Characterization of photosystem II heterogeneity in response to high salt stress in wheat leaves (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

The effect of high salt stress on PS II heterogeneity was investigated in wheat (Triticum aestivum) leaves. On the basis of antenna size, PS II has been classified into three forms, i.e., α, β, and γ centers while on the basis of electron transport properties of the reducing side of the reaction centers, two distinct forms of PS II have been suggested, i.e., Q_B reducing centers and Q_B non-reducing centers. The chlorophyll a (Chl a) fluorescence transients, which can quantify PS II behavior, were recorded using PEA to derive OJIP in vivo with high time resolution and further analyzed according to JIP test. Our results showed that with an increase in the salt concentration during growth, the number of Q_B non-reducing centers increased. In antenna size heterogeneity the number of β and γ centers increased while the number of α centers decreased. A change in the energetic connectivity between the PS II units was also observed. Recovery studies showed that antenna heterogeneity was completely recovered from damage at 0.5 M NaCl concentration and partially recovered at 1 M NaCl concentration while reducing side heterogeneity showed no recovery at all after 0.5 M onwards.     

Chlorophyll a fluorescence study revealing effects of high salt stress on Photosystem II in wheat leaves

In order to study the effects of high salt stress on PS II in detached wheat (Triticum aestivum) leaves, the seedlings were grown in Knop solution and temperature was 20 ± 2 °C. Detached leaves were exposed to high salt stress (0.1–0.5 M NaCl) for 1 h in dark and Chl a fluorescence induction kinetics was measured. Various parameters like Fv/Fm, ABS/RC, ETo/TRo, performance index and area over the florescence curve were measured and the energy pipeline model was deduced in response to salt stress. Our results show that the damage caused due to high salt stress is more prominent at the donor side rather than the acceptor side of PS II. Moreover the effects of high salt stress are largely reversible, as the acceptor side damage is completely recovered (～100%) while the recovery of the donor side is less than 85%. Based on our results we suggest that in response to high salt stress, the donor side of PS II is affected more as compared to the acceptor side of PS II.     

Effects of dual stress (high salt and high temperature) on the photochemical efficiency of wheat leaves (Triticum aestivum)

In this study, we have focused on those components of Photosystem (PS) II which are significantly affected by dual stress (high salt and temperature) on wheat as measured by Plant Efficiency Analyser (PEA). It was observed that some of the chlorophyll a fluorescence parameters were temperature dominated, while some other parameters were salt dominated. We have also observed additive effects for parameters like antenna size heterogeneity. An important observation was that in high temperature alone, the K-step was observed at 40 °C, while in case of dual stress, the K-step was observed at 45 °C, while the Chl a fluorescence transient of 40 °C?+?0.5 M?NaCl was quite similar to 35 °C transient curve. In the presence of salt, K-step was observed at higher temperature suggesting a protection of OEC by salt. Plants are under dual stress, but effect of temperature stress is less severe in presence of salt stress. Thus, we can say that salt stress caused partial prevention from high temperature stress but it did not cause complete protection of PS II.     

Computational analysis of fluorescence induction curves in intact spinach leaves treated at different pH

Effects of change in pH have been investigated on spinach leaf discs by measuring fluorescence induction kinetics using plant efficiency analyzer (PEA). On the basis of computational analysis of the results, we have reported that acidic pH causes a significant inhibition of the donor and the acceptor side of PS II. Energy flux models have been presented using the software Biolyzer HP 3. Effects of pH were investigated on the antenna size heterogeneity of PS II and a relative change in the proportions of α, β, and γ centers was observed.     

How does Lobaria pulmonaria regulate photosystem II during progressive desiccation and osmotic water stress?

The effects of decreasing water potential (Ψ) on O₂ evolution and fluorescence yield at room temperature and at 77 K were investigated using the lichen Lobaria pulmonaria. Changes in Ψ were created either by atmospheric desiccation or by osmotic dehydration, with either sucrose, sorbitol or NaCl as osmoticum. Independent of the method used to establish Ψ, similar inactivation patterns were obtained and were reversible after reincubation in pure water for 10 min. Our data indicate that exposure to increasing water stress acts at two levels. In the first phase, at ‘mild’ stress, i.e. at Ψ greater than ?13, ?16 and ?20 MPa for drying, NaCl and sucrose treatments, respectively, a progressive decline in O₂ production and the fluorescence yield (ΔF/F_m′ and F_v/F_m) was correlated with increases in non-photochemical quenching (q_N). At the same time the photochemical quenching (q_p) changed only sligthly, indicating the absence of overreduction. The F_o level remained relatively constant in this first stage of water loss. A ΔpH mediated down regulation and a donor side limitation of photosystem (PS) II are discussed. When the water stress was severe, a further decrease in the fluorescence yield was observed and correlated with a considerable decrease in F_o (second phase). Kinetic analysis of the 77 K emission showed that osmotic stress and atmospheric desiccation possibly lead to an increased spillover from PS II to PS I. In addition, a strong negative effect of NaF on the recovery from dehydration was found. This may indicate a state transition mediated by the displacement/recoupling of light harvesting complex (LHC) II from/to PS II. The photoprotective role of spatial rearrangements of antenna complexes during desiccation is discussed.     

