in light intensity.     

Non-photochemical quenching of chlorophyll fluorescence in Chlorella fusca acclimated to constant and dynamic light conditions

Non-photochemical quenching of chlorophyll fluorescence (NPQ) involves dissipation of light energy in the photosynthetic apparatus via a number of physiologically distinct processes. The relationships among NPQ, the (de)epoxidation state of the xanthophyll cycle pigments and state transitions was studied in the green alga Chlorella fusca, acquired from six differently light-acclimated continuous cultures. A 10 h light and 14 h darkness, periodicity was obeyed in all cultures. Three cultures received a high total daily irradiance, three others a low one. High and low irradiances were each dosed in three different modes at constant supply, with sine shape intensity modulation, or as a sine with superimposed oscillations. In the constant supply mode, but not for the sine and oscillating modes, high-light rendered a three-fold higher xantophyll cycle pigment content than low-light. Dynamic interconversion of xantophyll cycle pigments was restricted to high-light cultures. NPQ followed the kinetics of the light supply mode and was highest in high light cultures. In low-light cultures, NPQ correlated mainly to state transitions. These observations were supported by experiments with dithiothreithol-treated samples. The relative impact of xantophyll cycle operation and state transitions on NPQ in green algae from different light climates will be discussed with reference to higher plants. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effects of Exogenous Putrescine on Chlorophyll Fluorescence Imaging and Heat Dissipation Capacity in Cucumber (Cucumis sativus L.) Under Salt Stress

The objective of this study was to identify the effects of exogenous putrescine on photosynthetic performance and heat dissipation capacity in cucumber seedlings under salt stress. The stress of 75 mM NaCl for 7 days caused a significant decrease in net photosynthetic rate (P _N ). The experiment employed a chlorophyll fluorescence imaging technique and demonstrated that the maximal quantum yield of photosystem II photochemistry (Fv/Fm) and the actual photochemical efficiency of photosystem II (ΦPSII) were reduced by salt stress. Moreover, salt stress markedly reduced the photochemical quenching coefficient (qP) and non-photochemical quenching coefficient (qN), and significantly increased non-regulated heat dissipation (ΦNO). However, stressed plants supplied with exogenous putrescine exhibited higher P _N and ΦPSII, which indicated that putrescine can alleviate the detrimental effects on photosynthesis induced by salt stress. Putrescine sprayed on stressed plants significantly enhanced the regulated energy dissipation (ΦNPQ) and decreased ΦNO. Application of exogenous putrescine also changed the levels of xanthophyll cycle components and further enhanced the de-epoxidation state of xanthophyll cycle pigments under salt stress. Under control conditions, putrescine exerted little influence on the photosynthetic parameters in cucumber leaves. In conclusion, the application of exogenous putrescine may improve the heat dissipation capacity by promoting the xanthophyll cycle to reduce the damage caused by excess excitation energy, thus enhancing the salt tolerance of cucumber seedlings.     

Effects of ultraviolet (UV) exclusion on the seasonal concentration of photosynthetic and UV-screening pigments in Scots pine needles

The effects of ultraviolet (UV) radiation on the photosynthetic and UV‐screening pigments in needles of Scots pine (Pinus sylvestris L.) saplings were studied in a UV‐exclusion field chamber experiment in northern Finland (67°N) during 2001–2002. The chambers held filters that excluded both UVB and UVA, only UVB, transmitted all UV, or lacked filters. Analyses of control needles (no filter and polyethene filter) showed that the first changes to occur in spring (end of April) was an abrupt increase in the epoxidation state (EPS) of the xanthophyll cycle pigments, likely in relation with the beginning of the photosynthetic activity. The concentration of chlorophyll, lutein, neoxanthin, α‐carotene, β‐carotene, and the size of the xanthophyll cycle pool (violaxanthin+antheraxanthin+zeaxanthin=VAZ) changed only later when needles reached their summer photosynthesis state. Exclusion of UV radiation significantly affected the xanthophyll cycle but not the other photosynthetic pigments analysed. Interestingly, the effects on xanthophylls were dependent on the sampling date. Under UVA/B‐exclusion, the EPS was increased and VAZ pool size was unchanged in April, whereas EPS remained unchanged and the VAZ pool size was reduced in May and June. The existence of two sustained and active antenna modes during winter and summer could be an explanation for the specific UV‐exclusion effect in the different season. A high‐performance liquid chromatography analysis of soluble phenolics showed that the exclusion of UVA/B radiation caused a significant effect on five compounds out of 46 studied, without affecting the concentration of the total soluble phenolics. Under UVA/B‐exclusion, the concentration of three of them (secoisolariciresinol‐glucopyranoside, two unknown) was reduced while the concentration of dicoumaroyl‐astragalin and pinosylvin monomethylether was increased compared with both controls separately. In general, the exclusion of UVA/B caused a stronger effect than the exclusion of UVB on both photosynthetic and UV screening pigments. The effects of UV radiation on xanthophyll cycle pigments were season‐specific and detectable only under stressful spring conditions (freezing temperatures and high irradiance due to snow reflection). The effect on the xanthophyll cycle could be a direct consequence of UV treatments, or an indirect consequence of the changed flavonoid composition, or a combination of both.     

