State Of Xanthophyll Cycle Pigments Research Articles

The Amount ofZeaxanthin Epoxidase But Not the Amount ofViolaxanthin De-Epoxidase Is a Critical Determinant ofZeaxanthin Accumulation inArabidopsis thaliana andNicotiana tabacum.

The generation of violaxanthin (Vx) de-epoxidase (VDE), photosystem II subunit S (PsbS) and zeaxanthin (Zx) epoxidase (ZEP) (VPZ) lines, which simultaneously overexpress VDE, PsbS and ZEP, has been successfully used to accelerate the kinetics of the induction and relaxation of non-photochemical quenching (NPQ). Here, we studied the impact of the overexpression of VDE and ZEP on the conversion of the xanthophyll cycle pigments in VPZ lines of Arabidopsis thaliana and Nicotiana tabacum. The protein amount of both VDE and ZEP was determined to be increased to about 3- to 5-fold levels of wild-type (WT) plants for both species. Compared to WT plants, the conversion of Vx to Zx, and hence VDE activity, was only marginally accelerated in VPZ lines, whereas the conversion of Zx to Vx, and thus ZEP activity, was strongly increased in VPZ lines. This indicates that the amount of ZEP but not the amount of VDE is a critical determinant of the equilibrium of the de-epoxidation state of xanthophyll cycle pigments under saturating light conditions. Comparing the two steps of epoxidation, particularly the second step (antheraxanthin to Vx) was found to be accelerated in VPZ lines, implying that the intermediate Ax is released into the membrane during epoxidation by ZEP.

Correction of PRI for carotenoid pigment pools improves photosynthesis estimation across different irradiance and temperature conditions

We studied the influence of changing carotenoid pigments on the sensitivity of the photochemical reflectance index (PRI) to photosynthesis dynamics. The goal of the measurements was to examine how the introduction of ΔPRI into the working dataset can improve the estimation of photosynthesis. Spectral and photosynthetic characteristics of European beech and Norway spruce saplings were periodically measured in growth chambers with an adjustable irradiance and temperature. Patterns of environmental changes inside the growth chambers were created by periodic changes in irradiance and temperature. Four general irradiance periods lasting 10–12 days each were established. Introduced irradiance regimes varied in the sum of daily irradiance and amplitude of irradiance changes. Temperature was changed with more complex patterns to induce changes in xanthophyll cycle pigments at various time scales within these regimes. Our measurements confirmed the PRI linkage to photosynthetic light use efficiency (LUE). However, the strength of this connection was found to be dependent on changing pigment concentrations, specifically on the change in the ratio of chlorophylls to carotenoids. Furthermore, a negative interference in photosynthesis estimation from PRI was recorded if the temperature was lowered overnight to 12 °C. The differential PRI (ΔPRI), calculated as the simple difference between the PRI value measured during the daytime period and in early morning (PRI0), revealed a decreased effect from pigments and cold temperature on LUE estimation. The regression analysis among all measured data identified an increased association between PRI and LUE following the introduction of ΔPRI from R2 = 0.26 to 0.69 in beech and from R2 = 0.61 to 0.77 in spruce data. The analyses showed that both leaf carotenoid concentrations and the conversion state of xanthophyll cycle pigments played a significant role in determining PRI and PRI0 values and that the accurate assessment of these pigments in PRI across multiple levels of stress from irradiance and temperature might improve estimations of LUE through ΔPRI. In our data, ΔPRI appeared to be a good measure of photosynthesis, the dynamics of which differed between beech and spruce saplings upon switching temperatures.

