Steady State of Photosynthetic Electron Transport in Cells of the Cyanophyti Synechocystis PCC 6714 Having Different Stoichiometry between PS I and PS II: Analysis of Flash-Induced Oxidation-Reduction of Cytochrome f and P700 under Steady State of Photosynthesis

The steady state of photosynthetic electron transport drivenby two photosystems was studied with cells of the cyanophyteSynechocystis PCC 6714 by analyzing the flash-induced oxidation-reductionof Cyt f and P700 under continuous background illumination.We first analyzed the spectra and the kinetics of flash-inducedabsorption changes in the 400 to 440 nm wavelength region anddefined the absorption changes due to oxidation-reduction ofCyt f and P700. Results indicated that the flash-induced absorptionchanges at 420 and 435 nm are due to the oxidation-reductionof Cyt f and P700, respectively. Determination of the steadystate of Cyt f (420 nm) and P700 (435 nm) was made for the cellsgrown under a weak orange light exciting mainly PS II (PS IIlight) and having a high ratio of PS I to PS II (PS I/PS II),and those grown under a weak red light exciting preferentiallyPS I (PS I light) and having a low PS I/PS II. The steady stateof electron transport in cells of the two types were comparedunder PS I and PS II lights. The results indicated that: (1)under the light conditions used for growth (both red and orangelight), the intermediate electron pool between the two photosystemsremained in a redox state so as to keep both photosystems inthe open state. (2) When shifted to PS I light, the intermediatepool and PS I in cells of high PS I/PS II became extremely electron-poor,and so most of the PS I reaction centers were closed. (3) Theintermediate pool in cells of low PS I/PS II became extremelyelectron-rich when shifted to PS II light, and most of the PSII reaction centers were closed. The electron transport stateis released from such biased states by regulation of PS I/PSII. Results supported our previously proposed hypothesis thatthe stoichiometry between PS I and PS II is regulated so asto keep the two photosystems in the open state. The relationshipbetween the steady state of electron transport and the regulationof PS I/PS II is discussed. (Received August 2, 1990; Accepted December 10, 1990)     

Correlation of the cytochrome c 2 content of cyanobacterial Photosystem II with the EPR properties of the oxygen-evolving complex

The Mn₄ cluster of PS II advances through a series of oxidation states (S states) that catalyze the breakdown of water to dioxygen in the oxygen-evolving complex. The present study describes the engineering and purification of highly active PS II complexes from mesophilic His-tagged Synechocystis PCC 6803 and purification of PS II core complexes from thermophilic wild-type Synechococcus lividus with high levels of the extrinsic polypeptide, cytochrome c ₅₅₀. The g = 4.1 S₂ state EPR signal, previously not characterized in untreated cyanobacterial PS II, is detected in high yields in these PS II preparations. We present a complete characterization of the g = 4.1 state in cyanobacterial His-tagged Synechocystis PCC 6803 PS II and S. lividus PS II. Also presented are a determination of the stoichiometry of cytochrome c ₅₅₀ bound to His-tagged Synechocystis PCC 6803 PS II and analytical ultracentrifugation results which indicate that cytochrome c ₅₅₀ is a monomer in solution. The temperature-dependent multiline to g = 4.1 EPR signal conversion observed for the S₂ state in cyanobacterial PS II with high cytochrome c ₅₅₀ content is very similar to that previously found for spinach PS II. In spinach PS II, the formation of the S₂ state g = 4.1 EPR signal has been found to correlate with the binding of the extrinsic 17 and 23 kDa polypeptides. The finding of a similar correlation in cyanobacterial PS II with the binding of cytochrome c ₅₅₀ suggests a functional homology between cytochrome c ₅₅₀ and the 17 and 23 kDa extrinsic proteins of spinach PS II. This revised version was published online in June 2006 with corrections to the Cover Date.     