Close relationship between the state of the xanthophyll cycle pigments and photosystem II efficiency during recovery from winter stress

The potential involvement of the xanthophyll cycle in photoprotection of overwintering evergreen plants was investigated. Leaves from five evergreen species. Pseudotsuga menziesii, Pinus panderosa, Euonyums kiautschovicus. Mahonia repens and Malva neglecta, were collected from the field predawn during winter and transferred to the laboratory where chlorophyll fluorescence emission as well as the chlorophyll and carotenoid composition were ascertained periodically for 4.5 days. Leaves and needles from all species were found to have retained large amounts of the xanthophyll cycle pigments zeaxanthin and antheraxanthin, and they exhibited sustained low values of the intrinsic efficiency of photosystem II (PSII; measured as the ratio of variable to maximal fluorescence, F_v/F_m) upon collection. The increase in PSII efficiency was biphasic, with a rapid phase (requiring several hours) and a slow phase (requiring several days). Changes in the conversion state of the xanthophyll cycle were found to correlate with increases in PSII efficiency in both phases, with the latter phase involving large increases in both F_m (maximal fluorescence) and F_o (minimal fluorescence) throughout the period of recovery. The relationship between F_m quenching (expressed as nonphotochemical or Stern-Volmer quenching [NPQ] of F_m, i.e. F_m/ F_m–1) and F_o quenching (F_o/F_o–1) was linear, as expected for changes in xanthophyll cycle-dependent energy dissipation in the antenna complexes. Furthermore, the relationship between F_v/F_m and NPQ during recovery followed the theoretical relationship predicted for changes in the rate constant for energy dissipation in the antenna complexes. This fit between the theoretical relationship and the actual data indicates that all changes in NPQ or F_v/F_m can be accounted for by changes in this rate constant. The results suggest a role for the photoprotective xanthophyll cycle-dependent dissipation process in the lowered efficiency of PSII observed in coldstressed evergreen plants in the field.     

Cooperation of xanthophyll cycle with water-water cycle in the protection of photosystems 1 and 2 against inactivation during chilling stress under low irradiance

The xanthophyll cycle and the water-water cycle had different functional significance in chilling-sensitive sweet pepper upon exposure to chilling temperature (4 °C) under low irradiance (100 µmol m⁻² s⁻¹) for 6 h. During chilling stress, effects of non-photochemical quenching (NPQ) on photosystem 2 (PS2) in dithiothreitol (DTT) fed leaves remained distinguishable from that of the water-water cycle in diethyldithiocarbamate (DDTC) fed leaves. In DTT-fed leaves, NPQ decreased greatly accompanied by visible inhibition of the de-epoxidized ratio of the xanthophyll cycle, and maximum photochemical efficiency of PS2 (F_v/F_m) decreased markedly. Thus the xanthophyll cycle-dependent NPQ could protect PS2 through energy dissipation under chilling stress. However, NPQ had a slighter effect on photosystem 1 (PS1) in DTT-fed leaves than in DDTC-fed leaves, whereas effects of the water-water cycle on PS1 remained distinguishable from that of NPQ. Inhibiting superoxide dismutase (SOD) activity increased the accumulation of , the oxidation level of P700 (P700⁺) decreased markedly relative to the control and DTT-fed leaves. Both F_v/F_m and NPQ changed little in DDTC-fed leaves accompanied by little change of (A+Z)/(V+A+Z). This is the active oxygen species inducing PS1 photoinhibition in sweet pepper. The water-water cycle can be interrupted easily at chilling temperature. We propose that during chilling stress under low irradiance, the xanthophyll cycle-dependent NPQ has the main function to protect PS2, whereas the water-water cycle is not only the pathway to dissipate energy but also the dominant factor causing PS1 chilling-sensitivity in sweet pepper.This research was supported by the State Key Basic Research and Development Plan of China (G1998010100), the Natural Science Foundation of China (30370854), and the open project from Key Lab of Crop Biology of Shandong Province.     

