Purification and characterization of peridinin-chlorophyll a-proteins from the marine dinoflagellates Glenodinium sp. and Gonyaulax polyedra

Summary A peridinin-chlorophyll a-protein complex (PCP) was obtained in large quantity from the marine dinoflagellates, Glenodinium sp. and Gonyaulax polyedra. The chromoproteins have similar molecular weights, 35,500 for Glenodinium sp. and 34,500 for G. polyedra. The proteins from the PCP complex of Glenodinium sp. dissociated from the chromophore on treatment with 1% sodium dodecyl sulfate (SDS) at room temperature. The protein component was a single subunit with a molecular weight of 15,500. Proteins from the PCP complex of G. polyedra were composed of a single polypeptide with a molecular weight of about 32,000. Two peridinin-chlorophyll a-proteins from Glenodinium sp. accounted for 70% of the PCP complex and had isoelectric points of 7.4 and 7.3. The PCP complex from G. polyedra was dominated by a single chromoprotein with an isoelectric point of 7.2 Chromophore analysis indicated the presence of only peridinin and chlorophyll a in a molar ratio approaching 4:1. Other pigments characteristically found in dinoflagellates were absent. Fluorescence excitation spectra of purified PCP indicated an efficient energy transfer from peridinin to chlorophyll a, an observation that lends support to the reported role of peridinin as an accessory pigment in photosynthetic oxygen evolution. In several other brown colored dinoflagellates examined, PCP representtd less than 20% of the total peridinin. However, no PCP could be isolated from cultures of Amphidinium carterae (PY-1). This study provides further evidence that PCP is a normal component of most peridinin-containing dinoflagellates, and functions as a light-harvesting component of the dinoflagellate chloroplast. No fucoxanthin-containing analog of PCP was detected in the chrysophyte, Cricosphera carterae and the dinoflagellate Glenodinium foliaceum.Abbreviations PCP peridinin-chlorophyll a-protein complex - PCP's peridinin-chlorophyll a-proteins - SDS sodium dodecyl sulfate - pl isoelectric point - DEAE diethylaminoethyl cellulose - TLC thin layer chromatography - A optical absorbance at a designated wavelength - SIO (F.T. Haxo), Scripps Institution of Oceanography collection     

The role of peridinin-chlorophyll a-proteins in the photosynthetic light adaption of the marine dinoflagellate,Glenodinium sp.

Summary The marine dinoflagellate, Glenodinium sp., was cultured at a series of light levels and growth, pigmentation, and photosynthetic rates were compared. Under decreasing light conditions, growth rates decreased, cellular chlorophyll a and peridinin content per cell increased, and maximum cellular photosynthetic rates remained unchanged. Pigmentation changes were related to alterations in cellular concentrations of a peridinin-chlorophyll a-protein and an unidentified chlorophyll a component of the chloroplast membrane. Maintenance of photosynthetic rates with decreased irradiance is interpreted as an increase in the number of pigment molecules in the light-harvesting antenna associated with the reaction centers of the photosynthetic apparatus, thus increasing the potential for photon capture for photosynthesis.Abbreviations ASP 7 medium see Provasoli, 1963 - PCP peridinin-chlorophyll a-protein - PSU photosynthetic unit - P-I photosynthesis-irradiance curve - TLC thin layer chromatography - A optical absorbance at a designated wavelength - SIO (F.T. Haxo) Scripps Institution of Oceanography collection This paper represents a portion of a thesis submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy from the University of California, San Diego     

Isolation of membrane bound light-harvesting-complexes from the dinoflagellates Heterocapsa pygmaea and Prorocentrum minimum