Pigments, efficiency of photosynthesis and winter wheat productivity

Purpose. To analyze characteristics of pigment apparatus and photosynthetic efficiency of wheat in connection with the perspectives to increase yielding capacity. Methods. Field, small-plot, morphometric, spectrophotometric, high-performance liquid chromatography, statistical ones. Results. The results of comparative studies of the photosynthetic apparatus characteristics of two winter wheat varieties at different levels of its organization (chloroplast, leaf, and crop), with more than 40 years difference of breeding time, are presented. It was shown that under different growing conditions the modern variety ‘Favorytka’ differed both by higher content of chlorophyll as the main photosynthetic pigment and its gross amount in the leaves, as well as specific leaves weight and longer functioning of crop photosynthetic apparatus at the late stages of vegetation than old variety ‘Myronivska 808’. Based on the changes of the de-epoxidation state of xanthophyll cycle pigments, caused by variation of light conditions, the better photosynthetic apparatus efficiency of variety ‘Favorytka’ was established. All these changes ultimately contributed to more efficient use of absorbed light energy for biomass formation of this variety. Found on the analysis of the own data and literature, it is shown that increasing the efficiency of photosynthesis is a promising strategy for rai­sing plant productivity. Conclutions. It was found that the increase of yield of modern winter wheat variety ‘Favorytka’, as compared with variety ‘Myronivska 808’ to be bred in the 60s, was accompanied with a rise of content and gross amount of chlorophyll and a prolongation of functioning of crop photosynthetic apparatus during the reproductive period. In addition, the modern variety was characterized by an increase in photosynthetic productivity due to more efficient use of absorbed light energy.

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.

Using leaf optical properties to detect ozone effects on foliar biochemistry

Efficient methods for accurate and meaningful high-throughput plant phenotyping are limiting the development and breeding of stress-tolerant crops. A number of emerging techniques, specifically remote sensing methods, have been identified as promising tools for plant phenotyping. These remote sensing methods can be used to accurately and rapidly relate variations in leaf optical properties with important plant characteristics, such as chemistry, morphology, and photosynthetic properties at the leaf and canopy scales. In this study, we explored the potential to utilize optical (λ=500-2,400nm) near-surface remote sensing reflectance spectroscopy to evaluate the effects of ozone pollution on photosynthetic capacity of soybean (Glycine max Merr.). The research was conducted at the Soybean Free Air Concentration Enrichment (SoyFACE) facility where we subjected plants to ambient (44nL L(-1)) and elevated ozone (79-82nL L(-1) target) concentrations throughout the growing season. Exposure to elevated ozone resulted in a significant loss of productivity, with the ozone-treated plants displaying a ~30% average decrease in seed yield. From leaf reflectance data, it was also clear that elevated ozone decreased leaf nitrogen and chlorophyll content as well as the photochemical reflectance index (PRI), an optical indicator of the epoxidation state of xanthophyll cycle pigments and thus physiological status. We assessed the potential to use leaf reflectance properties and partial least-squares regression (PLSR) modeling as an alternative, rapid approach to standard gas exchange for the estimation of the maximum rates of RuBP carboxylation (V c,max), an important parameter describing plant photosynthetic capacity. While we did not find a significant impact of ozone fumigation on V c,max, standardized to a reference temperature of 25°C, the PLSR approach provided accurate and precise estimates of V c,max across ambient plots and ozone treatments (r (2)=0.88 and RMSE=13.4μmolm(-2) s(-1)) based only on the variation in leaf optical properties and despite significant variability in leaf nutritional status. The results of this study illustrate the potential for combining the phenotyping methods used here with high-throughput genotyping methods as a promising approach for elucidating the basis for ozone tolerance in sensitive crops.

Blue light is essential for high light acclimation and photoprotection in the diatom Phaeodactylum tricornutum

The objective of the present study was to test the hypothesis that the acclimation to different light intensities in the diatom Phaeodactylum tricornutum is controlled by light quality perception mechanisms. Therefore, semi-continuous cultures of P. tricornutum were illuminated with equal amounts of photosynthetically absorbed radiation of blue (BL), white (WL), and red light (RL) and in combination of two intensities of irradiance, low (LL) and medium light (ML). Under LL conditions, growth rates and photosynthesis rates were similar for all cultures. However, BL cultures were found to be in an acclimation state with an increased photoprotective potential. This was deduced from an increased capacity of non-photochemical quenching, a larger pool of xanthophyll cycle pigments, and a higher de-epoxidation state of xanthophyll cycle pigments compared to WL and RL cultures. Furthermore, in the chloroplast membrane proteome of BL cells, an upregulation of proteins involved in photoprotection, e.g. the Lhcx1 protein and zeaxanthin epoxidase, was evident. ML conditions induced increased photosynthesis rates and a further enhanced photoprotective potential for algae grown under BL and WL. In contrast, RL cultures exhibited no signs of acclimation towards increased irradiance. The data implicate that in diatoms the photoacclimation to high light intensities requires the perception of blue light.