Quantitative analysis of 77K fluorescence emission spectra in Synechocystis sp. PCC 6714 and Chlamydomonas reinhardtii with variable PS I/PS II stoichiometries

Low-temperature (77 K) fluorescence emission spectra of intact cells of a cyanobacterium, Synechocystis sp. PCC 6714, and a green alga, Chlamydomonas reinhardtii, were quantitatively analyzed to examine differences in PS I/PS II stoichiometries. Cells cultured under different spectral conditions had various PS I/PS II molar ratios when estimated by oxidation-reduction difference absorption spectra of P700 (for PS I) and Cyt b-559 (for PS II) with thylakoid membranes. The fluorescence emission spectra under the Chl a excitation at 435 nm were resolved into several component bands using curve-fitting methods and the relative band area between PS II (F685 and F695) and PS I (F710 or F720) emissions was compared with the PS I/PS II stoichiometries of the various cell types. The results indicated that the PS I/PS II fluorescence ratios correlated closely with photosystem stoichiometries both in Synechocystis sp. PCC 6714 and in C. reinhardtii grown under different light regimes. Furthermore, the correlation between the PS I/PS II fluorescence ratios and the photosystem stoichiometries is also applicable to vascular plants.     

Autotrophic cells of the <Emphasis Type="Italic">Synechocystis psbH</Emphasis> deletion mutant are deficient in synthesis of CP47 and accumulate inactive PS II core complexes

Cells of the psbH deletion mutant IC7 of the cyanobacterium Synechocystis PCC 6803 grown in the absence of glucose contain strongly reduced levels of chlorophyll when compared with cells grown in the presence of glucose, or compared with wild-type (WT) cells. Low-temperature fluorescence emission spectra revealed decreased content of both active PS II (Photosystem II) and PS I (Photosystem I) complexes. Analysis of thylakoid membrane complexes of IC7 by native electrophoresis showed a similar set of chlorophyll–proteins, namely a PS II core complex and trimeric and monomeric PS II complexes, as in WT. However, in contrast to WT, the ³⁵S-methionine protein labeling pattern of the mutant exhibited no preferential labeling of the D1 protein in the PS II core complexes, and the labeled D1 and D2 proteins accumulated predominantly in the PS II reaction center lacking CP47. The results show that in autotrophically grown cells of the psbH deletion mutant, selective D1 turnover is inhibited and synthesis of CP47 becomes a limiting step in the PS II assembly.     

Regulation of the Stoichiometry of Thylakoid Components in the Photosynthetic System of Cyanophytes: Model Experiments Showing that Control of the Synthesis or Supply of Chl a can Change the Stoichiometric Relationship between the Two Photosystems

Regulation of the assembly of the photosystem I (PS I) complexin response to the light regime in the photosynthetic systemof cyanophytes was studied in Synechocystis PCC 6714. The relationshipbetween the assembly of the PS I complex and synthesis of Chla was examined by model experiments in which synthesis of Chla was controlled by two inhibitors, gabaculine (GAB) and 2,2'-dipyridyl(DP). Both inhibitors caused a change to a lower ratio of PSI to PS II even under light that normally induces a high ratioof PS I to PS II. The change in stoichiometry induced by theseinhibitors was suppressed when protein synthesis was inhibitedby chloram-phenicol, similarly to the change in the stoichiometryinduced by light that excites mainly PS I (PS I light). Comparisonof the levels of PS I, PS II and Cyt b₆-f complexes per cellindicated that a selective suppression of the assembly of thePS I complex was induced by the inhibitors: the stoichiometricrelationship among PS I, PS II and Cyt b₆-f complexes was identicalto that induced by PS I light or white light of high intensity.GAB induced a decrease in size of the phycobilisome also, whileDP did not, similarly to PS I light. The results indicate thatthe ratio of PS I to PS II can be changed by the control ofsynthesis of Chl a. They also suggest that control of the synthesisor supply of Chl a probably exerted at site(s) in or after theprocess of the Mg-protoporphyrin branch, is involved in themechanism of regulation of the assembly of the PS I complexin cyanophytes. (Received September 7, 1989; Accepted November 20, 1989)     

The consequences of chlorophyll deficiency for photosynthetic light use efficiency in a single nuclear gene mutation of cowpea

