Evolution of PS IIα and PS IIβ centers during the greening of Euglena gracilis Z: Correlations with changes in lipid content

The building up of the two types of reaction centers, PS II and PS II, was investigated during the greening of Euglena gracilis Z cells in resting medium. The maximal values in the proportion of PS II centers (55%) and in the oxygen evolved per chlorophyll were reached at the outbreak of greening, when accumulation of galactolipids (MGDG and DGDG) rich in unsaturated fatty acids occurred, and when anionic lipids (SQDG and PG) emerged. As the greening progressed, the chlorophyll accumulation corresponded to a secondary enrichment in PS II centers, which built up more rapidly than PS II centers; correlatively, a general saturation of the fatty acids constitutive of all lipid classes took place.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DGDG digalactosyldiacylglycerol - FAME Tatty acid methyl esters - HEPES acide (N-[2-hydroxyethyl]piperazine-N-[2-ethane sulfonic] - MGDG monogalactosyldiacylglycerol - PC phosphatidylcholine - PE phosphatidylethanolamine - PG phosphatidylglycerol - PQ plastoquinone - PS I Photosystem I - PS II Photosystem II - Q_A primary quinone electron acceptor of PS II - Q_B secondary quinone electron acceptor of PS II - SQDG sulfoquinovosyldiacylglycerol     

High misses after odd flashes in oxygen evolution in thoroughly dark-adapted thylakoids from pea and Chenopodium album

In Photosystem II (PS II), water is oxidized to molecular oxygen and plastoquinone is reduced to plastoquinol. The oxidation of water requires the accumulation of four oxidizing equivalents, through the so-called S-states of the oxygen evolving complex; the production of plastoquinol requires the accumulation of two reducing equivalents on a bound plastoquinone, Q_B. It has been generally believed that during the flash-induced transition of each of the S-states (S_n S_n+1, where n=0, 1, 2 and 3), a certain small but equal fraction of the PS II reaction centers are unable to function and, thus, miss being turned over. We used thoroughly dark-adapted thylakoids from peas (Pisum sativum) and Chenopodium album (susceptible and resistant to atrazine) starting with 100% of the oxygen evolving complex in the S₁ state. Thylakoids were illuminated with saturating flashes, providing a double hit parameter of about 0.07. Our experimental data on flashnumber dependent oscillations in the amount of oxygen per flash fit very well with a binary pattern of misses: 0, 0.2, 0, 0.4 during S₀ S₁, S₁ S₂, S₂ S₃ and S₃ S₀ transitions. Addition of 2 mM ferricyanide appears to shift this pattern by one flash. These results are consistent with the bicycle model recently proposed by V. P. Shinkarev and C. A. Wraight (Oxygen evolution in photosynthesis: From unicycle to bicycle, 1993, Proc Natl Acad Sci USA 90: 1834–1838), where misses are due to the presence of P⁺ or Q_A ^- among the various equilibrium states of PS II centers.Abbreviations miss parameter - double hit parameter - PS II Photosystem II - Q_A primary one-electron acceptor of PS II, a plastoquinone molecule - Q_B secondary plastoquinone two-electron acceptor of PS II - S-states (S_n, where n=0, 1, 2, 3 or 4) redox states of the oxygen evolving complex     

Characterization of photosystem II in stroma thylakoid membranes

The functional state of the PS II population localized in the stroma exposed non-appressed thylakoid region was investigated by direct analysis of the PS II content of isolated stroma thylakoid vesicles. This PS II population, possessing an antenna size typical for PS II, was found to have a fully functional oxygen evolving capacity in the presence of an added quinone electron acceptor such as phenyl-p-benzoquinone. The sensitivity to DCMU for this PS II population was the same as for PS II in control thylakoids. However, under more physiological conditions, in the absence of an added quinone acceptor, no oxygen was evolved from stroma thylakoid vesicles and their PS II centers were found to be incapable to pass electrons to PS I and to yield NADPH. By comparison of the effect of a variety of added quinone acceptors with different midpoint potentials, it is concluded that the inability of PS II in the stroma thylakoid membranes to contribute to NADPH formation probably is due to that Q_A of this population is not able to reduce PQ, although it can reduce some artificial acceptors like phenyl-p-benzoquinone. These data give further support to the notion of a discrete PS II population in the non-appressed stroma thylakoid region, PS II, having a higher midpoint potential of Q_A than the PS II population in the appressed thylakoid region, PS II. The physiological significance of a PS II population that does not produce any NADPH is discussed.Abbreviations pBQ p-benzoquinone - Chl chlorophyll - DCBQ 2,6-dichloro-p-benzoquinone - DCIP 2,6-dichloroindophenol - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DMBQ 2,5-dimethyl-p-benzoquinone - DQ duroquinone(tetramethyl-p-benzoquinone) - FeCN ferricyanide (potassium hexacyanoferrat) - MV methylviologen - NADPH,NADP⁺ reduced or oxidized form of nicotinamide adenine dinucleotide phosphate respectively - PpBQ phenyl-p-benzoquinone - PQ plastoquinone - PS II photosystem II - PS I photosystem I - Q_A primary quinone acceptor of PS II - Q_B secondary quinone acceptor of PS II - E microEinstein     

