Changes In Fluorescence Yield Research Articles

Flash-kinetics as a complementary analytical tool in PAM fluorimetry

A new measuring system based on the already existing Multi-Color-PAM Fluorimeter (Schreiber et al. in Photosynth Res 113:127–144, 2012) was developed that in addition to standard PAM measurements enables pump-and-probe flash measurements and allows simultaneous measurements of the changes in chlorophyll fluorescence yield (F) during application of saturating flashes (ST). A high-power Chip-on-Board LED array provides ST flashes with close to rectangular profiles at wide ranges of widths (0.5 µs to 5 ms), intensities (1.3 mmol to 1.3 mol 440 nm quanta m−2 s−1) and highly flexible repetition times. Using a dedicated rising-edge profile correction, sub-µs time resolution is obtained for assessment of initial fluorescence and rise kinetics. At maximal to moderate flash intensities the flash-kinetics (changes of F during course of ST, STK) are strongly affected by ‘High Intensity Quenching’ (HIQ), consisting of Car-triplet quenching, TQ, and donor-side-dependent quenching, DQ. The contribution of TQ is estimated by application of a second ST after 20 µs dark-time. Upon application of flash trains (ST sequences with defined repetition times) typical period-4 oscillations in dark fluorescence yield (F0) and ST-induced fluorescence yield, FmST, are obtained which can be measured in vivo both with suspensions and from the surface of leaves. Examples of application with dilute suspensions of Chlorella and an intact dandelion leaf are presented. It is shown that weak far-red light (730–740 nm) advances the S-state distribution of the water-splitting system by one step, resulting in substantial lowering of FmST and also of the I1-level in the polyphasic rise of fluorescence yield induced by a multiple-turnover flash (MT). Based on comparative measurements of STK and the polyphasic rise kinetics with the same Chlorella sample, it is concluded that the generally observed lower values of maximal fluorescence yields using ST-protocols compared to MT-protocols are due to a higher extent of HIQ (mainly DQ) and the contribution of variable PSI fluorescence to FmST.

Hyperspectral and Photodiode Retrievals of Nighttime LED‐Induced Chlorophyll Fluorescence (LEDIF) for Tracking Photosynthetic Phenology in a Vineyard

AbstractThe magnitude of chlorophyll fluorescence emission represents both chlorophyll content and energy quenching processes enabling its application to serve as a proxy of photosynthetic activity. Thus, there is interest in advancing methods for canopy‐scale monitoring of chlorophyll fluorescence. Remotely sensed solar‐induced fluorescence (SIF) retrievals offer daytime monitoring of chlorophyll fluorescence, which can serve as an indicator of photosynthesis. However, it represents an instantaneous measurement during the day, which is strongly influenced by incoming radiation, solar angle, and sun/shade fraction—making it difficult to tease out baseline information on plant health and potential photosynthetic capacity—which could be tracked by changes in fluorescence yield (independent of sunlight). Recent advances have demonstrated the potential for inducing nighttime chlorophyll fluorescence via LED light sources at the canopy‐scale, which can be retrieved as LED‐induced chlorophyll fluorescence (LEDIF), potentially serving as a baseline indicator of plant health and photochemical capacity, independent of daytime conditions. In this study, we explored two methods of LEDIF retrievals: (a) hyperspectral sensor (1.33 nm full‐width half max) and (b) low‐cost Red‐Far‐Red photodiode sensor. LEDIF retrieved by the hyperspectral sensor demonstrated strong correlations with daytime SIF and gross primary productivity during mid to end of season phenology (R2 > 0.70). In contrast, phenological dynamics of LEDIF retrieved by the photodiode sensor was more subtle, likely due to weaker signal‐to‐noise ratio, but still demonstrated some potential. Overall, LEDIF offers a technique to monitor nighttime chlorophyll fluorescence emissions (and changes in its spectral shape with a hyperspectral sensor) to assess canopy‐scale phenology of photosynthetic potential.

Effects of Novel Photosynthetic Inhibitor [CuL2]Br2 Complex on Photosystem II Activity in Spinach.