We have isolated Chl a-Chl c-carotenoid binding proteins from the dinoflagellates Prorocentrum minimum and Heterocapsa pygmaea grown under high (500 mol m^–2 s^–1, HL) and low (35 mol m^–2 s^–1, LL) light conditions. We compared various isolation procedures of membrane bound light harvesting complexes (LHCs) and assayed the functionality of the solubilized proteins by determining the energy transfer efficiency from the accessory pigments to Chl a by means of fluorescence excitation spectra. The identity of the newly isolated protein-complexes were confirmed by immunological cross-reactions with antibodies raised against the previously described membrane bound Chl a-c proteins (Boczar et al. (1980) FEBS Lett 120: 243–247). Spectroscopic analysis demonstrated the relatedness of these proteins with the recently described Chl-a-c ₂-peridinin (ACP) binding protein (Hiller et al. (1993) Photochem Photobiol 57: 125–131; Iglesias Prieto et al. (1993) Phil Trans R Soc London B 338: 381–392). The water-soluble peridinin-Chl a binding-protein (PCP) was not detectable in P. minimum. Two functional forms of ACP with different pigmentation were isolated. A variant of ACP which was isolated from high-light grown cells, that specifically binds increased amounts of diadinoxanthin was compared to the previously described ACPs that bind proportionately more peridinin.Abbreviations ACP Chl a-Chl c-peridinin binding protein - AEBSF 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride - DDM dodecyl -d maltoside - Deriphat 160 N-lauryl-beta-iminopropionic acid - HEPES (N-2-hydroxyethylpiparizine-N-2-ethanesulphonic acid) - HL high light (500 mol m^–2 s^–1) - LL low light (35 mol m^–2 s^–1) - ₇₃₀ fluorescence yield (emission at 730 nm) - PCP peridinin-Chl a-binding protein - PMSF phenyl-methyl-sulfonyl-fluoride - PS I Photosystem I - PS II Photosystem II     

Photoinactivation of functional photosystem II and D1-protein synthesis in vivo are independent of the modulation of the photosynthetic apparatus by growth irradiance

To investigate whether the in-vivo photoinhibition of photosystem II (PSII) function by excess light is an intrinsic property of PSII, the maximal photochemical efficiency of PSII (Fv/Fm) and the content of functional PSII (measured by repetitive flash yield of oxygen evolution) were determined in leaves of pea (Pisum sativum L.), grown in 50 (low light), 250 (medium light), and 650 (high light) mol photons·m^–2·s^–1. The modulation of PSII functionality in vivo was induced in 1.1% CO₂ by varying either (i) the duration (0–2 h) of light treatment (fixed at 1800 mol photons· m^–2·s^–1) or (ii) irradiance (0–3200 mol photons·m^–2·s^–1) at a fixed duration (1 h), after infiltration of leaves with water (control), lincomycin (an inhibitor of chloroplast-encoded protein synthesis), or a combination of lincomycin with nigericin (an uncoupler), through the cut petioles of leaves of 22-to 24-d-old plants. The reciprocity law of irradiance and duration of illumination for PSII function in vivo (Park et al. 1995, Planta 196: 401–411) holds in all differently light-grown peas, demonstrating that inactivation of functional PSII depends on photon exposure (mol photons·m^–2), not on the rate of photon absorption. In vivo, PSII acts as an intrinsic photon counter and at higher photon exposures is inactivated following absorption of about 3 × 10⁷ photons. There is a functional heterogeneity of PSII in vivo with 25% less-stable PSIIs that are inactivated at low photon exposure, compared to 75% more-stable PSIIs regardless of modulation of the photosynthetic apparatus. We suggest that the less-stable PSIIs represent monomers located in the nonappressed granal margins, while the more-stable PSIIs are dimers located in the appressed grana membrane cores. The capacity for D1-protein synthesis was the same in all the light-acclimated peas and saturated at low light, indicating that D1-protein repair is also an intrinsic property of PSII. This accounts for the low intensity required for recovery of photoinhibition in sun and shade plants which is independent of light-harvesting antennae size or PSII/PSI stoichiometries.Abbreviations D1-protein psbA gene product - D2 protein psbD gene product - Fo chlorophyll fluorescence corresponding to open PSII reaction centres - Fv, Fm variable and maximum fluorescence after dark incubation, respectively - PS photosystem - Q_B secondary quinone electron acceptor Financial support for this research by the Department of Employment, Education and Training/Australian Research Council International Research Fellowships Program (Korea) is gratefully acknowledged.     