Electroluminescence

An overview is presented of research based on the observation by Arnold and Azzi (1971) (Photochem Photobiol 14: 233–240), that an electric field induces charge-recombination luminescence in a suspension of photosynthetic membrane vesicles. The electroluminescence signals from Photosystems I and II are discussed in relation to the shape of the vesicles and the membrane potentials generated by the externally applied electric field. The use of the electroluminescence amplitude as a probe to study the kinetics and energetics of charge separation, and of its kinetics to monitor the electric-field induced charge recombination process are reviewed. Currently unresolved issues regarding the emission yield of electroluminescence are briefly discussed and the properties are summarized of the unexplained Photosystem II luminescence which is not sensitive to the membrane potential.Abbreviations DCMU 3(3,4-dichlorophenyl)-1,1-dimethylurea - EL electroluminescence - PS I, II Photosystem I, II - TPB tetraphenylboron, an artificial electron donor for PS II - P primary electron donor - S_i Y_z P680 Pheo Q_A Q_B sequence of electron transfer components in PS II - plastocyanin P700 A₀ A₁ F_x F_A (or F_B) sequence of electron transfer components in PS I     

Chimaeric CP47 mutants of the cyanobacterium Synechocystis sp. PCC 6803 carrying spinach sequences: Construction and function

Chimaeric mutants of the cyanobacterium Synechocystis sp. PCC 6803 have been generated carrying part or all of the spinach psbB gene, encoding CP47 (one of the chlorophyll-binding core antenna proteins in Photosystem II). The mutant in which the entire psbB gene had been replaced by the homologous gene from spinach was an obligate photoheterotroph and lacked Photosystem II complexes in its thylakoid membranes. However, this strain could be transformed with plasmids carrying selected regions of Synechocystis psbB to give rise to photoautotrophs with a chimaeric spinach/cyanobacterial CP47 protein. This process involved heterologous recombination in the cyanobacterium between psbB sequences from spinach and Synechocystis 6803; which was found to be reasonably effective in Synechocystis. Also other approaches were used that can produce a broad spectrum of chimaeric mutants in a single experiment. Functional characterization of the chimaeric photoautotrophic mutants indicated that if a decrease in the photoautotrophic growth rates was observed, this was correlated with a decrease in the number of Photosystem II reaction centers (on a chlorophyll basis) in the thylakoid membrane and with a decrease in oxygen evolution rates. Remaining Photosystem II reaction centers in these chimaeric mutants appeared to function rather normally, but thermoluminescence and chlorophyll a fluorescence measurements provided evidence for a destabilization of Q_B ^–. This illustrates the sensitivity of the functional properties of the PS II reaction center to mild perturbations in a neighboring protein.Abbreviations diuron 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_v variable chlorophyll a fluorescence - HEPES N-(2-hydroxyethyl)piperazine-N-(2-ethanesulfonic acid) - (k)bp (kilo)base pairs - PS II Photosystem II - Q_A primary electron-accepting plastoquinone in Photosystem II - Q_B secondary electron-accepting plastoquinone in Photosystem II - SDS sodium dodecyl sulfate     

Temperature-dependent changes in Photosystem II heterogeneity support a cycle of Photosystem II during photoinhibition