Two different strategies for light utilization in photosynthesis in relation to growth and cold acclimation

Seedlings of Lodgepole pine (Pinus contorta L.) and winter wheat (Triticum aestivum L. cv. Monopol) were cold acclimated under controlled conditions to induce frost hardiness. Lodgepole pine responded to cold acclimation by partial inhibition of photosynthesis with an associated partial loss of photosystem II reaction centres, and a reduction in needle chlorophyll content. This was accompanied by a low daily carbon gain, and the development of a high and sustained capacity for non‐photochemical quenching of absorbed light. This sustained dissipation of absorbed light as heat correlated with an increased de‐epoxidation of the xanthophyll cycle pigments forming the quenching forms antheraxanthin and zeaxanthin. In addition, the PsbS protein known to bind chlorophyll and the xanthophyll cycle pigments increased strongly during cold acclimation of pine. In contrast, winter wheat maintained high photosynthetic rates, showed no loss of chlorophyll content per leaf area, and exhibited a high daily carbon gain and a minimal non‐photochemical quenching after cold acclimation. In accordance, cold acclimation of wheat neither increased the de‐epoxidation of the xanthophylls nor the content of the PsbS protein. These different responses of photosynthesis to cold acclimation are correlated with pine, reducing its need for assimilates when entering dormancy associated with termination of primary growth, whereas winter wheat maintains a high need for assimilates as it continues to grow and develop throughout the cold‐acclimation period. It appears that without evolving a sustained ability for controlled dissipation of absorbed light as heat throughout the winter, winter green conifers would not have managed to adapt and establish themselves so successfully in the cold climatic zones of the northern hemisphere.     

Allosteric regulation of the light-harvesting system of photosystem II

Non-photochemical quenching of chlorophyll fluorescence (NPQ) is symptomatic of the regulation of energy dissipation by the light-harvesting antenna of photosystem II (PS II). The kinetics of NPQ in both leaves and isolated chloroplasts are determined by the transthylakoid delta pH and the de-epoxidation state of the xanthophyll cycle. In order to understand the mechanism and regulation of NPQ we have adopted the approaches commonly used in the study of enzyme-catalysed reactions. Steady-state measurements suggest allosteric regulation of NPQ, involving control by the xanthophyll cycle carotenoids of a protonation-dependent conformational change that transforms the PS II antenna from an unquenched to a quenched state. The features of this model were confirmed using isolated light-harvesting proteins. Analysis of the rate of induction of quenching both in vitro and in vivo indicated a bimolecular second-order reaction; it is suggested that quenching arises from the reaction between two fluorescent domains, possibly within a single protein subunit. A universal model for this transition is presented based on simple thermodynamic principles governing reaction kinetics.     

Nocturnally retained zeaxanthin does not remain engaged in a state primed for energy dissipation during the summer in two Yucca species growing in the Mojave Desert

Differently oriented leaves of Yucca schidigera and Yucca brevifolia were characterized in the Mojave Desert with respect to photosystem II and xanthophyll cycle activity during three different seasons, including the hot and dry summer, the relatively cold winter, and the mild spring season. Photosynthetic utilization of a high percentage of the light absorbed in PSII was observed in all leaves only during the spring, whereas very high levels of photoprotective, thermal energy dissipation were employed both in the summer and the winter season in all exposed leaves of both species. Both during the summer and the winter season, when energy dissipation levels were high diurnally, xanthophyll cycle pools (relative to either Chl or other carotenoids) were higher relative to the spring, and a nocturnal retention of high levels of zeaxanthin and antheraxanthin (Z + A) occurred in all exposed leaves of both species. Although this nocturnal retention of Z + A was associated with nocturnal maintenance of a low PSII efficiency (F_v/F_m) on a cold winter night, pre‐dawn F_v/F_m was high in (Z + A)‐retaining leaves following a warm summer night. This indicates nocturnal engagement of Z + A in a state primed for energy dissipation throughout the cold winter night – while high levels of retained Z + A were not engaged for energy dissipation prior to sunrise on a warm summer morning. Possible mechanisms for a lack of sustained engagement of retained Z + A for energy dissipation at elevated temperatures are discussed.     