Culture of the marine phytoplankter,Dunaliella tertiolecta,with light-dark cycles

Summary Dunaliella tertiolecta was grown in continuous culture, maintained by a single daily dilution to a constant cell concentration, with photoperiods of duration 3 to 18 hours. Illumination was provided with filtered tungsten light having a maximum intensity at 580–590 m. Average intensity at the culture surface was maintained at 0.05 cal/cm²xmin. Temperature was regulated at 20° C. Daily cell production and the fraction of cells dividing daily are reported for each of the photoperiods. Cyclic diurnal variations were noted in cell pigment content, cell volume and photosynthetic rate. Corresponding variations in cell carbon/chlorophyll a ratios were small and may be unimportant for the estimation of biomass from chlorophyll a in field work. An equation for calculating daily growth rate in continuous light, from light absorption by cell chlorophyll a, derived previously, was modified to include the influence of providing light in discrete photoperiods.Supported by U. S. Atomic Energy Commission, Contract AT(11-1)-34 Project 108.     

Light Regulation of Peridinin-Chlorophyll a-Protein (PCP) Complexes in the Dinoflagellate, Glenodinium sp. : Use of Anti-Pcp Antibodies to Detect Pcp Gene Products in Cells Grown in Different Light Conditions

As a step toward developing the tools needed to study the molecular bases of light regulation of gene expression in dinoflagellates, light-harvesting peridinin-chlorophyll a-protein (PCP) complexes from Glenodinium sp. were purified and used to generate anti-PCP antibodies. Affinity purified anti-PCP antibodies were isolated from the crude anti-PCP antiserum resulting in improved specificity of immune reactions. The affinity purified anti-PCP antibodies were shown to react strongly and specifically with all major isoforms of PCP complexes in Glenodinium sp. cells, and were used to assess qualitative changes in the levels of PCP gene products in cells grown under different light conditions. Western blot analysis revealed a two- to three-fold increase in detectable PCP apoprotein in low light compared to high light grown cells. In vitro translation reactions supplied with total RNA from high and low light grown Glenodinium sp. cultures also showed an approximate twofold increase in translatable PCP mRNAs in low light grown cells as determined by immunoprecipitation of the primary translation products with affinity purified anti-PCP antibodies. In addition, PCP apoproteins appear to be encoded as larger pre-proteins, since the major immunoprecipitated products from in vitro translation are 23 and 22 kilodaltons, while mature PCP apoproteins are 15.5 kilodaltons. The parallel increases in PCP apoprotein and translatable PCP mRNAs indicate that light regulation of PCP complexes occurs at the level of PCP mRNA abundance.     

Effects of photon flux density on the morphology,photosynthesis and growth of a CAM orchid, <Emphasis Type="Italic">Doritaenopsis</Emphasis> during post-micropropagation acclimatization

Micropropagated plantlets are fragile and often lack sufficient vigour to survive the acclimatization shock during transplantation to the soil. Effects of photosynthetic photon flux densities (PPFDs) on growth, photosynthesis and anatomy of micropropagated Doritaenopsis were studied after 4 months of acclimatization in a greenhouse at 25 °C. The plantlets were transferred to three different PPFDs for four months, i.e. low light (175), intermediate light (270) and high light (450 mol m^–2 s^–1). For most of the growth parameters measured i.e. leaf length, leaf area, leaf width, fresh weight, dry weight, chlorophyll (Chl) a/b ratio, were greater for the intermediate light levels after 4 months of acclimatization. The only exception was leaf thickness, which was increased more under high light levels. Results showed that the survival of Doritaenopsis plantlets was greatest (90%) in low light and intermediate light (89%) compared with only (73%) at high light. However, at low light levels, pigment concentrations (chlorophyll a, b and total chlorophyll) were higher. Net CO₂ assimilation (A), stomatal conductance (g) and transpiration (E) were higher in plantlets grown at high level PPFD than at low after 4 months of acclimatization. Photosynthetic efficiency (Fv/Fm) decreased insignificantly; only at mid day for the high light treatment whereas leaf temperature and stomatal closure increased compared to low light. Scanning electron microscopic (SEM) images of leaves from acclimatized plantlets showed an increase in wax formation for the high light grown plantlets compared to those at low light. Microscopic analysis of acclimatized root sections showed highly developed multiseriate-velamen layers and higher root cell activity; while shoots had larger leaf air spaces than those of in vitro grown plantlets. These results suggest that physiological acclimation occurs at the intermediate PPFD (270 mol m^–2 s^–1) in Doritaenopsis compared to treatment at the high light level.     

