Chl Fluorescence Measurements Research Articles

Unravelling the different components of nonphotochemical quenching using a novel analytical pipeline.

Photoprotection in plants includes processes collectively known as nonphotochemical quenching (NPQ), which quench excess excitation-energy in photosystem II. NPQ is triggered by acidification of the thylakoid lumen, which leads to PsbS-protein protonation and violaxanthin de-epoxidase activation, resulting in zeaxanthin accumulation. Despite extensive study, questions persist about the mechanisms of NPQ. We have set up a novel analytical pipeline to disentangle NPQ induction curves measured at many light intensities into a limited number of different kinetic components. To validate the method, we applied it to Chl-fluorescence measurements, which utilised the saturating-pulse methodology, on wild-type (wt) and zeaxanthin-lacking (npq1) Arabidopsis thaliana plants. NPQ induction curves in wt and npq1 can be explained by four components ( , , and ). The fastest two ( and ) correlate with pH difference formed across the thylakoid membrane in wt and npq1. In wt, the slower component ( ) appears to be due to the formation of zeaxanthin-related quenching whilst for npq1, this component is 'replaced' by a slower component ( ), which reflects a photoinhibition-like process that appears in the absence of zeaxanthin-induced quenching. Expanding this approach will allow the effects of mutations and other abiotic-stress factors to be directly probed by changes in these underlying components.

Metabolomics combined with physiology and transcriptomics reveal how Nicotiana tabacum leaves respond to cold stress

Low temperature-induced cold stress is a major threat to plant growth, development and distribution. Unraveling the responses of temperature-sensitive crops to cold stress and the mechanisms of cold acclimation are critical for food demand. In this study, combined physiological, transcriptomic, and metabolomic analyses were conducted on Nicotiana tabacum suffering short-term 4 °C cold stress. Our results showed that cold stress destroyed cellular membrane stability, decreased the chlorophyll (Chl) and carotenoid contents, and closed stomata, resulting in lipid peroxidation and photosynthesis restriction. Chl fluorescence measurements revealed that primary photochemistry, photoelectrochemical quenching and photosynthetic electron transport in Nicotiana tabacum leaves were seriously suppressed upon exposer to cold stress. Enzymatic and nonenzymatic antioxidants, including superoxide dismutase, catalase, peroxidase, reduced glutathione, proline, and soluble sugar, were all profoundly increased to trigger the cold acclimation defense against oxidative damage. A total of 178 metabolites and 16,204 genes were differentially expressed in cold-stressed Nicotiana tabacum leaves. MEturquoise and MEblue modules identified by WGCNA were highly correlated with physiological indices, and the corresponding hub genes were significantly enriched in pathways related to photosynthesis - antenna proteins and flavonoid biosynthesis. Untargeted metabolomic analysis identified specific metabolites, including sucrose, phenylalanine, glutamine, glutamate, and proline, that enhance plant cold acclimation. Combined transcriptomics and metabolomic analysis highlight the vital roles of carbohydrate and amino acid metabolism in enhancing the cold tolerance of Nicotiana tabacum. Our comprehensive investigation provides novel insights for efforts to alleviate low temperature-induced oxidative damage to Nicotiana tabacum plants and proposes a breeding target for cold stress-tolerant cultivars.

Physiological Characteristics of Photosynthesis in Yellow-Green, Green and Dark-Green Chinese Kale (Brassica oleracea L. var. alboglabra Musil.) under Varying Light Intensities.

