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     

Complex formation in plant thylakoid membranes. Competition studies on membrane protein interactions using synthetic peptide fragments

Thylakoid membranes of pea were used to study competition between extra-membrane fragments and their parental membrane-bound proteins. Phosphorylated and unphosphorylated fragments of light harvesting complex II (LHC II) from higher plants were used to compete with LHC II for interactions with itself and with other thylakoid protein complexes. Effects of these peptide fragments of LHC II and of control peptides were followed by 80 K chlorophyll fluorescence spectroscopy of isolated thylakoids. The phosphorylated LHC II fragment competes with membrane-bound phosphoproteins in the phosphatase reaction. The same fragment accelerates the process of dark-to-light adaptation and decreases the rate of the light-to-dark adaptation when these are followed by fluorescence spectroscopy. In contrast, the non-phosphorylated LHC II peptide does not affect the rate of adaptation but produces results consistent with inhibition of formation of a quenching complex. In this quenching complex we propose that LHC II remains inaccessible to the LHC II kinase, explaining an observed decrease in LHC II phosphorylation in the later stages of the time-course of phosphorylation. The most conspicuous protein which is steadily phosphorylated during the time-course of phosphorylation is the 9 kDa (psbH) protein. The participation of the phosphorylated form of psbH in the quenching complex, where it is inaccessible to the phosphatase, may explain its anomalously slow dephosphorylation. The significance of the proposed complex of LHC II with phospho-psbH is discussed.Abbreviations LHC II light harvesting complex II - PS II Photosystem II - PS I Photosystem I     

Polyamines in the photosynthetic apparatus

The three main polyamines putrescine (Put), spermidine (Spd) and spermine (Spm) were characterized by HPLC in intact spinach leaf cells, intact chloroplasts, thylakoid membranes, Photosystem II membranes, the light-harvesting complex and the PS II complex. All contain the three polyamines in various ratios; the HPLC polyamine profiles of highly resolved PS II species (a Photosystem II core and the rection center) suggest an enrichment in the polyamine Spm.Abbreviations Chl chlorophyll - HPLC high performance liquid chromatography - LHC light-harvesting complex - PS II Photosystem II - PS II-RC Photosystem II reaction center - Put putrescine - Spd spermidine - Spm spermine - 10%S-core D₁-D₂-Cyt b559-47 kD-43 kD complex     

Evaluation of the role of State transitions in determining the efficiency of light utilisation for CO2 assimilation in leaves

Wheat leaves were exposed to light treatments that excite preferentially Photosystem I (PS I) or Photosystem II (PS II) and induce State 1 or State 2, respectively. Simultaneous measurements of CO₂ assimilation, chlorophyll fluorescence and absorbance at 820 nm were used to estimate the quantum efficiencies of CO₂ assimilation and PS II and PS I photochemistry during State transitions. State transitions were found to be associated with changes in the efficiency with which an absorbed photon is transferred to an open PS II reaction centre, but did not correlate with changes in the quantum efficiencies of PS II photochemistry or CO₂ assimilation. Studies of the phosphorylation status of the light harvesting chlorophyll protein complex associated with PS II (LHC II) in wheat leaves and using chlorina mutants of barley which are deficient in this complex demonstrate that the changes in the effective antennae size of Photosystem II occurring during State transitions require LHC II and correlate with the phosphorylation status of LHC II. However, such correlations were not found in maize leaves. It is concluded that State transitions in C₃ leaves are associated with phosphorylation-induced modifications of the PS II antennae, but these changes do not serve to optimise the use of light absorbed by the leaf for CO₂ assimilation.Abbreviations Fm, Fo, Fv maximal, minimal and variable fluorescence yields - 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 - _{PS I}, _{PS II} relative quantum efficiencies of PS I and PS II photochemistry - _{CO 2} quantum yield of CO₂ assimilation     