Acclimation of Arabidopsis thaliana to the light environment: changes in photosynthetic function

Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta was grown under light regimes of differing spectral qualities, which results in differences in the stoichiometries of the two photosynthetic reaction centres. The acclimative value of these changes was investigated by assessing photosynthetic function in these plants when exposed to two spectrally distinct actinic lights. Plants grown in an environment enriched in far-red light were better able to make efficient use of non-saturating levels of actinic light enriched in long-wavelength red light. Simultaneous measurements of chlorophyll fluorescence and absorption changes at 820 nm indicated that differences between plants grown under alternative light regimes can be ascribed to imbalances in excitation of photosystems I and II (PSI, PSII). Measurements of chlorophyll fluorescence emission and excitation spectra at 77 K provided strong evidence that there was little or no difference in the composition or function of PSI or PSII between the two sets of plants, implying that changes in photosynthetic stoichiometry are primarily responsible for the observed differences in photosynthetic function.Abbreviations Chl chlorophyll - FR far-red light - HF highirradiance FR-enriched light (400 mol·m^–2·s^–1, RFR = 0.72) - HW high-irradiance white light (400 mol·m^–2 1·1 s^–1RFR = 1.40) - LHCI, LHCII light-harvesting complex of PSI, PSII - qO quenching of dark-level chlorophyll fluorescence - qN non-photochemical quenching of variable chlorophyll fluorescence - qP photochemical quenching of variable chlorophyll fluorescence - R red light - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase We thank Dr. Sasha Ruban for assistance with the 77 K fluorescence measurements and for helpful discussions. This work was supported by Natural Environment Research Council Grant GR3/7571A.     

Prolonged incubation with low concentrations of mercury alters energy transfer and chlorophyll (Chl) a protein complexes in Synechococcus 6301: changes in Chl a absorption and emission characteristics and loss of the F695 emission band

Synechococcus PCC 6301 cells grown in the presence of low sublethal levels of (about 2 m) mercury induced alterations in chlorophyll (Chl) a absorption without significant alterations in phycocyanin. Chl a fluorescence emission in Hg²⁺ -raised cells showed a large (about 18 nm) blue shift in the peak emission. No major spectral changes in phycobilisome (PBsome) emission characteristic were noticed, indicating major structural alterations in Chl-protein complexes by incubation with Hg²⁺ ions. Low temperature (77 K) emission spectra of cells grown in the presence of Hg²⁺ showed a loss of the characteristic Chl a emission band at 695 nm (F695), which is known to be linked to photosystem II photochemistry and to originate from the Chl a of core antenna polypeptide CP 47 of photosystem II. The SDS-PAGE polypeptide profile of thylakoids indicates a loss of a polypeptide(s) with a molecular mass between 40 and 60 k Da by Hg²⁺ incubation of cells. Our results suggest that prolonged incubation of Synechococcus 6301 cells with low concentrations of Hg²⁺ affects the Chl a spectral properties and the structure of Chl-protein complexes.     

Changes in pigment content of the diatom Navicula pelliculosa (Bréb.) Hilse in silicon-starvation synchrony

Summary Exponentially grown cells of the freshwater diatom Navicula pelliculosa (Bréb) Hilse, contained chlorophyll a, chlorophyll c, fucoxanthin, diadinoxanthin, diatoxanthin, neofucoxanthin, -carotene, and an unknown pigment, the absorption spectrum of which is reported. Changes in amounts of chlorophyll a, fucoxanthin and diadinoxanthin were determined during the course of silicon-starvation synchrony carried out in the light or dark. Changes in the rate of chlorophyll a and fucoxanthin syntheses were similar. Synthesis ceased after 5–7 hr of silicon starvation, but recommenced in cultures kept in the light, once silicon was re-introduced. In cultures kept in the dark no significant synthesis was observed after re-introduction of silicon. Diadinoxanthin synthesis continued in the light at all times, although at a lower rate during the silicon-starvation period. In the dark, synthesis of this pigment ceased when cell division stopped, and the amount per unit volume of culture decreased. These results are discussed in relation both to the effect of silicon on the metabolism of the diatom and to the possible function of the carotenoids.Dedicated to Prof. C. B. van Niel on the occasion of his 70th birthday.     