Chlorophyll index, photochemical reflectance index and chlorophyll fluorescence measurements of rice leaves supplied with different N levels

Rapid and non-destructive diagnosis of plant N status is highly required in order to optimise N fertilizer management and use-efficiency. Additionally to handheld devices for measurements of chlorophyll indices (e.g., SPAD meter) parameters of canopy reflectance via remote sensing approaches are intensively investigated and the photochemical reflectance index (PRI) appears to be a reliable indicator for changes of the epoxidation state of xanthophyll cycle pigments. In order to assess the suitability of a handheld PRI as an additional tool for N diagnosis, rice plants were grown in a nutrient solution experiment with seven N-supply levels (0.18–5.71mM) and CI (SPAD) and PRI values and chlorophyll fluorescence parameters measured 20 and 28days after onset of treatments. N-supply had effects on both CI (SPAD) and PRI values with a more reliable differentiation between levels. Maximum quantum yield of PSII (Fv/Fm), actual efficiency of PSII photochemistry (ФPSII) and regulated non-photochemical quenching (ФNPQ) did not differ significantly between N levels. Non-photochemical quenching (NPQ) and fast- relaxing NPQ (NPQF) were significantly affected by N-supply. NPQ and NPQF, but not the slow-relaxing component (NPQS), were correlated with CI (SPAD) and PRI values. This finding which has not been reported for N-supply effects so far is indirect evidence that low N-supply induced xanthophyll cycle activity and that PRI values are able to indicate this at least in plants subject to severe N deficiency.

Reflectance continuum removal spectral index tracking the xanthophyll cycle photoprotective reactions in Norway spruce needles.

This laboratory experiment tested the ability of the spectral index called 'area under curve normalised to maximal band depth' (ANMB) to track dynamic changes in the xanthophyll cycle of Norway spruce (Picea abies (L.) Karsten) needles. Four-year-old spruce seedlings were gradually acclimated to different photosynthetic photon flux densities (PPFDs) and air temperature regimes. The measurements were conducted at the end of each acclimation period lasting for 11 days. A significant decline in the chlorophylls to carotenoids ratio and the increase of the amount of xanthophyll cycle pigments indicated a higher need for carotenoid-mediated photoprotection in spruce leaves acclimated to high PPFD conditions. Similarly, the photochemical reflectance index (PRI) changed from positive to negative values after changing light conditions from low to high intensity as a consequence of the increase in carotenoid content. Systematic responses of PRI to the de-epoxidation state of xanthophyll cycle pigments (DEPS) were, however, observed only during high temperature treatments and after the exposition of needles to high irradiance. The ANMB index computed from needle reflectance between 507 and 556nm was able to track dynamic changes in DEPS without any influence induced by changing the content of leaf photosynthetic pigments (chlorophylls, carotenoids).

Relation of chlorophyll fluorescence sensitive reflectance ratios to carbon flux measurements of montanne grassland and norway spruce forest ecosystems in the temperate zone.

We explored ability of reflectance vegetation indexes (VIs) related to chlorophyll fluorescence emission (R 686/R 630, R 740/R 800) and de-epoxidation state of xanthophyll cycle pigments (PRI, calculated as (R 531 − R 570)/(R 531 − R 570)) to track changes in the CO2 assimilation rate and Light Use Efficiency (LUE) in montane grassland and Norway spruce forest ecosystems, both at leaf and also canopy level. VIs were measured at two research plots using a ground-based high spatial/spectral resolution imaging spectroscopy technique. No significant relationship between VIs and leaf light-saturated CO2 assimilation (A MAX) was detected in instantaneous measurements of grassland under steady-state irradiance conditions. Once the temporal dimension and daily irradiance variation were included into the experimental setup, statistically significant changes in VIs related to tested physiological parameters were revealed. ΔPRI and Δ(R 686/R 630) of grassland plant leaves under dark-to-full sunlight transition in the scale of minutes were significantly related to A MAX (R 2 = 0.51). In the daily course, the variation of VIs measured in one-hour intervals correlated well with the variation of Gross Primary Production (GPP), Net Ecosystem Exchange (NEE), and LUE estimated via the eddy-covariance flux tower. Statistical results were weaker in the case of the grassland ecosystem, with the strongest statistical relation of the index R 686/R 630 with NEE and GPP.

