Changes In Photosynthetic Electron Transport Research Articles

Proteome analysis of response to different spectral light irradiation in Synechocystis sp. PCC 6803

In cyanobacteria, it is known that the excitation ratios of photosystem (PS) I and PSII changes with the wavelength of irradiated light due to mobile phycobilisome (PBS) and spillover, affecting the photosynthetic ATP/NADPH synthesis ratio and metabolic flux state. However, the mechanisms by which these changes are controlled have not been well studied. In this study, we performed a targeted proteomic analysis of Synechocystis sp. PCC 6803 under different spectral light conditions to clarify the regulation mechanisms of mobile PBS, spillover and metabolisms under different light qualities at the protein level. The results showed an increase in the amount of proteins mainly involved in CO2 fixation under Red1 light conditions with a high specific growth rate, suggesting that the rate of intracellular metabolism is controlled by the rate of carbon uptake, not by changes in the amount of each enzyme. Correlation analysis between protein levels and PSI/PSII excitation ratios revealed that PsbQUY showed high correlations and significantly increased under Blue and Red2 light conditions, where the PSI/PSII excitation ratio was higher due to spillover. In the strains lacking the genes encoding these proteins, a decrease in the PSI/PSII excitation ratio was observed, suggesting that PsbQUY contribute to spillover occurrence. SignificanceIn cyanobacteria, the photosynthetic apparatus's responses, such as state transition [mobile PBS and spillover], occur due to the intensity and wavelength of irradiated light, resulting in changes in photosynthetic electron transport and metabolic flux states. Previous studies have analyzed the response of Synechocystis sp. PCC 6803 to light intensity from various directions, but only spectroscopic analysis of the photosynthetic apparatus has been done on the response to changes in the wavelength of irradiated light. This study analyzed the response mechanisms of mobile PBS, spillover, photosynthetic, and metabolic systems in Synechocystis sp. PCC 6803 under six different spectral light conditions by a targeted proteomic analysis. As a result, many proteins were successfully quantified, and the metabolic enzymes and photosynthetic apparatus were analyzed using an integrated approach. Principal component and correlation analyses and volcano plots revealed that the PSII subunits PsbQ, PsbU, and PsbY have a strong correlation with the PSI/PSII excitation ratio and contribute to spillover occurrence. Thus, statistical analysis based on proteome data revealed that PsbQ, PsbU, and PsbY are involved in spillover, as revealed by spectroscopic analysis.

Changes in Photosynthetic Electron Transport during Leaf Senescence in Two Barley Varieties Grown in Contrasting Growth Regimes.

Leaf senescence is an important process for plants to remobilize a variety of metabolites and nutrients to sink tissues, such as developing leaves, fruits and seeds. It has been suggested that reactive oxygen species (ROS) play an important role in the initiation of leaf senescence. Flag leaves of two different barley varieties, cv. Lomerit and cv. Carina, showed differences in the loss of photosystems and in the production of ROS at a late stage of senescence after significant loss of chlorophyll (Krieger-Liszkay etal. 2015). Here, we investigated photosynthetic electron transport and ROS production in primary leaves of these two varieties at earlier stages of senescence. Comparisons were made between plants grown outside in natural light and temperatures and plants grown in temperature-controlled growth chambers under low light intensity. Alterations in the content of photoactive P700, ferredoxin and plastocyanin (PC) photosynthetic electron transport were analyzed using in vivo near-infrared absorbance changes and chlorophyll fluorescence, while ROS were measured with spin-trapping electron paramagnetic resonance spectroscopy. Differences in ROS production between the two varieties were only observed in outdoor plants, whereas a loss of PC was common in both barley varieties regardless of growth conditions. We conclude that the loss of PC is the earliest detectable photosynthetic parameter of leaf senescence while differences in the production of individual ROS species occur later and depend on environmental factors.