Occurrence of the carotenoid lactucaxanthin in higher plant LHC II

The pigment composition of the light-harvesting complexes of Photosystem II (LHC II) has been determined for lettuce (Lactuca sativa). In common with other members of the composite, the photosynthetic tissues of this species may contain large amounts of the carotenoid lactucaxanthin (, -carotene-3,3'-diol) in addition to their normal compliment of carotenoids. The occurrence and distribution of lactucaxanthin in LHC II has been examined using isoelectric focusing of BBY particles followed by reversed-phase HPLC analysis of the pigments. The major carotenoids detected in LHC IIb, LHC IIa (CP29) and LHC IIc (CP26) purified from dark-adapted lettuce were lutein, violaxanthin, neoxanthin and lactucaxanthin. Lactucaxanthin has been shown to be a major component of PS II, accounting for 26% of total xanthophyll in both LHC IIb (23% total xanthophyll) and in the minor complexes (12–16%). In this study, LHC IIb was clearly resolved into four bands and their carotenoid composition determined. These four bands proved to be very similar in their pigment content and composition, although the relative amounts of neoxanthin and lutein in particular were found to increase from bands 1 to 4 (i.e. with increasing electrophoretic mobility). The operation of the xanthophyll cycle has also been examined in the LHC of L. sativa following light treatment. The conversion efficiency for violaxanthinzeaxanthin was nearly identical for each light-harvesting complex examined at 58–61%. Nearly half of the zeaxanthin formed in PS II was associated with LHC IIb, although the molar ratio of zeaxanthin:chlorophyll a was highest in the minor LHC.Abbreviations HPLC high performance liquid chromatography - IEF isoelectric focusing - LHCII light-harvesting complex associated with Photosystem II - PS II Photosystem II - qE pH-dependent nonphotochemical quenching of chlorophyll fluorescence     

Heat-induced reversible changes in Photosystem 1 absorption cross-section of pea chloroplasts and sub-chloroplast preparations

Reversible changes in the room temperature fluorescence quenching at 685 nm and light scattering level at 577 nm, indicating about 15% of granal unstacking, induced by high temperature treatment (40°C, for 5 min) of pea chloroplasts were shown. Analysis of the low temperature excitation fluorescence spectra of the 735 nm Photosystem 1 (PS 1) band (F735), in the 635–725 nm region, has revealed the involvement of light-harvesting (LHC 2, maxima at 650 and 676 nm) and the proximal Photosystem 2 antenna (maxima 668, 687 nm) in heat-induced enhancement of the PS 1 long wavelength antenna absorption cross-section. It was found that the two PS 1 sub-chloroplast preparations, achieved by the digitonin method, possessed different characteristics of this enhancement. For the heavier fraction (100 000 g) the additional absorption cross-section was formed mostly at the expense of PS 2 antennas (apparently spillover), but for the lighter PS 1 fraction (145 000 g) the changes have indicated an -transfer mechanism, i.e., participation of only LHC 2 in the energy transfer towards PS 1. This may indicate the heterogeneous character of the temperature-induced energy redistribution across the PS 1-containing chloroplast membrane compartments. The model of heat-induced changes in the pigment-protein complex arrangement is discussed in terms of domain organisation of the thylakoid membrane.Abbreviations Chl a/b ratio between chlorophyll a and chlorophyll b concentrations - CP43 and CP47 proximal Photosystem 2 antenna complexes - D1/D2 complex Photosystem 2 reaction centre complex - EDTA ethylenediaminetetraacetic acid - F685 and F696 Photosystem 2 low temperature fluorescence bands - F735 Photosystem 1 low temperature fluorescence band - Fp free pigment band in green gel electrophoresis - LHC 2 light-harvesting chlorophyll a/b complex - LHCP I, II and III light-harvesting bands in green gel electrophoresis - Cp1 and Cpa bands in green gel electrophoresis which are associated with Photosystem 1 and 2 reaction centre complexes with internal antennas - P700 Photosystem 1 reaction centre - PPC pigment-protein complex - PS 1 and Photosystem 1 alpha and Photosystem 1 beta - PS 2 and Photosystem 2 alpha and Photosystem 2 beta - RC reaction centre - SDS-PAGE sodiumdodecylsulphate-polyacrylamide gel electrophoresis - St1-St2 state-1-state-2 transitions     

Selective extraction of CP 26 and CP 29 proteins without affecting the binding of the extrinsic proteins (33, 23 and 17 kDa) and the DCMU sensitivity of a Photosystem II core complex