Mechanism of xanthophyll-cycle-mediated photoprotection in Cerasus humilis seedlings under water stress and subsequent recovery

Periodic drought fluctuation is a common phenomenon in Northwest China. We analyzed the response of Chinese dwarf cherry (Cerasus humilis) seedlings, a dwarf shrub species with considerably strong adaptabilities, exposed to water stress (WS) by withholding water for 21 d, and subsequent recovery of 7 d. Leaf relative water content (LRWC), net photosynthetic rate (P N), maximal quantum yield of PSII photochemistry (Fv/Fm), and effective quantum yield of PSII photochemistry (ΦPSII) decreased with increasing water deficit. In contrast, the nonphotochemical quenching of Chl fluorescence (NPQ) significantly increased, as well as the amounts of violaxanthin (V) + antheraxanthin (A) + zeaxanthin (Z). In the whole levels, the photosynthetic pigment composition did not display significant changes in WS seedlings. However, the de-epoxidation state of xanthophyll cycle pigments [(Z+0.5A)/VAZ ] generally exhibited higher values in WS seedlings. The significant inhibition of de-epoxidation by dithiothreitol (DTT) and negligible changes of epoxidation of Z by glucosamine (Gla) were both observed; the slight but stably upregulated transcript level of violaxanthin de-epoxidase (VDE) and downregulated zeaxanthin epoxidase (ZEP) expression profile were found during WS period, indicating that they were regulated on post-transcript levels. VDE activity, via the accumulation of Z and A, which confers a greater capacity of photoprotection, appears to contribute to the survival of severely stressed plants.

Relationship between photochemical efficiency of photosystem II and the photochemical reflectance index of mango tree: merging data from different illuminations, seasons and leaf colors

In order to elucidate the effects of chlorophyll concentration and seasonal temperature on the relationship between photosystem II (PSII) efficiency and the photochemical reflectance index (PRI) of leaves under different light intensity, mango (Mangifera indica), a low-temperature-sensitive species, was used for the study. From early winter to summer, we selected several days to measure chlorophyll fluorescence and leaf spectral reflectance of mango leaves with dark green to yellow green colors, under natural sunlight from predawn to sunset and under six levels (0, 200, 400, 800, 1200 and 2000 mumol m(-2) s(-1)) of artificial illumination. When leaves were exposed to light, both PRI and PSII efficiency decreased with the increase in illumination, yet the PSII efficiency-PRI relationship varied with temperature and leaf color. Both predawn PRI and the X-intercept of the PSII efficiency-PRI regression equations were higher in dark green leaves and on the day with higher minimum air temperature, and lower on opposite conditions. These were due to the influence of chlorophyll on the reflection of wavebands for detecting PRI, and leaves retained a higher degree of epoxidation state of xanthophyll cycle pigments in cold predawn. Therefore, when data obtained at different seasons and with different leaf colors were pooled for analysis, PRI was not closely related to PSII efficiency. Yet, either in the darkness of predawn or under a given level of illumination, PSII efficiency always showed a significant positive correlation with PRI, with data from different leaf colors and seasons merged for statistics analysis. Because both the intercept and slope of the PSII efficiency-PRI equation showed a negative regression with photosynthetic photon flux (PPF), an empirical regression model, i.e., PSII efficiency = c + d . PPF + e . (PPF)(2) + f . PRI + g . PPF . PRI, could be fitted for multiple regression analysis. Based on the close correlation between the estimated and measured PSII efficiency (r(2) = 0.844-0.907, P < 0.001), using dynamic data obtained from leaves with yellow green to dark green colors, measurement was taken at predawn (F(v)/F(m)) and under any given strength of sunlight and artificial illumination (DeltaF/F(m)') through different seasons. We, thus, concluded that this empirical regression model could simulate both the seasonal and diurnal variations of PSII efficiency.