High light treatments were given to attached leaves of pumpkin (Cucurbita pepo L.) at room temperature and at 1°C where the diffusion- and enzyme-dependent repair processes of Photosystem II are at a minimum. After treatments, electron transfer activities and fluorescence induction were measured from thylakoids isolated from the treated leaves. When the photoinhibition treatment was given at 1°C, the Photosystem II electron transfer assays that were designed to require electron transfer to the plastoquinone pool showed greater inhibition than electron transfer from H₂O to paraphenyl-benzoquinone, which measures all PS II centers. When the light treatment was given at room temperature, electron transfer from H₂O to paraphenyl-benzoquinone was inhibited more than whole-chain electron transfer. Variable fluorescence measured in the presence of ferricyanide decreased only during room-temperature treatments. These results suggest that reaction centers of one pool of Photosystem II, non-Q_B-PS II, replace photoinhibited reaction centers at room temperature, while no replacement occurs at 1°C. A simulation of photoinhibition at 1°C supports this conclusion.Abbreviations BSA bovine serum albumin - Chl chlorophyll - DCMU 3-(3,4,-dichlorophenyl)-1,1,-dimethylurea - DCPIP dichlorophenol-indophenol (2,6-dichloro-4((4-hydroxyphenyl)imino)-2,5-cyclohexadien-1-one) - DPC diphenyl carbazide (2,2-diphenylcarbonic dihydrazide) - FeCN ferricyanide (hexacyanoferrate(III)) - _app apparent quantum yield of photosynthetic oxygen evolution - MV methyl viologen (1,1-dimethyl-4,4-bipyridinium dichloride) - PPBQ phenyl-p-benzoquinone - PPFD photosynthetic photon flux density - PQ pool plastoquinone - Q_B secondary quinone acceptor of PS II - RT room temperature - WC whole chain electron transfer     

Cyclic electron flow around Photosystem II in vivo

The oxygen flash yield (Y_O2) and photochemical yield of PS II (_{PS II}) were simultaneously detected in intact Chlorella cells on a bare platinum oxygen rate electrode. The two yields were measured as a function of background irradiance in the steady-state and following a transition from light to darkness. During steady-state illumination at moderate irradiance levels, Y_O2 and _{PS II} followed each other, suggesting a close coupling between the oxidation of water and Q_A reduction (Falkowski et al. (1988) Biochim. Biophys. Acta 933: 432–443). Following a light-to-dark transition, however, the relationship between Q_A reduction and the fraction of PS II reaction centers capable of evolving O₂ became temporarily uncoupled. _{PS II} recovered to the preillumination levels within 5–10 s, while the Y_O2 required up to 60 s to recover under aerobic conditions. The recovery of Y_O2 was independent of the redox state of Q_A, but was accompanied by a 30% increase in the functional absorption cross-section of PS II (_{PS II}). The hysteresis between Y_O2 and the reduction of Q_A during the light-to-dark transition was dependent upon the reduction level of the plastoquinone pool and does not appear to be due to a direct radiative charge back-reaction, but rather is a consequence of a transient cyclic electron flow around PS II. The cycle is engaged in vivo only when the plastoquinone pool is reduced. Hence, the plastoquinone pool can act as a clutch that disconnects the oxygen evolution from photochemical charge separation in PS II.Abbreviations ADRY acceleration of the deactivation reactions of the water-splitting enzyme (agents) - Chl chlorophyll - cyt cytochrome - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_O minimum fluorescence yield in the dark-adapted state - F_I minimum fluorescence yield under ambient irradiance or during transition from the light-adapted state - F_M maximum fluorescence yield in the dark-adapted state - F_M maximum fluorescence yield under ambient irradiance or during transition from light-adapted state - F_V, F_V variable fluorescence (F_V=F_M–F_O ; F_V=F_M–F_I) - FRR fast repetition rate (fluorometer) - _{PS II} quantum yield of Q_A reduction (_{PS II}=(F_M – F_O)/F_M or _{PS II})=(F_M= – F_I=)/F_M=) - LHCII Chl a/b light harvesting complexes of Photosystem II - OEC oxygen evolving complex of PS II - P680 reaction center chlorophyll of PS II - PQ plastoquinone - POH₂ plastoquinol - PS I Photosystem I - PS II Photosystem II - RC II reaction centers of Photosystem II - PS II the effective absorption cross-section of PHotosystem II - TL thermoluminescence - Y_O2 oxygen flash yield The US Government right to retain a non-exclusive, royalty free licence in and to any copyright is acknowledged.     

The role of light in formation of modified S2-state upon low pH-treatment of oxygen-evolving Photosystem II

An abnormal, structurally modified, kinetically stable S₂-state has been reported to be induced when Photosystem II was treated with NaCl-EGTA (or EDTA) in the light or with pH in darkness, both are assumed to release functional Ca²⁺. In order to compare the mechanism of induction of modified S₂-state between the two treatments, effects of illumination during or before low pH-treatment on formation of the abnormal S₂-state were investigated by means of thermoluminescence measurements and low temperature EPR spectroscopy. Following results have been obtained: Flash illumination during low pH-treatment did not practically induce the abnormal S₂-state, whereas preflash illumination given immediately before low pH-treatment efficiently induced the abnormal S₂-state, and its amplitude showed a period-four oscillation on varying the preflash number with maxima at the second and sixth flashes. The abnormal S₂-state thus induced by preflashes was identical with the modified S₂-state that could be induced in dark-low pH-treated Photosystem II by excitation at 0°C after neutralization to pH 6.5. It was inferred that in low pH-treatment, modified S₂-state can be formed from both S₂- and S₃-states, but its yield from the latter is much higher than from the former, consistent with the early results by Boussac et al. obtained for NaCl-EGTA-light or NaCl-citrate-light treatment.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - EDTA ethylenediaminetetraactate - EGTA ethylene glycol bis(-aminoethylether)-N,N,N,N-tetraacetic acid - Mes 2-(N-morpholino)ethanesulfonic acid - PS II Photosystem II     