The effects of the novel [CuL2]Br2 complex (L = bis{4H-1,3,5-triazino [2,1-b]benzothiazole-2-amine,4-(2-imidazole)}copper(II) bromide complex) on the photosystem II (PSII) activity of PSII membranes isolated from spinach were studied. The absence of photosynthetic oxygen evolution by PSII membranes without artificial electron acceptors, but in the presence of [CuL2]Br2, has shown that it is not able to act as a PSII electron acceptor. In the presence of artificial electron acceptors, [CuL2]Br2 inhibits photosynthetic oxygen evolution. [CuL2]Br2 also suppresses the photoinduced changes of the PSII chlorophyll fluorescence yield (FV) related to the photoreduction of the primary quinone electron acceptor, QA. The inhibition of both characteristic PSII reactions depends on [CuL2]Br2 concentration. At all studied concentrations of [CuL2]Br2, the decrease in the FM level occurs exclusively due to a decrease in Fv. [CuL2]Br2 causes neither changes in the F0 level nor the retardation of the photoinduced rise in FM, which characterizes the efficiency of the electron supply from the donor-side components to QA through the PSII reaction center (RC). Artificial electron donors (sodium ascorbate, DPC, Mn2+) do not cancel the inhibitory effect of [CuL2]Br2. The dependences of the inhibitory efficiency of the studied reactions of PSII on [CuL2]Br2 complex concentration practically coincide. The inhibition constant Ki is about 16 µM, and logKi is 4.8. As [CuL2]Br2 does not change the aromatic amino acids’ intrinsic fluorescence of the PSII protein components, it can be proposed that [CuL2]Br2 has no significant effect on the native state of PSII proteins. The results obtained in the present study are compared to the literature data concerning the inhibitory effects of PSII Cu(II) aqua ions and Cu(II)-organic complexes.

Combining near-infrared radiance of vegetation and fluorescence spectroscopy to detect effects of abiotic changes and stresses

Solar-induced chlorophyll fluorescence (SIF) shows great potential to assess plants physiological state and response to environmental changes. Recently the near-infrared reflectance of vegetation (NIRv) provides a promising way to quantify the confounding effect of canopy structure in SIF, while the difference between SIF and NIRv under varying environmental conditions has not been well explored. Here we developed a simple approach to extract the fluorescence yield (ΦF) by the combined use of SIF and the near-infrared radiance of vegetation (NIRvR). The proposed NIRvR approach was evaluated in multiple ways, including with the seasonal leaf-level steady-state fluorescence yield. Results indicate that NIRvR-derived ΦF well captured the seasonal variation of the fluorescence yield changes, and achieved similar results with the existing approach. Both SIF and NIRvR were derived from the airborne imaging fluorescence spectrometer HyPlant for three case studies to evaluate the impacts of light adaptation, heat stress and water limitation on ΦF. For the light adaptation case study, ΦF over the low-light adapted sugar beet field was about 1.3 times larger compared to an unaffected reference area while the difference in NIRvR was minimal, which clearly shows the short-term photosynthetic light induction effect and the ability of SIF to detect plant physiological responses. For the heat stress experiment, ΦF decreased during a natural heatwave in 2015 in the fields of rapeseed from 0.0150 to 0.0130, barley from 0.0152 to 0.0144, and wheat from 0.0146 to 0.0142 which showed signs of senescence, while slightly increased from 0.0125 to 0.0130 in the corn field which was still in growing. At the water-limited sugar beet field, ΦF first increased towards solar noon and then slightly decreased during the afternoon over the water-limited areas from 0.017 to 0.021 and 0.020, with high temperature and high light at noon. The advantages to use SIF/NIRvR as a proxy of ΦF to detect stress-induced limitations in photosynthesis include that the impacts of canopy structure and sun-sensor geometry on the ΦF estimation are explicitly cancelled, and photosynthetically active radiation (PAR) is not required as input. Finally, our approach is directly applicable to satellite-derived estimates of SIF, enabling the study of variations in ΦF to detect the effects of abiotic changes and stresses at large scale.

Stark absorption and Stark fluorescence spectroscopies: Theory and simulations.