Wavelength-dependent xanthophyll cycle activity in marine microalgae exposed to natural ultraviolet radiation

The wavelength dependency of xanthophyll cycling in two marine microalgae (Thalassiosira weissflogii and Dunaliella tertiolecta) was studied by establishing biological weighting functions (BWFs) during exposure to natural ultraviolet radiation. High-(HL) and low-(LL) light-acclimated cultures of both species were exposed outdoors for up to 60 min under a series of UVR (280–400 nm) cut-off filters, after which the de-epoxidation state of xanthophyll cycle pigments, radiocarbon assimilation and photochemical quantum yield were measured. Exposures were repeated 4–8 times during the daily cycle to create exposure–response curves for each wavelength condition. UVR affected the three target processes significantly in both species and biological weights increased with decreasing wavelength, particularly in the UVBR region (280–315 nm). Minor wavelength dependency was observed between 315 and 360 nm. After BWF normalization to 300 nm, the LL cultures showed highly similar responses when comparing the three target processes, while the BWFs for the HL cultures differed significantly. The observed enhanced xanthophyll cycling activity in the UVR region implied that xanthophylls had an active role in diminishing UVR stress. However, this enhancement seems to be an indirect effect of damage within the dark reactions of photosynthesis. Hence, another vital target process further downstream in the photosynthetic process, possibly involved in the dark reactions, seems to be responsible for the high similarity in BWFs.

Irradiance prior to and during desiccation improves the tolerance to excess irradiance in the desiccated state of the old forest lichen Lobaria pulmonaria

Hydrated thalli of the lichen Lobaria pulmonaria were either preconditioned to dim irradiance (DI, 5 µmol m−2 s−1) or medium irradiance (MI, 200 µmol m−2 s−1) for 6 h. After this 6 h period, the thalli were allowed to desiccate under the two respective irradiances. Thereafter, these dry lichens were exposed to high irradiance (HI, 1 000 µmol m−2 s−1) for 60 h. After this HI treatment, the maximal photochemical quantum yield (FV/FM) and the de-epoxidation state of xanthophyll cycle pigments (DEPS) were highest in thalli preconditioned to MI. Hence irradiance in the last hydrated period before sampling is significant for the physiological state of lichens. A standardized irradiance pre-treatment before start of experiments is recommended.

Alleviation of low-temperature photoinhibition in gamma-irradiated red pepper (Capsicum annuum L.) plants

We studied the radiation-induced stress resistance in red pepper leaves under conditions of low-temperature photoinhibition or artificially induced photo-oxidative stress. Plants irradiated with 4, 8, or 16-Gy gamma rays were more resistant to both stress factors than were the controls. However, exposure to a low temperature for 12 h with illumination or photo-oxidative treatment for 1 h differentially affected the irradiated leaves, although they had similar stress intensities as defined by their maximal photochemical efficiencies (Fv/Fm). Decreases in Fv/Fm induced by the two stress factors were instead alleviated, dose-dependently, by as much as 22 to 41% (low temperature) or 14 to 29% (photo-oxidation) in the irradiated groups. In contrast, non-photochemical quenching (NPQ) and the de-epoxidation state of xanthophyll cycle pigments could not be correlated with this enhanced stress resistance in the irradiated groups. These results suggest that the adaptive response of plants exposed to gamma radiation is more effective in protecting against low-temperature photoinhibition than against photo-oxidative stress. We also discuss here the involvement of antioxidative defense systems for increased resistance against low-temperature photoinhibition in irradiated red pepper.

