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PubMedDate not supplied

[Increase in the phosphorescence and delayed fluorescence of chlorophyll in barley leaves after selective suppression of carotene biosynthesis].

The action of inhibitors of carotenoid biosynthesis--herbicides SAN 9789 and SAN 6706 on the pigment composition and chlorophyll low temperature (-196 degrees) delayed luminescence (delayed fluorescence and phosphorescence) has been studied in the barley leaves. The herbicides were introduced into etiolated leaves before greening. It is shown that in the mature leaves which were developed in the presence of herbicides biosynthesis of alpha, beta and gamma carotenes is selectively blocked while the concentration of xanthophylls and chlorophylls is not essentially changed. The lack of carotenes is accompanied by an increase of the quantum yields of chlorophyll phosphorescence and delayed fluorescence by the factors of 10 and 6 respectively. The data suggest that carotenes and xanthophylls take an approximately equal part in quenching the chlorophyll triplet states in normal chloroplasts.

Carotenoids↗
PubMedDate not supplied

[Study of aggregation of chlorophyll and its analogs in artificial pigment--protein complexes].

Investigations of synthetic pigment-protein complexes by the spectral methods showed that during chlorophyll complex formation with casein self-assembly of different pigment packing that are analogous to the system of their native forms according to spectral properties, takes place. Under the condition of the links between protein and methylchlorophyllide, the latter has slighter and pheophytin has greater tendency to aggregation that chlorophyll. The conclusions are based on our results and data on the nature of chlorophyll-protein interaction and structure of spectral forms of pigment. Monomers are fixed on the protein by the coordination of Mg atoms. Oligomers are fixed electrostatically by polar sides of porphyrin cycles free of pigment-pigment interaction. Dimers and chlorophyll-water aggregates are fixed hydrophobically by phytol residuals.

Caseins↗
PubMed1977

Study on the biological effect of fast neutrons 1. Effect of fast neutrons on germinability of barley seeds and on the chlorophyll content of the seedlings according to the dose used.

Barley seeds were irradiated in the active zone of the reactor at 500 watt, at favourable flux of fast neutrons in the dose interval of 823--2144 rad. Seeds after irradiation were "cooled" for 48 hours, then germinated, and the length of shoots of the seven days old seedings was measured. The value of the half lethal dose is 1130 rad, estimated from the growth curve. There is a well-defined relation of chlorophyll content (calculated for fresh weight unit) to the dose used. It was concluded from the phenotype of the leaves that the decrease of chlorophyll content was due first of all to the presence of chlorophyll-less cells and cell lines and not to the decrease of chlorophyll content of the individual cells. Irradiation yielded genetically defected cells.

Chlorophyll↗
PubMed1997

Prediction of structure and organisation of chlorophyll a/b binding polypeptides in PS I and II.

Secondary structures, functionally important residues, antigenic sites, membrane spanning segments and hydropathicity of light harvesting chlorophyll a/b binding polypeptides (LHC) are predicted by theoretical methods from the amino acid sequence of the polypeptides. The reported structural features of the Pea LHC (Lhcb 1 gene product) from electron crystallographic studies have been compared by alignment with other types of chlorophyll a/b binding polypeptides for structural prediction. Fifteen conserved residues D85, D89, E113, H116, E/Q133, E/Q181, E189, D/N233, E252, N/H255, Q/E269, E/D/Q280, N281, H285, D288 (number indicates position in the aligned sequence), are identified which are potential ligands to Mg2+ of chlorophylls. Three amino acid residues D89, E/Q131 and D/N 233 are proposed as ligands to chlorophylls b2, a7 and b2 respectively, for which ligands are not identified in electron crystallographic study.

Amino Acid Sequence↗
PubMed1997

LhcaR1 of the red alga Porphyridium cruentum encodes a polypeptide of the LHCI complex with seven potential chlorophyll a-binding residues that are conserved in most LHCs.