Thylakoid membrane energization and swelling in photoinhibited Chlamydomonas cells is prevented in mutants unable to perform cyclic electron flow

Photoinhibition of Photosystem II in unicellular algae in vivo is accompanied by thylakoid membrane energization and generation of a relatively high pH as demonstrated by ¹⁴C-methylamine uptake in intact cells. Presence of ammonium ions in the medium causes extensive swelling of the thylakoid membranes in photoinhibited Chlamydomonas reinhardtii but not in Scenedesmus obliquus wild type and LF-1 mutant cells. The rise in pH and the related thylakoid swelling do not occur at light intensities which do not induce photoinhibition. The rise in pH and membrane energization are not induced by photoinhibitory light in C. reinhardtii mutant cells possessing an active Photosystem II but lacking cytochrome b₆/f, plastocyanin or Photosystem I activity and thus being unable to perform cyclic electron flow around Photosystem I. In these mutants the light-induced turnover of the D1 protein of Reaction Center II is considerably reduced. The high light-dependent rise in pH is induced in the LF-1 mutant of Scenedesmus which can not oxidize water but otherwise possesses an active Reaction Center II indicating that PS II-linear electron flow activity and reduction of plastoquinone are not required for this process. Based on these results we conclude that photoinhibition of Photosystem II activates cyclic electron flow around Photosystem I which is responsible for the high membrane energization and pH rise in cells exposed to excessive light intensities.Abbreviations cyt b₆/f cytochrome b₆/f - Diuron 3-(3,4-dichlorophenyl)-1 dimethyl urea - Q_B the secondary quinone acceptor of reaction center II - DNP 2,4,Dinitrophenol - FCCP carbonyl cyanide trifluoromethoxy phenylhydrazone - SDS-PAGE sodium dodecylsulfate polyacrylamide gel electrophoresis     

Functional size of Photosystem II determined by radiation inactivation

The functional size of Photosystem II (PS II) was investigated by radiation inactivation. The technique provides an estimate of the functional mass required for a specific reaction and depends on irradiating samples with high energy -rays and assaying the remaining activity. The analysis is based on target theory that has been modified to take into account the temperature dependence of radiation inactivation of proteins. Using PS II enriched membranes isolated from spinach we determined the functional size of primary charge separation coupled to water oxidation and quinone reduction at the Q_B site: H₂O (Mn)₄ Y_z P680 Pheophytin Q phenyl-p-benzoquinone. Radiation inactivation analysis indicates a functional mass of 88 ± 12 kDa for electron transfer from water to phenyl-p-benzoquinone. It is likely that the reaction center heterodimer polypeptides, D1 and D2, contribute approximately 70 kDa to the functional mass, in which case polypeptides adding up to approximately 20 kDa remain to be identified. Likely candidates are the and subunits of cytochrome b ₅₅₉and the 4.5 kDa psbI gene product.Abbreviations Cyt cytochrome - PS Photosystem - P680 primary electron donor of Photosystem II - Q_A primary quinone acceptor of Photosystem II - Q_B secondary quinone acceptor of Photosystem II - Y_z tyrosine donor to P680     

Evidence for two types of electron transfer processes through Photosystem II

Inhibition of electron flow from H₂O to methylviologen by 3-(34 dichlorophenyl)-1,1 dimethyl urea (DCMU), yields a biphasic curve — an initial high sensitivity phase and a subsequent low sensitivity phase. The two phases of electron flow have a different pH dependence and differ in the light intensity required for saturation.Preincubation of chloroplasts with ferricyanide causes an inhibition of the high sensitivity phase, but has no effect on the low sensitivity phase. The extent of inhibition increases as the redox potential during preincubation becomes more positive. Tris-treatment, contrary to preincubation with ferricyanide, affects, to a much greater extent, the low sensitivity phase.Trypsin digestion of chloroplasts is known to block electron flow between Q _A and Q _B, allowing electron flow to ferricyanide, in a DCMU insensitive reaction. We have found that in trypsinated chloroplasts, electron flow becomes progressively inhibited by DCMU with increase in pH, and that DCMU acts as a competitive inhibitor with respect to [H⁺]. The sensitivity to DCMU rises when a more negative redox potential is maintained during trypsin treatment. Under these conditions, only the high sensitivity, but not the low sensitivity phase is inhibited by DCMU.The above results indicate the existence of two types of electron transport chains. One type, in which electron flow is more sensitive to DCMU contains, presumably Fe in a Q _A Fe complex and is affected by its oxidation state, i.e., when Fe is reduced, it allows electron flow to Q _B in a DCMU sensitive step; and a second type, in which electron transport is less sensitive to DCMU, where Fe is either absent or, if present in its oxidized state, is inaccessible to reducing agents.Abbreviations DCMU 3-(34 dichlorophenyl)-1, 1 Dimethyl urea - MV methyl viologen - PS II Photosystem II - Tris tris (hydroxymethyl)aminomethane     

