Quinone Electron Acceptor Research Articles

Phe265 of the D1 protein is required to stabilize plastoquinone binding in the QB-binding site of photosystem II in Synechocystis sp. PCC 6803

In Photosystem II electrons from water splitting pass through a primary quinone electron acceptor (QA) to the secondary plastoquinone (QB). The D2 protein forms the QA-binding site and the D1 protein forms the QB-binding site. A non-heme iron sits between QA and QB resulting in a quinone-Fe-acceptor complex that must be activated before assembly of the oxygen-evolving complex can occur. An extended loop (residues 223–266) between the fourth (helix D) and fifth (helix E) helices of the D1 protein activates forward electron transfer via a conformational change that stabilizes a bidentate bicarbonate ligand to the non-heme iron while simultaneously stabilizing the binding of QB. We show that positioning of D1:Phe265 to provide a hydrogen bond to the distal oxygen of QB is required for forward electron transfer. In addition, mutations targeting D1:Phe265, resulted in a 50 mV decrease in the QB/QB– midpoint potential.

Rice (Oryza sativa L.) exhibits differential responses to salt stress and salt shock in electron transfer efficiency

The study of crop response to soil salinity has traditionally focused on the effects of varying salt concentrations, with less attention given to the methodologies of salt application. This research aimed to elucidate the differential impacts of two distinct salinity exposure methods on the photosynthetic characteristics of rice. Plants subjected to a gradual increase in salinity experience salt stress, whereas those immediately exposed to high salinity levels encounter a salt shock condition. Our findings revealed that rice plants accumulated comparable levels of shoot Na+ under both salt stress and salt shock at 4 and 10 days after the onset of stress. No significant differences were observed in the steady-state photosynthetic rate and electron transport rate (ETR). In contrast, salt stress markedly reduced the relative variable fluorescence at the ‘J’ step (Vj), thereby enhancing the probability of an exciton transferring an electron beyond the primary quinone electron acceptor (QA−) (ΨEo). This led to a notable increase in the activity of leaf photosystem II (PSII), as evidenced by the Performance Index on absorption basis (PIabs), which was twice as high as that of the control group. This mechanism is believed to facilitate the activation of alternative metabolic pathways. Additionally, kinetic measurements of light potential indicated that salt stress improved the initial induction of ETR after a brief exposure to high photosynthetically active radiation (PAR) compared to the control. This effect was not observed in the salt shock treatment group. Our results suggest that plants exhibit distinct physiological responses to salt stress versus salt shock, and the potential positive influence on leaf photosynthetic activity during salt stress may have been previously underestimated.

Transcriptome analysis of the bloom-forming dinoflagellate Prorocentrum donghaiense exposed to Ginkgo biloba leaf extract, with an emphasis on photosynthesis.

Ginkgo biloba leaf extract (GBE) can effectively treat bloom-forming freshwater algae. However, there is limited information about the underlying suppression mechanism of the marine bloom-forming Prorocentrum donghaiense-the most dominant algal bloom species in the East China Sea. We investigated the effect of GBE on P. donghaiense in terms of its response to photosynthesis at the molecular/omic level. In total, 93,743 unigenes were annotated using six functional databases. Furthermore, 67,203 differentially expressed genes (DEGs) were identified in algae treated with 1.8 g∙L-1 GBE. Among these DEGs, we identified the genes involved in photosynthesis. PsbA, PsbB and PsbD in photosystem II, PsaA in photosystem I, and PetB and PetD in the cytochrome b6/f complex were downregulated. Other related genes, such as PsaC, PsaE, and PsaF in photosystem I; PetA in the cytochrome b6/f complex; and atpA, atpD, atpH, atpG, and atpE in the F-type H+-ATPase were upregulated. These results suggest that the structure and activity of the complexes were destroyed by GBE, thereby inhibiting the electron flow between the primary and secondary quinone electron acceptors, primary quinone electron acceptor, and oxygen-evolving complex in the PSII complex, and interrupting the electron flow between PSII and PSI, ultimately leading to a decline in algal cell photosynthesis. These findings provide a basis for understanding the molecular mechanisms underlying P. donghaiense exposure to GBE and a theoretical basis for the prevention and control of harmful algal blooms.