Nuclear proteome analysis of Chlamydomonas with response to CO2 limitation

Chlamydomonas reinhardtii is a unicellular green alga that can survive at a wide range of inorganic carbon (Ci) concentrations by regulating the activity of a CO2-concentrating mechanism (CCM) as well as other cellular functions. Under CO2 limited conditions, C. reinhardtii cells display a wide range of adaptive responses including changes in photosynthetic electron transport, mitochondria localization in the cells, the structure of the pyrenoid starch sheath, and primary metabolism. In addition to these functional and structural changes, gene and protein expression are also affected. Several physiological aspects of the CO2 response mechanism have been studied in detail. However, the regulatory components (transcription factors and transcriptional regulators) involved in this process are not fully characterized. Here we report a comprehensive analysis of the C. reinhardtii nuclear proteome using liquid chromatography electrospray ionization spectrometry (LC-ESI-MS). The study aims to identify the proteins that govern adaptation to varying CO2 concentrations in Chlamydomonas. The nuclear proteome of C. reinhardtii cells grown in the air at high (5%) and low (0.04%) CO2 concentrations were analyzed. Using this approach, we identified 1378 proteins in total, including 90 putative transcription factors and 27 transcriptional regulators. Characterization of these new regulatory components could shed light on the molecular mechanisms underlying acclimation to CO2 stress.

Interchloroplast communications in Chara are suppressed under the alkaline bands and are relieved after the plasma membrane excitation

Immobile chloroplasts in Chara internodal cells release photometabolites into the streaming cytoplasm that distributes the exported solutes and provides metabolic connectivity between spatially remote plastids. The metabolite transmission by fluid flow is evident from chlorophyll fluorescence changes in shaded chloroplasts upon local illumination applied upstream of the analyzed area. The connectivity correlates with the pH pattern on cell surface: it is strong in cell regions with high H+-pump activity and is low in regions featuring large passive H+ influx (OH– efflux). One explanation for low connectivity under the alkaline bands is that H+ influx lowers the cytoplasmic pH, thus retarding metabolic conversions of solutes carried by the microfluidic transporter. The cessation of H+ influx across the plasma membrane by eliciting the action potential and by adding NH4Cl into the medium greatly enhanced the amplitude of cyclosis-mediated fluorescence transients. The transition from latent to the transmissive state after the dark pretreatment was paralleled by the temporary increase in chlorophyll fluorescence, reflecting changes in photosynthetic electron transport. It is proposed that the connectivity between distant chloroplasts is controlled by cytoplasmic pH.

Drought-induced changes in photosynthetic electron transport in maize probed by prompt fluorescence, delayed fluorescence, P700 and cyclic electron flow signals

The effect of drought on the prompt chlorophyll a fluorescence (PF) transient (OJIP), delayed chlorophyll a fluorescence (DF), modulated 820-nm reflection (MR), energy conversion efficiencies in photosystems (PS) I and II, and cyclic electron flow (CEF) activity in two maize hybrids with contrasting drought tolerance was investigated. Our aim was to identify the target site of drought stress on the photosynthetic electron transport chain and investigate the relevance of the CEF pathway to the drought tolerance of maize plants. The OJIP analysis showed that drought stress, depending on its duration, decreased FP, increased FJ, and induced a pronounced K-band and a positive L-band. Moreover, OJIP parameters, including PIABS, RC/CSO, TRO/ABS, and ETO/TRO, were significantly reduced. The DF analysis showed that the values of I1 and I2 in the induction curve and L1 and L2 derived from the decay curve decreased progressively with the duration of drought stress. The MR analysis showed that drought stress inactivated both the fast decrease and slow increase phases of the MR transient, resulting in a gradual decrease in both VPSI and VPSII-PSI. The energy conversion analysis showed that drought stress decreased the PSI photochemical quantum yield Y(I) and PSII photochemical quantum yield Y(II). Compared to the tolerant hybrid, the drought-induced changes in the sensitive hybrid were stronger and appeared at an earlier treatment stage. The CEF activity analysis showed that the CEF pathway under drought stress operated for a longer time in the tolerant hybrid than that in the sensitive hybrid. The above results indicate that drought stress damaged the donor and acceptor sides of PSII, the PSII reaction center and the acceptor side of PSI and decreased the efficiency of both PSI and PSII and the capacity of electron transfer. The CEF pathway might play an important role in the tolerance of the maize photosynthetic electron transport chain to drought stress.