A highly purified oxygen evolving Photosystem II core complex was isolated from PS II membranes solubilized with the non-ionic detergent n-octyl--D-thioglucoside. The three extrinsic proteins (33, 23 and 17 kDa) were functionally bound to the PS II core complex. Selective extraction of the 22, 10 kDa, CP 26 and CP 29 proteins demonstrated that these species are not involved in the binding of the extrinsic proteins (33, 23 and 17 kDa) or the DCMU sensitivity of the Photosystem II complex.Abbreviations Chl chlorophyll - DCBQ 2,6-dichloro-p-benzoquinone - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - LHC light-harvesting complex - MES 2-(N-morpholino)ethanesulfonic acid - OGP n-octyl--d-glucoside - OTG n-octyl--d-thioglucoside - PAGE polyacrylamide gel electrophoresis - PS II Photosystem II - SDS sodium dodecyl sulfate     

Characterization of a 160 kD Photosystem II reaction center complex isolated from photoinhibited Dunaliella salina thylakoids

Photoinhibition in the green alga Dunaliella salina is accompanied by the formation of inactive Photosystem II reaction centers. In SDS-PAGE analysis, the latter appear as 160 kD complexes. These complexes are structurally stable, enough to withstand re-electrophoresis of excised gel slices from the 160 kD region. Western blot analyses with specific polyclonal antibodies raised against the D1 or D2 reaction center proteins provided evidence for the presence of both of these polypeptides in the re-electrophoresed 160 kD complex. Incubation of excised gel slices from the 160 kD region, under aerobic conditions at 4°C for a prolonged period of time, caused a break-up of the 160 kD complex into a 52 kD D1-containing and 80 and 26 kD D2-containing pieces. Western blot analysis with polyclonal antibodies raised against the apoproteins of CPI (reaction center proteins of PS I) did not show cross-reaction either with the 160 kD complex or with the 52, 80 and 26 kD pieces. The results show the presence of both D1 and D2 in the 160 kD complex and strengthen the notion of a higher molecular weight D1- and D2-containing complex that forms upon disassembly of photodamaged PS II units.Abbreviations Chl chlorophyll - PS II Photosystem II - D1 the 32 kD reaction center protein of PS II, encoded by the chloroplast psbA gene - D2 the 34 kD reaction center protein of PS II, encoded by the chloroplast psbD gene - CPI the 82 and 83 kD reaction center proteins of PS I, encoded by the chloroplast psaA and psaB genes - HL high light - LL low light This publication is dedicated to the memory of the late Professor Daniel Arnon, whom the first author will fondly remember for his many accounts of past scientific discovery and debate.     

The resolution and biochemical characterization of subcomplexes of the main light-harvesting chlorophyll a/b-protein complex of Photosystem II (LHC II)

LHC II isolated from carnation leaves has been solubilized and resolved by a newly developed, vertical-bed non-denaturing isoelectric focusing in polyacrylamide slab gels to yield three trimeric subcomplexes focusing at pH 4.52, 4.42 and 4.37 (designated a, b and c, respectively), comprising approximately 38%, 24% and 38% of the chlorophyll. The spectroscopic data demonstrated a close similarity among LHC II subcomplexes concerning their chlorophyll content and organization. The most alkaline and the most acidic subcomplex contained the 27 kDa polypeptide of LHC II while the intermediate pI fraction contained both LHC II polypeptides, i.e. 27 kDa and 26 kDa ones associated at 2:1 stoichiometry. The 27 kDa polypeptide could be resolved by denaturing isoelectrofocusing into 10 pI molecular isoforms covering 5.90–4.20 pH range. Three of the isoforms were found in the subcomplexes a and b and eight in the subcomplex c. The 26 kDa polypeptide comprised the unique pI molecular isoform focusing at pH 5.61.Abbreviations CBB G-250 Coomassie Brilliant Blue G-250 - chl chlorophyll - DM n-dodecyl--d-maltoside - EDTA ethylendiaminotetraacetic acid - IEF isoelectric focusing - LHC II the main light-harvesting chlorophyll a/b-protein complex of Photosystem II - LHCP II apoprotein of the main light-harvesting chlorophyll a/b-protein complex of Photosystem II - NP-40 polyethyleneglycol-p-isooctylphenyl ether - pI isoelectric point - OG octyl--d-glucopyranoside - PS II Photosystem II - SDS-PAGE sodium dodecylsulphate polyacrylamide gel electrophoresis - TCA trichlorooacetic acid     

Fluorescence studies on interaction between phospho-LHC II and subchloroplast Photosystem 1 preparations