Proline Dehydrogenase and Pyrroline 5 Carboxylate Dehydrogenase from Mycobacterium tuberculosis: Evidence for Substrate Channeling.

In Mycobacterium tuberculosis, proline dehydrogenase (PruB) and ∆1-pyrroline-5-carboxylate (P5C) dehydrogenase (PruA) are monofunctional enzymes that catalyze proline oxidation to glutamate via the intermediates P5C and L-glutamate-γ-semialdehyde. Both enzymes are essential for the replication of pathogenic M. tuberculosis. Highly active enzymes were expressed and purified using a Mycobacterium smegmatis expression system. The purified enzymes were characterized using natural substrates and chemically synthesized analogs. The structural requirements of the quinone electron acceptor were examined. PruB displayed activity with all tested lipoquinone analogs (naphthoquinone or benzoquinone). In PruB assays utilizing analogs of the native naphthoquinone [MK-9 (II-H2)] specificity constants Kcat/Km were an order of magnitude greater for the menaquinone analogs than the benzoquinone analogs. In addition, mycobacterial PruA was enzymatically characterized for the first time using exogenous chemically synthesized P5C. A Km value of 120 ± 0.015 µM was determined for P5C, while the Km value for NAD+ was shown to be 33 ± 4.3 µM. Furthermore, proline competitively inhibited PruA activity and coupled enzyme assays, suggesting that the recombinant purified monofunctional PruB and PruA enzymes of M. tuberculosis channel substrate likely increase metabolic flux and protect the bacterium from methylglyoxal toxicity.

Photosynthetic response mechanism to polybrominated diphenyl ether exposure in Chlorella pyrenoidosa

Polybrominated diphenyl ether (PBDE) contamination is common in aquatic environments and can severely damage aquatic organisms. However, there is a lack of information on the response and self-adaptation mechanisms of these organisms. Chlorella pyrenoidosa was treated with 2,2′,4,4′-tetrabromodiphenyl ether (BDE47), causing significant growth inhibition, pigment reduction, oxidative stress, and chloroplast atrophy. Photosynthetic damage contributed to inhibition, as indicated by Fv/Fm, Chl a fluorescence induction, photosynthetic oxygen evolution activity, and photosystem subunit stoichiometry. Here, Chl a fluorescence induction and quinone electron acceptor (QA−) reoxidation kinetics showed that the PSII donor and acceptor sides were insensitive to BDE47. Quantitative analyses of D1 and PsaD proteins illustrated that PSII and PSI complexes were the main primary targets of photosynthesis inhibition by BDE47. Significant modulation of PSII complex might have been caused by the potential binding of BDE47 on D1 protein, and molecular docking was performed to investigate this. Increased activation of antioxidant defense systems and photosystem repair as a function of exposure time indicated a positive resistance to BDE47. After a 5-day exposure, 23 % of BDE47 was metabolized. Our findings suggest that C. pyrenoidosa has potential as a bioremediator for wastewater-borne PBDEs and can improve our understanding of ecological risks to microalgae.

Potassium alleviates over-reduction of the photosynthetic electron transport chain and helps to maintain photosynthetic function under salt-stress.

Potassium ions enhance photosynthetic tolerance to salt stress. We hypothesized that potassium ions, by minimizing the trans-thylakoid proton diffusion potential difference, can alleviate over-reduction of the photosynthetic electron transport chain and maintain the functionality of the photosynthetic apparatus. This study investigated the effects of exogenous potassium on the transcription level and activity of proteins related to the photosynthetic electron-transport chain of tobacco seedlings under salt stress. Salt stress retarded the growth of seedlings and caused an outflow of potassium ions from the chloroplast. It also lowered qP (indicator of the oxidation state of QA , the primary quinone electron acceptor in Photosystem II (PSII) and YPSII (average photochemical yield of PSII in the light-adapted state) while increasing YNO+NF (nonregulatory energy dissipation in functional and nonfunctional PSII), accompanied by decreased expression of most light-harvesting, energy-transduction, and electron-transport genes. However, exogenous potassium prevented these effects due to NaCl. Interestingly, lincomycin (an inhibitor of the synthesis of chloroplast-encoded proteins in PSII) significantly diminished the alleviation effect of exogenous potassium on salt stress. We attribute the comprehensive NaCl-induced downregulation of transcription and photosynthetic activities to retrograde signaling induced by reactive oxygen species. There probably exist at least two types of retrograde signaling induced by reactive oxygen species, distinguished by their sensitivity to lincomycin. Exogenous potassium appears to exert its primary effect by ameliorating the trans-thylakoid proton diffusion potential difference via a potassium channel, thereby accelerating ATP synthesis and carbon assimilation, alleviating over-reduction of the photosynthetic electron transport chain, and maintaining the functionality of photosynthetic proteins.