The accessory light-harvesting polypeptides associated with photosystem I (LHCI) in Porphyridium cruentum bind chlorophyll a, zeaxanthin and beta-carotene. A cDNA library of P. cruentum was screened with an antiserum specific to the LHCI polypeptides, and an 0.9 kb fragment was identified as coding for an LHCI polypeptide. This cDNA, which we named LhcaR1, has an open reading frame encoding 222 amino acid residues including a putative transit peptide of 28 amino acids. Hydropathy analysis suggests that there are three transmembrane helices in the mature polypeptide. Each of the amino acid residues that bind chlorophyll (six residues) and serve in stabilizing the helices in higher-plant LHCs are conserved in helices 1 and 3 of P. cruentum LhcaR1. The N-terminal flanking regions of these two helices also show high sequence conservation with other LHCs. Helix 2 contains a seventh putative chlorophyll-binding site, but resembles helix 2 of higher-plant LHCs to a lesser degree. A sequence motif of 11 residues found near the N-terminus and in each of the three helices suggests the possibility that the red algal LhcaR1 derives from a gene duplication. Polypeptides of the expected molecular weight in six other red algae (Achrochaetium, Bangia, Callithamnion, Cyanidium, Polysiphonia, Spermothamnion) were recognized by the antiserum to P. cruentum LHCI, indicating a wide distribution of LHCI in rhodophytes.

Algal Proteins↗
PubMed1987

Effects of coal-smoke pollutants from different sources on the growth, chlorophyll content, stem anatomy and cuticular traits of Euphorbia hirta L.

Variations occurred in the growth, assimilate partitioning, chlorophyll content, stem anatomy and leaf cuticular traits of Euphorbia hirta L. on long-term exposure to coal-smoke pollutants prevailing at two sites, one situated close to a railway loco shed (site B) and another in the vicinity of a thermal power plant (site C). The Botanical Garden of Aligarh Muslim University, Aligarh, was considered as a control site (A). Site C possessed a greater load of coal-smoke pollutants than site B. The present study had shown that coal-smoke pollutants have led to a decrease in plant height, jeopardised the production of leaves and enhanced their fall, and caused a reduction in leaf area, leading to decreases of the total photosynthetic area of the plants, with increasing pollution load. The losses incurred in chlorophyll a were relatively more than chlorophyll b and, as a result, the total chlorophyll contents of leaves were decreased in polluted plants. The dry weights of stems, roots and leaves were decreased to different degrees, whereas the shoot/root dry weight ratio was found to increase in the polluted environment. The growth of stem cortex and pith were slightly affected on site B, but showed significant decreases on site C, due to a greater load of pollutants. Decreased area of xylem tissue was found to couple with an increasing number of vessels of reduced sizes. The stomatal density, pore size and index showed decreases, while the epidermal cells were larger and trichomes longer, on both surfaces of polluted leaves.

Journal Article↗
PubMed1990

Chlorophyll, carotenoids and the activity of the xanthophyll cycle.

Clone spruce trees (Picea abies L. Karst.) were exposed in the Hohenheim open-top chambers to low levels of O(3) and SO(2), singly and in combination, and to simulated precipitation of two pH treatments (Seufert et al., this volume). At the end of five years of continuous exposure, needles from the 13-year-old trees were sampled and analysed for pigments content by means of HPLC (high pressure liquid chromatography). The pigment content was determined for three needle age classes. Chlorophyll a content, measured on a dry weight basis, was similar for all needle age classes in the control chambers receiving only the simulated rain treatments at pH 5.0 or 4.0, and the chamber receiving O(3) and the rain treatment at pH 4.0. Also, no differences were noted in one-year-old needles in the chambers with SO(2) and simulated precipitation at pH 4.0 and SO(2) + O(3) and simulated precipitation at pH 4.0. Reductions of approximately 10 and 35% were measured in two-year-old needles from the chambers with SO(2) and precipitation at pH 4.0, and SO(2) + O(3) and precipitation at pH 4.0. The three-year-old needles from these chambers had 40% lower chlorophyll a content compared to the control chambers. No treatment effects were seen on the molar ratios of chlorophyll b, the carotenes, lutein, neoxanthin, and the sum of carotenoids involved in the xanthophyll cycle, violaxanthin + antheraxanthin + zeaxanthin, to chlorophyll [Formula: see text]. The xanthophyll cycle, assayed in one-year-old needles under defined light conditions (520 microE m(-2) s(-1), while light) was active in all samples. Needles from the control chambers and the chambers with SO(2) and with O(3) behaved similarly and differed from the SO(2) + O(3) treated needles by a 50% higher zeaxanthin content reached under light.

Journal Article↗
PubMed2001

Energy transfer in the peridinin chlorophyll-a protein of Amphidinium carterae studied by polarized transient absorption and target analysis.