Stark spectroscopy experiments are widely used to study the properties of molecular systems, particularly those containing charge-transfer (CT) states. However, due to the small transition dipole moments and large static dipole moments of the CT states, the standard interpretation of the Stark absorption and Stark fluorescence spectra in terms of the Liptay model may be inadequate. In this work, we provide a theoretical framework for calculations of Stark absorption and Stark fluorescence spectra and propose new methods of simulations that are based on the quantum-classical theory. In particular, we use the forward-backward trajectory solution and a variant of the Poisson bracket mapping equation, which have been recently adapted for the calculation of conventional (field-free) absorption and fluorescence spectra. For comparison, we also apply the recently proposed complex time-dependent Redfield theory, while exact results are obtained using the hierarchical equations of motion approach. We show that the quantum-classical methods produce accurate results for a wide range of systems, including those containing CT states. The CT states contribute significantly to the Stark spectra, and the standard Liptay formalism is shown to be inapplicable for the analysis of spectroscopic data in those cases. We demonstrate that states with large static dipole moments may cause a pronounced change in the total fluorescence yield of the system in the presence of an external electric field. This effect is correctly captured by the quantum-classical methods, which should therefore prove useful for further studies of Stark spectra of real molecular systems. As an example, we calculate the Stark spectra for the Fenna-Matthews-Olson complex of green sulfur bacteria.

SafePod: A respiration chamber to characterise apple fruit response to storage atmospheres

During long-term storage of apples, detection of low O2 stress is used to optimise low O2 storage regimes for dynamic controlled atmosphere (DCA) storage. Monitoring respiratory characteristics, specifically the respiratory quotient (RQ), provides a tool to achieve this. The objectives of this study were to evaluate protocols to monitor response of apple consignments to low O2 using a respiration chamber, the SafePod, designed for use in commercial stores and research laboratories, and to compare the RQ response to changes in chlorophyll fluorescence (CF) yield from the fruit skin as used in DCA-CF.Protocols to identify the lowest oxygen limit (LOL), the O2 concentration below which RQ rises, were tested using ‘Braeburn’ (sensitive to low O2) and ‘Gala’ (less sensitive to low O2). A protocol that allows fruit to acclimatise at each O2 concentration takes several weeks and is therefore not practical for commercial use. A rapid profile without fruit acclimatisation can be completed in 2–3 days. Although this underestimates RQ values, and results in an increase in RQ at a higher O2 concentration than observed for acclimatised fruit, the rapid RQ protocol provides a practical method to compare response of apple consignments between cultivars, orchards and seasons. By the rapid protocol, the LOL of ‘Braeburn’ consignments was near 0.6 kPa and of ‘Gala’ consignments was near 0.2 kPa, consistent with detection of alcoholic taints below the LOL in each case. The RQ response using the SafePod was consistent with increase in CF yield using HarvestWatch™.Fruit respiration rates change through the storage season, including a substantial decrease over the first 2 months after harvest. As RQ response is affected by respiration rate, accurate comparison of consignments depends on profiles being measured at the same stage in the storage season. It is more difficult to determine the LOL by RQ profiling later in the season when respiration rates are lower.

Detection and Quantification of Photosystem II Inhibitors Using the Freshwater Alga Desmodesmus subspicatus in Combination with High-Performance Thin-Layer Chromatography.

We present a novel tool for detecting and monitoring photosystem II (PSII) inhibitors, using the freshwater alga Desmodesmus subspicatus, in environmental samples fractionated by high-performance thin-layer chromatography (HPTLC). After chromatographic separation of a sample on a HPTLC plate, the algal suspension is sprayed homogeneously on the plate, and PSII-inhibition by specific sample components is detected based on changes in fluorescence yield, viewed by a maxi Imaging-Pulse-Amplitude-Modulation fluorometer. Dose-dependent responses to the PSII-inhibitor herbicides atrazine and diuron, frequently detected in water bodies, are demonstrated without and with chromatographic separation. The limits of quantification for atrazine and diuron with chromatographic separation were 1.94 ng and 99 pg, respectively, allowing the detection of environmentally relevant concentrations of these herbicides. The developed method was also employed to analyze sample extracts collected during a passive sampling campaign in surface waters. The obtained data correlated well with results from LC-MS/MS chemical analysis but also revealed unknown PSII-inhibiting activities. The proposed methodology represents a rapid and sensitive screening tool for the simultaneous effect-based detection of PSII-inhibitors in environmental samples.