Probing the nucleotide-binding activity of a redox sensor: two-component regulatory control in chloroplasts

Two-component signal transduction systems mediate adaptation to environmental changes in bacteria, plants, fungi, and protists. Each two-component system consists of a sensor histidine kinase and a response regulator. Chloroplast sensor kinase (CSK) is a modified sensor histidine kinase found in chloroplasts—photosynthetic organelles of plants and algae. CSK regulates the transcription of chloroplast genes in response to changes in photosynthetic electron transport. In this study, the full-length and truncated forms of Arabidopsis CSK proteins were overexpressed and purified in order to characterise their kinase and redox sensing activities. Our results show that CSK contains a modified kinase catalytic domain that binds ATP with high affinity and forms a quinone adduct that may confer redox sensing activity.

Light intensity-dependent retrograde signalling in higher plants

Plants are able to acclimate to highly fluctuating light environment and evolved a short- and long-term light acclimatory responses, that are dependent on chloroplasts retrograde signalling. In this review we summarise recent evidences suggesting that the chloroplasts act as key sensors of light intensity changes in a wide range (low, high and excess light conditions) as well as sensors of darkness. They also participate in transduction and synchronisation of systemic retrograde signalling in response to differential light exposure of distinct leaves. Regulation of intra- and inter-cellular chloroplast retrograde signalling is dependent on the developmental and functional stage of the plastids. Therefore, it is discussed in following subsections: firstly, chloroplast biogenic control of nuclear genes, for example, signals related to photosystems and pigment biogenesis during early plastid development; secondly, signals in the mature chloroplast induced by changes in photosynthetic electron transport, reactive oxygen species, hormones and metabolite biosynthesis; thirdly, chloroplast signalling during leaf senescence. Moreover, with a help of meta-analysis of multiple microarray experiments, we showed that the expression of the same set of genes is regulated specifically in particular types of signals and types of light conditions. Furthermore, we also highlight the alternative scenarios of the chloroplast retrograde signals transduction and coordination linked to the role of photo-electrochemical signalling.

Drought and Salinity Stress: Changes in Hydratase and Dehydratase Activities of Thylakoid-Associated Carbonic Anhydrase in Pea Seedlings

Changes in hydratase and dehydratase activity of thylakoid-associated carbonic anhydrase (tCA) was examined and compared with the changes in photosynthetic electron transport in pea seedlings exposed to drought and salinity stresses (for 24 h and for 48 h) in combination with additional nonstressed (recovery) time (48 h). It was established that drought and salinity stresses provoked different effects on tCA activity. The hydratase reaction increased by ∼2.5 times in plants exposed to 24 h of dehydration, while the salt treatment caused a sharp increase in hydratase reaction only after 48 h of exposure. There was a decrease in dehydratase activity in drought-stressed plants, while in salt-stressed plants it increased after 48 h of exposure. As for recovery, all CA activities returned to control levels after 48 h except hydratase activity after the salinity treatment, where 25% higher values were observed. Electron-transfer rates through the whole electron-transport chain displayed constant behavior under ...

PSI-Driven Cyclic Electron Flow Allows Intertidal Macro-Algae Ulva sp. (Chlorophyta) to Survive in Desiccated Conditions

Ulva sp. (Chlorophyta) is a representative species of the intertidal macro-algae responsible for the green tides that occurred along the shores of Qingdao in 2008 and had detrimental effects on the preparation for the 2008 Beijing Olympic Games sailing competition. In view of its significance, we have investigated the photosynthetic performance of the photosystems and the changes in photosynthetic electron transport that occur during desiccation and rehydration of Ulva sp. The PSII activity in Ulva sp. declined gradually during the course of desiccation, which was reflected by the decreased maximum quantum yield and effective quantum yield, whereas the PSI activity fluctuated significantly. In contrast, the electron transport rates of PSII approached zero at severe levels of desiccation, but the electron transport of PSI, which still operated, could be suppressed effectively by a specific inhibitor. Furthermore, the electron transport of PSI during rehydration of desiccated thalli was recovered faster than that of PSII. All these results implied that the linear electron flow was abolished in desiccated Ulva sp., whereas the cyclic PSI activity was significantly elevated, was still active at severe levels of desiccation and could be restored faster than PSII activity. Based on these results, we concluded the PSI-driven cyclic electron flow might provide desiccation tolerance and additional flexibility for the cell physiology of Ulva sp. under desiccation conditions, which might be one of the most important factors that make Ulva sp. well suited to experience daily cycles of desiccation at low tide and rehydration at high tide.