Senescence-induced alterations in the photosystem II functions of Cucumis sativus cotyledons: probing of senescence driven alterations of photosystem II by chlorophyll a fluorescence induction O-J-I-P transients

Senescence-induced alterations in photosystem II (PS II) structure and photofunctions were probed in cucumber (Cucumis sativus) cotyledons, using fast O-J-I-P Chlorophyll a (Chl a) fluorescence transients. Analysis of measured and derived parameters of the fast fluorescence O-J-I-P transient revealed senescence-induced alterations in (i), PS II acceptor side electron transfer equilibrium between QA and QB, the primary stable and secondary acceptors of PS II; (ii), intersystem PQ pool size and (iii), affected electron transfer from PS II to PS I. Also, senescence of cotyledons triggered conversion of QA-reducing (fully active) to non- QA-reducing PS II (heat sink) centres. Further, some of the remaining active PS II centres showed a high apparent trapping efficiency due to clustering and energetic connectivity (grouping) between the antennae of active and inactive centers. The overall density of active PS II reaction centers showed a temporal decrease due to the onset of foliar senescence. Thus, the fast Chl a fluorescence transients, with a time resolution of at least 50 mircosec and use of the equations of JIP-test, provide a valuable, non-invasive rapid biophysical probe to study the ageing in plants in terms of detecting photosynthetic activities and the heterogeneity of different types of photosynthetic units. Further, these results were found to be in agreement with the earlier in vitro studies using thylakoids isolated from senescing cotyledons where it was shown that senescence induced heterogeneity in PS II centers affected acceptor side QA<-->QB equilibrium.     

Photosystem I shows a higher tolerance to sorbitol‐induced osmotic stress than photosystem II in the intertidal macro‐algae Ulva prolifera (Chlorophyta)

The photosynthetic performance of the desiccation‐tolerant, intertidal macro‐algae Ulva prolifera was significantly affected by sorbitol‐induced osmotic stress. Our results showed that photosynthetic activity decreased significantly with increases in sorbitol concentration. Although the partial activity of both photosystem I (PS I) and photosystem II (PS II) was able to recover after 30 min of rehydration, the activity of PS II decreased more rapidly than PS I. At 4 M sorbitol concentration, the activity of PS II was almost 0 while that of PS I was still at about one third of normal levels. Following prolonged treatment with 1 and 2 M sorbitol, the activity of PS I and PS II decreased slowly, suggesting that the effects of moderate concentrations of sorbitol on PS I and PS II were gradual. Interestingly, an increase in non‐photochemical quenching occurred under these conditions in response to moderate osmotic stress, whereas it declined significantly under severe osmotic stress. These results suggest that photoprotection in U. prolifera could also be induced by moderate osmotic stress. In addition, the oxidation of PS I was significantly affected by osmotic stress. P700⁺ in the thalli treated with high concentrations of sorbitol could still be reduced, as PS II was inhibited by 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea (DCMU), but it could not be fully oxidized. This observation may be caused by the higher quantum yield of non‐photochemical energy dissipation in PS I due to acceptor‐side limitation (Y(NA)) during rehydration in seawater containing DCMU.     

Salt stress inhibits photosystems II and I in cyanobacteria

Recent studies of responses of cyanobacterial cells to salt stress have revealed that the NaCl-induced decline in the photosynthetic activities of photosystems II and I involves rapid and slow changes. The rapid decreases in the activities of both photosystems, which occur within a few minutes, are reversible and are associated with osmotic effects, which induce the efflux of water from the cytosol through water channels and rapidly increase intracellular concentrations of salts. Slower decreases in activity, which occur within hours, are irreversible and are associated with ionic effects that are due to the influx of Na(+) and Cl(-) ions through K(+)(Na(+)) channels and, probably, Cl(-) channels, with resultant dissociation of extrinsic proteins from photosystems. In combination with light stress, salt stress significantly stimulates photoinhibition by inhibiting repair of photodamaged photosystem II. Tolerance of photosystems to salt stress can be enhanced by genetically engineered increases in the unsaturation of fatty acids in membrane lipids and by intracellular synthesis of compatible solutes, such as glucosylglycerol and glycinebetaine. In this review, we summarize recent progress in research on the effects of salt stress on photosynthesis in cyanobacteria.     