Sustained Nonphotochemical Quenching Shapes the Seasonal Pattern of Solar‐Induced Fluorescence at a High‐Elevation Evergreen Forest

AbstractTraditional methods of carbon monitoring in mountainous regions are challenged by complex terrain. Recently, solar‐induced fluorescence (SIF) has been found to be an indicator of gross primary production (GPP), and the increased availability of remotely sensed SIF provides an opportunity to estimate GPP across the Western United States. Although the empirical linkage between SIF and GPP is strong, the current mechanistic understanding of this linkage is incomplete and depends upon changes in leaf biochemical processes in which absorbed sunlight leads to photochemistry, heat (via nonphotochemical quenching [NPQ]), fluorescence, or tissue damage. An improved mechanistic understanding is necessary to leverage SIF observations to improve representation of ecosystem processes within land surface models. Here we included an improved fluorescence model within the Community Land Model, Version 4.5 (CLM 4.5), to simulate seasonal changes in SIF at a subalpine forest in Colorado. We found that when the model accounted for sustained NPQ, this provided a larger seasonal change in fluorescence yield leading to simulated SIF that more closely resembled the observed seasonal pattern (Global Ozone Monitoring Experiment‐2 [GOME‐2] satellite platform and a tower‐mounted spectrometer system). We found that an acclimation model based on mean air temperature was a useful predictor for sustained NPQ. Although light intensity was not an important factor for this analysis, it should be considered before applying the sustained NPQ and SIF to other cold climate evergreen biomes. More leaf‐level fluorescence measurements are necessary to better understand the seasonal relationship between sustained and reversible components of NPQ and to what extent that influences SIF.

Nocturnal Light Emitting Diode Induced Fluorescence (LEDIF): A new technique to measure the chlorophyll a fluorescence emission spectral distribution of plant canopies in situ

-Solar-induced chlorophyll a Fluorescence (SIF), which is distributed over a relatively broad (~200 nm) spectral range, is a signal intricately connected to the efficiency of photosynthesis and is now observable from space. Variants of the Fraunhofer Line Depth/Discriminator (FLD) method are used as the basis of retrieval algorithms for estimating SIF from space. Although typically unobserved directly, recent advances in FLD-based algorithms now facilitate the prediction (by model inversion) of the canopy emitted fluorescence spectrum from the discrete-feature FLD retrievals.-Here we present first canopy scale measurements of chlorophyll a fluorescence spectra emitted from Scots pine at two times of year, and also from a lingonberry dominated understory. We used a high power multispectral Light Emitting Diode (LED) array to illuminate the respective canopies at night and measured under standardised conditions using a field spectrometer mounted in the nadir position above the canopy. We refer to the technique, which facilitates the in situ upscaling of a commonly measured leaf scale quantity to the canopy, as nocturnal LED-Induced chlorophyll a Fluorescence (LEDIF).-The shape of the LEDIF spectra was dependant on the colour of the excitation light and also on the dominant species. Because we measured pine at two different times of year we were also able to show an increase in the canopy scale apparent quantum yield of fluorescence which was consistent with leaf-level increase in fluorescence yield recorded with a monitoring PAM fluorometer.-The automation of the LEDIF technique could be used to estimate seasonal changes in canopy fluorescence spectra and yield from fixed or mobile platforms and provide a window into functional traits across species and architectures. LEDIF could also be used to evaluate FLD and inversion-based retrievals of canopy spectra, as well as different irradiance normalisation schemes typically applied to SIF data to account for the dependence of SIF on ambient light conditions.

Hydroxyectoine protects Mn-depleted photosystem II against photoinhibition acting as a source of electrons.