Photosynthetic activity of the calyx, green shoulder, pericarp, and locular parenchyma of tomato fruit

Photosynthesis of tomato fruit was studied using green fruit from six heritage cultivars of Lycopersicon esculentum Mill. and one of Lycopersicon pimpinellifolium. Chlorophyll concentrations in the green shoulder, pericarp and locular parenchyma of the fruit were determined and the apparent photosynthetic electron transport activity (ETR) and chlorophyll fluorescence quenching characteristics of these tissues and the calyx were compared. In all cultivars, green shoulder formation, apparent as intense pigmentation of the proximal pericarp shoulder, was positively related to the degree of shading of the fruit during development. Appearing as a photosynthetic adaptive trait for increasing the photoautotrophic capacity of fruit grown under low light, the green shoulder contained 17-57% of the total pericarp chlorophyll content. The pericarp below the green shoulder had lower chlorophyll a+b. At a photon flux density (PFD) of 1200 mol m^-2 s^-1, different fruit tissues were found to have different levels of ETR. In 'Yellow Pear', the upper surface of the calyx had an ETR of 154 mol m^-2 s^-1, while the lower surface had an ETR of 88 mol m^-2 s^-1. On the green shoulder, ETR was 203 mol m^-2 s^-1, whereas in the pericarp distal to the green shoulder, ETR was 97 mol m^-2 s^-1. In the locular parenchyma, ETR was 66 mol m^-2 s^-1. This trend towards a lower ETR in distal and internal fruit tissues appeared to indicate a shift towards a more shade-type photosynthesis. Concomitant with this shift were changes in chlorophyll fluorescence quenching characteristics. Generally when tissues displayed reduced levels of ETR they also displayed a faster decrease in the photochemical quenching coefficient qp and a more rapid diversion of absorbed photon energy to non-photosynthetic activity found in the calyx, green shoulder, pericarp, and locular parenchyma suggest that all of these tissues have significant roles in CO2 scavenging and the provision of carbon assimilates. The potential role of fruit photosynthesis in influencing the fruit acid to sugar ratio and hence fruit quality is discussed.     

Nucleotide sequence of a cDNA clone encoding the precursor of the peridinin-chlorophyll a-binding protein from the dinoflagellate Symbiodinium sp.

mRNA from the dinoflagellate Symbiodinium sp. isolated from the staghorn coral Acropora formosa was used for the construction of cDNA libraries. A cDNA clone was identified which encoded the precursor of peridinin-chlorophyll a-binding protein (PCP), including a 52 amino acid transit peptide and the 313 amino acid mature protein. The deduced amino acid sequence clearly contains an internal duplication, implying that amongst dinoflagellates the M _r 35 000 form of PCP has arisen by duplication and fusion of genes encoding the M _r 15 000 form. This is the first reported sequence of a dinoflagellate light-harvesting protein. The anatomy of the mature protein and the transit peptide are discussed.Abbreviations PCP peridinin-chlorophyll a-binding protein; cab, chlorophyll a/b-binding protein - LHC light-harvesting complex - FCP fucoxanthin-chlorophyll a/c-binding protein     

Functional organization and plasticity of the photosynthetic unit of the cyanobacterium Anacystis nidulans

Anacystis nidulans grown under high and low light, 100 and 10 μE m^?2 s^?1, respectively, was analyzed with respect to chlorophyll/P700, phycobiliproteins/P700, chlorophyll/cell, and oxygen evolution parameters. The photosynthetic unit sizes of this cyanobacterium, measured as the ratio of total chromophores (chlorophyll and bilin) to P700, were shown to be similar to those of higher plants and green algae. High light grown cells possessed a photosynthetic unit consisting of a core of 157 ± 6 chlorophyll a molecules per P700 associated with a light harvesting system of 95 ± 3.5 biliprotein chromophores. Low light grown cells had substantially more biliprotein chromophores per P700 (125 ± 3.1) than high light cells, but showed no significant difference in the numbers of chlorophyll a molecules per P700 (149 ± 4). Analyses of aqueous biliprotein extracts indicate that low light grown cells produce proportionately more phycocyanin relative to allophycocyanin than high light cells. Calculations of the molecular weight of biliproteins per P700 suggest that there is less than one phycobilisome per reaction center I under both growth conditions. Differences in chlorophyll/cell ratios and oxygen evolution characteristics were also observed. High light cells contain 6.3 × 10^?12 mg chlorophyll cell^?1, while low light grown cells contain 12.8 × 10^?12 mg chlorophyll cell^?1. Photosynthetic oxygen evolution rate vs. light intensity curves indicate that high light grown cells reach maximal levels of oxygen evolution at higher light intensity than low light grown cells. Maximal rates of oxygen evolution were 16.6 μmol oxygen min^?1 (mg chlorophyll)^?1 for high and 8.4 μmol oxygen min^?1 (mg chlorophyll)^?1 for low light cells. Maximal oxygen evolution rates per cell were equivalent for both cell types, although the amount of P700 per cell was lower in high light cells. High light grown cells are therefore capable of producing more oxygen per reaction center I than low light grown cells.     