Photoinduced electron transfer from spinach chloroplasts to carbon paste electrodes through an exogenous electron acceptor

The photocurrent generated from chloroplasts of spinach is investigated using a carbon paste electrode. Chloroplasts extracted from spinach are shown to interact with a carbon paste electrode through five types of redox mediators. Among these mediators, 2,5-dichloro-1,4-benzoquinone (2,5-DCBQ) is found to be the best electron-transfer mediator, generating the highest photocurrent of approximately 310 nA cm−2 per μg chlorophyll. In addition, the photocurrent density is investigated as a function of the photon flux density, concentration of 2,5-DCBQ, and the amount of chloroplasts deposited onto the surface of the electrode. Photoreduction of 2,5-DCBQ exhibited Michaelis–Menten-like kinetics, and the reduction rates were dependent on the amount of 2,5-DCBQ and the light intensity. Even 10 days after the chloroplasts were extracted, the electrodes with the extracted spinach chloroplasts exhibited ∼90% of the photoinduced current that they exhibited immediately after the chloroplast extraction. The source of photocurrent was confirmed in experiments using 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), an inhibitor of electron transfer from primary quinone electron acceptor QA to secondary quinone electron acceptor QB in photosystem II, and KCN, an inhibitor of electron transfer from cytochrome b6f complex to photosystem I.

Mechanism of Asparagine-Mediated Proton Transfer in Photosynthetic Reaction Centers.

In photosynthetic reaction centers from purple bacteria (PbRCs), light-induced charge separation leads to the reduction of the terminal electron acceptor quinone, QB. The reduction of QB to QB•- is followed by protonation via Asp-L213 and Ser-L223 in PbRC from Rhodobacter sphaeroides. However, Asp-L213 is replaced with nontitratable Asn-L222 and Asn-L213 in PbRCs from Thermochromatium tepidum and Blastochloris viridis, respectively. Here, we investigated the energetics of proton transfer along the asparagine-involved H-bond network using a quantum mechanical/molecular mechanical approach. The potential energy profile for the H-bond between H3O+ and the carbonyl O site of Asn-L222 shows that the proton is predominantly localized at the Asn-L222 moiety in the T. tepidum PbRC protein environment, easily forming the enol species. The release of the proton from the amide -NH2 site toward Ser-L232 via tautomerization suffers from the energy barrier. Upon reorientation of Asn-L222, the enol -OH site forms a short low-barrier H-bond with Ser-L232, facilitating protonation of QB•- in a Grotthuss-like mechanism. This is a basis of how asparagine or glutamine side chains function as acceptors/donors in proton transfer pathways.

Effects of Mycotoxin Fumagillin, Mevastatin, Radicicol, and Wortmannin on Photosynthesis of Chlamydomonas reinhardtii.