The objective of this work was to study physiological characteristics and photosynthetic apparatus in differentially pigmented leaves of three Chinese kale cultivars. Chlorophyll (Chl) fluorescence and photochemical reflectance index (PRI) measurements in green, yellow-green, and dark-green cultivars in response to varying light intensities. As light intensity increased from 200 to 2000 photosynthetic photon flux density (PPFD), fraction of light absorbed in photosystem (PS) II and PRI values in all plants were strongly lowered, but fraction of light absorbed in PSII dissipated via thermal energy dissipation and non-photochemical quenching (NPQ) values in all plants wereremarkably elevated.When plants were exposed to 200 PPFD, the values of fraction of light absorbed in PSII, utilized in photosynthetic electron transport(p), andfraction of light absorbed excitation energy in PSII dissipated via thermal energy dissipation (D), remained stable regardless of the changes in levels of Chla + b. Under 800 and 1200 PPFD, the values of p and electron transport rate (ETR) decreased, but D and NPQ increased as Chla + bcontent decreased, suggesting that decrease inChla + bcontent led to lower PSII efficiency and it became necessary to increase dissipate excess energy. On the contrary, in 2000 PPFD, leaves with lower Chla + bcontent had relatively higher p and electron transport rate (ETR) values and lower D level, as well as tended to increase more in NPQ but decrease more in PRI values. The consistent relations between PRI and NPQ suggest that NPQ is mainly consisted ofthe xanthophyll cycle-dependentenergy quenching.Yellow-green cultivar showed lower Chla + bcontent but high carotenoids/Chla + b ratio and had high light protection ability under high PPFD. The precise management of photosynthetic parameters in response to light intensity can maximize the growth and development of Chinese kale plants.

Whole Irradiated Plant Leaves Showed Faster Photosynthetic Induction Than Individually Irradiated Leaves via Improved Stomatal Opening.

Rapid photosynthetic induction is crucial for plants under fluctuating light conditions in a crop canopy as well as in an understory. Most previous studies have focused on photosynthetic induction responses in a single leaf, whereas the systemic responses of the whole plant have not been considered. In a natural environment, however, both single leaves and whole plants are exposed to sunlight, since the light environment is not uniform even within a given plant. In the present study, we examined whether there is any difference between the photosynthetic induction response of a leaf of a whole irradiated plant and an individually irradiated leaf in Arabidopsis thaliana to consider photosynthetic induction as the response of a whole plant. We used two methods, the visualization of photosynthesis and direct measurements of gas-exchange and Chl fluorescence, to demonstrate that whole irradiated plant promoted its photosynthetic induction via improved stomatal opening compared with individually irradiated leaf. Furthermore, using two Arabidopsis knockout mutants of abscisic acid transporter, abcg25 and abcg40, the present study suggests that abscisic acid could be involved in this systemic response for stomatal opening, allowing plants to optimize the use of light energy at minimal cost in plants in a dynamic light environment.

Optical Properties of Corals Distort Variable Chlorophyll Fluorescence Measurements.

Pulse-amplitude-modulated (PAM) fluorimetry is widely used in photobiological studies of corals, as it rapidly provides numerous photosynthetic parameters to assess coral ecophysiology. Coral optics studies have revealed the presence of light gradients in corals, which are strongly affected by light scattering in coral tissue and skeleton. We investigated whether coral optics affects variable chlorophyll (Chl) fluorescence measurements and derived photosynthetic parameters by developing planar hydrogel slabs with immobilized microalgae and with bulk optical properties similar to those of different types of corals. Our results show that PAM-based measurements of photosynthetic parameters differed substantially between hydrogels with different degrees of light scattering but identical microalgal density, yielding deviations in apparent maximal electron transport rates by a factor of 2. Furthermore, system settings such as the measuring light intensity affected F 0, Fm , and Fv /Fm in hydrogels with identical light absorption but different degrees of light scattering. Likewise, differences in microalgal density affected variable Chl fluorescence parameters, where higher algal densities led to greater Fv /Fm values and relative electron transport rates. These results have important implications for the use of variable Chl fluorimetry in ecophysiological studies of coral stress and photosynthesis, as well as other optically dense systems such as plant tissue and biofilms.

Light Quality Affects Chloroplast Electron Transport Rates Estimated from Chl Fluorescence Measurements.