Chloroplast proteins were phosphorylated under two test conditions: white light irradiance alone and white light irradiance with the addition of glucose and glucose oxidase, used to produce an anaerobic medium. The interaction of phospho-LHC II with Photosystem 1 (PS 1) was studied for two types of PS I preparation. Changes in the chlorophyll a/b ratio and the ratio of 650 and 680 nm band intensities (E650/E680) in fluorescence excitation spectra were used in calculating the phospho-LHC II portion which became associated with PS 1. It is shown that the associated portion of phospho-LHC II varies for each of the PS 1 preparations and phosphorylation procedures. Possible conclusions as regards the transfer of various sets of LHC II subpopulations under different phosphorylation procedures and the differences of interaction with PS 1 are discussed.Abbreviations PS 1 Photosystem 1 - PS 2 Photosystem 2 - LHC II light-harvesting chlorophyll a/b protein complex II - Chl chlorophyll - fluorescence quantum yield - _f life time of fluorescence at =685 nm - F735 fluorescence band with a maximum at 735 nm - F685 fluorescence band with a maximum at 685 nm - E650/E680 ratio of amplitudes in excitation fluorescence spectrum at 650 and 680 nm     

Competition between annihilation and trapping leads to strongly reduced yields of photochemistry under ps-flash excitation

Excitation of photosynthetic systems with short intense flashes is known to lead to exciton-exciton annihilation processes. Here we quantify the effect of competition between annihilation and trapping for Photosystem II, Photosystem I (thylakoids from peas and membranes from the cyanobacterium Synechocystis sp.), as well as for the purple bacterium Rhodospirillum rubrum. In none of the cases it was possible to reach complete product saturation (i.e. closure of reaction centers) even with an excitation energy exceeding 10 hits per photosynthetic unit. The parameter introduced by Deprez et al. ((1990) Biochim. Biophys. Acta 1015: 295–303) describing the competition between exciton-exciton annihilation and trapping was calculated to range between 4.5 (PS II) and 6 (Rs. rubrum). The rate constants for bimolecular exciton-exciton annihilation ranged between (42 ps)^-1 and (2.5 ps)^-1 for PS II and PS I-membranes of Synechocystis, respectively. The data are interpreted in terms of hopping times (i.e. mean residence time of the excited state on a chromophore) according to random walk in isoenergetic antenna.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - LHC II light harvesting complex II - P primary donor - PS I Photosystem I - PS II Photosystem II - PSU photosynthetic unit - RC reaction center     

Changes in lipid composition of Photosystem 1 particles from thylakoids phosphorylated under reductive or anaerobic conditions

Changes in lipid composition of Photosystem 1 (PS 1) particles isolated from thylakoids phosphorylated under reductive or anaerobic conditions have been studied. Under reductive conditions, there was an increase in monogalactosyldiacylglycerol containing highly saturated fatty acids and phosphatidylglycerol containing transhexadecenoic fatty acid. Under anaerobic conditions, the amount of all lipid classes was increased. As we have shown earlier (S. V. Manuilskaya, O. I. Volovik, A. I. Mikhno, A. I. Polischuk and S. M. Kochubey (1990) Photosynthetica 24: 419–423) these changes were due to a co-migration of some lipid species and light-harvesting chlorophyll a/b complex LHC II from PS 2 to PS 1. These data allow us to conclude that LHC II consists of the lipoproteins containing specific lipids. Different composition of lipids co-migrating with LHC II under various conditions of phosphorylation might be caused by the variety of LHC II subpopulations transferred under each reductive condition.Abbreviations PS 1 Photosystem 1 - PS 2 Photosystem 2 - LHC II light-harvesting chlorophyll a/b protein complex II - Chl chlorophyll - MGDG monogalactosyldiacylglycerol - DGDG digalactosyldiacylglycerol - PG phosphatidylglycerol - SQDG sulfoquinovosyldiacylglycerol     

Progress towards structural elucidation of Photosystem II

In recent years Photosystem II, and in particular the oxygen evolving component of the enzyme, have been the subject of intense biochemical and biophysical analysis. To date no high resolution structural model of the complex has been produced. As a consequence unambiguous interpretation of much experimental data has proven difficult, leading to a lack of consensus over many basic questions regarding the mechanisms involved, the oligomerization state of the enzyme in vivo and even the exact biochemical composition.This review is a summary of the progress towards the production of a structural model of PS II-derived from either X-ray crystallography or electron microscopy based techniques-and the current opinions, which have arisen from these structural analyses, on the structural topology and assemblage of the various subunits that constitute the complex.Abbreviations C₁₂-M dodecyl maltoside - CP chlorophyll protein - cyt b-559 cytochrome b-559 - DMPC dimyristoyl phosphatidyl choline - EC electron crystallography - EM electron microscopy - LHC II light harvesting complex II - OEC oxygen evolving complex - OG octyl--glucopyranoside - PS I Photosystem I - PS II Photosystem II - Tris N-tris (hydroxymethyl) amino ethane     