In the present study, we have investigated the effect of hydroxyectoine (Ect-OH), a heterocyclic amino acid, on oxygen evolution in photosystem II (PS II) membrane fragments and on photoinhibition of Mn-depleted PS II (apo-WOC-PS II) preparations. The degree of photoinhibition of apo-WOC-PS II preparations was estimated by the loss of the capability of exogenous electron donor (sodium ascorbate) to restore the amplitude of light-induced changes of chlorophyll fluorescence yield (∆F). It was found that Ect-OH (i) stimulates the oxygen-evolving activity of PS II, (ii) accelerates the electron transfer from exogenous electron donors (K4[Fe(CN)6], DPC, TMPD, Fe2+, and Mn2+) to the reaction center of apo-WOC-PS II, (iii) enhances the protective effect of exogenous electron donors against donor-side photoinhibition of apo-WOC-PS II preparations. It is assumed that Ect-OH can serve as an artificial electron donor for apo-WOC-PS II, which does not directly interact with either the donor or acceptor side of the reaction center. We suggest that the protein conformation in the presence of Ect-OH, which affects the extent of hydration, becomes favorable for accepting electrons from exogenous donors. To our knowledge, this is the first study dealing with redox activity of Ect-OH towards photosynthetic pigment-protein complexes.

Is carbonic anhydrase activity of photosystem II required for its maximum electron transport rate?

It is known, that the multi-subunit complex of photosystem II (PSII) and some of its single proteins exhibit carbonic anhydrase activity. Previously, we have shown that PSII depletion of HCO3−/CO2 as well as the suppression of carbonic anhydrase activity of PSII by a known inhibitor of α‑carbonic anhydrases, acetazolamide (AZM), was accompanied by a decrease of electron transport rate on the PSII donor side. It was concluded that carbonic anhydrase activity was required for maximum photosynthetic activity of PSII but it was not excluded that AZM may have two independent mechanisms of action on PSII: specific and nonspecific. To investigate directly the specific influence of carbonic anhydrase inhibition on the photosynthetic activity in PSII we used another known inhibitor of α‑carbonic anhydrase, trifluoromethanesulfonamide (TFMSA), which molecular structure and physicochemical properties are quite different from those of AZM. In this work, we show for the first time that TFMSA inhibits PSII carbonic anhydrase activity and decreases rates of both the photo-induced changes of chlorophyll fluorescence yield and the photosynthetic oxygen evolution. The inhibitory effect of TFMSA on PSII photosynthetic activity was revealed only in the medium depleted of HCO3−/CO2. Addition of exogenous HCO3− or PSII electron donors led to disappearance of the TFMSA inhibitory effect on the electron transport in PSII, indicating that TFMSA inhibition site was located on the PSII donor side. These results show the specificity of TFMSA action on carbonic anhydrase and photosynthetic activities of PSII. In this work, we discuss the necessity of carbonic anhydrase activity for the maximum effectiveness of electron transport on the donor side of PSII.

Mechanisms of drought-induced dissipation of excitation energy in sun- and shade-adapted drought-tolerant mosses studied by fluorescence yield change and global and target analysis of fluorescence decay kinetics.

Some mosses stay green and survive long even under desiccation. Dissipation mechanisms of excess excitation energy were studied in two drought-tolerant moss species adapted to contrasting niches: shade-adapted Rhytidiadelphus squarrosus and sun-adapted Rhytidium rugosum in the same family. (1) Under wet conditions, a light-induced nonphotochemical quenching (NPQ) mechanism decreased the yield of photosystem II (PSII) fluorescence in both species. The NPQ extent saturated at a lower illumination intensity in R. squarrosus, suggesting a larger PSII antenna size. (2) Desiccation reduced the fluorescence intensities giving significantly lower F 0 levels and shortened the overall fluorescence lifetimes in both R. squarrosus and R. rugosum, at room temperature. (3) At 77K, desiccation strongly reduced the PSII fluorescence intensity. This reduction was smaller in R. squarrosus than in R. rugosum. (4) Global and target analysis indicated two different mechanisms of energy dissipation in PSII under desiccation: the energy dissipation to a desiccation-formed strong fluorescence quencher in the PSII core in sun-adapted R. rugosum (type-A quenching) and (5) the moderate energy dissipation in the light-harvesting complex/PSII in shade-adapted R. squarrosus (type-B quenching). The two mechanisms are consistent with the different ecological niches of the two mosses.

Kinetics of photosystem II electron transport: a mathematical analysis based on chlorophyll fluorescence induction.