Acclimation of Arabidopsis thaliana to the light environment: Changes in composition of the photosynthetic apparatus

Acclimation to changes in the light environment was investigated in Arabidopsis thaliana (L.) Heynh. cv. Landsberg erecta. Plants grown under four light regimes showed differences in their development, morphology, photosynthetic performance and in the composition of the photosynthetic apparatus. Plants grown under high light showed higher maximum rates of oxygen evolution and lower levels of light-harvesting complexes than their low light-grown counterparts; plants transferred to low light showed rapid changes in maximum photosynthetic rate and chlorophyll-a/b ratio as they became acclimated to the new environment. In contrast, plants grown under lights of differing spectral quality showed significant differences in the ratio of photosystem II to photosystem I. These changes are consistent with a model in which photosynthetic metabolism provides signals which regulate the composition of the thylakoid membrane.Abbreviations Aac1 gene encoding actin - Chl chlorophyll - F far-red-enriched light (R:FR = 0.72) - FR far-red light - H high light (400 mol · m^–2 · s^–1) - L low light (100 ml · m^–2 · s^–1) - LHCII light-harvesting complex of PSII - Lhcb genes encoding the proteins of LHCII - R red light - Rbcs genes encoding the small subunit of Rubisco - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - W white light (R:FR = 1.40) This work was supported by Natural Environment Research Council Grant No. GR3/7571A. We would like to thank H. Smith (Botany Department, University of Leicester) and E. Murchie (University of Sheffield) for helpful discussions.     

Toxicity of carbofuran to soil isolates of Chlorella vulgaris,Nostoc linckia and N. muscorum

A microalga, Chlorella vulgaris, and two diazotrophic cyanobacteria, Nostoc linckia and N. muscorum, all isolated from a rice soil, were compared for their response in terms of growth and metabolic activities to the application of carbofuran. The toxicity criteria included cell constituents (chlorophyll a, total protein, carbohydrate), ¹⁴CO₂ uptake and nitrate reductase, besides nitrogenase activity (acetylene reduction) in the cyanobacteria. C. vulgaris and N. muscorum were more sensitive to carbofuran than was N. linckia. The significant toxicity of the insecticide, observed with higher concentrations of 20 and 50 g ml^–1, to nitrogenase activity in N. linckia was reversed by the addition of ATP at 10 M. Transmission electron microscopy of the cultures, exposed to 50 g carbofuran ml^–1 showed certain cellular abnormalities, indicating interference of the insecticide with membrane properties. Correspondence to: K. Venkateswarlu     

Distribution of Chlorophyll between the Two Photoreactions in Photosynthesis of the Blue-Green Alga Anacystis nidulans Grown at Two Different Light Intensities

Anacystis nidulans was grown in white light of two different intensities, 7 and 50 W ·m^?2. The in vivo pigmentations of the two cultures were compared. The ratio phycocyanin/chlorophyll a was 0.96 for cells grown at 7 W · m^?2 and 0.37 for cells grown at 50 W · m^?2. Phycocyanin-free photosynthetic lamellae (PSI-particles) were prepared, using French press treatment and fractionated centrifugation. Algae grown in the irradiance of 50 W · m^?2 showed a chlorophyll a/P700 ratio of 260, while algae grown at 7 W · m^?2 had a value of 140. Corresponding PSI-particles showed values of 122 and 109 respectively. Light-induced absorption difference spectra measured between 400–450nm indicated different ratios between cytochrome f and P700 in the two algal cultures. Enhancement studies of photosynthetic oxygen evolution were carried out. When a background beam of 691 nm was superimposed upon a signal beam of 625 nm, good enhancement was observed for both cultures. With the wavelengths 675 and 691 nm together a pronounced enhancement could be detected only in algae grown at the higher light level. Absorption spectra recorded on whole cells at 77°K revealed a small shift of the main red chlorophyll a absorption peak caused by light intensity. It is proposed that the reduction of the phycocyanin/chlorophyll a ratio in high light-grown cells is accompanied by an increased energy distribution by chlorophyll a into PSII.     

