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PubMed1996

Cloning and characterization of the chlorophyll biosynthesis gene chlM from Synechocystis PCC 6803 by complementation of a bacteriochlorophyll biosynthesis mutant of Rhodobacter capsulatus.

A bacteriochlorophyll a biosynthesis mutant of the purple photosynthetic bacterium Rhodobacter capsulatus was functionally complemented with a cosmid genomic library from Synechocystis sp. PCC 6803. The complemented R. capsulatus strain contains a defined mutation in the bchM gene that codes for Mg-protoporphyrin IX methyltransferase, the enzyme which converts Mg-protoporphyrin IX to Mg-protoporphyrin IX methylester using S-adenosyl-L-methionine as a cofactor. Since chlorophyll biosynthesis also requires the same methylation reaction, the Synechocystis genome should similarly code for a Mg-protoporphyrin IX methyltransferase. Sequence analysis of the complementing Synechocystis cosmid indicates that it contains an open reading frame exhibiting 29% sequence identity to BchM. In addition, expression of the Synechocystis gene in the R. capsulatus bchM mutant via the strong R. capsulatus puc promoter was shown to support nearly wild-type levels of bacteriochlorophyll a synthesis. To our knowledge, the Synechocystis sequence thus represents the first chlorophyll biosynthesis gene homolog of bchM. The complementing Synechocystis cosmid was also shown to code for a gene product that is a member of a highly conserved family of RNA binding proteins, the function of which in cyanobacteria remains undetermined.

Amino Acid Sequence↗
PubMed1975

Effect of vanadium on growth, chlorophyll formation and iron metabolism in unicellular green algae.

In the presence of vanadium, growth of Scenedesmus obliquus and Chlorella pyrenoidosa was increased five to sixfold as determined by dry weight, when cultured under autotrophic conditions for 7 days. The stimulation by vanadium decreased with increasing stability towards hydrolysis of the iron(III)-compounds added. Pentavalent vanadium (20 mug V/1 as NH4VO3) was able to overcome completely a limited iron-deficiency in the algae following growth in presence of 1.8 - 10(-5) m ferric chloride. Vanadium did not alter the iron uptake into the algal cells. 90% of offered 48V was taken up by Scenedesmus obliquus during 5 days of growth, and 21% thereof were found in the chloroplast fraction. In presence of vanadium, the chlorophyll formation was stimulated in Scenedesmus obliquus. This stimulation by vanadium was found to be light-dependent but occurred to a certain extent in the dark also. The main porphyrin of the yellow mutant 211-11h/20 of Chlorella vulgaris was identified as protoporphyrin-IX. The formation of this compound was stimulated by vanadium within 10 days up to 83%. The role of vanadium in the biosynthesis of chlorophylls is discussed.

Chlorella↗
PubMed1978

Concentration quenching in chlorophyll-alpha and relation to functional charge transfer in vivo.

Chlorophyll-alpha in ordinary solvents exhibits concentration quenching. Dimeric chlorophyll is reasonably well confirmed as the quenching species, by a critical reanalysis of available data on concentration dependence and on spectral features, in ordinary solvents, and in several analogous quenching environments. This quenching in the dimer in vitro is somewhat less firmly analyzed as due to a new fast internal conversion. Much peripheral evidence supports transient charge transfer as the cause of internal conversion. The same evidence points to a strong similarity to functional charge transfer in vivo. I suggest that inability to extract P680 may be due to its conversion to a form resembling P700 by addition of water. A number of straightforward experiments are suggested to test these proposals. In particular, it is desirable to test for the existence of a vibronic perturbation (from a higher npi* state) in the dimer, as an alternative to charge transfer for explaining the "observed" internal conversion. Such a vibronic cause would raise interesting problems for phototrap function in vivo.

Chlorophyll↗
PubMed1976

Photoelectrochemical effects in the electrolyte-pigment-metal system. III. Chlorophyll films short-circuit photocurrent transients light energy conversion efficiency.