The light harvesting and photosynthetic characteristics of a chlorophyll-deficient mutant of cowpea (Vigna unguilata), resulting from a single nuclear gene mutation, are examined. The 40% reduction in total chlorophyll content per leaf area in the mutant is associated with a 55% reduction in pigment-proteins of the light harvesting complex associated with Photosystem II (LHC II), and to a lesser extent (35%) in the light harvesting complex associated with Photosystem I (LHC I). No significant differences were found in the Photosystem I (PS I) and Photosystem II (PS II) contents per leaf area of the mutant compared to the wildtype parent. The decreases in the PS I and PS II antennae sizes in the mutant were not accompanied by any major changes in quantum efficiencies of PS I and PS II in leaves at non-saturating light levels for CO₂ assimilation. Although the chlorophyll deficiency resulted in an 11% decrease in light absorption by mutant leaves, their maximum quantum yield and light saturated rate of CO₂ assimilation were similar to those of wildtype leaves. Consequently, the large and different decreases in the antennae of PS II and PS I in the mutant are not associated with any loss of light use efficiency in photosynthesis.Abbreviations LHC I, LHC II light harvesting chlorophyll a/b protein complexes associated with PS I and PS II - A₈₂₀ light-induced absorbance change at 820 nm - ø_{PS I}, ø_{PS II} relative quantum efficiencies of PS I and PS II photochemistry     

Prolonged high light treatment of plant cells results in changes of the amount,the localization and the electrophoretic behavior of several thylakoid membrane proteins

The effect of a 30 h high light treatment on the amount and the localization of thylakoid proteins was analysed in low light grown photoautotrophic cells of Marchantia polymorpha and Chenopodium rubrum. High light treatment resulted in a net loss of D1 protein which was accompanied by comparable losses of other proteins of the PS II core (reaction center with inner antenna). LHC II proteins were not reduced correspondingly, indicating that these complexes are less affected by prolonged high light. High light influenced the distribution of PS II components between the grana and the stroma region of the thylakoid membrane, probably by translocation of the respective PS II proteins. Additionally, modifications of several thylakoid proteins were detected in high light treated cells of C. rubrum. These effects are discussed in relation to photoinhibitory damage and repair processes.Abbreviations BCA bioinchonic acid - chl chlorophyll - CF₁ coupling factor - CYC cycloheximide - GT grana thylakoids - HL high light - LL low light - PAGE polyacrylamide gel electrophoresis - PFD photon flux density - PS I Photosystem I - PS II Photosystem II - RC reaction center - SDS sodium dodecylsulfate - ST stroma thylakoids - Thyl unfractionated thylakoids     

Decrease of polypeptides in the PS I antenna complex with increasing growth irradiance in the red alga Porphyridium cruentum

Thylakoids isolated from cells of the red alga Porphyridium cruentum exhibit an increased PS I activity on a chlorophyll basis with increasing growth irradiance, even though the stoichiometry of Photosystems I and II in such cells shows little change (Cunningham et al. (1989) Plant Physiol 91: 1179–1187). PS I activity was 26% greater in thylakoids of cells acclimated at 280 mol photons · m^–2 · s^–1 (VHL) than in cells acclimated at 10 mol photons · m^–2 · s^–1 (LL), indicating a change in the light absorbance capacity of PS I. Upon isolating PS I holocomplexes from VHL cells it was found that they contained 132±9 Chl/P700 while those obtained from LL cells had 165±4 Chl/P700. Examination of the polypeptide composition of PS I holocomplexes on SDS-PAGE showed a notable decrease of three polypeptides (19.5, 21.0 and 22 kDa) in VHL-complexes relative to LL-complexes. These polypeptides belong to a novel LHC I complex, recently discovered in red algae (Wolfe et al. (1994a) Nature 367: 566–568), that lacks Chl b and includes at least six different polypeptides. We suggest that the decrease in PS I Chl antenna size observed with increasing irradiance is attributable to changes occurring in the LHC I-antenna complex. Evidence for a Chl-binding antenna complex associated with PS II core complexes is lacking at this point. LHC II-type polypeptides were not observed in functionally active PS II preparations (Wolfe et al. (1994b) Biochimica Biophysica Acta 1188: 357–366), nor did we detect polypeptides that showed immunocross-reactivity with LHC II specific antisera (made to Chlamydomonas and Euglena LHC II).Abbreviations Bis-Tris bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane - DCPIP 2,6-dichlorophenol indophenol - -dm dodecyl--d-maltoside - HL high light of 150 mol photons · m^–2 · s^–1 - LGB lower green band - LHC I light-harvesting complex of PS I - LHC II light-harvesting complex of PS II - LL low light of 10 mol photons · m^–2 · s^–1 - ML medium light of 50 mol photons · m^–2 · s^–1 - MES 2-(N-morpholino) ethanesulfonic acid - P700 reaction center of PS I - PFD photon flux density - Trizma tris(hydroxymethyl)aminomethane - UGB upper green band - VHL very high light of 280 mol photons · m^–2 · s^–1     