Selective extraction of 22 kDa and 10 kDa polypeptides from Photosystem II without removal of 23 kDa and 17 kDa extrinsic proteins

Selective solubilization of Photosystem II membranes with the non-ionic detergent octyl thioglucopyranoside has allowed the isolation of a PS II system which has been depleted of the 22 and 10 kDa polypeptides but retains all three extrinsic proteins (33, 23 and 17 kDa). The PS II membranes which have been depleted of the 22 and 10 kDa species show high rates of oxygen evolution activity, external calcium is not required for activity and the manganese complex is not destroyed by exogenous reductants. When we compared this system to control PS II membranes, we observed a minor modification of the reducing side, and a conversion of the high-potential to the low-potential form of cytochrome b ₅₅₉.Abbreviations Chl- chlorophyll - DCBQ- 2,5-dichloro-p-benzoquinone - DCMU- 3-(3,4-dichlorophenyl)-1,1-dimethylurea - ESR- electron spin resonance - MES- 2-(N-morpholino)ethanesulfonic acid - OTG- octyl--d-thioglucopyranoside - PS II- Photosystem II - PEG- polyethylene glycol, M_r=6000 - Tris- 2-amino-2-hydroxyethylpropane-1,3-diol     

Energy transfer for low temperature fluorescence in PS II mutant thylakoids

The Chl-protein complexes of three maize (Zea mays L.) mutants and one barley (Hordeum vulgare L.) mutant were analyzed using low temperature Chl fluorescence emissions spectroscopy and LDS-polyacrylamide gel electrophoresis. The maize mutants hcf-3, hcf-19, and hcf-114 all exhibited a high Chl fluorescence (hcf) phenotype indicating a disruption of the energy transfer within the photosynthetic apparatus. The mutations in each of these maize mutants affects Photosystem II. The barley mutant analyzed was the well characterized Chl b-less mutant chlorina-f2, which did not exhibit the hcf phenotype. Chlorina-f2 was used because no complete Chl b-less mutant of maize is available. Analysis of hcf-3, hcf-19, and hcf-114 revealed that in the absence of CP43, LHC II can still transfer excitation energy to CP47. These results suggest that in mutant membranes LHC II can interact with CP47 as well as CP43. This functional interaction of LHC II with CP47 may only occur in the absence of CP43, however, it is possible that LHC II is positioned in the thylakoid membranes in a manner which allows association with both CP43 and CP47.Abbreviations hcf high chlorophyll fluorescence - LDS lithium dodecyl sulfate - LHC II light-harvesting complex of Photosystem II - LHC I light-harvesting complex of Photosystem I - CPIa chlorophyll-protein complex consisting of LHC I and the PS I core complex - CPI chlorophyll-protein complex consisting of the PS I core complex - CP47 47 kDa chlorophyll-protein of the Photosystem II core - CP43 43 kDa chlorophyll-protein of the Photosystem II core - CP29 29 kDa chlorophyll-protein of Photosystem II - CP26 26 kDa chlorophyll-protein of Photosystem II - CP24 24 kDa chlorophyll-protein of Photosystem II - fp free pigments     

Photoinhibition in Chlamydomonas reinhardtii: Effect on state transition,intersystem energy distribution and Photosystem I cyclic electron flow

The energy distribution, state transitions and photosynthetic electron flow during photoinhibition of Chlamydomonas reinhardtii cells have been studied in vivo using photoacoustics and modulated fluorescence techniques. In cells exposed to 2500 W/m² light at 21 °C for 90 min, 90% of the oxygen evolution activity was lost while photochemical energy storage as expressed by the parameter photochemical loss (P.L.) at 710–720 nm was not impaired. The energy storage vs. modulation frequency profile indicated an endothermic step with a rate constant of 2.1 ms. The extent of the P.L. was not affected by DCMU but was greatly reduced by DBMIB. The regulatory mechanism of the state 1 to state 2 transition process was inactivated and the apparent light absorption cross section of photosystem II increased during the first 20 min of photoinhibition followed by a significant decrease relative to that of photosystem I. These results are consistent with the inactivation of the LHC II kinase and the presence of an active cyclic electron flow around photosystem I in photoinhibited cells.Abbreviations PS I, PS II Photosystem I and Photosystem II respectively - P.L. photochemical loss - DCMU 3-(3,4-dichlorophenyl-1,1-dimethyl urea - LHC II light harvesting chlorophyll a,b-protein complex of PS II - DBMIB 2,5 dibromo-3-methyl-6-isopropyl-p-benzoquinone     