Short-circuit photocurrents produced by a semitransparent metallic electrode covered with multilayers of chlorophyll and immersed in an electrolyte have been studied. The action spectrum of maximum photocurrent is identical to the absorption spectrum of the film of pigment. The kinetics of rise of the photocurrent are photoindependent for thin multilayers but dependent on thickness. Comparison of efficiency of light energy conversion on the basis of short-circuit photocurrent seems to show that the stack of Chl a monolayers is the more efficient chlorophyll solid system. Introduction for the same number of Chl a molecules of vitamin K1 and Phytol shows that vitamin K1 acts probably like an excitation quencher as in monolayers at the water-air interface.

Biological Transport↗
PubMed1985

Inhibition of chlorophyll synthesis in Hordeum vulgare by 3-amino 2,3-dihydrobenzoic acid (gabaculin).

Gabaculin (3-amino 2,3-dihydrobenzoic acid) is shown to be a very potent inhibitor of chlorophyll formation in Hordeum vulgare. Exposure of leaf segments to 30 microM gabaculin results in an 80% inhibition of chlorophyll synthesis, and this is paralleled by a decrease in carotenoid. Dual-inhibitor studies with dioxoheptanoic acid, which is an inhibitor of delta-amino-laevulinic acid dehydratase, show that gabaculin inhibits an earlier step than dioxoheptanoic acid and affects delta-amino-laevulinic acid synthesis rather than its subsequent metabolism.

Aminolevulinic Acid↗
PubMed1992

Chlorophyll fluorescence lifetime determination of waterstressed C3- and C4-plants.

This article describes the effect of water stress on the room temperature chlorophyll fluorescence lifetime of plants of wheat (C3) and maize (C4). In addition, net CO2 assimilation rate (PN), stomatal conductance and the fluorescence quenching coefficients qP and qNP at steady state conditions were recorded. The overall fluorescence decay of the control plants can be described by an average decay time of 1 ns for both plant types. Water stress did not modify this parameter in the case of wheat, whereas a shortening of the decay was observed for waterstressed maize plants (tau = 0.45 ns). This shortening in the chlorophyll fluorescence decay was accompanied by an increase in the non-photochemical quenching (qNP). The photochemical quenching (qP) and therefore the electron transport via photosystem II remains unaffected by water stress. The most pronounced effect of the stress for both plant types was a decrease in PN due to a closure of the stomata.

Chlorophyll↗
PubMed1976

Toxicity of polychlorinated biphenyls (PCBs) to Euglena gracilis: cell population growth, carbon fixation, chlorophyll level, oxygen consumption, and protein and nucleic acid synthesis.

Populations of Euglena gracilis in exponential growth under light were exposed to 2.5, 5.0, 7.5, and 10 ppm of Aroclor 1221. The ID50/48 of Aroclor 1221 was estimated to be 4.4 ppm, while Aroclor 1232 tested at 20, 35, 50, and 100 ppm resulted in an id50/48 of 55 ppm. With Aroclor 1242, no inhibition of growth was observed with up to 100 ppm exposure. Cell cultures exposed to 4.4 ppm of Aroclor 1221 for 48 hrs had a significantly reduced rate of carbon fixation and reduced levels of chlorophyll after correction for cell density. Oxygen consumption was not affected at the ID50 level of the Aroclor. Uptake of [3H]-leucine in treated cultures was twice that of controls, and [3H]-uridine uptake was significantly lower. Uptake of [3H]-thymidine, and incorporation of [3H]-leucine, [3H]-thymidine, and [3H]-uridine were not significantly different in treated and control cultures. Thes results suggest that at the ID50 level, polychlorinated biphenyls (PCBs) reduce cell population growth in Euglena gracilis by inhibition of photosynthesis and/or chlorophyll production.

Animals↗
PubMed1991

Sequence analysis and phylogenetic reconstruction of the genes encoding the large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase from the chlorophyll b-containing prokaryote Prochlorothrix hollandica.

