PQ Pool Research Articles

The function of PROTOPORPHYRINOGEN IX OXIDASE in chlorophyll biosynthesis requires oxidised plastoquinone in Chlamydomonas reinhardtii

In the last common enzymatic step of tetrapyrrole biosynthesis, prior to the branching point leading to the biosynthesis of heme and chlorophyll, protoporphyrinogen IX (Protogen) is oxidised to protoporphyrin IX (Proto) by protoporphyrinogen IX oxidase (PPX). The absence of thylakoid-localised plastid terminal oxidase 2 (PTOX2) and cytochrome b6f complex in the ptox2 petB mutant, results in almost complete reduction of the plastoquinone pool (PQ pool) in light. Here we show that the lack of oxidised PQ impairs PPX function, leading to accumulation and subsequently uncontrolled oxidation of Protogen to non-metabolised Proto. Addition of 3(3,4-Dichlorophenyl)-1,1-dimethylurea (DCMU) prevents the over-reduction of the PQ pool in ptox2 petB and decreases Proto accumulation. This observation strongly indicates the need of oxidised PQ as the electron acceptor for the PPX reaction in Chlamydomonas reinhardtii. The PPX-PQ pool interaction is proposed to function as a feedback loop between photosynthetic electron transport and chlorophyll biosynthesis.

Salinity tolerance of quinoa (Chenopodium quinoa Willd) as assessed by chloroplast ultrastructure and photosynthetic performance

Quinoa has been widely used as a model crop for understanding salt-tolerance in halophyte plants. However, a salt tolerance mechanism regarding the photosynthetic activity is poorly evaluated. Here we report the effects of salt stress on photosynthesis of the halophyte Chenopodium quinoa cultivated under different NaCl concentrations (0, 100, and 300 mM). Our results revealed no apparent effect of moderate salinity (100 mM NaCl) on plant growth while, a reduction of plant biomass production was detected under 300 mM NaCl without any symptom of toxicity. No significant effect on the chloroplasts ultrastructure was observed under moderate salinity. Some morphological deformations of chloroplasts such as swelling of thylakoids, disruption of envelope, accumulation of starch grains and plastoglobuli were observed following prolonged exposure to severe stress. Under moderate salinity, no considerable change was detected on the rate of primary photochemistry (fluorescence O-J phase) and the reduction of the PQ pool (J-I phase), no apparent effects on the minimal (F0), the maximal fluorescence (Fm) and the maximal photochemical efficiency (Fv/Fm) showing a high stability of PSII. The high PSII efficiency was also confirmed by the donor side intactness (constant Fk/Fj ratio) and stable antenna size. The high stability of PSII efficiency was also demonstrated by enhanced communication between antenna complex and PSII reaction centre and the stability of OEC complex (PsbQ and PsbO proteins). However, high salinity (300 mM NaCl) results in a swelling of thylakoids and disappearance of grana and in a decrease of Fm and (Fv/Fm) leading to the down-regulation of PSII activity. In addition, a significant increase in F0 occurred and could be associated to the presence of some QA− in darkness in equilibrium with a partially reduced PQ pool. High salt treatment could be responsible for the thermal phase which induced an increase in chl a fluorescence (JIP rise with oxidized PQ pool).

Control of the maximal chlorophyll fluorescence yield by the Q<sub>B</sub> binding site

Differences in maximal yields of chlorophyll variable fluorescence (Fm) induced by single turnover (ST) and multiple turnover (MT) excitation are as great as 40%. Using mutants of Chlamydomonas reinhardtii we investigated potential mechanisms controlling Fm above and beyond the QA redox level. Fm was low when the QB binding site was occupied by PQ and high when the QB binding site was empty or occupied by a PSII herbicide. Furthermore, in mutants with impaired rates of plastoquinol reoxidation, Fm was reached rapidly during MT excitation. In PSII particles with no mobile PQ pool, Fm was virtually identical to that obtained in the presence of PSII herbicides. We have developed a model to account for the variations in maximal fluorescence yields based on the occupancy of the QB binding site. The model predicts that the variations in maximal fluorescence yields are caused by the capacity of secondary electron acceptors to reoxidize QA-.

Bioenergetic reprogramming plasticity under nitrogen depletion by the unicellular green alga Scenedesmus obliquus.