Chl fluorescence has been used widely to calculate photosynthetic electron transport rates. Portable photosynthesis instruments allow for combined measurements of gas exchange and Chl fluorescence. We analyzed the influence of spectral quality of actinic light on Chl fluorescence and the calculated electron transport rate, and compared this with photosynthetic rates measured by gas exchange in the absence of photorespiration. In blue actinic light, the electron transport rate calculated from Chl fluorescence overestimated the true rate by nearly a factor of two, whereas there was closer agreement under red light. This was consistent with the prediction made with a multilayer leaf model using profiles of light absorption and photosynthetic capacity. Caution is needed when interpreting combined measurements of Chl fluorescence and gas exchange, such as the calculation of CO2 partial pressure in leaf chloroplasts.

Connecting active to passive fluorescence with photosynthesis: a method for evaluating remote sensing measurements of Chl fluorescence.

Recent advances in the retrieval of Chl fluorescence from space using passive methods (solar-induced Chl fluorescence, SIF) promise improved mapping of plant photosynthesis globally. However, unresolved issues related to the spatial, spectral, and temporal dynamics of vegetation fluorescence complicate our ability to interpret SIF measurements. We developed an instrument to measure leaf-level gas exchange simultaneously with pulse-amplitude modulation (PAM) and spectrally resolved fluorescence over the same field of view - allowing us to investigate the relationships between active and passive fluorescence with photosynthesis. Strongly correlated, slope-dependent relationships were observed between measured spectra across all wavelengths (Fλ , 670-850nm) and PAM fluorescence parameters under a range of actinic light intensities (steady-state fluorescence yields, Ft ) and saturation pulses (maximal fluorescence yields, Fm ). Our results suggest that this method can accurately reproduce the full Chl emission spectra - capturing the spectral dynamics associated with changes in the yields of fluorescence, photochemical (ΦPSII), and nonphotochemical quenching (NPQ). We discuss how this method may establish a link between photosynthetic capacity and the mechanistic drivers of wavelength-specific fluorescence emission during changes in environmental conditions (light, temperature, humidity). Our emphasis is on future research directions linking spectral fluorescence to photosynthesis, ΦPSII, and NPQ.

Impact of intraspecific competition on photosynthetic apparatus efficiency in potato (Solanum tuberosum) plants

Unfavourable growth conditions significantly determine the yield of crop plants. Intraspecific competition is a condition in which plants compete with each other for environmental resources. An excessive density contributes to increased competition within species, which results in disruption of photosynthesis process. According to this idea, experiments were conducted to investigate the photosynthetic response of potato (Solanum tuberosum) plants to excessive congestion. Two potato varieties of different earliness classes (Vineta and Satina) were used to evaluate the efficiency of the photosynthetic apparatus based on chlorophyll (Chl) fluorescence measurements under stress conditions. Changes in Chl contents of the tested plants were also analysed. In relation to intraspecific competition, we can conclude that the Vineta variety was less sensitive to this stress factor. In contrast, the photosynthetic apparatus of the Satina variety showed less efficient functioning under these conditions. In this study, the application of Chl fluorescence was presented for the first time in order to analyse the effects of intraspecific competition in plants.

Investigating role of Triton X-100 in ameliorating deleterious effects of anthracene in wheat plants

This study focused on the deleterious effect of anthracene (ANT) and role of a surfactant, Triton (TX-100), in recovery from inhibitory effect of ANT. Fast chlorophyll (Chl) fluorescence measurements were performed in wheat plants. Results revealed that maximum quantum yield of PSII, area over the fluorescence curve, performance index (PI), and reaction centre density was negatively affected by ANT treatment. The effects on PSII quantum efficiency, reaction centre density, absorption, and trapping were partially recovered by TX-100. PSII heterogeneity in terms of PSII antenna heterogeneity, corresponding to PSII α, β, and γ centres, and reducing side, corresponding to QB-reducing and QB-nonreducing centres, were also investigated. The damage caused by ANT to PSII antenna heterogeneity was recovered almost by 100% owing to TX-100.