Prochlorophytes similar to Prochloron sp. and Prochlorothrix hollandica have been suggested as possible progenitors of the plastids of green algae and land plants because they are prokaryotic organisms that possess chlorophyll b (chl b). We have sequenced the Prochlorothrix genes encoding the large and small subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase(rubisco), rbcL and rbcS, for comparison with those of other taxa to assess the phylogenetic relationship of this species. Length differences in the large subunit polypeptide among all sequences compared occur primarily at the amino terminus, where numerous short gaps are present, and at the carboxy terminus, where sequences of Alcaligenes eutrophus and non-chlorophyll b algae are several amino acids longer. Some domains in the small subunit polypeptide are conserved among all sequences analyzed, yet in other domains the sequences of different phylogenetic groups exhibit specific structural characteristics. Phylogenetic analyses of rbcL and rbcS using Wagner parsimony analysis of deduced amino acid sequences indicate that Prochlorothrix is more closely related to cyanobacteria than to the green plastid lineage. The molecular phylogenies suggest that plastids originated by at least three separate primary endosymbiotic events, i.e., once each leading to green algae and land plants, to red algae, and to Cyanophora paradoxa. The Prochlorothrix rubisco genes show a strong GC bias, with 68% of the third codon positions being G or C. Factors that may affect the GC content of different genomes are discussed.

Amino Acid Sequence↗
PubMed1977

Culture and morphoogenetic response of a lethal, chlorophyll-deficient mutant of tobacco to hormones, amino acids and sucrose.

This report describes the culture of Su/Su, Su/su and su/su tissue in vitro. High levels of auxin and low levels of cytokinin increase growth of the cells. The cells do not need exogenous amino acids for rapid growth and the chlorophyll deficiency cannot be overcome by amino acids. Reduced levels of auxin and sucrose enhance differentiation, whereas cysteine in the autoclaved medium inhibits differentiation. The Su chlorophyll mutant of tobacco provides a marker for in vitro studies on photosynthesis and photorespiration, chloroplast genetics and cell fusion techniques.

Cell Differentiation↗
PubMed1980

The effects of intravenously administered chlorophyll-A on naturally occurring serum protease inhibitors in rabbits.

Effects of intravenously administered protease inhibitors on naturally occurring serum inhibitors were investigated in rabbits. Water-dispersed chlorophyll-a, trasylol and leupeptin were tested as exogenous protease inhibitors in the experiments. From the results of experiments, it was concluded that: 1) Pretreatment with chlorophyll-a infusion into rabbits, most effectively prevented the rapid consumption of naturally occurring serum protease inhibitors after successive trypsin infusion, and the duration of its action was observed longest when compared with other exogenous inhibitors, such as trasylol or leupeptin. 2) Final therapeutic effects of these exogenous protease inhibitors seem to have depended upon the disappearing way of the administered inhibitors from the blood in certain period of time, as well as upon biochemical potency of their inhibiting activity.

Animals↗
PubMed2005

Thermostability of photosynthesis in two new chlorophyll b-less rice mutants.