The OJDIP rise in chlorophyll fluorescence during induction at different light intensities was mathematically modeled using 24 master equations describing electron transport through photosystem II (PSII) plus ordinary differential equations for electron budgets in plastoquinone, cytochrome f, plastocyanin, photosystem I, and ferredoxin. A novel feature of the model is consideration of electron in- and outflow budgets resulting in changes in redox states of Tyrosine Z, P680, and QA as sole bases for changes in fluorescence yield during the transient. Ad hoc contributions by transmembrane electric fields, protein conformational changes, or other putative quenching species were unnecessary to account for primary features of the phenomenon, except a peculiar slowdown of intra-PSII electron transport during induction at low light intensities. The lower than F m post-flash fluorescence yield F f was related to oxidized tyrosine Z. The transient J peak was associated with equal rates of electron arrival to and departure from QA and requires that electron transfer from QA- to QB be slower than that from QA- to QB-. Strong quenching by oxidized P680 caused the dip D. Reduced plastoquinone, a competitive product inhibitor of PSII, blocked electron transport proportionally with its concentration. Electron transport rate indicated by fluorescence quenching was faster than the rate indicated by O2 evolution, because oxidized donor side carriers quench fluorescence but do not transport electrons. The thermal phase of the fluorescence rise beyond the J phase was caused by a progressive increase in the fraction of PSII with reduced QA and reduced donor side.

Loss of LHCI system affects LHCII re-distribution between thylakoid domains upon state transitions.

LHCI, the peripheral antenna system of Photosystem I, includes four light-harvesting proteins (Lhca1-Lhca4) in higher plants, all of which are devoid in the Arabidopsis thaliana knock-out mutant ΔLhca. PSI absorption cross-section was reduced in the mutant, thus affecting the redox balance of the photosynthetic electron chain and resulting in a more reduced PQ with respect to the wild type. ΔLhca plants developed compensatory response by enhancing LHCII binding to PSI. However, the amplitude of state transitions, as measured from changes of chlorophyll fluorescence in vivo, was unexpectedly low than the high level of PSI-LHCII supercomplex established. In order to elucidate the reasons for discrepancy, we further analyzed state transition in ΔLhca plants. The STN7 kinase was fully active in the mutant as judged from up-regulation of LHCII phosphorylation in state II. Instead, the lateral heterogeneity of thylakoids was affected by lack of LHCI, with LHCII being enriched in stroma membranes with respect to the wild type. Re-distribution of this complex affected the overall fluorescence yield of thylakoids already in state I and minimized changes in RT fluorescence yield when LHCII did connect to PSI reaction center. We conclude that interpretation of chlorophyll fluorescence analysis of state transitions becomes problematic when applied to mutants whose thylakoid architecture is significantly modified with respect to the wild type.

Correlation between Strawberry (Fragaria ananassa Duch.) Productivity and Photosynthesis-Related Parameters under Various Growth Conditions.

In the present study, we investigated changes in chlorophyll fluorescence, photosynthetic parameters and fruit yields, as well as fruit phytochemical accumulation of strawberry (Fragaria ananassa Duch.) that had been cultivated in a greenhouse under different combinations of light intensity and temperature. In plants grown with low light (LL) photosystem II chlorophyll fluorescence was found to increase as compared with those grown under high light (HL). When strawberry plants were grown with temperature higher than 5°C in addition to LL, they showed decrease in non-photochemical quenching (NPQ), photochemical quenching (qP), as well as chlorophyll fluorescence decrease ratio (RFd) when compared with other combinations of light and temperature. Moreover, fruit yield of strawberry was closely correlated with chlorophyll fluorescence-related parameters such as NPQ, qP, and RFd, but not with the maximum efficiency of PS II (Fv/Fm). Although plant groups grown under different combinations of light and temperature showed almost comparable levels of photosynthesis rates (Pr) when irradiated with low-intensity light, they displayed clear differences when measured with higher irradiances. Plants grown under HL with temperature above 10°C showed the highest Pr, in contrast to the plants grown under LL with temperature above 5°C. When the stomatal conductance and the transpiration rate were measured, plants of each treatment showed clear differences even when analyzed with lower irradiances. We also found that fruit production during winter season was more strongly influenced by growth temperature than light intensity. We suggest that fruit productivity of strawberry is closely associated with chlorophyll fluorescence and photosynthesis-related parameters during cultivation under different regimes of temperature and light.