Mycotoxins are one of the most important sources for the discovery of new pesticides and drugs because of their chemical structural diversity and fascinating bioactivity as well as unique novel targets. Here, the effects of four mycotoxins, fumagillin, mevastatin, radicicol, and wortmannin, on photosynthesis were investigated to identify their precise sites of action on the photosynthetic apparatus of Chlamydomonas reinhardtii. Our results showed that these four mycotoxins have multiple targets, acting mainly on photosystem II (PSII). Their mode of action is similar to that of diuron, inhibiting electron flow beyond the primary quinone electron acceptor (QA) by binding to the secondary quinone electron acceptor (QB) site of the D1 protein, thereby affecting photosynthesis. The results of PSII oxygen evolution rate and chlorophyll (Chl) a fluorescence imaging suggested that fumagillin strongly inhibited overall PSII activity; the other three toxins also exhibited a negative influence at the high concentration. Chl a fluorescence kinetics and the JIP test showed that the inhibition of electron transport beyond QA was the most significant feature of the four mycotoxins. Fumagillin decreased the rate of O2 evolution by interrupting electron transfer on the PSII acceptor side, and had multiple negative effects on the primary photochemical reaction and PSII antenna size. Mevastatin caused a decrease in photosynthetic activity, mainly due to the inhibition of electron transport. Both radicicol and wortmannin decreased photosynthetic efficiency, mainly by inhibiting the electron transport efficiency of the PSII acceptor side and the activity of the PSII reaction centers. In addition, radicicol reduced the primary photochemical reaction efficiency and antenna size. The simulated molecular model of the four mycotoxins' binding to C. reinhardtii D1 protein indicated that the residue D1-Phe265 is their common site at the QB site. This is a novel target site different from those of commercial PSII herbicides. Thus, the interesting effects of the four mycotoxins on PSII suggested that they provide new ideas for the design of novel and efficient herbicide molecules.

Cold priming improves chilling resistance in wheat seedlings: Changing of photosystem II imprints during recovery from priming

Chilling stress oriented from frequent extreme events has seriously depressed growth and productivity of wheat. Cold priming can effectively improve cold tolerance of crops. However, it is still unclear about the changes of stress memory imprints during recovery from priming and the potential function of the imprints on improved cold tolerance. Here we subjected wheat plants to cold priming (10 °C/6 °C, 1 d) at the trifoliate stage, and then to chilling stresses (2 °C/0 °C, 1 d) at 3, 6, 9, 12, 15 and 18 days of recovery from priming, respectively. Compared with non-primed plants (NL), primed plants (PL) showed the most obvious resistance phenotype at 9 days after priming. Its terminal spreading leaves exhibit temporally specific physiological memory and persistent a high photochemical quenching capacity of photosystem II after priming. In addition, it appeared faster photosynthetic performance in the post-stress rewarming phase during 3–12 days after priming. Further analysis of different leaf positions revealed that priming increased light energy absorption by the augmented antennal pigments content and synthesis of light-harvesting proteins, and a higher percentage of primary and secondary quinone electron acceptors (QAQB) active reaction centers (RCs) which enhanced energy capture and transfer performance in primed control plants (PC). Meanwhile, the increased antenna pigments in PC indirectly affected PL. Compared with NL, priming increased electron flux into the inter-system by alleviating the degradation of antenna pigments and increased Photosystem II Subunit S (PsbS) gene expression to promote the occurrence of non-photochemical quenching (NPQ), lowered the excitation pressure per RC, and increased the percentage of QA-RCs, it finally improved the light energy absorption and utilization capacity in PL. In summary, priming-induced memory is mainly present in primed leaves (3rd and 4th leaves during R6-R15) and diminishes with spatial and temporal changes, it exists even no obvious difference in phenotype. This study contributes to a deeper understanding of the long-term effects of fluctuating temperatures on the plant.

Insights into melatonin-induced photosynthetic electron transport under low-temperature stress in cucumber.