Leaves of the two new chlorophyll b-less rice mutants VG28-1, VG30-5 and the wild type rice cv. Zhonghua 11 were subjected to temperatures 28, 36, 40, 44 and 48 degrees C in the dark for 30 min or gradually elevated temperature from 30 degrees C to 80 degrees C at 0.5 degrees C/min. The thermostability of photosynthetic apparatus was estimated by the changes in chlorophyll fluorescence parameters, photosynthetic rate and pigment content, chloroplast ultrastructure and tissue location of H2O2 accumulation. There were different patterns of F(o)-temperature curves between the Chl b-less mutants and the wild type plant, and the temperature of F(o) rising threshold was shifted 3 degrees C lower in the Chl b-less mutants (48 degrees C) than in the wild type (51 degrees C). At temperature up to about 45 degrees C, chloroplasts were swollen and thylakoid grana became misty accompanied with the complete loss of photosynthetic oxygen evolution in the two Chl b-less mutants, but chloroplast ultrastructure in the wild type showed no obvious alteration. After 55 degrees C exposure, the disordered thylakoid and significant H2O2 accumulation in leaves were found in the two Chl b-less mutants, whereas in the wild type plant, less H2O2 was accumulated and the swollen thylakoid still maintained a certain extent of stacking. A large extent of the changes in qP, NPQ and Fv/Fm was consistent with the Pn decreasing rate in the Chl b-less mutants during high temperature treatment as compared with the wild type. The results indicated that the Chl b-less mutants showed a tendency for higher thermosensitivity, and loss of Chl b in LHC II could lead to less thermostability of PSII structure and function. Heat damage to photosynthetic apparatus might be partially attributed to the internal oxidative stress produced at severely high temperature.

Chlorophyll↗
PubMed1988

Primary structure of barley genes encoding quinone and chlorophyll a binding proteins of photosystem II.

The psbA, psbD and psbC genes encoding the quinone binding D-1 and D-2 apoproteins and the 44 kD chlorophyll a-apoprotein 3 have been located in the chloroplast genome of barley. They are found on a 23 kbp SalI restriction endonuclease fragment in the large single copy region of the chloroplast DNA adjacent to the inverted repeat. As in other species the psbD and psbC genes have reading frames which overlap by 53 bp. They are transcribed in the same direction but translated with a frameshift of one nucleotide. Ten amino acid substitutions are found among the 18 N-terminal residues of the D-2 polypeptide if barley, spinach, tobacco, pea and the liverwort Marchantia are compared. Only 8 substitutions are present among the 335 other residues of the D-2 polypeptide. The amino acid residues located in the putative binding site for the special pair reaction center chlorophyll and the residues probably serving as ligands to non-heme iron in the D-1 and D-2 proteins of barley are strictly conserved when compared to those of purple bacteria and of other higher plants. Identity is also observed for the residues of importance in the binding of quinones.

Amino Acid Sequence↗
PubMed2006

Rapid and quantitative extraction method for the determination of chlorophylls and carotenoids in olive oil by high-performance liquid chromatography.

Colour is an organoleptic characteristic of virgin olive oil and an important attribute that affects the consumer perception of quality. Chlorophylls and carotenoids are the main pigments responsible for the colour of virgin olive oil. A simple analytical method for the quantitative determination of chlorophylls and carotenoids in virgin olive oils has been developed. The pigments were isolated from small samples of oil (1.0 g) by solid-phase extraction using diol-phase cartridges (diol-SPE), and the extract was analysed by reverse-phase HPLC with diode-array UV detection. Chromatographic peak resolution, reproducibility (coefficient of variation (C.V.) <4.5%) and recovery (>98.4%) for each component were satisfactory. A comparative study of the proposed method was performed versus classical liquid-liquid extraction (LLE) with N,N'-dimethylformamide and solid-phase extraction using a C18 column (C18-SPE). While 96.4% of the pigments were recovered by LLE, only 51.3% were isolated by C18-SPE in comparison to diol-SPE. Likewise, a higher alteration of pigment composition was observed when such LLE and C18-SPE procedures were used. In this sense, a higher ratio of pheophytin in comparison to that isolated by the diol-SPE procedure was achieved with both extraction procedures, indicating a greater extent of the pheophytinization reaction. Therefore, quantification of pigments from virgin olive oil by diol-SPE followed by RP-HPLC was found to be rapid, simple, required only a small amount of sample, consumed only small amounts of organic solvents, and provided high recoveries, accuracy and precision.

Carotenoids↗
PubMed2006

Light and dark modulation of chlorophyll biosynthetic genes in response to temperature.