Changes in Chlorophyll Fluorescence During the Course of Photoperiod and in Response to Drought in Casuarina equisetifolia Forst. and Forst.

The effects of drought and the diurnal changes in photosynthetic electron transport were studied in non-nodulated plants of Casuarina equisetifolia. The induction of fluorescence showed a slightly higher I step in water-stressed than control plants, and the time from the start of irradiation to the P step of induction was significantly shortened by drought. The quantum efficiency of photosystem 2 (PS2) in the dark-adapted state (Fv/Fm) was generally not affected by drought, whereas it decreased during the central hours of the day. The decrease in quantum yield of PS2 electron transport (Φ2) in water-stressed plants was associated with decreases in the photochemical efficiency of open (oxidised) PS2 centres (Fv'/Fm') and increases in non-photochemical quenching (qN) rather than with increased closure of PS2 centres (lowered photochemical quenching, qP). In contrast, the changes in quantum yield of electron transport during the day were related to changes in qP rather than in Fv'/Fm'. When chlorophyll fluorescence was measured at the same irradiance during the day, a greater qN was observed at the end of the drying cycle than after watering, and early and late in the photoperiod than in the central hours of the day. The greater qN at the beginning and end of the day did not prevent an increase in energy not used photochemically nor dissipated non-photochemically. Drought did not affect this excess of photon energy.

Roles of chemosensory pathways in transient changes in swimming speed of Rhodobacter sphaeroides induced by changes in photosynthetic electron transport.

The response of free-swimming Rhodobacter sphaeroides to increases and decreases in the intensity of light of different wavelengths was analyzed. There was a transient (1 to 2 s) increase in swimming speed in response to an increase in light intensity, and there was a similar transient stop when the light intensity decreased. Measurement of changes in membrane potential and the use of electron transport inhibitors showed that the transient increase in swimming speed, following an increase in light intensity, and the stop following its decrease were the result of changes in photosynthetic electron transport. R. sphaeroides has two operons coding for multiple homologs of the enteric chemosensory genes. Mutants in the first chemosensory operon showed wild-type photoresponses. Mutants with the cheA gene of the second operon (cheAII) deleted, either with or without the first operon present, showed inverted photoresponses, with free-swimming cells stopping on an increase in light intensity and increasing swimming speed on a decrease. These mutants also lacked adaptation. Transposon mutants with mutations in cheAII, which also reduced expression of downstream genes, however, showed no photoresponses. These results show that (i) free-swimming cells respond to both an increase and a decrease in light intensity (tethered cells only show the stopping on a step down in light intensity), (ii) the signal comes from photosynthetic electron transfer, and (iii) the signal is primarily channelled through the second chemosensory pathway. The different responses shown by the cheAII deletion and insertion mutants suggest that CheWII is required for photoresponses, and a third sensory pathway can substitute for CheAII as long as CheWII is present. The inverted response suggests that transducers are involved in photoresponses as well as chemotactic responses.

Ferredoxin-1 mRNA is destabilized by changes in photosynthetic electron transport.

In transgenic tobacco, pea Ferredoxin-1 (Fed-1) mRNA accumulates rapidly in response to photosynthesis even when the transgene is driven by a constitutive promoter. To investigate the role of photosynthesis on Fed-1 mRNA stability, we used the tetracycline repressible Top10 promoter system to specifically shut off transcription of the Fed-1 transgene. The Fed-1 mRNA has a half-life of approximately 2.4 hr in the light and a half-life of only 1.2 hr in the dark or in the presence of the photosynthetic electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). These data indicate that cessation of photosynthesis, either by darkness or DCMU results in a destabilization of the Fed-1 mRNA. Furthermore, the Fed-1 mRNA half-life is reduced immediately upon transfer to darkness, suggesting that Fed-1 mRNA destabilization is a primary response to photosynthesis rather than a secondary response to long-term dark adaptation. Finally, the two different methods for efficient tetracycline delivery reported here generally should be useful for half-life measurements of other mRNAs in whole plants.