Thermoluminescence properties of the isolated photosystem two reaction centre

Formation of thermoluminescence signals is characteristics of energy- and charge storage in Photosystem II. In isolated D1/D2/cytochrome b-559 Photosystem II reaction centre preparation four thermoluminescence components were found. These appear at -180 (Z band), between -80 and -50 (Z_v band), at -30 and at +35°C. The Z band arises from pigment molecules but not correlated with photosynthetic activity. The Z_v and -30°C bands arise from the recombination of charge pairs stabilized in the Photosystem II reaction centre complex. The +35°C band probably corresponds to the artefact glow peak resulting from a pigment-protein-detergent interaction in subchloroplast preparations (Rózsa Zs, Droppa M and Horváth G (1989) Biochim Biophys Acta 973, 350–353).Abbreviations Chl chlorophyll - Cyt cytochrome - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - D1 psbA gene product - D2 psbD gene product - P680 primary electron donor of PS II - Pheo pheophytin - PS II Photosystem II - Q_A primary quinone acceptor of PS II - Q_B secondary quinone acceptor of PS II - RC reaction centre of PS II - TL thermoluminescence     

Implication of D1 degradation in phosphorylation-induced state transitions

State transitions and lateral migration of phosphorylated mobile-LHC II upon thylakoid unstacking have been reported as being interdependent. However, now the thyakoid unstacking event can be separated from the thyakoid phosphorylation and the associated F730/F685 enhancement by using the serine-type-protease inhibitor benzamidine. Thus, lateral migration appears not be necessary, and it can be shown that LHC II-rich fragments, originating in peripheral granal membranes, can be released by digitonin although in reduced amounts. On the other hand, phosphorylation of thylakoid proteins greatly stimulates the light-induced D1 degradation, which is observed in chloroplasts phosphorylated even at very low light (15 µmol m^–2s^–1). Thylakoid pretreatment with FSBA (the PS II protein-kinase inhibitor) blocks the light-induced and ATP-stimulated D1 degradation, and the F730/F685 ratio increase; this suggests that the dissociation of the PS II unit, resulting from the introduction of repulsive negative charges ( ATP groups) into LHC II and PS II core proteins, leads to D1 degradation. In chloroplast samples transferred to darkness following short-time phosphorylation, the D1 level is recovered. The results suggest that disassembly of PS II and D1 degradation occur parallel to State transitions. The removal of outer phospho-LHC II from PS II and its association with PS I at the periphery of grana may allow D1 degradation and increased light utilization by PS I, while net de novo synthesis of D1, stimulated by ATP, may lead to the assembly of new PS II units which could bind dephosphorylated LHC II in the dark, resulting in increased light utilization by PS II.     

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     

Identification of a novel light-harvesting complex II protein (LHC IIc′)

The caroteno-chlorophyll-protein, LHC IIc, is a relatively minor component of the PS II antenna. Isolated LHC IIc contains a major protein of 28 kDa along with a 26 kDa subunit in lower abundance. Previously, it was not known if the 26 kDa protein was closely related to the 28 kDa LHC IIc protein or if it was a comigrating LHC IIb contaminating subunit. A sequence of 20 amino acid residues was obtained by direct protein micro-sequencing of an internal cyanogen bromide-derived peptide fragment of the 26 kDa protein isolated from barley. The sequence shows, and antibody reactions confirm, that the 26 kDa protein is similar but distinct from both the 28 kDa LHC IIc and LHC IIb protein sequences, indicating that there remains at least one more cab gene to be identified in higher plants. Furthermore, it is difficult to interpret the data in any way other than that there is a novel LHC II pigment-protein (LHC IIc) that co-migrates with LHC IIc.Abbreviations CC core complex - LHC light-harvesting complex - PVDF polyvinylidene fluoride