Investigation of deleterious effects of chromium phytotoxicity and photosynthesis in wheat plant

Increasing human and industrial activities lead to heavy metal pollution. Heavy metal chromium (Cr) is considered to be a serious environmental contaminant for the biota. Phytotoxic effects of Cr were studied in wheat plants. Growth parameters were largely inhibited as a result of disturbances in the plant cell metabolism in response to Cr toxicity. Chromium toxicity led to decline in a number of active reaction centres of PSII, rate of electron transport, and change in PSII heterogeneity. Chromium did not cause any change in heterogeneity of the reducing side. A significant change in antenna size heterogeneity of PSII occurred in response to Cr toxicity. Chromium seems to have extensive effects on the light harvesting complex of PSII.     

Alteration of photosystem II properties with non-photochemical excitation quenching

Oxygen yield from single turnover flashes and multiple turnover pulses was measured in sunflower leaves differently pre-illuminated to induce either 'energy-dependent type' non-photochemical excitation quenching (qE) or reversible, inhibitory type non-photochemical quenching (qI). A zirconium O2 analyser, combined with a flexible gas system, was used for these measurements. Oxygen yield from saturating single turnover flashes was the equivalent of 1.3-2.0 micromole(-) m(-2) in leaves pre-adapted to low light. It did not decrease when qE quenching was induced by a 1 min exposure to saturating light, but it decreased when pre-illumination was extended to 30-60 min. Oxygen evolution from saturating multiple turnover pulses behaved similarly: it did not decrease with the rapidly induced qE but decreased considerably when exposure to saturating light was extended or O2 concentration was decreased to 0.4%. Parallel recording of chlorophyll fluorescence and O2 evolution during multiple turnover pulses, interpreted with the help of a mathematical model of photosystem II (PS II) electron transport, revealed PS II donor and acceptor side resistances. These experiments showed that PS II properties depend on the type of non-photochemical quenching present. The rapidly induced and rapidly reversible qE type (photoprotective) quenching does not induce changes in the number of active PS II or in the PS II maximum turnover rate, thus confirming the antenna mechanism of qE. The more slowly induced but still reversible qE type quenching (photoinactivation) induced a decrease in the number of active PS II and in the maximum PS II turnover rate. Modelling showed that, mainly, the acceptor side resistance of PS II increased in parallel with the reversible qI.     

Characterization of the fast and slow reversible components of non-photochemical quenching in isolated pea thylakoids by picosecond time-resolved chlorophyll fluorescence analysis.

The fast and slow reversible components of non-photochemical chlorophyll fluorescence quenching commonly assigned to the qE and the qI mechanism have been studied in isolated pea thylakoids which were prepared from leaves after a moderate photoinhibitory treatment. Chlorophyll fluorescence decays were measured at picosecond resolution and analyzed on the basis of the heterogeneous exciton/radical pair equilibrium model. Our results show that the fast reversible non-photochemical quenching is completely assigned to the PS II antenna and is related to zeaxanthin. The slow reversible qI type quenching is located at the PS II reaction center and involves enhanced nonradiative decay of the primary charge separated state to its ground state and/or triplet excited state. Apart from its independence from the proton gradient, the qI quenching shows striking similarities to a particular form of qE quenching which is also located at the PS II reaction center and has resently been resolved in isolated thylakoids from dark-adapted leaves [Wagner, B., et al. (1996) J. Photochem. Photobiol., B 36, 339-350]. Our data suggest that during exposure to the supersaturating light the reaction center qE component was replaced by qI quenching. This qE to qI transition is supposed to be part of the mechanism of the long-term downregulation of PS II during photoinhibition. It is also evident that under the conditions used in our study zeaxanthin-dependent antenna quenching is not involved in the slow reversible downregulation of PS II but that it retains its dependence on the proton gradient during exposure to strong light.     

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 Q_B-nonreducing centres, were also investigated. The damage caused by ANT to PSII antenna heterogeneity was recovered almost by 100% owing to TX-100.     