Temperature and light significantly influence chloroplast development and chlorophyll biosynthesis. To understand the mechanism of the modulation of chlorophyll biosynthesis, the levels of transcripts and proteins of many enzymatic steps of tetrapyrrole biosynthesis in wheat and cucumber were simultaneously examined. The effect of low (chill-stress) as well as high (heat-stress) temperatures on dark- and light-grown seedlings was monitored. The protochlorophyllide oxidoreductase (POR) content was greatly reduced in response to light in control and heat-stressed seedlings. However, the POR level was not reduced in light-exposed chill-stressed seedlings. The genes for glutamate semialdehyde aminotransferase (gsa; cucumber), glutamyl-tRNA reductase (GluTR; cucumber), 5-aminolevulinic acid dehydratase (Ala D; cucumber and wheat) and for a subunit of Mg-chelatase (Chl I; wheat) showed a reduced expression in cold stress compared to controls and heat-stress conditions. Although expression of the ferrochelatase gene (Fch) and geranylgeranyl reductase gene (Chl P) was upregulated in light, they were downregulated by both chill- and heat-stress. Interestingly, gsa and uroporphyrinogen decarboxylase gene (UroD) and gene product abundance was stimulated by light and heat-stress implying the presence of both light and heat-inducible elements in their promoters. This observation corroborates with the previous report of increased enzymatic activity of UroD in heat-stressed cucumber seedlings. The gsa and Uro D may play an important role in tolerance of the greening process of plants to heat-stress.

Blotting, Northern↗
PubMed2001

Energy dissipation in photosynthesis: does the quenching of chlorophyll fluorescence originate from antenna complexes of photosystem II or from the reaction center?

Dissipation of light energy was studied in the moss Rhytidiadelphus squarrosus (Hedw.) Warnst., and in leaves of Spinacia oleracea L. and Arabidopsis thaliana (L.) Heynh., using chlorophyll fluorescence as an indicator reaction. Maximum chlorophyll fluorescence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU)-treated spinach leaves, as produced by saturating light and studied between and -20 degrees C, revealed an activation energy delta E of 0.11 eV. As this suggested recombination fluorescence produced by charge recombination between the oxidized primary donor of photosystem II and reduced pheophytin, a mathematical model explaining fluorescence, and based in part on known characteristics of primary electron-transport reactions, was developed. The model permitted analysis of different modes of fluorescence quenching, two localized in the reaction center of photosystem II and one in the light-harvesting system of the antenna complexes. It predicted differences in the relationship between quenching of variable fluorescence Fv and quenching of basal, so-called F0 fluorescence depending on whether quenching originated from antenna complexes or from reaction centers. Such differences were found experimentally, suggesting antenna quenching as the predominant mechanism of dissipation of light energy in the moss Rhytidiadelphus, whereas reaction-center quenching appeared to be important in spinach and Arabidopsis. Both reaction-center and antenna quenching required activation by thylakoid protonation but only antenna quenching depended on or was strongly enhanced by zeaxanthin. De-protonation permitted relaxation of this quenching with half-times below 1 min. More slowly reversible quenching, tentatively identified as so-called qI or photoinhibitory quenching, required protonation but persisted for prolonged times after de-protonation. It appeared to originate in reaction centers.

Bryopsida↗
PubMed2000

Influence of ascorbate and the Mehler peroxidase reaction on non-photochemical quenching of chlorophyll fluorescence in maize mesophyll chloroplasts.