Rapid regulation of excitation energy in two pennate diatoms from contrasting light climates

Non-photochemical quenching (NPQ) is a fast acting photoprotective response to high light stress triggered by over excitation of photosystem II. The mechanism for NPQ in the globally important diatom algae has been principally attributed to a xanthophyll cycle, analogous to the well-described qE quenching of higher plants. This study compared the short-term NPQ responses in two pennate, benthic diatom species cultured under identical conditions but which originate from unique light climates. Variable chlorophyll fluorescence was used to monitor photochemical and non-photochemical excitation energy dissipation during high light transitions; whereas whole cell steady state 77 K absorption and emission were used to measure high light elicited changes in the excited state landscapes of the thylakoid. The marine shoreline species Nitzschia curvilineata was found to have an antenna system capable of entering a deeply quenched, yet reversible state in response to high light, with NPQ being highly sensitive to dithiothreitol (a known inhibitor of the xanthophyll cycle). Conversely, the salt flat species Navicula sp. 110-1 exhibited a less robust NPQ that remained largely locked-in after the light stress was removed; however, a lower amplitude, but now highly reversible NPQ persisted in cells treated with dithiothreitol. Furthermore, dithiothreitol inhibition of NPQ had no functional effect on the ability of Navicula cells to balance PSII excitation/de-excitation. These different approaches for non-photochemical excitation energy dissipation are discussed in the context of native light climate.     

Xanthophylls as modulators of membrane protein function

This review discusses the structural aspect of the role of photosynthetic antenna xanthophylls. It argues that xanthophyll hydrophobicity/polarity could explain the reason for xanthophyll variety and help to understand their recently emerging function - control of membrane organization and the work of membrane proteins. The structure of a xanthophyll molecule is discussed in relation to other amphiphilic compounds like lipids, detergents, etc. Xanthophyll composition of membrane proteins, the role of their variety in protein function are discussed using as an example for the major light harvesting antenna complex of photosystem II, LHCII, from higher plants. A new empirical parameter, hydrophobicity parameter (H-parameter), has been introduced as an effective measure of the hydrophobicity of the xanthophyll complement of LHCII from different xanthophyll biosynthesis mutants of Arabidopsis. Photosystem II quantum efficiency was found to correlate well with the H-parameter of LHCII xanthophylls. PSII down-regulation by non-photochemical chlorophyll fluorescence quenching, NPQ, had optimum corresponding to the wild-type xanthophyll composition, where lutein occupies intrinsic sites, L1 and L2. Xanthophyll polarity/hydrophobicity alteration by the activity of the xanthophyll cycle explains the allosteric character of NPQ regulation, memory of illumination history and the hysteretic nature of the relationship between the triggering factor, ΔpH, and the energy dissipation process.     

Photoprotection in a zeaxanthin- and lutein-deficient double mutant of Arabidopsis

When light absorption by a plant exceeds its capacity for light utilization, photosynthetic light harvesting is rapidly downregulated by photoprotective thermal dissipation, which is measured as nonphotochemical quenching of chlorophyll fluorescence (NPQ). To address the involvement of specific xanthophyll pigments in NPQ, we have analyzed mutants affecting xanthophyll metabolism in Arabidopsis thaliana. An npq1 lut2 double mutant was constructed, which lacks both zeaxanthin and lutein due to defects in the violaxanthin de-epoxidase and lycopene -cyclase genes. The npq1 lut2 strain had normal Photosystem II efficiency and nearly wild-type concentrations of functional Photosystem II reaction centers, but the rapidly reversible component of NPQ was completely inhibited. Despite the defects in xanthophyll composition and NPQ, the npq1 lut2 mutant exhibited a remarkable ability to tolerate high light.This revised version was published online in October 2005 with corrections to the Cover Date.