The peridinin chlorophyll-a protein (PCP) of dinoflagellates differs from the well-studied light-harvesting complexes of purple bacteria and green plants in its large (4:1) carotenoid to chlorophyll ratio and the unusual properties of its primary pigment, the carotenoid peridinin. We utilized ultrafast polarized transient absorption spectroscopy to examine the flow of energy in PCP after initial excitation into the strongly allowed peridinin S2 state. Global and target analysis of the isotropic and anisotropic decays reveals that significant excitation (25-50%) is transferred to chlorophyll-a directly from the peridinin S2 state. Because of overlapping positive and negative features, this pathway was unseen in earlier single-wavelength experiments. In addition, the anisotropy remains constant and high in the peridinin population, indicating that energy transfer from peridinin to peridinin represents a minor or negligible pathway. The carotenoids are also coupled directly to chlorophyll-a via a low-lying singlet state S1 or the recently identified SCT. We model this energy transfer time scale as 2.3 +/- 0.2 ps, driven by a coupling of approximately 47 cm(-1). This coupling strength allows us to estimate that the peridinin S1/SCT donor state transition moment is approximately 3 D.

Absorption↗
PubMed1997

Photoelectric properties of chlorophyll and carotene solutions in nematic liquid crystal located between semiconducting electrodes.

The photopotential and photocurrent generation for chlorophyll a, beta-carotene and a mixture of these pigments dissolved in nematic liquid crystal and located between transparent semiconducting electrodes were measured. Both pigments exhibit photopotential and photocurrent generation. From the photocurrent amplitudes it follows that the efficiency of electron transfer to a semiconducting electrode from beta-carotene is higher than from chlorophyll alpha. The photocurrent amplitude of the pigment mixture is slightly lower than that calculated as a sum of amplitudes of pigments located in separated cells. This difference can be explained by secondary effects, such as competition between carotene and chlorophyll molecules in a process of adsorption on a semiconducting electrode. Therefore it seems that no charge transfer complexes of chlorophyll and carotene are formed in the investigated model system.

Journal Article↗
PubMed1998

Differences in Chlorophyll Loss at 13 degrees C for Two Broccoli (Brassica oleracea L.) Cultivars Associated with Antioxidant Enzyme Activities.

Two broccoli cultivars which have been previously found to have large differences in yellowing susceptibility were studied to determine if there was a relationship between antioxidant capability and chlorophyll loss at a simulated retail display temperature. Greenbelt cultivar retained a stable chlorophyll content over 4 days at 13 degrees C, while Emperor cultivar showed a constant decline in chlorophyll content. These differences were not related to differences in water loss. Superoxide dismutase (SOD) and peroxidase (POD) activities were approximately 30% higher in Greenbelt than in Emperor. The ratio of superoxide dismutase to peroxidase activity was also lower in Greenbelt. The susceptibility of Emperor to oxygen radical damage and lipid peroxidation is expected to be higher since an excessive SOD activity, with respect to POD activity, can potentially lead to enhanced hydroxyl radical formation. Catalase (CAT) activity was higher in Emperor, suggesting that catalase is not important in providing resistance to chlorophyll loss in broccoli. Water soluble, nonenzymatic antioxidant activity was similar for both cultivars. These results support the hypothesis that antioxidant protection offered by superoxide dismutase and peroxidase is important to retention of green color in broccoli.

Journal Article↗
PubMed2005

Theoretical determination of standard oxidation and reduction potentials of chlorophyll-a in acetonitrile.