The role of ApcD and ApcF in energy transfer from phycobilisomes to PS I and PS II in a cyanobacterium

The role of the phycobilisome core components, ApcD and ApcF, in transferring energy from the phycobilisome to PS I and PS II in the cyanobacterium Synechocystis sp. PCC6803 has been investigated. The genes encoding these proteins have been disrupted in the genomes of wild type Synechocystis sp. PCC6803 and a PS II deficient mutant, PsbD1CD2^-, by inserting antibiotic resistance genes into their coding regions. Data from fluorescence emission spectra and pigment content analysis for these inactivation mutants is presented. These data suggest that both ApcD and ApcF are involved in the energy transfer route to PS II and PS I. In both cases, the energy transfer may to the reaction centres may be via the chromophore of ApcE (the L cm) or anchor polypeptide). The major route of energy transfer to both kinds of reaction centre appears to involve ApcF rather than ApcD. When both ApcF and ApcD are absent, the phycobilisomes are unable to transfer energy to either reaction centre. We suggest a model for the pathways of energy transfer from the phycobilisomes to PS I and PS II.     

低温光抑制恢复过程中黄瓜叶片PSⅡ活性及其电子传递对PSⅠ的影响

以“津春4号”黄瓜为试材,通过测定黄瓜叶片叶绿素荧光快速诱导动力学曲线和对820 nm光的吸收曲线,结合叶绿素荧光淬灭分析,研究低温光胁迫(4℃,200 μmol·m-2·s-1)6 h后,黄瓜叶片在常温(25℃)不同光强(0、15、200μmol·m-2·s-1)下PS Ⅰ和PS Ⅱ活性的恢复,以及恢复过程中PS Ⅰ与PS Ⅱ的相互作用.结果表明:低温光胁迫6h后,PS Ⅰ和PS Ⅱ发生不同程度的光抑制.在常温恢复阶段,PS Ⅱ活性快速恢复且对光强不敏感;PS Ⅰ活性在弱光下(15 μmol·m-2·s-1)快速恢复,在较强光(200 μmol·m-2·s-1)下恢复较慢.在低温光抑制恢复过程中,常温下PS Ⅱ活性恢复较快可能导致PS Ⅱ向PS Ⅰ的线性电子传递过快,进而抑制PS Ⅰ的活性恢复.因此,在进行黄瓜抗冷性育种时,不应该仅追求较高的PS Ⅱ抗性和较快的PS Ⅱ恢复速度,还应该注意两个光系统活性的协调.在生产中,应当在低温逆境发生及其之后较长一段时间内采取措施降低叶表面光照强度,以利于对植株光合机构的保护和光合活性的恢复.     