Protective cytokinin action switches to damaging during senescence of detached wheat leaves in continuous light

The effect of exogenous application of the cytokinin meta‐topolin [mT; N6‐(meta‐hydroxybenzyl)adenine] on artificial senescence of detached wheat leaves (Triticum aestivum L. cv. Hereward) was studied and compared in leaves senescing under continuous light (100 µmol photons m−2 s−1) and darkness. Senescence‐induced deterioration in structure and function of the photosynthetic apparatus was characterized by reduction in chlorophyll content, maximal efficiency of photosystem (PS) II photochemistry (Fv/Fm) and the rate of CO2 assimilation, by increase in the excitation pressure on PSII (1 − qP) and a level of lipid peroxidation and by modifications in chloroplast ultrastructure. While in darkened leaf segments mT effectively slowed senescence‐induced changes in all measured parameters, in light‐senescing segments the effect of mT changed into opposite a few days after detachment. We observed an overexcitation of photosynthetic apparatus, as indicated by pronounced increases in the excitation pressure on PSII and in a deepoxidation state of xanthophyll cycle pigments, marked starch grain accumulation in chloroplasts and stimulation of lipid peroxidation in light‐senescing leaf segments in mT. Possible mechanisms of acceleration of senescence‐accompanying decrease in photosynthetic function and increase in lipid peroxidation during mT influence are discussed. We propose that protective mT action in darkness becomes damaging during artificial senescence in continuous light due to overexcitation of photosynthetic apparatus resulting in oxidative damage.

The Photosynthetic Properties of Rice Leaves Treated with Low Temperature and High Irradiance

Photosynthetic characteristics in rice (Oryza sativa L.) leaves were examined after treatment with low temperature (15 degrees C) and high irradiance (1,500 micromol quanta m(-2) s(-1)). Decreases in quantum efficiencies in PSII (PhiPSII) and PSI (PhiPSI) and in the rate of CO2 assimilation were observed with a decrease in the maximal quantum efficiency of PSII (F(v)/F(m)) by simultaneous measurements of Chl fluorescence, P700+ absorbance and gas exchange. The decreases in PhiPSII were most highly correlated with those in CO2 assimilation. Although the initial (the activity immediately measured upon extraction) and total (the activity following pre-incubation with CO2 and Mg2+) activities of ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco) decreased slightly, the maximal activity (the activity following treatment with SO4(2-)) of Rubisco remained almost constant. These results indicate that the decrease in CO2 assimilation rate with the decreasing F(v)/F(m) was not caused by a decrease in Rubisco activity but rather by a decrease in RuBP regeneration capacity which resulted from the decrease in the rate of the linear electron transport. On the other hand, the decrease in PhiPSI was very small and the ratio of PhiPSI to PhiPSII increased. The de-epoxidation state of xanthophyll cycle pigments also increased. Thus, the cyclic electron transport around PSI occurred in photoinhibited leaves.

Towards elucidating the energy of the first excited singlet state of xanthophyll cycle pigments by X-ray absorption spectroscopy

The first excited singlet state (S 1) of carotenoids (also termed 2A g −) plays a key role in photosynthetic excitation energy transfer due to its close proximity to the S 1 (Q y ) level of chlorophylls. The determination of carotenoid 2A g − energies by optical techniques is difficult; transitions from the ground state (S 0, 1A g −) to the 2A g − state are forbidden (“optically dark”) due to parity (g ← //→ g) as well as pseudo-parity selection rules (− ← //→ −). Of particular interest are S 1 energies of the so-called xanthophyll-cycle pigments (violaxanthin, antheraxanthin and zeaxanthin) due to their involvement in photoprotection in plants. Previous determinations of S 1 energies of violaxanthin and zeaxanthin by different spectroscopic techniques vary considerably. Here we present an alternative approach towards elucidation of the optically dark states of xanthophylls by near-edge X-ray absorption fine structure spectroscopy (NEXAFS). The indication of at least one π* energy level (about 0.5 eV below the lowest 1B u + vibronic sublevel) has been found for zeaxanthin. Present limitations and future improvements of NEXAFS to study optically dark states of carotenoids are discussed. NEXAFS combined with simultaneous optical pumping will further aid the investigation of these otherwise hardly accessible states.