Transient state characteristics of adaptation to changes in light conditions for the cyanobacterium Oscillatoria agardhii

Transitions in growth irradiance level from 92 to 7 Em^-2 s^-1 and vice versa caused changes in the pigment contents and photosynthesis of Oscillatoria agardhii. The changes in chlorophyll a and C-phycocyanin contents during the transition from high to low irradiance (HL) were reflected in photosynthetic parameters. In the LH transition light utilization efficiencies of the cells changed faster than pigment contents. This appeared to be related to the lowering of light utilization efficiencies of photosynthesis. As a possible explanation it was hypothesized that excess photosynthate production led to feed back inhibition of photosynthesis. Time-scales of changes in the maximal rate of O₂ evolution were discussed as changes in the number of reaction centers of photosystem II in relation to photosynthetic electron transport. Parameters that were subject to change during irradiance transitions obeyed first order kinetics, but hysteresis occurred when comparing HL with LH transients. Interpretation of first order kinetic analysis was discussed in terms of adaptive response vs changes in growth rate.Non-standard abbreviations Chla chlorophyll a - CPC C-phycocyanin - PS II photosystem II - PS I photosystem I - RC II reaction center of photosystem II - P photosynthetic O₂-evolution - I irradiance, Em^-2 s^-1 - light utilization efficiency of cells, mmol O₂·mg dry wt^-1·h^-1/Em^-2 s^-1 - light utilization efficiency of photosynthetic apparatus, mol O₂·mol Chla ^-1·h^-1/Em^-2 s^-1 - P_max maximal rate of O₂ evolution by cells, mol O₂·mg dry wt^-1·h^-1 - P_max maximal rate of O₂ evolution by photosynthetic apparatus, mol O₂·mol·Chla ^-1·h^-1 - LL low light, E m^-2 s^-1 - HL high light, E m^-2 s^-1 - LH low to high light transition - HL high to low light transition - k specific rate of adaptation, h^-1 - specific growth rate, h^-1 - Q pool size of cell constituent, mol·mg dry wt^-1 - q net synthesis rate of cell constituent, mol·mg dry wt^-1·h^-1     

X-ray structures of the peridinin-chlorophyll-protein reconstituted with different chlorophylls

The peridinin-chlorophyll a-protein (PCP) from dinoflagellates is a soluble light harvesting antenna which gathers incoming photons mainly by the carotenoid peridinin. In PCPs reconstituted with different chlorophylls, the peridinin to chlorophyll energy transfer rates are well predicted by a Förster-like theory, but only if the pigment arrangements are identical in all PCPs. We have determined the X-ray structures of PCPs reconstituted with Chlorophyll-b (Chl-b), Chlorophyll-d (Chl-d) and Bacteriochlorophyll-a (BChl-a) to resolutions ?2 Å. In all three cases the pigment arrangements are essentially the same as in native PCP. Hydrogen bonding is not responsible for preferential incorporation of “non-native” chlorophylls over Chl-a.     

Carotenoid composition and photon-use efficiency of photosynthesis inGossypium hirsutum L. grown under conditions of slightly suboptimum leaf temperatures and high levels of irradiance