Assessment of cadmium phytotoxicity alleviation by silicon using chlorophyll a fluorescence

The aim of this study was to investigate the effects of silicon in alleviating cadmium stress in maize plants grown in a nutrient solution and to evaluate the potential of the spectral emission parameters and the ratio of red fluorescence (Fr) to far-red fluorescence (Ffr) in assessing the beneficial effects of Si. An experiment was carried out using a nutrient solution with a toxic dose of Cd and six doses of Si; biomass, Cd, Si, and photosynthetic pigments of the plants were measured. Chlorophyll (Chl) a fluorescence analysis demonstrated that Si alleviated Cd toxicity in plants. Chl fluorescence measurements were sensitive in detecting such effects even when significant changes in biomass production and concentrations of photosynthetic pigments were not observed. The spectral emission and the Fr/Ffr ratio were sensitive to the effects of Si. Silicon caused a reduction in the translocation of Cd to the shoots of maize plants.

Effects of Bleaching by Nitrogen Deficiency on the Quantum Yield of Photosystem II in Synechocystis sp. PCC 6803 Revealed by Chl Fluorescence Measurements.

Estimation of photosynthesis by Chl fluorescence measurement of cyanobacteria is always problematic due to the interference from respiratory electron transfer and from phycocyanin fluorescence. The interference from respiratory electron transfer could be avoided by the use of DCMU or background illumination by blue light, which oxidizes the plastoquinone pool that tends to be reduced by respiration. On the other hand, the precise estimation of photosynthesis in cells with a different phycobilisome content by Chl fluorescence measurement is difficult. By subtracting the basal fluorescence due to the phycobilisome and PSI, it becomes possible to estimate the precise maximum quantum yield of PSII in cyanobacteria. Estimated basal fluorescence accounted for 60% of the minimum fluorescence, resulting in a large difference between the 'apparent' yield and 'true' yield under high phycocyanin conditions. The calculated value of the 'true' maximum quantum yield of PSII was around 0.8, which was similar to the value observed in land plants. The results suggest that the cause of the apparent low yield reported in cyanobacteria is mainly ascribed to the interference from phycocyanin fluorescence. We also found that the 'true' maximum quantum yield of PSII decreased under nitrogen-deficient conditions, suggesting the impairment of the PSII reaction center, while the 'apparent' maximum quantum yield showed a marginal change under the same conditions. Due to the high contribution of phycocyanin fluorescence in cyanobacteria, it is essential to eliminate the influence of the change in phycocyanin content on Chl fluorescence measurement and to evaluate the 'true' photosynthetic condition.

Relationship Between Photochemical Quenching and Non-Photochemical Quenching in Six Species of Cyanobacteria Reveals Species Difference in Redox State and Species Commonality in Energy Dissipation.

Although the photosynthetic reaction center is well conserved among different cyanobacterial species, the modes of metabolism, e.g. respiratory, nitrogen and carbon metabolism and their mutual interaction, are quite diverse. To explore such uniformity and diversity among cyanobacteria, here we compare the influence of the light environment on the condition of photosynthetic electron transport through Chl fluorescence measurement of six cyanobacterial species grown under the same photon flux densities and at the same temperature. In the dark or under weak light, up to growth light, a large difference in the plastoquinone (PQ) redox condition was observed among different cyanobacterial species. The observed difference indicates that the degree of interaction between respiratory electron transfer and photosynthetic electron transfer differs among different cyanobacterial species. The variation could not be ascribed to the phylogenetic differences but possibly to the light environment of the original habitat. On the other hand, changes in the redox condition of PQ were essentially identical among different species at photon flux densities higher than the growth light. We further analyzed the response to high light by using a typical energy allocation model and found that ‘non-regulated’ thermal dissipation was increased under high-light conditions in all cyanobacterial species tested. We assume that such ‘non-regulated’ thermal dissipation may be an important ‘regulatory’ mechanism in the acclimation of cyanobacterial cells to high-light conditions.

Low PSI content limits the photoprotection of PSI and PSII in early growth stages of chlorophyll b-deficient wheat mutant lines.