In this study, the differences in chlorophyll fluorescence transient (OJIP) and modulated 820 nm reflection (MR820) of cucumber leaves were probed to demonstrate an insight into the precise influence of melatonin (MT) on cucumber photosystems under low temperature stress. We pre-treated cucumber seedlings with different levels of MT (0, 25, 50, 100, 200, and 400 μmol · L-1) before imposing low temperature stress (10 °C/6 °C). The results indicated that moderate concentrations of MT had a positive effect on the growth of low temperature-stressed cucumber seedlings. Under low temperature stress conditions, 100 μmol · L-1 (MT 100) improved the performance of the active photosystem II (PSII) reaction centers (PIabs), the oxygen evolving complex activity (OEC centers) and electron transport between PSII and PSI, mainly by decreasing the L-band, K-band, and G-band, but showed differences with different duration of low temperature stress. In addition, these indicators related to quantum yield and energy flux of PSII regulated by MT indicated that MT (MT 100) effectively protected the electron transport and energy distribution in the photosystem. According to the results of WO-I ≥ 1 and MR820 signals, MT also affected PSI activity. MT 100 decreased the minimal value of MR/MRO and the oxidation rate of plastocyanin (PC) and PSI reaction center (P700) (Vox ), while increased △MRslow/MRO and deoxidation rates of PC+ and P700 + (Vred ). The loss of the slow phase of MT 200 and MT 400-treated plants in the MR820 kinetics was due to the complete prevention of electron movement from PSII to re-reduce the PC+ and P700 +. These results suggest that appropriate MT concentration (100 μmol · L-1) can improve the photosynthetic performance of PS II and electron transport from primary quinone electron acceptor (QA) to secondary quinone electron acceptor (QB), promote the balance of energy distribution, strengthen the connectivity of PSI and PSII, improve the electron flow of PSII via QA to PC+ and P700 + from reaching PSI by regulating multiple sites of electron transport chain in photosynthesis, and increase the pool size and reduction rates of PSI in low temperature-stressed cucumber plants, All these modifications by MT 100 treatment promoted the photosynthetic electron transfer smoothly, and further restored the cucumber plant growth under low temperature stress. Therefore, we conclude that spraying MT at an appropriate concentration is beneficial for protecting the photosynthetic electron transport chain, while spraying high concentrations of MT has a negative effect on regulating the low temperature tolerance in cucumber.

Formal Glycosylation of Quinones with exo-Glycals Enabled by Iron-Mediated Oxidative Radical-Polar Crossover.

The intermolecular C-O coupling reaction of 1,4-quinones with exo-glycals under iron hydride hydrogen atom transfer (HAT) conditions is described. This method provides a direct and regioselective access to a wide range of phenolic O-ketosides related to biologically relevant natural products in diastereomeric ratios up to >98:2 in the furanose and pyranose series. No trace of the corresponding C-glycosylated products that might have resulted from the radical alkylation of 1,4-quinones was observed. The results of mechanistic experiments suggest that the key C-O bond-forming event proceeds through an oxidative radical-polar crossover process involving a single-electron transfer between the HAT-generated glycosyl radical and the electron-acceptor quinone.

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.

Conformational Changes and H-Bond Rearrangements during Quinone Release in Photosystem II.

In photosystem II (PSII) and photosynthetic reaction centers from purple bacteria (PbRC), the electron released from the electronically excited chlorophyll is transferred to the terminal electron acceptor quinone, QB. QB accepts two electrons and two protons before leaving the protein. We investigated the molecular mechanism of quinone exchange in PSII, conducting molecular dynamics (MD) simulations and quantum mechanical/molecular mechanical (QM/MM) calculations. MD simulations suggest that the release of QB leads to the transformation of the short helix (D1-Phe260 to D1-Ser264), which is adjacent to the stromal helix de (D1-Asn247 to D1-Ile259), into a loop and to the formation of a water-intake channel. Water molecules enter the QB binding pocket via the channel and form an H-bond network. QM/MM calculations indicate that the H-bond network serves as a proton-transfer pathway for the reprotonation of D1-His215, the proton donor during QBH-/QBH2 conversion. Together with the absence of the corresponding short helix but the presence of Glu-L212 in PbRC, it seems likely that the two type-II reaction centers undergo quinone exchange via different mechanisms.

Effects of Stromal and Lumenal Side Perturbations on the Redox Potential of the Primary Quinone Electron Acceptor QA in Photosystem II.