Simultaneous nitrogen depletion and 3,4-dichlorophenol addition induce a bioenergetic microalgal reprogramming, through strong Cyt b 6 f synthesis, that quench excess electrons from dichlorophenol's biodegradation to an overactivated photosynthetic electron flow and H 2 -productivity. Cellular energy management includes "rational" planning and operation of energy production and energy consumption units. Microalgae seem to have the ability to calculate their energy reserves and select the most profitable bioenergetic pathways. Under oxygenic mixotrophic conditions, microalgae invest the exogenously supplied carbon source (glucose) to biomass increase. If 3,4-dichlorophenol is added in the culture medium, then glucose is invested more to biodegradation rather than to growth. The biodegradation yield is enhanced in nitrogen-depleted conditions, because of an increase in the starch accumulation and a delay in the establishment of oxygen-depleted conditions in a closed system. In nitrogen-depleted conditions, starch cannot be invested in PSII-dependent and PSII-independent pathways for H2-production, mainly because of a strong decrease of the cytochrome b 6 f complex of the photosynthetic electron flow. For this reason, it seems more profitable for the microalga under these conditions to direct the metabolism to the synthesis of lipids as cellular energy reserves. Nitrogen-depleted conditions with exogenously supplied 3,4-dichlorophenol induce reprogramming of the microalgal bioenergetic strategy. Cytochrome b 6 f is strongly synthesized (mainly through catabolism of polyamines) to manage the electron bypass from the dichlorophenol biodegradation procedure to the photosynthetic electron flow (at the level of PQ pool) and consequently through cytochrome b 6 f and PSI to hydrogenase and H2-production. All the above showed that the selection of the appropriate cultivation conditions is the key for the manipulation of microalgal bioenergetic strategy that leads to different metabolic products and paves the way for a future microalgal "smart" biotechnology.

Differential sensitivity of light-harnessing photosynthetic events in wheat and sunflower to exogenously applied ionic and nanoparticulate silver

Potential impacts of inevitable leaks of silver nanoparticles (AgNPs) into environment on human beings need attention. Owing to the vitality of photosynthesis in maintaining life and ecosystem functioning, impacts of exogenously applied nanoparticulate and Ag+ on photosystem (PS)II function, which governs overall photosynthesis, in wheat and sunflower were evaluated. PSII efficiency and related Chl a fluorescence kinetics of these two plants remained unaffected by AgNPs. However, Ag+ caused a significant decline in the PSII activity and related fluorescence steps in wheat, but not in sunflower. Electron flow between QA and PQ pool was found most sensitive to Ag+. Number of active reaction centers, electron transport, trapping of absorbed light for photochemistry, and performance index declined, while dissipation of absorbed light energy as heat significantly increased in wheat exposed to Ag+. Total antioxidant activity in sunflower was least affected by both Ag and AgNPs. In contrast, in the case of wheat, the antioxidant activity was declined by Ag+ but not by AgNPs. Further, the amount of silver absorbed by plants exposed to Ag+ was higher than that absorbed by plants exposed to AgNPs. While wheat retained majority of Ag in its roots, sunflower showed major Ag accumulation in stem. Photosynthetic events in sunflower, unlike wheat, were least affected as no detectable Ag levels was recorded in their leaves. Our findings revealed that AgNPs seemed non/less-toxic to light harnessing photosynthetic machinery of wheat, compared to Ag+. Photosynthetic events in sunflower were not affected by Ag+, either, as its translocation to leaves was restricted.

Salt stress effects on the photosynthetic electron transport chain in two chickpea lines differing in their salt stress tolerance.

The main objective of this study was to evaluate the effects of salt stress on the photosynthetic electron transport chain using two chickpea lines (Cicer arietinum L.) differing in their salt stress tolerance at the germination stage (AKN 87 and AKN 290). Two weeks after sowing, seedlings were exposed to salt stress for 2weeks and irrigated with 200ml of 200mM NaCl every 2days. The polyphasic OJIP fluorescence transient and the 820-nm transmission kinetics (photosystem I) were used to evaluate the effects of salt stress on the functionality of the photosynthetic electron transport chain. It was observed that a signature for salt stress was a combination of a higher J step (VJ), a smaller IP amplitude, and little or no effect on the primary quantum yield of PSII (φPo). We observed for AKN 290 a shorter leaf life cycle, which may represent a mechanism to cope with salt stress. For severely salt-stressed leaves, an inhibition of electron flow between the PQ pool and P700 was found. The data also suggest that the properties of electron flow beyond PSI are affected by salt stress.