In vivo analyses of electron and proton transport-related processes as well as photoprotective responses were carried out at different stages of growth in chlorophyll b (Chl b)-deficient mutant lines (ANK-32A and ANK-32B) and wild type (WT) of wheat (Triticum aestivum L.). In addition to a high Chl a-b ratio, ANK mutants had a lower content of photo-oxidizable photosystem I (PSI, P m), and several parameters indicated a low PSI/PSII ratio. Moreover, simultaneous measurements of Chl fluorescence and P700 indicated a shift of balance between redox poise of the PSII acceptor side and the PSII donor side, with preferential reduction of the plastoquinone pool, resulting in an over reduced PSI acceptor side (high Φ NA values). This was the probable reason for PSI inactivation observed in the ANK mutants, but not in WT. In later growth phases, we observed partial relief of "chlorina symptoms," toward WT. Measurements of ΔA 520 decay confirmed that, in early growth stages, the ANK mutants with low PSI content had a limited capacity to build up the transthylakoid proton gradient (ΔpH) needed to trigger non-photochemical quenching (NPQ) and to regulate the electron transport by cytochrome b 6/f. Later, the increase in the PSI/PSII ratio enabled ANK mutants to reach full NPQ, but neither over reduction of the PSI acceptor side nor PSI photoinactivation due to imbalance between the activity of PSII and PSI was mitigated. Thus, our results support the crucial role of proper regulation of linear electron transport in the protection of PSI against photoinhibition. Moreover, the ANK mutants of wheat showing the dynamic developmental changes in the PSI/PSII ratio are presented here as very useful models for further studies.

Effects of drought stress on the evergreen Quercus ilex L., the deciduous Q. robur L. and their hybrid Q. × turneri Willd.

Five-year-old trees of deciduous Quercus robur L., evergreen Q. ilex L., and their semideciduous hybrid, Q. × turneri Willd. (var. pseudoturneri), growing in pots, were subjected to drought stress by withholding water for 18–22 days, until leaf water potentials decreased below −2 MPa. Gas-exchange rates, oxygen evolution, and modulated chlorophyll (Chl) fluorescence measurements revealed that by strong stomata closure and declining photosynthetic capacity down to approximately 50%, all three taxa responded with strongly reduced photosynthesis rates. In Q. robur, photochemical quenching of the drought-stressed plants was much lower than in nonstressed controls. Dissection of the occurring events in the photosynthetic electron transport chain by fast Chl fluorescence induction analysis with the JIP-test were discussed.

Genotypic differentiation in the stomatal response to salinity and contrasting photosynthetic and photoprotection responses in five olive (Olea europaea L.) cultivars

Olive tree is moderately tolerant to salinity. Differences in salt tolerance among olive tree cultivars are documented. Salt-mediated decreases of stomatal conductance and photosynthesis are common in the studied cultivars, but other photosynthetic characteristics in response to increasing salinity are much less investigated. Greenhouse experiments were conducted to investigate the responses to increasing salinity up to 200mM NaCl of five olive tree cultivars: ‘Chemlali’, ‘Chetoui’, ‘Arbequina I18’, ‘Arbosana I43’ and ‘Koroneiki’, widely used in Tunisia. Techniques used include determinations of leaf concentrations of the toxic ions Na and Cl, photosynthetic pigment analyses, gas exchange and chlorophyll (Chl) fluorescence measurements. Relationships between thermal energy dissipation and changes in pigment composition were investigated. With 100mM NaCl water, the most readily available water in arid regions of Tunisia, ‘Chemlali’, ‘Chetoui’ and ‘Arbosana’ grew similarly to control plants, which is a valuable information for horticultural purposes. In all cultivars, leaf Na and Cl concentrations increased and net CO2 assimilation rates decreased as salt concentration increased. A negative correlation was observed between photosynthesis and leaf thickness. Data revealed genotypic differentiation in the stomatal response to salinity, decreasing in ‘Koroneiki’, ‘Arbequina’ and ‘Arbosana’ but remaining fairly constant in ‘Chemlali’ and ‘Chetoui’. In those cultivars where the amount of energy thermally dissipated increased in response to salinity, it was well correlated with increases of (zeaxanthin+antheraxanthin)/(violaxanthin+antheraxanthin+zeaxanthin) ((Z+A)/(V+A+Z)) and lutein to Chl molar ratios. In summary, salinity caused marked reductions in photosynthetic rates well correlated with leaf Na and Cl accumulation. Photosynthesis limitation was partially attributed to stomata closure, except in the cultivars ‘Chemlali’ and ‘Chetoui’ in which stomatal conductances were unaltered. Possible decrease of CO2 mesophyll conductance mediated by increases of leaf thickness is discussed. Salinity increased Z+A− and/or lutein-mediated thermal energy dissipation of some of the olive tree cultivars investigated.