Binding affinity of bicarbonate and formate in herbicide-resistant D1 mutants of Synechococcus sp. PCC 7942

We examined the effects of mutations at amino acid residues S264 and F255 in the D1 protein on the binding affinity of the stimulatory anion bicarbonate and inhibitory anion formate in Photosystem II (PS II) in Synechococcus sp. PCC 7942. Measurements on the rates of oxygen evolution in the wild type and mutant cells in the presence of different concentrations of formate with a fixed bicarbonate concentration and vice versa, analyzed in terms of an equilibrium activator-inhibitor model, led to the conclusion that the equilibrium dissociation constant for bicarbonate is increased in the mutants, while that of the formate remains unchanged (11±0.5 mM). The hierarchy of the equilibrium dissociation constant for bicarbonate (highest to lowest, ±2 M) was: D1-F255L/S264A (46 M)>D1-F255Y/ S264A (31 M)D1-S264A (34 M)D1-F255Y (33 M)>wild type (25 M). The data suggest the importance of D1-S264 and D1-F255 in the bicarbonate binding niche. A possible involvement of bicarbonate and these two residues in the protonation of Q_B ^-, the reduced secondary plastoquinone of PS II, in the D1 protein is discussed.Abbreviations Chl a chlorophyll a - DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DMQ 2,5-dimethyl-p-benzoquinone - HEPES N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid - MES 2-[N-morpholino]ethanesulfonic acid - PSI Photosystem I - PS II Photosystem II - Q_A bound plastoquinone, a one-electron acceptor in Photosystem II - Q_B another bound plastoquinone, a two-electron acceptor in Photosystem II This paper is dedicated to the memory of my dear friend Robin Hill-Govindjee.     

Interaction between photosynthetic and respiratory electron-transfer chains in the membranes of Anabaena variabilis

The rate of CO₂- and p-benzoquione-dependent photosynthetic O₂ evolution by Anabaena variabilis cells remained unaltered and the rate of O₂ uptake observed after switching off the light (endogenous respiration) was enhanced by a factor of 6–8 when the O₂ concentration was increased from 200 to 400 M. Photosystem-I-linked O₂ uptake and respiration of the cells incubated with ascorbate and N,N,NN-tetramethyl-p-phenylenediamine was not appreciable influenced by the O₂ concentration. 2-Iodo-6-isopropyl-3-methyl-2,4,4-trinitrodiphenyl ether, blocking electron transfer at the plastoquinone level, suppressed O₂ evolution and had no influence on endogenous respiration. 2-n-Heptyl-4-hydroxyquinoline-N-oxide, an inhibitor of electron transfer between photosystems II and I, as well as the cytochrome-oxidase inhibitors N ₃ ^- , CN^- and NH₂OH, caused a 35–50% retardation of endogenous respiration and blocked photosynthetic O₂ evolution. The molar ratio of cytochromes b₆, f, c-553, aa₃ and photosystem-I reaction centers in the isolated membranes equalled approx. 2:1:2:0.7:2. It is inferred that endogenous respiration of A. variabilis cells is inhibited by the light-induced electron flow through both photosystems at the level of the plastoquinone-plastocyanin-oxidoreductase complex.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DNP-INT 2-iodo-6-isopropyl-3-methyl-2,4,4-trinitrodiphenyl ether - Hepes 4-(2-hydroxyethyl)-1-piperazine ethansulfonic acid - TMPD N,N,NN-tetramethyl-p-phenylenediamine     

Thermoluminescence investigations on the site of action of o-phthalaldehyde in photosynthetic electron transport