Regulation of Photosystem Composition in Cyanobacterial Photosynthetic System: Evidence Indicating that Photosystem I Formation is Controlled in Response to the Electron Transport State

The quantitative composition of thylakoid components such asthe photosystem (PS) I/PS II ratio in the cyanobacterial photosyntheticsystem is regulated in response to the light regime. The regulationoccurs as changes in PS I content due to control of either PSI formation or decomposition. In order to determine which ofthese two is controlled in this regulation, experiments wereperformed to determine the light-induced PS I decrease in cellsof Synechocystis PCC 6714 under conditions where protein synthesiswas suppressed, i.e. the incubation without a nitrogen sourcefor cell growth or with chloramphenicol. The results revealedthat light-induced PS I decrease did not occur when synthesisof the thylakoid system was suppressed by incubation withouta nitrogen source or by the addition of chloramphenicol, indicatingthat (1) the thylakoid composition is regulted in the processof thylakoid formation and (2) the regulation is achieved bythe control of PS I formation. (Received November 6, 1987; Accepted March 2, 1988)     

Oxygen evolving membranes and particles from the transformable cyanobacterium Synechocystis sp. PCC6803

Membranes and PS II particles retaining high rates of O₂-evolving activity have been isolated from the transformable cyanobacterium, Synechocystis sp. PCC6803. Membranes from cells grown under red light exhibit rates of O₂-evolution ranging from 500–700 mole O₂/mg chl/h. PS II particles are prepared by a simple procedure involving DEAE column chromatography of detergent extracts obtained by simultaneous treatment of membranes with octylglucoside and dodecylmaltoside. The isolated PS II fraction is enriched in polypeptides immunologically cross-reactive with polypeptides present in core reaction center preparations of spinach, exhibits 77 K fluorescence emission maxima at 685 and 696 nm, but not emission and absorption due to phycobilines and is capable of rates of O₂-evolution exceeding 1000 mole O₂/mg chl/h.Abbreviations DM dodecyl--D-maltoside - OG octyl--D-glucoside     

Consequences of LHC II deficiency for photosynthetic regulation in chlorina mutants of barley

The light-harvesting chlorophyll a/b proteins associated with PS II (LHC II) are often considered to have a regulatory role in photosynthesis. The photosynthetic responses of four chlorina mutants of barley, which are deficient in LHC II to varying degrees, are examined to evaluate whether LHC II plays a regulatory role in photosynthesis. The efficiencies of light use for PS I and PS II photochemistry and for CO₂ assimilation in leaves of the mutants were monitored simultaneously over a wide range of photon flux densities of white light in the presence and absence of supplementary red light. It is demonstrated that the depletions of LHC II in these mutants results in a severe imbalance in the relative rates of excitation of PS I and PS II in favour of PS I, which cannot be alleviated by preferential excitation of PS II. Analyses of xanthophyll cycle pigments and fluorescence quenching in leaves of the mutants indicated that the major LHC II components are not required to facilitate the light-induced quenching associated with zeaxanthin formation. It is concluded that LHC II is important to balance the distribution of excitation energy between PS I and PS II populations over a wide range of photon flux densities. It appears that LHC II may also be important in determining the quantum efficiency of PS II photochemistry by reducing the rate of quenching of excitation energy in the PS II primary antennae.Abbreviations Fm, Fv maximal and variable fluorescence yields in a light adapted state - LHC II light harvesting chlorophyll a/b protein complex associated with PS II - q_p photochemical quenching - A₈₂₀ light-induced absorbance change at 820 nm - ø_PSI, ø_PSII relative quantum efficiencies of PS I and PS II photochemistry - øCO₂ quantum yield of CO₂ assimilation     

Low concentrations of NaHSO3 increase cyclic photophosphorylation and photosynthesis in cyanobacterium Synechocystis PCC6803