Unraveling salt stress signaling in plants

Salt stress is a major environmental factor limiting plant growth and productivity. A better understanding of the mechanisms mediating salt resistance will help researchers design ways to improve crop performance under adverse environmental conditions. Salt stress can lead to ionic stress, osmotic stress and secondary stresses, particularly oxidative stress, in plants. Therefore,to adapt to salt stress, plants rely on signals and pathways that re-establish cellular ionic, osmotic, and reactive oxygen species(ROS) homeostasis. Over the past two decades, genetic and biochemical analyses have revealed several core stress signaling pathways that participate in salt resistance. The Salt Overly Sensitive signaling pathway plays a key role in maintaining ionic homeostasis,via extruding sodium ions into the apoplast. Mitogenactivated protein kinase cascades mediate ionic, osmotic,and ROS homeostasis. SnR K2(sucrose nonfermenting1-related protein kinase 2) proteins are involved in maintaining osmotic homeostasis. In this review, we discuss recent progress in identifying the components and pathways involved in the plant's response to salt stress and their regulatory mechanisms. We also review progress in identifying sensors involved in salt-induced stress signaling in plants.     

Ionic and osmotic effects of NaCl-induced inactivation of photosystems I and II in Synechococcus sp

We report here that osmotic effects and ionic effects are both involved in the NaCl-induced inactivation of the photosynthetic machinery in the cyanobacterium Synechococcus sp. PCC 7942. Incubation of the cyanobacterial cells in 0.5 M NaCl induced a rapid and reversible decline and subsequent slow and irreversible loss of the oxygen-evolving activity of photosystem (PS) II and the electron transport activity of PSI. An Na(+)-channel blocker protected both PSII and PSI against the slow, but not the rapid, inactivation. The rapid decline resembled the effect of 1.0 M sorbitol. The presence of both an Na(+)-channel blocker and a water-channel blocker protected PSI and PSII against the short- and long-term effects of NaCl. Salt stress also decreased cytoplasmic volume and this effect was enhanced by the Na(+)-channel blocker. Our observations suggested that NaCl had both osmotic and ionic effects. The osmotic effect decreased the amount of water in the cytosol, rapidly increasing the intracellular concentration of salts. The ionic effect was caused by an influx of Na(+) ions through potassium/Na(+) channels that also increased concentrations of salts in the cytosol and irreversibly inactivated PSI and PSII.     

模拟干旱和盐分胁迫对沙枣幼苗PSⅡ活力的影响

通过PEG-6000和NaCl模拟实验研究了干旱和盐分胁迫对沙枣幼苗叶片光系统Ⅱ(PSⅡ)活力的影响。结果显示:PEG胁迫使沙枣幼苗叶片PSⅡ在300μs时相对于FJ-Fo的可变荧光比值(Wk)升高,却降低了单位反应中心密度(RC/CSo)和最大量子产量(Fv/Fm),导致效能指数(PIABS)随水势降低而显著下降,阻碍了电子传递链中供体和受体侧的电子传递,也抑制了叶绿素的合成,从而全面抑制PSⅡ的活力;NaCl胁迫对沙枣叶片PSⅡ活力没有显著影响。比较两种处理等渗溶液下的结果发现,盐离子对沙枣叶片PSⅡ活力具有正效应,它抵消了渗透效应对沙枣叶片PSⅡ活力的抑制作用,这可能与盐离子进入叶片细胞,减轻了渗透胁迫有关。     

Adaptation of photosynthesis under iron deficiency in maize

This paper explores the effects of high light stress on Fe-deficient plants. Maize (Zea mays) plants were grown under conditions of Fe deficiency and complete nutrition. Attached, intact leaves of Fe-deficient and control plants were used for gas exchange experiments under suboptimal, optimal and photoinhibitory illumination. Isolated chloroplasts were used to study photosynthetic electron transport system, compromised by the induction of Fe deficiency. The reaction centers of PS II (measured as reduction of Q, the primary electron acceptor of P 680) and PS I (measured as oxidation of P 700) were estimated from the amplitude of light induced absorbance change at 320 and 700 nm, respectively. Plants were subjected to photoinhibitory treatment for different time periods and isolated chloroplasts from these plants were used for electron transport studies. Carbon dioxide fixation in control as well as in Fe-deficient plants decreased in response to high light intensities. Total chlorophyll, P 700 and Q content in Fe-deficient chloroplasts decreased, while Chl a/b ratio and Q/P 700 ratio increased. However, electron transport through PS II suffered more after photoinhibitory treatment as compared to electron transport through PS I or whole chain. Electron transfer through PS I+PS II, excluding the water oxidation complex showed a decrease in Fe-deficient plants. However, electron transport through this part of the chain did not suffer much as a result of photoinhibition, suggesting a defect in the oxidising side of PS II.     