QM/MM calculations were performed on ethyl chlorophyllide-a and its radical cation and anion, by using the density functional (DF) B3LYP method to determine the molecular characteristics, and a molecular mechanics (MM) method to simulate the solvating medium. The presence of the solvent was accounted for during the optimization of the geometry of the 85-atom chlorophyll-a system by using an ONIOM methodology. A total of 24 solvent molecules were explicitly considered during the optimization process, and these were treated by the universal force field (UFF) method. Initially, the split-valence 3-21G basis set was used for optimizing the geometry of the 85-atom species, neutral, cation and anion. Electronic energies were then determined for the optimized species by making use of the polarized 6-31G(d) basis set. The ionization energy calculated (6.0 eV) is in very good agreement with the observed one (6.1 eV). The MM+ force field was used to investigate the dynamics of the acetonitrile molecules around the neutral species as well as the radical ions of chlorophyll. The required atomic charges on all the atoms were obtained from calculations on all involved molecules at the DFT/6-31G(d) level. Randomly sampled configurations were used to determine the first solvation layer contribution to the free energy of solvation of various species. A truncated 46-atom model of ethyl chlorophyllide-a was used to evaluate the thermal energies of neutral chlorophyll molecule relative to its two radical ions in the gas phase. Born energy, Onsager energy, and the Debye-Huckel energy of the chlorophyll-solvent aggregate were added as perturbative corrections to the free energy of solvation that was initially obtained through molecular dynamics method for the same complex. These calculations yield the oxidation potential as 0.75 +/- 0.32 V and the reduction potential -1.18 +/- 0.31 V at 298.15 K. The calculated values are in good agreement with the experimental midpoint potentials of +0.76 and -1.04 V, respectively.

Journal Article↗
PubMed2002

;Low-waves' in chlorophyll fluorescence kinetics indicate deprivation of bicarbonate.

A brief reversible lowering of chlorophyll fluorescence yield (so called low-waves) immediately after application of a saturating light pulse in parallel with a short-time enhancement of the P700 oxidation level was observed in the green alga Haematococcus pluvialis. The phenomenon occurred in the steady-state time region of fluorescence induction kinetics under mild acidic conditions, and was eliminated by bicarbonate. Shortly after expression of low-waves, the photosynthetic oxygen evolution rate decreased and the non-photochemical chlorophyll fluorescence quenching component increased. The enhancement of the non-photochemical chlorophyll fluorescence quenching component was nigericin-sensitive indicating its dependence on the transthylakoid proton gradient. On the other hand, the formation of low-waves was not removed by the uncoupler. Only when bicarbonate was applied additionally, the reversible short-term decrease in fluorescence yield following each saturating light flash was abolished. Dimethyl-4-nitroso-aniline as an artificial electron acceptor of Photosystem I did not limit the brief drops in fluorescence. However, formate as a competitive inhibitor of bicarbonate binding in Photosystem II induced low-wave formation. Therefore, our results suggest that low-waves in chlorophyll fluorescence kinetics indicate deprivation of bicarbonate in the reaction centre of Photosystem II.

Journal Article↗
PubMed2002

Regulation of the tetrapyrrole biosynthetic pathway leading to heme and chlorophyll in plants and cyanobacteria.

Photosynthetic organisms synthesize chlorophylls, hemes, and bilin pigments via a common tetrapyrrole biosynthetic pathway. This review summarizes current knowledge about the regulation of this pathway in plants, algae, and cyanobacteria. Particular emphasis is placed on the regulation of glutamate-1-semialdehyde formation and on the channelling of protoporphyrin IX into the heme and chlorophyll branches. The potential role of chlorophyll molecules that are not bound to photosynthetic pigment-protein complexes ('free chlorophylls') or of other Mg-containing porphyrins in regulation of tetrapyrrole synthesis is also discussed.

Journal Article↗
PubMed2001

A giant chlorophyll-protein complex induced by iron deficiency in cyanobacteria.

Cyanobacteria are abundant throughout most of the world's water bodies and contribute significantly to global primary productivity through oxygenic photosynthesis. This reaction is catalysed by two membrane-bound protein complexes, photosystem I (PSI) and photosystem II (PSII), which both contain chlorophyll-binding subunits functioning as an internal antenna. In addition, phycobilisomes act as peripheral antenna systems, but no additional light-harvesting systems have been found under normal growth conditions. Iron deficiency, which is often the limiting factor for cyanobacterial growth in aquatic ecosystems, leads to the induction of additional proteins such as IsiA (ref. 3). Although IsiA has been implicated in chlorophyll storage, energy absorption and protection against excessive light, its precise molecular function and association to other proteins is unknown. Here we report the purification of a specific PSI-IsiA supercomplex, which is abundant under conditions of iron limitation. Electron microscopy shows that this supercomplex consists of trimeric PSI surrounded by a closed ring of 18 IsiA proteins binding around 180 chlorophyll molecules. We provide a structural characterization of an additional chlorophyll-containing, membrane-integral antenna in a cyanobacterial photosystem.