Glow curves from spinach leaf discs infiltrated with o-phthalaldehyde (OPA) show significant similarity to those obtained by DCMU treatment which is known to block the electron flow from Q_A, the stable acceptor of Photosystem II (PS II). In both the cases, the thermoluminescence (TL) peak II (Q band) was intensified significantly, whereas peaks III and IV (B band) were suppressed. Total TL yield of the glow curve remained constant even when the leaf discs were infiltrated with high concentrations of OPA (4 mM) or with DCMU (100 M), indicating that even at these high concentrations no significant change in the number of species undergoing charge recombination in PS II occurred. However, studies with thylakoids revealed significant differences in the action of OPA and DCMU on PS II. Although OPA, at a certain concentration and time of incubation, reduced the B band intensity by about 50–70%, and completely abolished the detectable oxygen evolution, it still retained the TL flash yield pattern, and, thus, S state turnover. OPA is known to inhibit the oxidoreductase activity of in vitro Cyt b₆/f (Bhagwat et al. (1993) Arch Biochem Biophys 304: 38–44). However, in the OPA treated thylakoids the extent of inhibition of O₂ evolution was not reduced even in the presence of oxidized tetramethyl-p-phenylenediamine which accepts electrons from plastoquinol and feeds then directly to Photosystem I. This suggests that OPA inhibition is at a site prior to plastoquinone pool in the electron transport chain, in agreement with it being between Q_A and Q_B. However, an unusual feature of OPA inhibition is that even though all oxygen evolution was completely suppressed, a significant fraction of PS II centers were functional and turned over with the same periodicity of four in the absence of any added electron donor, an observation which appears to be similar to that reported by Wydrzynski (Wydrzynski et al. (1985) Biochim Biophys Acta 809: 125–136) with lauroylcholine chloride, a lipid analogue compound. The detailed chemistry of OPA inhibition remains to be studied. Since we dedicate this paper to William A. Arnold, discoverer of delayed light and TL in photosynthesis, we have also included in the Introduction, a brief history of how TL work was initiated at BARC (Bombay, India).Abbreviations Chl chlorophyll - Cyt b₆/f Cytochrome b₆/f - DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCIP 2,6-dichloropenolindophenol - DCMU 3-(3,4-dichlorophenyl-) 1,1-dimethyl urea - HEPES (N-[2-hydroxyethyl]piperazine-N-[2-ethanesulfonic acid]) - LCC lauroylcholine chloride - OPA o-phthalaldehyde - PS I Photosystem I - PS II Photosystem II - TL thermoluminescence - TMPD 2,3,5,6-tetramethyl-p-phenylenediamine     

Reduced levels of cytochrome b 6/f in transgenic tobacco increases the excitation pressure on Photosystem II without increasing sensitivity to photoinhibition in vivo

We have examined tobacco transformed with an antisense construct against the Rieske-FeS subunit of the cytochromeb ₆ f complex, containing only 15 to 20% of the wild-type level of cytochrome f. The anti-Rieske-FeS leaves had a comparable chlorophyll and Photosystem II reaction center stoichiometry and a comparable carotenoid profile to the wild-type, with differences of less than 10% on a leaf area basis. When exposed to high irradiance, the anti-Rieske-FeS leaves showed a greatly increased closure of Photosystem II and a much reduced capacity to develop non-photochemical quenching compared with wild-type. However, contrary to our expectations, the anti-Rieske-FeS leaves were not more susceptible to photoinhibition than were wild-type leaves. Further, when we regulated the irradiance so that the excitation pressure on photosystem II was equivalent in both the anti-Rieske-FeS and wild-type leaves, the anti-Rieske-FeS leaves experienced much less photoinhibition than wild-type. The evidence from the anti-Rieske-FeS tobacco suggests that rapid photoinactivation of Photosystem II in vivo only occurs when closure of Photosystem II coincides with lumen acidification. These results suggest that the model of photoinhibition in vivo occurring principally because of limitations to electron withdrawal from photosystem II does not explain photoinhibition in these transgenic tobacco leaves, and we need to re-evaluate the twinned concepts of photoinhibition and photoprotection.Abbreviations Chl chlorophyll - DCMU 3-(3,4-dichlophenyl)-1,-dimethylurea - Fo and Fo minimal fluorescence when all PS II reaction centers are open in dark- and light-acclimated leaves, respectively - Fm and Fm maximal fluorescence when all PS II reaction centers are closed in dark- and light-acclimated leaves, respectively - Fv variable fluorescence (Fm-Fo) in dark acclimated leaves - Fv variable fluorescence (Fm-Fo) in lightacclimated leaves - NPQ non-photochemical quenching of fluorescence - PS I and PS II Photosystem I and II - P680 primary electron donor of the reaction center of PS II - PFD photosynthetic flux density - Q_A primary acceptor quinone of PS II - q_p photochemical quenching of fluorescence - V+A+Z violaxanthin+antheraxanthin+zeaxanthin     