Photosystem II-cyclic electron flow powers exceptional photoprotection and record growth in the microalga Chlorella ohadii

The desert microalga Chlorella ohadii was reported to grow at extreme light intensities with minimal photoinhibition, tolerate frequent de/re-hydrations, yet minimally employs antenna-based non-photochemical quenching for photoprotection. Here we investigate the molecular mechanisms by measuring Photosystem II charge separation yield (chlorophyll variable fluorescence, Fv/Fm) and flash-induced O2 yield to measure the contributions from both linear (PSII-LEF) and cyclic (PSII-CEF) electron flow within PSII. Cells grow increasingly faster at higher light intensities (μE/m2/s) from low (20) to high (200) to extreme (2000) by escalating photoprotection via shifting from PSII-LEF to PSII-CEF. This shifts PSII charge separation from plastoquinone reduction (PSII-LEF) to plastoquinol oxidation (PSII-CEF), here postulated to enable proton gradient and ATP generation that powers photoprotection. Low light-grown cells have unusually small antennae (332 Chl/PSII), use mainly PSII-LEF (95%) and convert 40% of PSII charge separations into O2 (a high O2 quantum yield of 0.06mol/mol PSII/flash). High light-grown cells have smaller antenna and lower PSII-LEF (63%). Extreme light-grown cells have only 42 Chl/PSII (no LHCII antenna), minimal PSII-LEF (10%), and grow faster than any known phototroph (doubling time 1.3h). Adding a synthetic quinone in excess to supplement the PQ pool fully uncouples PSII-CEF from its natural regulation and produces maximum PSII-LEF. Upon dark adaptation PSII-LEF rapidly reverts to PSII-CEF, a transient protection mechanism to conserve water and minimize the cost of antenna biosynthesis. The capacity of the electron acceptor pool (plastoquinone pool), and the characteristic times for exchange of (PQH2)B with PQpool and reoxidation of (PQH2)pool were determined.

Relative functional and optical absorption cross-sections of PSII and other photosynthetic parameters monitored in situ, at a distance with a time resolution of a few seconds, using a prototype light induced fluorescence transient (LIFT) device.

The prototype light-induced fluorescence transient (LIFT) instrument provides continuous, minimally intrusive, high time resolution (~2s) assessment of photosynthetic performance in terrestrial plants from up to 2m. It induces a chlorophyll fluorescence transient by a series of short flashes in a saturation sequence (180 ~1μs flashlets in <380μs) to achieve near-full reduction of the primary acceptor QA, followed by a relaxation sequence (RQA; 90 flashlets at exponentially increasing intervals over ~30ms) to observe kinetics of QA re-oxidation. When fitted by the fast repetition rate (FRR) model (Kolber et al. 1998) the QA flash of LIFT/FRR gives smaller values for FmQA from dark adapted leaves than FmPAM from pulse amplitude modulated (PAM) assays. The ratio FmQA/FmPAM resembles the ratio of fluorescence yield at the J/P phases of the classical O-J-I-P transient and we conclude that the difference simply is due to the levels of PQ pool reduction induced by the two techniques. In a strong PAM-analogous WL pulse in the dark monitored by the QA flash of LIFT/FRR φPSIIWL ≈ φPSIIPAM. The QA flash also tracks PQ pool reduction as well as the associated responses of ETR QA → PQ and PQ → PSI, the relative functional (σPSII) and optical absorption (aPSII) cross-sections of PSII in situ with a time resolution of ~2s as they relax after the pulse. It is impractical to deliver strong WL pulses at a distance in the field but a longer PQ flash from LIFT/FRR also achieves full reduction of PQ pool and delivers φPSIIPQ ≈ φPSIIPAM to obtain PAM-equivalent estimates of ETR and NPQ at a distance. In situ values of σPSII and aPSII from the QA flash with smaller antenna barley (chlorina-f2) and Arabidopsis mutants (asLhcb2-12, ch1-3 Lhcb5) are proportionally similar to those previously reported from in vitro assays. These direct measurements are further validated by changes in antenna size in response to growth irradiance. We illustrate how the QA flash facilitates our understanding of photosynthetic regulation during sun flecks in natural environments at a distance, with a time resolution of a few seconds.