Phosphorylation of spinach chlorophyll-protein complexes. CPII, but not CP29, CP27, or CP24, is phosphorylated in vitro

Previous studies have indicated that the reversible phosphorylation of a population of antenna complexes that can donate energy to PS II ('mobile LHC II') plays a regulatory role in the state 1-state 2 transition in thylakoid membranes. The relationship of phosphorylated LHC II to the multiple PS II-associated chlorophyll a/b-proteins resolvable on green gels is currently unclear. We have used a high resolution gel system to analyze thylakoids phosphorylated in vitro. The only PS II-associated antenna complex to become phosphorylated is CPII, indicating that this complex represents the mobile LHC II. The other putative PS II antenna complexes, CP29, CP24, and the new complex designated CP27 which comigrates with CPII, are not phosphorylated and are probably components of the bound 'LHC II' antenna.     

Energy transfer and quantum yield in Photosystem II

The photoreduction and dark reoxidation of Q_α and Q_β, the primary electron acceptors of Photosystems (PS) IIα and IIβ, respectively, in the presence of 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea (DCMU) were studied in tobacco chloroplasts by means of fluorescence and absorbance measurements. The magnitude of a correction for an absorbance change by the oxidizing side of PS II needed in our previous study of the quantum yield of Q reduction (Biochim. Biophys. Acta 635 (1981), 111–120) has been determined. The absorbance change occurs in PS IIα mainly. The maximum fluorescence yield was found to be the same as in the mutant Su/su, which has a 3-fold higher reaction center concentration and a lower PS IIα to PS IIβ ratio. The kinetics of the light-induced fluorescence increase were measured after various pretreatments and the corresponding kinetics of the integrated fluorescence deficit were analyzed into their α and β components. From the results the contribution to the minimum fluorescence level, the degree of energy transfer between units, and the quantum efficiency of Q reduction were calculated for both types of PS II. This led to the following conclusions. The absence of energy between PS IIβ antennae is confirmed. Fluorescence quenching in PS IIα was adequately described by the matrix model, except for a decrease in the energy transfer between units during photoreduction of Q_α, possibly due to the formation of ‘islets’ of closed centers. PS II reaction centers in which Q is reduced do not significantly quench fluorescence. The ratio of variable to maximum fluorescence, 0.77 in PS IIα and 0.92 in PS IIβ, multiplied by the fraction of Q remaining in the reduced state after one saturating flash, 0.88 in PS IIα and greater than 0.95 in PS IIβ, leads to a net quantum efficiency of Q reduction in the presence of DCMU and NH₂OH of 0.68 in PS IIα and about 0.90 in PS IIβ. These values are in good agreement with the measured overall quantum efficiency of Q reduction.     

Thermodynamics of electron transfer in oxygenic photosynthetic reaction centers: volume change, enthalpy, and entropy of electron-transfer reactions in manganese-depleted photosystem II core complexes

We have previously reported the thermodynamic data of electron transfer in photosystem I using pulsed time-resolved photoacoustics [Hou et al. (2001) Biochemistry 40, 7109-7116]. In the present work, using preparations of purified manganese-depleted photosystem II (PS II) core complexes from Synechocystis sp. PCC 6803, we have measured the DeltaV, DeltaH, and estimated TDeltaS of electron transfer on the time scale of 1 micros. At pH 6.0, the volume contraction of PS II was determined to be -9 +/- 1 A3. The thermal efficiency was found to be 52 +/- 5%, which corresponds to an enthalpy change of -0.9 +/- 0.1 eV for the formation of the state P680+Q(A-) from P680*. An unexpected volume expansion on pulse saturation of PS II was observed, which is reversible in the dark. At pH 9.0, the volume contraction, the thermal efficiency, and the enthalpy change were -3.4 +/- 0.5 A3, 37 +/- 7%, and -1.15 +/- 0.13 eV, respectively. The DeltaV of PS II, smaller than that of PS I and bacterial centers, is assigned to electrostriction and analyzed using the Drude-Nernst equation. To explain the small DeltaV for the formation of P680+Q(A-) or Y(Z*)Q(A-), we propose that fast proton transfer into a polar region is involved in this reaction. Taking the free energy of charge separation of PS II as the difference between the energy of the excited-state P680* and the difference in the redox potentials of the donor and acceptor, the apparent entropy change (TDeltaS) for charge separation of PS II is calculated to be negative, -0.1 +/- 0.1 eV at pH 6.0 (P680+Q(A-)) and -0.2 +/- 0.15 eV at pH 9.0 (Y(Z*)Q(A-)). The thermodynamic properties of electron transfer in PS II core reaction centers thus differ considerably from those of bacterial and PS I reaction centers, which have DeltaV of approximately -27 A3, DeltaH of approximately -0.4 eV, and TDeltaS of approximately +0.4 eV.