Photochemical activities,pigment distribution and photosynthetic unit size of subchloroplast particles isolated from synchronized cells of Scenedesmus obliquus

Photosystem II (PS II) reactions of chloroplast particles show the same variations during the synchronous life cycle of Scenedesmus obliquus, strain D₃ (Gaffron Biol. Zbl. 59, 302 1939), as the whole cells they derived from. Photosystem I (PS I) reactions of whole cells and of subchloroplast particles show little or no variation in their activity, whereas PS I reactions of chloroplast particles vary like PS II reactions during the life cycle. The variation in chloroplast particles could be attributed to the change in the reoxidation capacity of plastoquinone still attached to PS I. Digitonin-treatment of chloroplast particles from Scenedesmus and subsequent sucrose density gradient separation yielded 3 distinct fractions: Fraction I contained pure PS I particles with the most efficient PS I-mediated methylviologen (MV) reduction with subsequent oxygen uptake (3 mmol O₂/mg Chl·h); no Hill reaction; and a high chlorophyll a/b ratio, and a vast amount of unbound protein xanthophyll complexes. Fraction II is enriched in PS II particles, with little PS I activity (less than 10% of the PS I particles) and a low chlorophyll a/b ratio. The activity of the water-splitting system was completely lost. This fraction must also contain most of the light-harvesting pigment system. Fraction III is also enriched in PS II with even less PS I activity, but the ratio of chlorophyll a/b is slightly higher than in whole cells and the water-splitting system is intact. -carotene was part of all fractions whereas functional xanthophylls seemed to be restricted to the PS II particles. From the constant chlorophyll P/700 ratio we had to conclude that size of the photosynthetic unit does not change during the life cycle of a synchronized Scenedesmus obliquus culture.Abbreviations DBMIB 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethyl-urea - DCPIP dichlorphenolindophenol - MV methylviologen (paraquat) - PS I photosystem I - PS II photosystem II - DPC diphenyl-carbazide     

Mechanisms for controlling balance between light input and utilisation in the salt tolerant alga Dunaliella C9AA

The yield of photosynthetic O₂ evolution was measured in cultures of Dunaliella C9AA over a range of light intensities, and a range of low temperatures at constant light intensity. Changes in the rate of charge separation at Photosystem I (PS I) and Photosystem II (PS II) were estimated by the parameters _{PS I} and _{PS II} . _{PS I} is calculated on the basis of the proportion of centres in the correct redox state for charge separation to occur, as measured spectrophotometrically. _{PS II} is calculated using chlorophyll fluorescence to estimate the proportion of centres in the correct redox state, and also to estimate limitations in excitation delivery to reaction centres. With both increasing light intensity and decreasing temperature it was found that O₂ evolution decreased more than predicted by either _{PS I} or _{PS II}. The results are interpreted as evidence of non-assimilatory electron flow; either linear whole chain, or cyclic around each photosystem.Abbreviations F₀ dark level of chlorophyll fluorescence yield (PS II centres open) - F_m maximum level of chlorophyll fluorescence yield (PS II centres closed) - F_v variable fluorescence (F_m-F₀) - PS I Photosystem I - PS II Photosystem II - P₇₀₀ reaction centre chlorophyll(s) of PS I - q_N coefficient of non-photochemical quenching of chlorophyll fluorescence - q_P coefficient of photochemical quenching of fluorescence yield - q_E high-energy-state quenching coefficient - _{PS I} yield of PS I - _{PS II} yield of PS II - _S yield of photosynthetic O₂ evolution - _P intrinsic yield of open PS II centres     

Binary oscillations in the Kok model of oxygen evolution in oxygenic photosynthesis