Changes in glutathione and xanthophyll cycle pigments in the high light-stressed lichens Umbilicaria antarctica and Lasallia pustulata

Hydrated thalli of two lichen species – Umbilicaria antarctica and Lasallia pustulata – were exposed to high light (1800 μmol m −2 s −1) for 30 min. High light exposure led to a decrease of total glutathione in both species, while de-epoxidation state of xanthophyll cycle pigments and non-photochemical quenching increased. In the subsequent recovery, the values of de-epoxidation state of xanthophyll cycle pigments decreased towards initial values. Glutathione (GSH) was resynthetised slowly. In conclusion, zeaxanthin-related protection is probably more involved than GSH-related protection in short-term response to high light stress in U. antarctica and L. pustulata. Faster recovery from photoinhibition in L. pustulata than U. antarctica is mainly due to faster conversion of zeaxanthin to violaxanthin and larger GSH pool of former species.

Dynamic light use and protection from excess light in upper canopy and coppice leaves of Nothofagus cunninghamii in an old growth, cool temperate rainforest in Victoria, Australia

Responses to simulated sunflecks were examined in upper canopy and coppice leaves of Nothofagus cunninghamii growing in an old-growth rainforest gully in Victoria, Australia. Shaded leaves were exposed to a sudden increase in irradiance from 20 to 1500 micromol m(-2) s(-1). Gas exchange and chlorophyll fluorescence were measured during a 10 min simulated sunfleck and, in the ensuing dark treatment, we examined the recovery of PS II efficiency and the conversion state of xanthophyll cycle pigments. Photosynthetic induction was rapid compared with tropical and northern hemisphere species. Stomatal conductance was relatively high in the shade and stomata did not directly control photosynthetic induction under these conditions. During simulated sunflecks, zeaxanthin was formed rapidly and photochemical efficiency was reduced. These processes were reversed within 30 min in coppice leaves, but this took longer in upper canopy leaves. Poor drought tolerance and achieving a positive carbon balance in a shaded canopy may be functionally related to high stomatal conductance in the shade in N. cunninghamii. The more persistent reduction in photochemical efficiency of upper canopy leaves, which means less efficient light use in subsequent shade periods, but stronger protection from high light, may be related to the generally higher irradiance and longer duration of sunflecks in the upper canopy, but potentially reduces carbon gain during shade periods by 30%.

Knock-out of the genes coding for the Rieske protein and the ATP-synthase delta-subunit of Arabidopsis. Effects on photosynthesis, thylakoid protein composition, and nuclear chloroplast gene expression.

In Arabidopsis, the nuclear genes PetC and AtpD code for the Rieske protein of the cytochrome b(6)/f (cyt b(6)/f) complex and the delta-subunit of the chloroplast ATP synthase (cpATPase), respectively. Knock-out alleles for each of these loci have been identified. Greenhouse-grown petc-2 and atpd-1 mutants are seedling lethal, whereas heterotrophically propagated plants display a high-chlorophyll (Chl)-fluorescence phenotype, indicating that the products of PetC and AtpD are essential for photosynthesis. Additional effects of the mutations in axenic culture include altered leaf coloration and increased photosensitivity. Lack of the Rieske protein affects the stability of cyt b(6)/f and influences the level of other thylakoid proteins, particularly those of photosystem II. In petc-2, linear electron flow is blocked, leading to an altered redox state of both the primary quinone acceptor Q(A) in photosystem II and the reaction center Chl P700 in photosystem I. Absence of cpATPase-delta destabilizes the entire cpATPase complex, whereas residual accumulation of cyt b(6)/f and of the photosystems still allows linear electron flow. In atpd-1, the increase in non-photochemical quenching of Chl fluorescence and a higher de-epoxidation state of xanthophyll cycle pigments under low light is compatible with a slower dissipation of the transthylakoid proton gradient. Further and clear differences between the two mutations are evident when mRNA expression profiles of nucleus-encoded chloroplast proteins are considered, suggesting that the physiological states conditioned by the two mutations trigger different modes of plastid signaling and nuclear response.