Impacts of genetically engineered alterations in carbon sink pathways on photosynthetic performance

Genetic engineering of photosynthetic organisms typically redirects native metabolism towards desirable products, which thereby represent new metabolic sinks. There is limited information on how these modifications impact the evolved mechanisms of photosynthetic energy metabolism and cellular growth. Two engineered strains of Synechocystis sp. PCC 6803 with altered carbon sink capacity were assayed for their photosynthetic and CO2 concentrating mechanism properties in conditions of high and low inorganic carbon (Ci) availability. In the ΔglgC mutant, glycogen cannot be synthesized and a carbon sink pathway has been effectively removed. The JU547 strain has been engineered by integration of the Pseudomonas syringae ethylene forming enzyme and provides a new sink. When cultured under high carbon conditions, ΔglgC displayed diminished photochemical efficiency, a more reduced NADPH pool, delayed initiation of the Calvin-Benson-Bassham cycle, and impairment of linear and cyclic electron flows. It also exhibited a large decrease in photochemical quenching indicative of the accumulation of QA−, normally associated with a reduced PQ pool, but appears instead to be the result of an undefined dissipative mechanism to spill excess energy. In the case of carbon sink integration, JU547 displayed slightly more oxidized PQ and NADPH pools and increased rates of cyclic electron flow and an enhanced demand for inorganic carbon as suggested by increase in the expression of the bicarbonate transporter, SbtA. Overall, the results highlight the importance of the native regulatory network of autotrophic metabolism in governing photosynthetic performance and provide cogent examples of both predicable and difficult to predict phenotypic consequences upon installation of new pathways in autotrophs.

Assessment of photosynthesis regulation in mixotrophically cultured microalga Chlorella sorokiniana

Mixotrophic growth of microalgae offers great potential as an efficient strategy for biofuel production. In this study, photosynthetic regulation of mixotrophically cultured Chlorella sorokiniana cells was systematically evaluated. Mixotrophic cells in the exponential growth phase showed the highest photosynthetic activity, where maximum photosynthetic O2 evolution was approximately 3- and 4-fold higher than cells in the same phase grown photoautotrophically in 1% CO2 (in air) and air, respectively. Additionally, characteristic chlorophyll fluorescence parameters demonstrated that no limitation in electron transport downstream of PSII was detected in mixotrophic cells. Up-regulation of photosynthetic activity was associated with high total ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylase activity and expression level of phosphoribulokinase (PRK). After 3days, photosynthetic O2 evolution of mixotrophic cells that went to the stationary phase, was strongly reduced, with reduced photochemical efficiency and reorganization of the PSII complex. Simultaneously, enzymatic activity for Rubisco carboxylase and mRNA levels of Rubisco and PRK diminished. Importantly, there was almost no non-photochemical quenching for mixotrophic cells, whether grown in log or stationary phase. A decline in the quantum efficiency of PSII and an oxidized plastoquinone pool (PQ pool) was observed under N-depleted conditions during mixotrophic growth. These results demonstrate that photosynthesis is regulated differently in mixotrophically cultured C. sorokiniana cells than in cells grown under photoautotrophic conditions, with a particularly strong impact by nitrogen levels in the cells.

Changes in the Photosystem II complex associated with hydrogen formation in sulfur deprived Chlamydomonas reinhardtii

Redox properties of the acceptor side of Photosystem II were studied during H2 gas production in cells of Chlamydomonas reinhardtii. Flash-induced variable fluorescence changes and thermoluminescence measurements were performed in wild type and Stm6 mutant cells during different stages of sulfur (S)-deprivation. Analysis of the fluorescence decay kinetics indicated that the forward electron transfer on the acceptor side of Photosystem II was dramatically slowed down during the O2 evolution and O2 consumption stages and was completely blocked in the anaerobic stage of S-deprivation, thus, indicating a complete reduction of the PQ-pool. During the H2 formation stage, the forward electron transfer kinetics in the μsec and msec time scale re-appeared indicating partially restored electron flow from QA− to QB and the PQ-pool. Thermoluminescese measurements fully confirmed the fluorescence kinetic analysis. Activation of hydrogenase in the H2 formation stage is responsible for re-oxidation of the PQ pool and reactivation of the electron flow which was found to be faster and more efficient on the Stm6 mutant due to the higher amount of functionally preserved Photosystem II.