The primary quinone electron acceptor QA is a key component in the electron transfer regulation in photosystem II (PSII), and hence accurate estimation of its redox potential, Em(QA-/QA), is crucial in understanding the regulatory mechanism. Although fluorescence detection has been extensively used for monitoring the redox state of QA, it was recently suggested that this method tends to provide a higher Em(QA-/QA) estimate depending on the sample status due to the effect of measuring light [Kato et al. (2019) Biochim. Biophys. Acta 1860, 148082]. In this study, we applied the Fourier transform infrared (FTIR) spectroelectrochemistry, which uses non-reactive infrared light to monitor the redox state of QA, to investigate the effects of stromal- and lumenal-side perturbations on Em(QA-/QA) in PSII. It was shown that replacement of bicarbonate bound to the non-heme iron with formate upshifted Em(QA-/QA) by ∼55 mV, consistent with the previous fluorescence measurement. In contrast, an Em(QA-/QA) difference between binding of 3-(3,4-dichlorophenyl)-1,1-dimethylurea and bromoxynil was found to be ∼30 mV, which is much smaller than the previous estimate, ∼100 mV, by the fluorescence method. This ∼30 mV difference was verified by the decay kinetics of the S2QA- recombination. On the lumenal side, Mn depletion hardly affected the Em(QA-/QA), confirming the previous FTIR result. However, removal of the extrinsic proteins by NaCl or CaCl2 wash downshifted the Em(QA-/QA) by 14-20 mV. These results suggest that electron flow through QA is regulated by changes both on the stromal and lumenal sides of PSII.

Cryo-EM structure of the dimeric Rhodobacter sphaeroides RC-LH1 core complex at 2.9 Å: the structural basis for dimerisation.

The dimeric reaction centre light-harvesting 1 (RC-LH1) core complex of Rhodobacter sphaeroides converts absorbed light energy to a charge separation, and then it reduces a quinone electron and proton acceptor to a quinol. The angle between the two monomers imposes a bent configuration on the dimer complex, which exerts a major influence on the curvature of the membrane vesicles, known as chromatophores, where the light-driven photosynthetic reactions take place. To investigate the dimerisation interface between two RC-LH1 monomers, we determined the cryogenic electron microscopy structure of the dimeric complex at 2.9 Å resolution. The structure shows that each monomer consists of a central RC partly enclosed by a 14-subunit LH1 ring held in an open state by PufX and protein-Y polypeptides, thus enabling quinones to enter and leave the complex. Two monomers are brought together through N-terminal interactions between PufX polypeptides on the cytoplasmic side of the complex, augmented by two novel transmembrane polypeptides, designated protein-Z, that bind to the outer faces of the two central LH1 β polypeptides. The precise fit at the dimer interface, enabled by PufX and protein-Z, by C-terminal interactions between opposing LH1 αβ subunits, and by a series of interactions with a bound sulfoquinovosyl diacylglycerol lipid, bring together each monomer creating an S-shaped array of 28 bacteriochlorophylls. The seamless join between the two sets of LH1 bacteriochlorophylls provides a path for excitation energy absorbed by one half of the complex to migrate across the dimer interface to the other half.

Oxidation of the reaction center chlorophyll of photosystem I is induced via close cooperation of photosystems II and I with progress of drought stress in soybean seedlings

ABSTRACT Drought is a serious abiotic stress that impairs plant productivity. We previously suggested that in rice (Oryza sativa L.) plants, the reaction center chlorophyll of photosystem I (PSI) (P700) is oxidized in response to drought stress via close cooperation of photosystem II (PSII) and PSI to protect PSI, whose fitness is crucial for photosynthesis and plant growth. It has rarely been reported whether such a defense system functions in other plant species. To collect another example, this subject was examined using soybean [Glycine max (L.)]. Irrigation of soil-cultured soybean seedlings was restricted to gradually impose drought stress. A decrease in the quantum efficiency of PSII [Y(II)], induction of non-photochemical quenching (NPQ) of excess light energy at PSII, reduction of the primary quinone electron acceptor of PSII (QA), and P700 oxidation proceeded before severe drought stress led to severe leaf wilting and photoinhibition of PSII and PSI. P700 oxidation, Y(II), and reduction of QA were strongly correlated in this period. These results are in accordance with those observed in rice. In contrast, such correlations disappeared when severely drought-stressed seedlings were included, as P700 oxidation was hampered while Y(II) further decreased in such seedlings. These results suggest that close cooperation of PSII and PSI leading to P700 oxidation functions to protect PSI against drought stress in soybean, and that similar defense system functions in soybean and rice unless drought stress are severe. The drought-stress responses of PSII and PSI are suggested to be regulated by acidification of thylakoid lumen in soybean, according to the changing patterns in the PSII and PSI parameters. Based on the similarity of the photochemistry of soybean and rice in response to drought stress, the effectiveness of P700 oxidation as an index for drought stress diagnosis is discussed. P700 oxidation is affected by factors other than drought stress such as N nutrition levels. However, effects of N nutrition levels are expected to be canceled by using another parameter obtained in P700 measurement as reference, suggesting the suitability of P700 oxidation for drought stress diagnosis.