Application of NaHSO₃ solution at low concentrations (20–200 μM) to the culture medium enhanced photosynthetic oxygen evolution in cyanobacterium Synechocystis PCC6803 by more than 10%. The slow phase of ms-DLE was strengthened, showing that the transmembrane proton motive force related to photophosphorylation was enhanced. It was also observed that dry weight as well as ATP content under illuminated conditions were both increased after the treatment, indicating that low concentrations of NaHSO₃ could enhance the supply of ATP and thus increase biomass accumulation. In accord with the promotion in the photosynthetic oxygen evolution and ATP content, the transient increase in chlorophyll fluorescence after the termination of actinic light was increased; and meanwhile, the half-time of re-reduction of P700⁺ in the presence of DCMU after a pulse light under background far-red light was shortened by approximately 30%, indicating that cyclic electron flow around PS I was accelerated by the treatment. Based on these results it is suggested that the increase in photosynthesis in Synechocystis PCC6803 caused by low concentrations of NaHSO₃ solution might be due to the stimulation of the cyclic electron flow around PS I and thus the increase in photophosphorylation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Oxidizing side of the cyanobacterial Photosystem I: Mutational analysis of the luminal H loop of the PsaB subunit

PS I core proteins are expected to interact with the electron donor proteins plastocyanin or cytochrome c ₆. To investigate the role of the luminal H loop of PsaB in the assembly and function of the PS I complex, we generated 15 deletion and repetition mutations in the H loop of the PsaB protein from Synechocystis sp. PCC 6803. The mutant strains differed in their photoautotrophic growth. The PS I proteins could not be detected in the membranes of mutants in which the N438–E448, I453–T464, or S500–G512 region was deleted from the PsaB protein, indicating the essential role of these segments in proper folding of the PsaB protein. Mutants with partial or complete deletion of the L469–D496 segment contained the PS I proteins. These results indicate that the regions near the transmembrane helices are more important for the assembly of PsaB than the middle region of the H loop. The L469-D496 segment in the H loop of PsaB is dispensable in the interaction between the PS I complex and the soluble donor proteins. These results suggested that sections of the H loop of PsaB are crucial for the structural integrity of the PsaB protein.     

Thf1 interacts with PS I and stabilizes the PS I complex in Synechococcus sp. PCC7942

Thylakoid formation1 protein (Thf1) is a multifunctional protein that is conserved in all photosynthetic organisms. In this study, we used the model cyanobacterium Synechococcus sp. PCC7942 (hereafter Synechococcus) to show that the level of Thf1 is altered in response to various stress conditions. Although this protein has been reported to be involved in thylakoid formation, the thylakoid membrane in the thf1 deletion strain (ΔThf1) was not affected. Compared with the WT, ΔThf1 showed reduced PS II activity, with increased levels of D1 under high light (HL) conditions, which was resulted from blocked D1 degradation by the FtsH protease and thus inhibits PS II repair. PS I was found to be more seriously affected than PS II in ΔThf1, even under low light conditions, suggesting that PS I damage could be the primary effect of thf1 deletion in Synechococcus. Further analysis revealed that the ΔThf1 mutant had a lower PS I subunit content and lower PS I stability under HL conditions. Further sucrose gradient fractionation of the membrane protein complexes and crosslinking and immunoblot analysis indicated that Thf1 interacts with PS I. Together, our results reveal that Thf1 interacts with PS I and thereby stabilizes PS I in Synechococcus.     

Changes in the photosynthetic apparatus in the cyanobacterium Synechocystis sp. PCC 6714 following light-to-dark and dark-to-light transitions

The photosynthetic apparatus of Synechocystis sp. PCC 6714 cells grown chemoheterotrophically (dark with glucose as a carbon source) and photoautotrophically (light in a mineral medium) were compared. Dark-grown cells show a decrease in phycocyanin content and an even greater decrease in chlorophyll content with respect to light-grown cells. Analysis of fluorescence emission spectra at 77 K and at 20 °C, of dark- and light-grown cells, and of phycobilisomes isolated from both types of cells, indicated that in darkness the phycobiliproteins were assembled in functional phycobilisomes (PBS). The dark synthesized PBS, however, were unable to transfer their excitation energy to PS II chlorophyll. Upon illumination of dark-grown cells, recovery of photosynthetic activity, pigment content and energy transfer between PBS and PS II was achieved in 24–48 h according to various steps. For O₂ evolution the initial step was independent of protein synthesis, but the later steps needed de novo synthesis. Concerning recovery of PBS to PS II energy transfer, light seems to be necessary, but neither PS II functioning nor de novo protein synthesis were required. Similarly, light, rather than functional PS II, was important for the recovery of an efficient energy transfer in nitrate-starved cells upon readdition of nitrate. In addition, it has been shown that normal phycobilisomes could accumulate in a Synechocystis sp. PCC 6803 mutant deficient in Photosystem II activity.Abbreviations APC allophycocyanin - CAP chloroamphenicol - Chl chlorophyll - DCMU 3(3,4-dichlorophenyl)-1,1-dimethylurea - CP-47 chlorophyll-binding Photosystem II protein of 47 kDa - EF exoplasmic face - PBS phycobilisome - PC phycocyanin - PS Photosystem     