The flash-induced kinetics of various characteristics of Photosystem II (PS II) in the thylakoids of oxygenic plants are modulated by a period of two, due to the function of a two-electron gate in the electron acceptor side, and by a period of four, due to the changes in the state of the oxygen-evolving complex. In the absence of inhibitors of PS II, the assignment of measured signal to the oxygen-evolving complex or to quinone acceptor side has frequently been done on the basis of the periodicity of its flash-induced oscillations, i.e. four or two. However, in some circumstances, the period four oscillatory processes of the donor side of PS II can generate period two oscillations. It is shown here that in the Kok model of oxygen evolution (equal misses and equal double hits), the sum of the concentrations of the S ₀ and S ₂ states (as well as the sum of concentrations of S ₁ and S ₃ states) oscillates with period of two: S ₀+S ₂S ₁+S ₃S ₀+S ₂S ₁+S ₃. Moreover, in the generalized Kok model (with specific miss factors and double hits for each S-state) there always exist such ₀, ₁, ₂, ₃ that the sum ₀[S₀] + ₁[S₁] + ₂[S₂] + ₃[S₃] oscillates with period of two as a function of flash number. Any other coefficients which are linearly connected with these coefficients, % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabiGaciaacaqabeaadaqaaqaaaOqaaiqbew7aLzaaja% aaaa!3917!\[\hat \varepsilon \]_i = c_1i + c₂, also generate binary oscillations of this sum. Therefore, the decomposition of the flash-induced oscillations of some measured parameters into binary oscillations, depending only on the acceptor side of PS II, and quaternary oscillations, depending only on the donor side of PS II, becomes practically impossible when measured with techniques (such as fluorescence of chlorophyll a, delayed fluorescence, electrochromic shift, transmembrane electrical potential, changes of pH and others) that could not spectrally distinguish the donor and acceptor sides. This property of the Kok cycle puts limits on the simultaneous analysis of the donor and acceptor sides of the RC of PS II in vivo and suggests that binary oscillations are no longer a certain indicator of the origin of a signal in the acceptor side of PS II.Abbreviations PS II Photosystem II - P680 primary electron donor of reaction center of PS II - Q_A one electron acceptor plastoquinone - Q_B two electron acceptor plastoquinone - S _n redox state of the oxygen evolving complex, where n=0,1,2,3 and 4 - Chl a chlorophyll a This paper is dedicated to the memory of Alexander Kononenko.     

The non-photochemical reduction of plastoquinone in leaves

Although it is generally assumed that the plastoquinone pool of thylakoid membranes in leaves of higher plants is rapidly oxidized upon darkening, this is often not the case. A multiflash kinetic fluorimeter was used to monitor the redox state of the plastoquinone pool in leaves. It was found that in many species of plants, particularly those using the NAD-malic enzyme C₄ system of photosynthesis, the pool actually became more reduced following a light to dark transition. In some Amaranthus species, plastoquinone remained reduced in the dark for several hours. Far red light, which preferentially drives Photosystem I turnover, could effectively oxidize the plastoquinone pool. Plastoquinone was re-reduced in the dark within a few seconds when far red illumination was removed. The underlying mechanism of the dark reduction of the plastoquinone pool is still uncertain but may involve chlororespiratory activity.Abbreviations apparent F_o observed fluorescence yield after dark adaptation - F_m maximum fluorescence when all Q_A is fully reduced - F_o minimum fluorescence yield when Q_A is fully oxidized and non-photochemical quenching is fully relaxed - F_s steady state fluorescence yield - PPFD photosynthetic photon flux density - PQ plastoquinone - Q_A primary quinone acceptor of the Photosystem II reaction center - Q_B secondary quinone acceptor to the Photosystem II reaction center - F F_m minus F_s     

The catalysis of nucleotide polymerization by compounds of divalent lead

Summary Soluble lead salts and a number of lead-containing minerals catalyze the formation of oligonucleotides from nucleoside 5-phosphorimidazolides. The effectiveness of lead compounds correlates strongly with their solubility. Under optimal conditions we were able to obtain 18% of pentamer and higher oligomers from ImpA. Reactions involving ImpU gave smaller yields.Abbreviations A adenosine - U uridine - Im imidazole - MeIm 1-methyl-imidazole - EDTA ethylenediaminetetraacetic acid - pA adenosine 5-phosphate - pU uridine 5-phosphate - Ap adenosine cyclic 2:3-phosphate - ATP adenosine 5-triphosphate - AppA P₁,P₂-diadenosine 5-diphosphate - pNp (N = A,U) nucleotide 2(3), 5-diphosphate - ImpA adenosine 5-phosphoreimidazolide - ImpU uridine 5-phosphorimidazolide - A ²pA adenylyl-[25]-adenosine - A ³pA adenylyl-[35]-adenosine - pA ²pA 5-phospho-adenylyl-[25]-adenosine - pA ³pA 5-phospho-adenylyl-[35]-adenosine - pUpU 5-phospho-uridylyl-uridine - pApU 5-phospho-adenylyl-uridine - pUpA 5-phospho-uridylyladenine - (pA)n (n, 2,3,4,) oligoadenylates with 5 terminal phosphate - ImpApA 5-phosphorimidazolide of adenylyl adenosine - (pA) ₅₊ pentamer and higher oligoadenylates with 5 terminal phosphate - (Ap)nA (n = 2,3,4) oligoadenylates without terminal phosphates In the following we do not specify the nature of the internucleotide linkageIn the following we do not specify the nature of the internucleotide linkage