Thylakoid membrane model of the Chl a fluorescence transient and P700 induction kinetics in plant leaves.

A new Thylakoid model is presented, which describes in detail the electron/proton transfer reactions between membrane protein complexes including photosystems II and I (PSII, PSI), cytochrome (Cyt) b 6 f, mobile plastoquinone PQ pool in the thylakoid membrane, plastocyanin in lumen and ferredoxin in stroma, reduction of NADP via FNR and cyclic electron transfer. The Thylakoid model parameters were fitted both to Chl fluorescence induction data (FI) and oxido-reductions of P700 (ΔA 810) measured from 20μs up to 20s in pea leaves. The two-wave kinetics of FI and ΔA 810 (O(JI)PSM and OABCDE) were described quantitatively, provided that the values of membrane electrochemical potential components ΔΨ(t), pHL(t)/pHS(t) are in physiologically relevant ranges. The time courses on the time scale from nanoseconds to tens of seconds of oxido-reduction changes of ET components as well as concentrations of proton/ions (K+, Cl-) were calculated. We assume a low constant FNR activity over this period. Charge movements across the thylakoid membrane by passive leakage and active ATPase transport and proton buffer reactions are simulated. The dynamics of charge fluxes during photosynthetic induction under low light (PFD 200μmol photons m-2s-1) were analyzed. The initial wave of P700 oxidation within 20ms during independent operation of PSI and PSII was followed after 50ms by PSI donor-side reduction from reduced PQ pool via Cyt b 6 f site. The Cyt b 6 f reactions contribute to the stabilization of fluxes in the time range 1s<t<10s. The detailed analysis of Chl a fluorescence at the PSM stage (t>10s) would need the investigation of FNR activation effect in order to explain the transitions between cyclic and linear electron transport.

The chlorophyll a fluorescence induction curve in the green microalga Haematococcus pluvialis: further insight into the nature of the P-S-M fluctuation and its relationship with the "low-wave" phenomenon at steady-state.

Chlorophyll fluorescence is an information-rich signal which provides an access to the management of light absorbed by PSII. A good example of this is the succession of fast fluorescence fluctuations during light-induced photosynthetic induction after dark-adaptation. During this period, the fluorescence trace exhibits several inflexion points: O-J-I-P-S-M-T. Whereas the OJIP part of this kinetics has been the subject of many studies, the processes that underly the PSMT transient are less understood. Here, we report an analysis of the PSMT phase in the green microalga Haematococcus pluvialis in terms of electron acceptors and light use by photochemistry, fluorescence and non-photochemical quenching (NPQ). We identify additional sub-phases between P and S delimited by an inflexion point, that we name Q, found in the second time scale. The P-Q phase expresses a transient photochemical quenching specifically due to alternative electron transport to oxygen. During the transition from Q to S, the NPQ increases and then relaxes during the S-M phase in about 1min. It is suggested that this transient NPQ observed during induction is a high energy state quenching (qE) dependent on the alternative electron transport to molecular oxygen. We further show that this NPQ is of the same nature than the NPQ, known as the low-wave phenomenon, which is transiently observed after a saturating light pulse given at steady-state. In both cases, the NPQ is oxygen-dependent. This NPQ is observed at external pH 6.0, but not at pH 7.5, which seems correlated with faster saturation of the PQ pool at pH 6.0.

The Dual Role of the Plastid Terminal Oxidase PTOX: Between a Protective and a Pro-oxidant Function.

The Dual Role of the Plastid Terminal Oxidase PTOX: Between a Protective and a Pro-oxidant Function.

Effect of Chlamydomonas plastid terminal oxidase 1 expressed in tobacco on photosynthetic electron transfer.