Bacterial Proteins↗
PubMed1970

The pi-Cation Radical of Chlorophyll a.

Chlorophyll a undergoes reversible one-electron oxidation in dichloromethane and butyronitrile. Removal of the electron by controlled potential electrolysis or by stoichiometric charge transfer to a known cation radical yields a radical (epr line width = 9 gauss, g = 2.0025 +/- 0.0001) whose optical spectrum is bleached relative to that of chlorophyll. Upon electrophoresis this bleached species behaves as a cation. By comparison with the known properties of pi-cation radicals of porphyrins and chlorins, the chlorophyll radical is also identified as a pi-cation. Further correlation of optical and epr properties with published studies on photosynthesis leads to the conclusion that oxidized P700, the first photochemical product of photosystem I in green plants, contains a pi-cation radical of the chlorin component of chlorophyll a. This radical is the likely source of the rapidly-decaying, narrow epr signal of photosynthesis.

Journal Article↗
PubMed1979

Ligated chlorophyll cation radicals: Their function in photosystem II of plant photosynthesis.

Magnesium tetraphenylchlorin, a synthetic model for chlorophyll, exhibits significant variations in the unpaired spin densities of its cation radicals with concomitant changes in oxidation potentials as a function of solvent and axial ligand. Similar effects are observed for chlorophyll (Chl) a and its cation radicals. Oxidation potentials for Chl --> Chl(+.) as high as +0.9 V (against a normal hydrogen electrode) are observed in nonaqueous solvents, with linewidths of the electron spin resonance signals of monomeric Chl(+.) ranging between 9.2 and 7.8 G in solution. These changes in electronic configuration and ease of oxidation are attributed to mixing of two nearly degenerate ground states of the radicals theoretically predicted by molecular orbital calculations. Comparison of the properties of chlorophyll in vitro with the optical, redox, and magnetic characteristics attributed to P-680, the primary donor of photosystem II which mediates oxygen evolution in plant photosynthesis, leads us to suggest that P-680 may be a ligated chlorophyll monomer whose function as a phototrap is determined by interactions with its (protein?) environment.

Journal Article↗
PubMed1986

Genetically engineered mutant of the cyanobacterium Synechocystis 6803 lacks the photosystem II chlorophyll-binding protein CP-47.

CP-47 is absent in a genetically engineered mutant of cyanobacterium Synechocystis 6803, in which the psbB gene [encoding the chlorophyll-binding photosystem II (PSII) protein CP-47] was interrupted. Another chlorophyll-binding PSII protein, CP-43, is present in the mutant, and functionally inactive PSII-enriched particles can be isolated from mutant thylakoids. We interpret these data as indicating that the PSII core complex of the mutant still assembles in the absence of CP-47. The mutant lacks a 77 K fluorescence emission maximum at 695 nm, suggesting that the PSII reaction center is not functional. The absence of primary photochemistry was indicated by EPR and optical measurements: no chlorophyll triplet originating from charge recombination between P680(+) and Pheo(-) was observed in the mutant, and there were no flash-induced absorption changes at 820 nm attributable to chlorophyll P680 oxidation. These observations lead us to conclude that CP-47 plays an essential role in the activity of the PSII reaction center.

Journal Article↗
PubMed1988

Determination of the aggregate size in detergent solution of the light-harvesting chlorophyll a/b-protein complex from chloroplast membranes.

The molecular mass of an oligomeric integral membrane protein, the light-harvesting chlorophyll a/b-protein complex from the photosynthetic membranes of chloroplasts, has been determined in detergent solution by analytical ultracentrifugation and measurement of the density increment at constant chemical potential of all diffusible solutes. The technique used eliminates any problems resulting from detergent binding to the protein, is independent of the particular detergent used (in this case the nonionic n-octyl beta-D-glucopyranoside), and gives the apparent weight-average molecular mass at different protein concentrations, allowing extrapolation to zero concentration. It means that the solutions of the complex must be brought to dialysis equilibrium with the solvent detergent solution and also requires a reliable method for measuring the protein concentration, for which amino acid analysis was used. The detergent-solubilized complex was a trimer that dissociated into monomers and dimers at low protein concentration. The accurate concentration determinations also allowed the molar chlorophyll-to-protein ratio to be measured as 15, corresponding to 8 chlorophyll a and 7 chlorophyll b molecules.

Journal Article↗