Trehalose protects Mn-depleted photosystem 2 preparations against the donor-side photoinhibition

Recently, it has been shown that the addition of 1M trehalose leads to the increase of the rate of oxygen photoconsumption associated with activation of electron transport in the reaction center of photosystem 2 (PS2) in Mn-depleted PS2 membranes (apo-WOC-PS2) [37]. In the present work the effect of trehalose on photoinhibition of apo-WOC-PS2 preparations (which are characterized by a high sensitivity to the donor side photoinhibition of PS2) was investigated. The degree of photoinhibition was estimated by the loss of the capability of exogenous electron donor (sodium ascorbate) to reactivate the electron transport (measured by light-induced changes of chlorophyll fluorescence yield (∆F)) in apo-WOC-PS2. It was found that 1M trehalose enhanced the Mn2+-dependent suppression of photoinhibition of apo-WOC-PS2: in the presence of trehalose the addition of 0.2μM Mn2+ (corresponding to 2 Mn2+ per one reaction center) was sufficient for an almost complete suppression of the donor side photoinhibition of the complex. In the absence of trehalose it was necessary to add 100μM Mn2+ to achieve a similar result. The effect of trehalose was observed during photoinhibition of apo-WOC-PS2 at low (15μmolphotons−1m−2) and high (200μmolphotons−1m−2) light intensity. When Mn2+ was replaced by other PS2 electron donors (ferrocyanide, DPC) as well as by Ca2+ the protective effect of trehalose was not observed. It was also found that 1M trehalose decreased photoinhibition of apo-WOC-PS2 if the samples contained endogenous manganese (1–2 Mn ions per one RC was enough for the maximum protection effect). It is concluded that structural changes in PS2 caused by the addition of trehalose enhance the capability of photochemical reaction centers of apo-WOC-PS2 to accept electrons from manganese (both exogenous and endogenous), which in turn leads to a considerable suppression of the donor side photoinhibition of PS2.

Fluorescence and phosphorescence of tryptophan in peptides of different length and sequence

To interpret accurately protein fluorescence and phosphorescence, it is essential to achieve a better understanding of the luminescence properties of tryptophan (Trp, or W) in peptides. In published literature data on luminescence of peptides of varied length are scarce. This article describes studies of fluorescence and phosphorescence properties of the eight Trp-containing synthetic peptides: WAK, AWK, SWA, KYLWE, AVSWK, WVSWAK, WAKLAWE, and AVSWAKLARE. The aim was to investigate which factors influence the fluorescence yield and phosphorescence-spectra and lifetimes. Absorption spectra, room temperature fluorescence emission and corresponding excitation spectra and time-resolved phosphorescence spectra (77K) have been recorded; the dependence of the fluorescence quantum yield on the specific peptide and its variation with the wavelength of excitation has been studied. The changes in fluorescence yield and shape of phosphorescence spectra are explained in terms of internal electron and proton transfer. The structured phosphorescence spectrum originates from proton transfer occurring upon excitation of Trp, while electron transfer gives rise to a non-structured luminescence spectrum. There is also electron transfer from higher vibronic S1 states. In the peptides there is higher probability of electron transfer than in Trp alone. The obtained data are interpreted in light of the peptides' sequence, length and conformation.

Involvement of molecular oxygen in the donor-side photoinhibition of Mn-depleted photosystem II membranes.