The plastid terminal oxidase PTOX is a plastohydroquinone:oxygen oxidoreductase that is important for carotenoid biosynthesis and plastid development. Its role in photosynthesis is controversially discussed. Under a number of abiotic stress conditions, the protein level of PTOX increases. PTOX is thought to act as a safety valve under high light protecting the photosynthetic apparatus against photodamage. However, transformants with high PTOX level were reported to suffer from photoinhibition. To analyze the effect of PTOX on the photosynthetic electron transport, tobacco expressing PTOX-1 from Chlamydomonas reinhardtii (Cr-PTOX1) was studied by chlorophyll fluorescence, thermoluminescence, P700 absorption kinetics and CO2 assimilation. Cr-PTOX1 was shown to compete very efficiently with the photosynthetic electron transport for PQH2 . High pressure liquid chromatography (HPLC) analysis confirmed that the PQ pool was highly oxidized in the transformant. Immunoblots showed that, in the wild-type, PTOX was associated with the thylakoid membrane only at a relatively alkaline pH value while it was detached from the membrane at neutral pH. We present a model proposing that PTOX associates with the membrane and oxidizes PQH2 only when the oxidation of PQH2 by the cytochrome b6 f complex is limiting forward electron transport due to a high proton gradient across the thylakoid membrane.

Antimycin A sensitive pathway independent from PGR5 cyclic electron transfer triggers non-photochemical reduction of PQ pool and state transitions in Arabidopsis thaliana

We investigated the mechanism involved in triggering state transitions at 40°C in Arabidopsis thaliana. Leaves (1–6week old) exposed to 40°C exhibited state II transition indicating its role as one of the earliest stress responsive mechanism apart from regulation of light energy distribution between photosystem (PS)II and PSI. Post illumination transients (rise in Fo′) revealed that non-photochemical reduction of PQ pool at 40°C in dark is responsible for activation of STN7 kinase, consequently light harvesting complex (LHC)II phosphorylation leading to state II condition. Later, in pgr5 mutant, non-photochemical reduction of PQ pool was observed indicating the involvement of alternative electron transfer routes. In chlororespiratory mutant crr2-2, state II transition occurred signifying that the reduction of PQ pool is independent from NDH mediated cyclic electron transfer. Further, antimycin A inhibitor studies in wt and mutants revealed its inhibitory action on non-photochemical reduction of PQ pool affecting both LHCII phosphorylation and migration to PSI which leads to state I. Thus, our study showed that antimycin A sensitive pathway independent from PGR5 dependent cyclic electron transfer, is responsible for inducing non-photochemical reduction of PQ pool and state transitions at 40°C.

Dissection of respiration and photosynthesis in the cyanobacterium Synechocystis sp. PCC6803 by the analysis of chlorophyll fluorescence

In cyanobacteria, photosynthesis and respiration share some components of electron transport chain. To explore the interaction between photosynthesis and respiration, we monitored the change in the yield of chlorophyll fluorescence due to state transition in ndh genes disruptants, deficient in NAD(P)H dehydrogenase (NDH-1) complexes serving for respiration or for carbon concentrating mechanism (CCM). The disruption of ndh genes essential for respiration resulted in low levels of chlorophyll fluorescence quenching in the dark (NPQDark) as well as in the low light (NPQLL). The lowered NPQDark and NPQLL in these ndh genes disruptants could be ascribed to the oxidation of the PQ pool due to the poor electron supply from NDH-1 complexes in respiratory electron transport. On the other hand, only NPQLL decreased upon disruption of the ndh genes essential for CCM. We propose that, in the disruptants of these ndh genes, the PQ pool is oxidized in the light through the increased photosystem I content, resulting in the lowered NPQLL. Apparently, the two different subsets of ndh genes affect photosynthetic electron transport although in totally different manners. It is also suggested that monitoring state transition is a simple method to evaluate the condition of photosynthesis, respiration and CCM.

Photosystem activity and state transitions of the photosynthetic apparatus in cyanobacterium Synechocystis PCC 6803 mutants with different redox state of the plastoquinone pool.