Disruption of the ndhF1 Gene Affects Chl Fluorescence through State Transition in the Cyanobacterium Synechocystis sp. PCC 6803, Resulting in Apparent High Efficiency of Photosynthesis

In Synechocystis sp. PCC 6803, the disruption of the ndhF1 gene (slr0844), which encodes a subunit of one of the NDH-1 complexes (NDH-1L complex) serving for respiratory electron transfer, causes the largest change in Chl fluorescence induction kinetics among the kinetics of 750 disruptants searched in the Fluorome, the cyanobacterial Chl fluorescence database. The cause of the explicit phenotype of the ndhF1 disruptant was examined by measurements of the photosynthetic rate, Chl fluorescence and state transition. The results demonstrate that the defects in respiratory electron transfer obviously have great impact on Chl fluorescence in cyanobacteria. The inactivation of NDH-1L complexes involving electron transfer from NDH-1 to plastoquinone (PQ) would result in the oxidation of the PQ pool, leading to the transition to State 1, where the yield of Chl fluorescence is high. Apparently, respiration, although its rate is far lower than that of photosynthesis, could affect Chl fluorescence through the state transition as leverage. The disruption of the ndhF1 gene caused lower oxygen-evolving activity but the estimated electron transport rate from Chl fluorescence measurements was faster in the mutant than in the wild-type cells. The discrepancy could be ascribed to the decreased level of non-photochemical quenching due to state transition. One must be cautious when using the Chl fluorescence parameter to estimate photosynthesis in mutants defective in state transition.

Gymnosperms Have Increased Capacity for Electron Leakage to Oxygen (Mehler and PTOX reactions) in Photosynthesis Compared with Angiosperms

Oxygen plays an important role in photosynthesis by participating in a number of O2-consuming reactions. O2 inhibits CO2 fixation by stimulating photorespiration, thus reducing plant production. O2 interacts with photosynthetic electron transport in the chloroplasts' thylakoids in two main ways: by accepting electrons from PSI (Mehler reaction); and by accepting electrons from reduced plastoquinone (PQ) mediated by the plastid terminal oxidase (PTOX). In this study, we show, using 101 plant species, that there is a difference in the potential for photosynthetic electron flow to O2 between angiosperms and gymnosperms. We found, from measurements of Chl fluorescence and leaf absorbance at 830 nm, (i) that electron outflow from PSII, as determined by decay kinetics of Chl fluorescence after application of a saturating light pulse, is more rapid in gymnosperms than in angiosperms; (ii) that the reaction center Chl of PSI (P700) is rapidly and highly oxidized in gymnosperms during induction of photosynthesis; and (iii) that these differences are dependent on oxygen. Finally, rates of O2 uptake measured by mass spectrometry in the absence of photorespiration were significantly promoted by illumination in dark-adapted leaves of gymnosperms, but not in those of angiosperms. The light-stimulated O2 uptake was around 10% of the maximum O2 evolution in gymnosperms and 1% in angiosperms. These results suggest that gymnosperms have increased capacity for electron leakage to oxygen in photosynthesis compared with angiosperms. The involvement of the Mehler reaction and PTOX in the electron flow to O2 is discussed.