Non-photochemical quenching of chlorophyll fluorescence (NPQ) and quantum yield of photosystem II (PSII) were studied with intact mesophyll chloroplasts of maize (Zea mays L.) during the initial minutes of illumination using the pulse-modulated chlorophyll fluorescence technique. Non-photochemical quenching was rapidly reversible in the dark at any point during illumination, which is indicative of energy-dependent dissipation of energy (mediated via thylakoid delta pH changes and ascorbate-dependent synthesis of zeaxanthin). In chloroplasts suspensions including 15 mM ascorbate in the medium, with addition of oxaloacetate and pyruvate, the PSII yield, rate of reduction of oxaloacetate and phosphorylation of pyruvate reached a maximum after approximately 2 min of illumination. Under these conditions, which promote phosphorylation and a decreased delta pH across the thylakoid membrane, NPQ rose to a maximum after 2-3 min of illumination, dropped to a minimum after about 6 min, and then increased to a steady-state level. A rather similar pattern was observed when leaves were illuminated following a 30-min dark period. Providing chloroplasts with higher levels of ascorbate (60 mM), prevented the transient drop in NPQ. Anaerobic conditions or addition of potassium cyanide caused a decrease in PSII yield, providing evidence for operation of the ascorbate-dependent Mehler-peroxidase reaction. These conditions also strongly suppressed the transient drop in NPQ. Dithiothreitol, an inhibitor of violaxanthin de-epoxidase, caused a large drop in NPQ even in the presence of high levels of ascorbate. The results suggest that the decline of NPQ occurs in response to an increase in lumen pH after initiation of phosphorylation, that this decline can be suppressed by conditions where ascorbate is not limiting for violaxanthin de-epoxidase, and that the increase of NPQ after such a decline is the result of development of energy dissipation in PSII reaction centers.

Anaerobiosis↗
PubMed2005

Inhibitory effect of fluoranthene on photosynthetic processes in lichens detected by chlorophyll fluorescence.

The effect of increasing concentration (0.01, 1 and 5 mg l(-1)) of fluoranthene (FLT) and the duration of exposure (12, 24, 36, 48 and 60 h) on the chlorophyll fluorescence parameters (F0, F(V)/F(M), Phi II and (F(M) - F(S))/F (S)) of symbiotic algae in thalli of two foliose lichens Lasallia pustulata and Umbilicaria hirsuta was investigated. Also the FLT concentration in thalli of both lichen species and the proportion of symbiotic algae (photobionts) in thalli was studied. The results obtained demonstrated that the applied concentration of FLT (1 and 5 mg l(-1)) affected primary photochemical processes of photosynthesis of algae in both lichen species. The F0 values increased and the F(V)/F(M) and Phi II values decreased. The fluoranthene content in thallus of both lichen species was increased with increasing FLT concentration in the environment. Chlorophyll fluorescence parameters showed different sensitivity to the content of FLT in Lasallia pustulata and Umbilicaria hirsuta, respectively. Higher sensitivity, detected in U. hirsuta, can be related to its diverse anatomical structure (higher portion of symbiotic algae in thalli) and to the different (nearly 2.5 times higher) content of FLT after exposure.

Chlorophyll↗
PubMed2006

Photosynthetic parameters as indicators of trinitrotoluene (TNT) inhibitory effect: change in chlorophyll a fluorescence induction upon exposure of lactuca sativa to TNT.

Trinitrotoluene, as a compound of conventional explosive, may cause inhibitory effect on terrestrial plants. When Lactuca sativa was exposed to different concentrations of trinitrotoluene (32-1000 mg/kg), photosynthetic process was investigated by using rapid chlorophyll fluorescence kinetic and pulse modulated fluorometry. The decrease of chlorophyll a variable fluorescence was seen to be caused by the deactivation of photosystem II reaction centers. We found for rapid variable fluorescence to be a useful indicator to evaluate the inhibitory effect of trinitrotoluene on photosystem II primary photochemistry and electron transport. The fluorescence parameters, related to the reduction state of photosystem II and to non-photochemical dissipation of light energy, showed a strong relation between the inhibitory effect of photosystem II activity and concentration of trinitrotoluene. The change of photosynthetic fluorescence parameters induced by trinitrotoluene was a reliable indication of the plant physiological state. We proposed for the reduction state of photosystem II and the non-photochemical energy dissipation to be a useful tool in bioassay toxicity testing of trinitrotoluene polluted soil.

Biomass↗