Changes in Photosynthetic Electron Transport in Potato Cultivars with Different Field Resistance after Infection with Phytophthora infestans

AbstractThe estimation of field resistance of potato cultivars to Phytophthora infestans are usually carried out in expensive and time consuming field experiments over several years. Therefore, a procedure is required for the fast and objective determination of qualitative and quantitative field resistance of new cultivars. This study correlated level of field resistance to P. infestans in leaf discs or leaflets of various potato cultivars to characteristic changes of Chlorophyll‐a fluorescence (CF) parameters Fm (maximal fluorescence) and Fv (maximal variable fluorescence). Two different inocula, both containing virulence genes 1‐11 were tested. The results were achieved when leaf discs from greenhouse or field plants were each inoculated with a P. infestans spore suspension and incubated for 24 h. In field measurements, comparable results were obtained 48 h after inoculation. The estimation of field resistance by measuring specific CF parameters could be an economical and rapid procedure to reduce or substitute visual lesion assessment for determining cultivar field resistance.

Curve-fit analysis of chlorophyll fluorescence spectra: Application to nutrient stress detection in sunflower

The in vivo laser-induced chlorophyll fluorescence (LICF) spectra of healthy and nutrient-deficient sunflower plants were measured on a Jobin Yvon monochromator with HeNe laser excitation. To correctly determine the peak center and to evaluate the relative contributions of the bands in the total fluorescence spectrum, the steady state LICF spectra were analyzed with a nonlinear iterative procedure using Gaussian, Lorentzian, Pearson, Voigt, and exponential Gaussian spectral functions. It was observed that curve fitting performed by using two Gaussian peaks centered at 690 and 730 nm usually fits well to the chlorophyll fluorescence spectra. After curve fitting, the mean peak centers of the red and far-red chlorophyll bands of control plants were observed at 688.2 and 725.4 nm, respectively. A blue shift of as much as 9 nm in the peak position of the far-red band was observed with nutrient stress, whereas the shift in position of the red band was only of the order of a few nanometers. Further, the width at half maximum of the far-red band was found to increase by as much as 20 nm with nutrient stress. Curve fitting could thus separate out the red and far-red fluorescence spectra from a pair of normally distributed curves centered at 690 and 730 nm wavelengths, thereby differentiating the effects due to reabsorption from those due directly to changes in photosynthetic electron transport. The F690F730 fluorescence intensity ratio obtained from curve-fitted parameters was found to be more sensitive to plant stress than were fluorescence values alone. Results indicate that a curve-fitting analysis of LICF spectra using Gaussian spectral functions is a very useful and sensitive method of evaluating spectral features from a statistical point of view and for accurate determination of contributions from constituent bands in the whole leaf fluorescence spectrum.

Senescence-related changes in photosynthetic electron transport are not due to alterations in thylakoid fluidity

During senescence of primary bean leaves, non-cyclic electron transport declines to a greater degree than the activities of Photosystem I and Photosystem II, and intersystem electron transport becomes rate-limiting. Paramagnetic and fluorescent lipid-soluble probes have been used to measure rotational motion and lateral motion in thylakoid membranes to determine if this decrease in intersystem electron transport reflects changes in lipid fluidity that could impede the translational motion of electron carriers between the two photosystems. Probes that partition deep in the bilayer as well as those that remain anchored at the membrane surface revealed no change in rotational motion with advancing age. Translational motion was measured by determining the excimer-to-monomer ratio for pyrene partitioned into the bilayer, and this ratio also showed no change with advancing senescence, nor were there changes in phase properties or fatty-acid composition of the membranes. During in vitro aging of chloroplasts, there was also a dramatic decrease in non-cyclic electron transport that was not accompanied by changes in membrane fluidity. Thus the decline in intersystem electron transport during senescence cannot be attributed to decreased translational diffusion of mobile electron carriers. Moreover, the mechanism of thylakoid membrane deterioration during senescence appears to differ from that operative in plasma and microsomal membranes for which large decreases in lipid fluidity with advancing age are well documented.

Aging induced changes in photosynthetic electron transport of detached barley leaves

Aging induced changes in photosynthetic electron transport of detached barley leaves