Monitoring moderate Cu and Cd toxicity by chlorophyll fluorescence and P<sub>700</sub> absorbance in pea leaves

We investigated the effect of moderate Cu2+ and Cd2+ stress by applying chlorophyll (Chl) fluorescence and P700 absorbance measurements to monitor the photosynthetic electron transport activity of 3-week-old Pisum sativum L. cv. Petit Provencal plants grown in a modified Hoagland solution containing 50 μM CuSO4 or 5 μM CdCl2. Both heavy metals caused a slight inhibition in PSII photochemistry as indicated by the decrease in the effective quantum efficiency of PSII (ΦPSII), the maximum electron transport capacity (ETRmax), and the maximum quantum yield for electron transport (α). PSI photochemistry was also affected by these heavy metals. Cu2+ and Cd2+ decreased the quantum efficiency of PSI (ΦPSI) as well as the number of electrons in the intersystem chain, and the Cu2+ treatment significantly reduced the number of electrons from stromal donors available for PSI. These results indicate that PSII and PSI photochemistry of pea plants are both sensitive to moderate Cu2+ and Cd2+ stress, which in turn is easily detected and monitored by Chl fluorescence and P700 absorbance measurements. Therefore, monitoring the photochemistry of pea plants with these noninvasive, yet sensitive techniques offers a promising strategy to study heavy metal toxicity in the environment.

Laser-Induced Chlorophyll Fluorescence Measurement System to Assess Photosynthetic Status within Leaves

Chlorophyll (Chl) fluorescence measured on a leaf surface only provides the photosynthetic status of chloroplasts near the surface due to self-shading effect of Chl. Here, we report a laser-induced Chl fluorescence measurement system, which enables measurement of fluorescence induction kinetics at different tissues within a leaf as well as on both leaf surfaces, to assess photosynthetic status within leaves. The logarithmic time-scaled Chl fluorescence induction kinetics obtained from Chenopodium album leaves showed polyphasic transients in which four inflection points designated as O, J, I and P, were observed. Adaxial surface and palisade mesophyll showed significantly higher fluorescence intensities at O than abaxial surface and spongy mesophyll, respectively. In contrast, fluorescence intensities at J, I and P were significantly higher at abaxial surface and spongy mesophyll. Using these fluorescence intensities, the JIP test was performed. The results of JIP test indicated that adaxial surface and palisade mesophyll are characterized by lower maximum quantum yield of photosystem II (PSII) and net rate of PSII closure, but higher rate of energy transfer into electron transport chain than abaxial surface and spongy mesophyll. Considering these dorsiventral and intra-leaf variations in Chl fluorescence parameters, this study suggested the necessity of Chl fluorescence measurements on tissues as well as both surfaces to evaluate photosynthetic status of a whole leaf.

The role of the PsbU subunit in the light sensitivity of PSII in the cyanobacterium Synechococcus 7942

In the present study we investigated the role of the PsbU subunit in the electron transport characteristics and light sensitivity of the Photosystem II complex. The experiments were performed by using an earlier characterized PsbU-less mutant of the cyanobacterium Synechococcus PCC 7942, which has enhanced antioxidant capacity (Balint et al. FEBS Lett. 580 (2006) 2117–2122). Flash induced Chl fluorescence measurements in the presence and absence of the electron transport inhibitor DCMU showed that both the S 2 Q A - and the S 2 Q B - recombination is slowed down in the PsbU mutant relative to the WT strain. Thermoluminescence measurements confirmed the increased stability of the S 2 Q A - and S 2 Q B - charge pairs by showing an increased peak temperature of Q and B bands, which were measured in the presence and absence of DCMU, respectively. In addition, the intensity of the TL bands is also increased in the PsbU mutant (≈1.7 times for the B band), as compared to the WT. The PsbU mutant shows enhanced loss of Photosystem II activity under exposure to high light intensity both in the absence and presence of the protein synthesis inhibitor lincomycin. It is concluded from the data that the lack of the PsbU subunit in Synechococcus PCC 7942 affects the energetic stability of the S 2 Q A - and S 2 Q B - charge pairs by modifying both the PSII donor and acceptor side components. This effect is most likely caused by structural changes in the vicinity of the Mn cluster and in the inner part of the PSII complex, which are induced by the lack of the PsbU subunit from the lumenal part of the complex. The light sensitivity of Photosystem II in Synechococcus 7942 in the absence of the PsbU subunit is likely due to reactive oxygen species, which are produced as a consequence of disturbed donor side structure and/or due to the modified energetic properties of the primary radical pair.