It has been shownby Khorobrykh et al. (Biochemistry (Moscow)67:683-688, 2002); Yanykin et al. (Biochim Biophys Acta 1797:516-523, 2010); Khorobrykh et al. (Biochemistry 50:10658-10665, 2011) that Mn-depleted photosystem II (PSII) membrane fragments are characterized by an enhanced oxygen photoconsumption on the donor side of PSII which is accompanied with hydroperoxide formation and it was suggested that the events are related to the oxidative photoinhibition of PSII. Experimental confirmation of this suggestion is presented in this work. The degree of photoinhibition was determined by the loss of the capability of exogenous electron donors (Mn(2+) or sodium ascorbate) to the reactivation of electron transport [measured by the light-induced changes of chlorophyll fluorescence yield (∆F)] in Mn-depleted PSII membranes. The transition from anaerobic conditions to aerobic ones significantly activated photoinhibition of Mn-depleted PSII membranes both in the absence and in the presence of exogenous electron acceptor, ferricyanide. The photoinhibition of Mn-depleted PSII membranes was suppressed upon the addition of exogenous electron donors (Mn(2+), diphenylcarbazide, and ferrocyanide). The addition of superoxide dismutase did not affect the photoinhibition of Mn-depleted PSII membranes. It is concluded that the interaction of molecular oxygen (rather than superoxide anion radical formed on the acceptor side of PSII) with the oxidized components of the donor side of PSII reflects the involvement of O2 in the donor-side photoinhibition of Mn-depleted PSII membranes.

The non-foliar hypoxic photosynthetic syndrome: evidence for enhanced pools and functionality of xanthophyll cycle components and active cyclic electron flow in fruit chlorenchyma.

Green fruits display a high engagement in CEF and enhanced VAZ cycle activity as a response to the demands imposed by their internal aerial conditions, particularly low O 2 , due to gas exchange limitations. In the present study, we used HPLC analysis, post-illumination changes in fluorescence yield under varying O2 and CO2 partial pressures and absorbance changes at 820 nm induced by far-red light to assess the carotenoid composition, the functionality of the xanthophyll cycle (VAZ) and the possibility of an active cyclic e (-) flow (CEF) in the fully exposed green fruits from Nerium oleander and Rosa sp. Equally exposed, mature leaves served as controls. Compared to leaves, fruits display less total chlorophylls and carotenoids but higher Car/Chl ratio, mainly shaped by the increased pools of the VAZ cycle components, in both species. The enhanced VAZ pool size in fruits is combined with a higher mid-day de-epoxidation state (DEPS). Moreover, fruits exhibit considerably lower levels of oxidizable P700, a faster re-reduction of PSI and significantly higher relative magnitude of CEF, irrespective of the O2/CO2 levels applied. We conclude that the higher VAZ investment may serve the enhanced heat dissipation needs in fruits, in the presence of a suppressed linear e (-) flow. In addition, the elevated potential of CEF may replenish the ATP lost due to hypoxia and concurrently facilitate the development of adequate non-photochemical quenching (NPQ), through its contribution to ΔpH increase. Since other non-foliar green organs exhibit a similar photosynthetic pattern, we argue that this may reflect a common strategy for green tissues under similar micro-environmental conditions, particularly hypoxia.

Effects of fully open-air [CO2] elevation on leaf photosynthesis and ultrastructure of Isatis indigotica fort.

Traditional Chinese medicine relies heavily on herbs, yet there is no information on how these herb plants would respond to climate change. In order to gain insight into such response, we studied the effect of elevated [CO2] on Isatis indigotica Fort, one of the most popular Chinese herb plants. The changes in leaf photosynthesis, chlorophyll fluorescence, leaf ultrastructure and biomass yield in response to elevated [CO2] (550±19 µmol mol–1) were determined at the Free-Air Carbon dioxide Enrichment (FACE) experimental facility in North China. Photosynthetic ability of I. indigotica was improved under elevated [CO2]. Elevated [CO2] increased net photosynthetic rate (P N), water use efficiency (WUE) and maximum rate of electron transport (J max) of upper most fully-expended leaves, but not stomatal conductance (gs), transpiration ratio (Tr) and maximum velocity of carboxylation (V c,max). Elevated [CO2] significantly increased leaf intrinsic efficiency of PSII (Fv’/Fm’) and quantum yield of PSII(ΦPS II), but decreased leaf non-photochemical quenching (NPQ), and did not affect leaf proportion of open PSII reaction centers (qP) and maximum quantum efficiency of PSII (Fv/Fm). The structural chloroplast membrane, grana layer and stroma thylakoid membranes were intact under elevated [CO2], though more starch grains were accumulated within the chloroplasts than that of under ambient [CO2]. While the yield of I. indigotica was higher due to the improved photosynthesis under elevated [CO2], the content of adenosine, one of the functional ingredients in indigowoad root was not affected.