Summary Cotton (Gossypium hirsutum L. var. DP 61) was grown at different temperatures during 12-h light periods, with either 1800–2000 mol photons m^–2 s^–1 (high photon flux density, PFD) or 1000–1100 mol m^–2 s^–1 (medium PFD) incident on the plants. Night temperature was 25°C in all experiments. Growth was less when leaf temperatures were below 30°C during illumination, the effect being greater in plants grown with high PFD (Winter and Königer 1991). Leaf pigment composition and the photon-use efficiency of photosynthesis were analysed to assess whether plants grown with high PFD and suboptimal temperatures experienced a higher degree of high irradiance stress during development than those grown with medium PFD. The chlorophyll content per unit area was 3–4 times less, and the content of total carotenoids about 2 times less, with the proportion of the three xanthophylls zeaxanthin + antheraxanthin + violaxanthin being greater in leaves grown at 20–21°C than in leaves grown at 33–34°C. In leaves from plants grown at 21°C and 1800–2000 mol photons m^–2 s^–1, zeaxanthin accounted for as much as 34% of total carotenoids in the middle of the photoperiod, the highest level recorded in this study. This finding is consistent with a protective role of zeaxanthin under conditions of excess light. At the lower temperatures, the photochemical efficiency of photosystem II, measured as the ratio of variable to maximum fluorescence yield (F _V/F _M) after 12-h dark adaptation, was 0.76 in medium PFD plants and 0.75 in high PFD plants compared with 0.83 and 0.79, respectively, at the higher temperatures. The photon-use efficiency of O₂ evolution () based on absorbed light between 630 and 700nm, decreased with decrease in temperature from 0.102 to 0.07 under conditions of high PFD, but remained above 0.1 at medium PFD. Owing to compensatory reactions in these long-term growth experiments, sustained differences inF _V/F _M and were much less pronounced than the differences in chlorophyll content and dry matter, particularly in plants which had developed at high PFD and low temperature. In fact, in these plants, which exhibited pronounced photobleaching, a largely functional photosynthetic apparatus was still maintained in cells adjacent to the lower leaf surfaces. This was indicated by measurements of photon use efficiencies of photosynthetic O₂ evolution with leaves illuminated first at the upper, and then at the lower surface.Abbreviations F _O yield of dark level fluorescence - F _M maximum yield of fluorescence, induced in a pulse of saturating light - F _V yield of variable fluorescence (=F _M-F _o) - PFD photon flux density - _iw photon use efficiency of O₂ evolution based on white (400–700 nm) incident light - _ir photon use efficiency based on red (630–700 nm) incident light - _aw photon use efficiency based on white absorbed light - _ar photon use efficiency based on red absorbed light     

Functional analysis of the iron-stress induced CP 43′ polypeptide of PS II in the cyanobacterium Synechococcus sp. PCC 7942

Under conditions of iron-stress, the Photosystem II associated chlorophyll a protein complex designated CP 43, which is encoded by the isiA gene, becomes the major pigment-protein complex in Synechococcus sp. PCC 7942. The isiB gene, which is located immediately downstream of isiA, encodes the protein flavodoxin, which can functionally replace ferredoxin under conditions of iron stress. We have constructed two cyanobacterial insertion mutants which are lacking (i) the CP 43 apoprotein (designated isiA ^–) and (ii) flavodoxin (designated isiB ^–). The function of CP 43 was studied by comparing the cell characteristics, PS II functional absorption cross-sections and Chl a fluorescence parameters from the wild-type, isiA ^– and isiB ^– strains grown under iron-stressed conditions. In all strains grown under iron deprivation, the cell number doubling time was maintained despite marked changes in pigment composition and other cell characteristics. This indicates that iron-starved cells remained viable and that their altered phenotype suggests an adequate acclimation to low iron even in absence of CP 43 and/or flavodoxin. Under both iron conditions, no differences were detected between the three strains in the functional absorption crossection of PS II determined from single turnover flash saturation curves of Chl a fluorescence. This demonstrates that CP 43 is not part of the functional light-harvesting antenna for PS II. In the wild-type and the isiB ^– strain grown under iron-deficient conditions, CP 43 was present in the thylakoid membrane as an uncoupled Chl-protein complex. This was indicated by (1) an increase of the yield of prompt Chl a fluorescence (F_o) and (2) the persistence after PS II trap closure of a fast fluorescence decay component showing a maximum at 685 nm.Abbreviations Chl chlorophyll - CP 43, CP 47 and CP 43 Chl a binding protein complexes of indicated molecular mass - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - F_m and F_m fluorescence when all PS II reaction centers are dosed in dark- and light-acclimated cells, respectively - F_o fluorescence when all PS II reaction centers are open in dark acclimated cells - F_v variable fluorescence after dark acclimation (F_m–F_o)