Regulation of Photosystem Stoichiometry in the Photosynthetic System of the Cyanophyte Synechocystis PCC 6714 in Response to Light-Intensity

Light-induced changes in stoichiometry among three thylakoidcomponents, PS I, PS II and Cyt b₆-f complexes, were studiedwith the cyanophyte Synechocystis PCC 6714. Special attentionwas paid to two aspects of the stoichiometric change; first,a comparison of the patterns of regulation in response to differencesin light-intensity with those induced by differences in light-quality,and second, the relationship between regulation of the stoichiometryand the steady state of the electron transport system. Resultsfor the former indicated that (1) the abundance of PS I on aper cell basis was reduced under white light at the intensityas high as that for light-saturation of photosynthesis, butPS I per cell was increased under low light-intensity, (2) PSII and Cyt b₆-f complexes remained fairly constant, and (3)changes in the abundance of PS I depended strictly on proteinsynthesis. The pattern was identical with that of chromaticregulation. For the second problem, the redox steady-statesof Cyt f and P700 under white light of various intensities weredetermined by flash-spectroscopy. Results indicated that (1)Cyt f and P700 in cells grown under low light-intensity [highratio of PS I to PS II (PS I/PS II)] were markedly oxidizedwhen the cells were exposed to high light-intensity, while theyremained in the reduced state under low light-intensity. (2)After a decrease in the abundance of PS I, most of P700 remainedin the reduced state even under high light-intensity, whilethe level of reduced Cyt f remained low. (3) Both Cyt f andP700 in cells of low PS I/PS II were fully reduced under lowlight-intensity, and Cyt f reduction following the flash wasrapid, which indicates that the turnover of PS I limits theoverall rate of electron flow. After an increase in the abundanceof PS I, the electron transport recovered from the biased state.(4) The redox steady-state of the Cyt b₆-f complex correlatedwell with the regulation of PS I/PS II while the state of thePQ pool did not. Based on these results, a working model ofthe regulation of assembly of the PS I complex, in which theredox steady-state of the Cyt b₆-f complex is closely relatedto the primary signal, is proposed. (Received August 2, 1990; Accepted December 10, 1990)     

A New 4.8-kDa Polypeptide Intrinsic to the PS II Reaction Center, as Revealed by Modified SDS-PAGE with Improved Resolution of Low-Molecular-Weight Proteins

Low-molecular-weight polypeptides in various PS II preparationsfrom spinach and wheat were analyzed by modified SDS-PAGE, whichgave good resolution of low-molecular-weight proteins with minimizedinterference by lipids. PS II membrane fragments contained atleast nine low-molecular-weight polypeptides of between 3.9kDa and 11 kDa, and all of them were identified in thylakoidmembranes. Of these nine polypeptides, the 10-kDa phosphoprotein,the 5-kDa, 4.8-kDa, and 4.1-kDa polypeptides, and the two subunitsof cytochrome b₅₅₉ were commonly found in O₂-evolving core complexesof wheat and spinach. In contrast, PS II reaction center complexesthat consisted of D1, D2 and two cytochrome b₅₅₉ polypeptidesretained only the 4.8- kDa polypeptide. Analysis by Westernblotting revealed that the 4.8-kDa polypeptide is an intrinsiccomponent of the PS II reaction center. (Received May 30, 1988; Accepted August 19, 1988)