PSII photochemistry responses to drought stress in autochthonous and modern sweet cherry cultivars

In this study, the JIP test was used to assess the drought tolerance of two sweet cherry cultivars (Prunus avium L.) (modern and autochthonous). Plants were exposed to progressive drought by withholding water and their fast (< 1 s) chlorophyll fluorescence kinetics was evaluated. JIP test analysis showed that drought stress caused a greater decrease in performance indices (PIABS and PItotal) in a modern cultivar, as compared to an autochthonous one. Our results suggest that limited reoxidation of primary quinone electron acceptor (QA), higher amount of secondary quinone electron acceptor (QB-) nonreducing reaction centres, or inhibition of the electron transport between QA and QB, decreased more seriously the photosynthetic performance of the modern cultivar. Further, higher positive L- and K-bands observed for the modern cultivar also suggest lower energetic connectivity between PSII units and increased inhibition of oxygen-evolving complex over autochthonous cultivar. Our results suggest that the autochthonous cultivar Crveni hruĹĄt had better photosynthetic performance under drought conditions, compared to the modern cultivar New Star.

Photochemistry of Photosystems II and I in Rice Plants Grown under Different N Levels at Normal and High Temperature.

Although N levels affect leaf photosynthetic capacity, the effects of N levels on the photochemistry of photosystems II and I (PSII and PSI, respectively) are not well-understood. In the present study, we examined this aspect in rice (Oryza sativa L. 'Hitomebore') plants grown under three different N levels at normal or high temperatures that can occur during rice culture and do not severely suppress photosynthesis. At both growth temperatures, the quantum efficiency of PSII [Y(II)] and the fraction of the primary quinone electron acceptor in its oxidized state were positively correlated with the amount of total leaf-N, whereas the quantum yields of non-photochemical quenching and donor-side limitation of PSI [Y(ND)] were negatively correlated with the amount of total leaf-N. These changes in PSII and PSI parameters were strongly correlated with each other. Growth temperatures scarcely affected these relationships. These results suggest that the photochemistry of PSII and PSI is coordinately regulated primarily depending on the amount of total leaf-N. When excess light energy occurs in low N-acclimated plants, oxidation of the reaction center chlorophyll of PSI is thought to be stimulated to protect PSI from excess light energy. It is also suggested that PSII and PSI normally operate at high temperature used in the present study. In addition, as the relationships between Y(II) and Y(ND) were found to be almost identical to those observed in osmotically stressed rice plants, common regulation is thought to be operative when excess light energy occurs due to different causes.

The Mechanisms of Electrogenic Reactions in Bacterial Photosynthetic Reaction Centers: Studies in Collaboration with Alexander Konstantinov.

In this review, we discuss our studies conducted in 1985-1988 in collaboration with A. A. Konstantinov, one of the top scientists in the field of membrane bioenergetics. Studying fast kinetics of membrane potential generation in photosynthetic reaction centers (RCs) of purple bacteria in response to a laser flash has made it possible to examine in detail the mechanisms of electrogenic reactions at the donor and acceptor sides of RCs. Electrogenesis associated with the intraprotein electron transfer from the exogenous secondary donors, redox dyes, and soluble cytochrome (cyt) c to the photooxidized dimer of bacteriochlorophyll P870 was studied using proteoliposomes containing RCs from the non-sulfur purple bacterium Rhodospirillum rubrum. It was found that reduction of the secondary quinone electron acceptor QB accompanied by its protonation in the chromatophores from R. rubrum in response to every second light flash was electrogenic. Spectral characteristics and redox potentials of the four hemes in the tightly bound cyt c in the RC of Blastochloris viridis and electrogenic reactions associated with the electron transfer within the RC complex were identified. For the first time, relative amplitudes of the membrane potential generated in the course of individual electrogenic reactions were compared with the distances between the redox cofactors determined based on the three-dimensional structure of the Bl. viridis RC.