To better understand how photosystem (PS) activity is regulated during state transitions in cyanobacteria, we studied photosynthetic parameters of photosystem II (PSII) and photosystem I (PSI) in Synechocystis PCC 6803 wild type (WT) and its mutants deficient in oxidases (Ox(-)) or succinate dehydrogenase (SDH(-)). Dark-adapted Ox(-) mutant, lacking the oxidation agents, is expected to have a reduced PQ pool, while in SDH(-) mutant the PQ pool after dark adaptation will be more oxidized due to partial inhibition of the respiratory chain electron carriers. In this work, we tested the hypothesis that control of balance between linear and cyclic electron transport by the redox state of the PQ pool will affect PSII photosynthetic activity during state transition. We found that the PQ pool was reduced in Ox(-) mutant, but oxidized in SDH(-) mutant after prolonged dark adaptation, indicating different states of the photosynthetic apparatus in these mutants. Analysis of variable fluorescence and 77K fluorescence spectra revealed that the WT and SDH(-) mutant were in State 1 after dark adaptation, while the Ox(-) mutant was in State 2. State 2 was characterized by ~1.5 time lower photochemical activity of PSII, as well as high rate of P700 reduction and the low level of P700 oxidation, indicating high activity of cyclic electron transfer around PSI. Illumination with continuous light 1 (440nm) along with flashes of light 2 (620nm) allowed oxidation of the PQ pool in the Ox(-) mutant, thus promoting it to State 1, but it did not affect PSII activity in dark adapted WT and SDH(-) mutant. State 1 in the Ox(-) mutant was characterized by high variable fluorescence and P700(+) levels typical for WT and the SDH(-) mutant, indicating acceleration of linear electron transport. Thus, we show that PSII of cyanobacteria has a higher photosynthetic activity in State 1, while it is partially inactivated in State 2. This process is controlled by the redox state of PQ in cyanobacteria through enhancement/inhibition of electron transport on the acceptor side of PSII.

The role of the high potential form of the cytochrome b559: Study of Thermosynechococcus elongatus mutants

Cytochrome b559 is an essential component of the photosystem II reaction center in photosynthetic oxygen-evolving organisms, but its function still remains unclear. The use of photosystem II preparations from Thermosynechococcus elongatus of high integrity and activity allowed us to measure for the first time the influence of cytochrome b559 mutations on its midpoint redox potential and on the reduction of the cytochrome b559 by the plastoquinone pool (or QB). In this work, five mutants having a mutation in the α-subunit (I14A, I14S, R18S, I27A and I27T) and one in the β-subunit (F32Y) of cytochrome b559 have been investigated. All the mutations led to a destabilization of the high potential form of the cytochrome b559. The midpoint redox potential of the high potential form was significantly altered in the αR18S and αI27T mutant strains. The αR18S strain also showed a high sensitivity to photoinhibitory illumination and an altered oxidase activity. This was suggested by measurements of light induced oxidation and dark re-reduction of the cytochrome b559 showing that under conditions of a non-functional water oxidation system, once the cytochrome is oxidized by P680+, the yield of its reduction by QB or the PQ pool was smaller and the kinetic slower in the αR18S mutant than in the wild-type strain. Thus, the extremely positive redox potential of the high potential form of cytochrome b559 could be necessary to ensure efficient oxidation of the PQ pool and to function as an electron reservoir replacing the water oxidation system when it is not operating.

Derivatives of diterpen labdane-8α,15-diol as photosynthetic inhibitors in spinach chloroplasts and growth plant inhibitors

In a search of new efficient herbicides of natural origin, four derivatives were prepared from labdane-8α,15-diol (1) and 15-O-acetyl-8α-hydroxy labdane (2) isolated from Croton ciliatoglanduliferus. Their inhibitory activity on photosynthetic electron transport on fresh, broken spinach chloroplasts and on the growth of plants were determined. Derivative 15-O-benzoyl-8α-hydroxy labdane (5) was seven times more active than 2 as reaction Hill inhibitor. Complex of 5 with the adjuvant 2-hydroxypropyl-β-cyclodextrin (5:HPB) (200μM) was sprayed on Physalys ixocarpa (green tomato) plants; 48h later the complex inhibited PS II by transforming the active reaction centers to silent reaction centers or “heat sinks”. After 72h this effect disappeared, probably 5:HPB was metabolized by the plant. Chlorophyll a fluorescence of Trifolium alexandrinum (clover) leaves was affected with 5:HPB at the level of PQ pool reduction. 5:HPB decreases the tomato and clover dry-biomass, without affecting Lolium perenne (grass) plants, suggesting that complex 5 acts as selective herbicide for dicotyledonous plants.