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CO(2) Assimilation and Activities of Photosynthetic Enzymes in High Chlorophyll Fluorescence Mutants of Maize Having Low Levels of Ribulose 1,5-Bisphosphate Carboxylase.

Photosynthetic properties were examined in several hcf (high chlorophyll fluorescence 11, 21, 42 and 45) nuclear recessive mutants of maize which were previously found to have normal photochemistry and low CO(2) fixation. Mutants usually either died after depletion of seed reserves (about 18 days after planting), or survived with slow growth up to 7 or 8 weeks. Both the activity and quantity of ribulose 1,5-bisphosphate carboxylase (Rubisco) were low in the mutants (5-25% of the normal siblings on a leaf area basis) and the loss of Rubisco tended to parallel the reduction in photosynthetic capacity. The Rubisco content in the mutants was often marginal for photosynthetic carbon gain, with some leaves and positions along a leaf having no net photosynthesis, while other leaves had a low carbon gain. Conversely, the activities of C(4) cycle enzymes, phosphoenolpyruvate carboxylase, pyruvate, Pi dikinase, NADP-malate dehydrogenase, and NADP-malic enzyme, were the same or only slightly reduced compared to the normal siblings. The mutants had about half as much chlorophyll content per leaf area as the normal green plants. However, the Rubisco activity in the mutants was low on both a leaf area and chlorophyll basis. Low Rubisco activity and lower chlorophyll content may both contribute to the low rates of photosynthesis in the mutants on a leaf area basis.

Journal Article↗

Detection and partial characterization of activity of chlorophyll synthetase in etioplast membranes.

The esterification of chlorophyllide a was investigated an irradiated etioplast-membrane fractions ('broken etioplasts') from oat seedlings (Avena sativa L.). As a substrate, [1(-3)H]geranylgeraniol and its monophosphate and diphosphate derivatives were prepared by chemical synthesis. Geranylgeraniol and its monphosphate derivative are incorporated into chlorophyll only in the presence of ATP whereas the diphosphate derivative is incorporated also without ATP. The yield of esterified chlorophyll is 80-90% of chlorophyllide with saturating substrate concentrations. The term 'chlorophyll synthetase' is used to describe the enzyme activity which is different from chlorophyllase. Other substrates are phytol and farnesol either with ATP or as the diphosphate derivatives. The relative specificity of 'chlorophyll synthetase' for thse substrates is geranylgeraniol:phytol:farnesol = 6:3:1. In these experiments in vitro, a new chlorphyll esterified with farnesol was detected which does not occur in intact plants. Geraniol and n-pentadecanol are no substrates for the enzyme. Protochlorphyllide which is present in non-irradiated etioplast membrane fractions is not esterified under the same conditions.

Carbon-Oxygen Ligases↗

The iron-regulated isiA gene of Fischerella muscicola strain PCC 73103 is linked to a likewise regulated gene encoding a Pcb-like chlorophyll-binding protein.

The expression of the chlorophyll a-binding, iron stress-induced protein IsiA is part of the cyanobacterial response to iron deficiency. A new isiA gene from the filamentous heterocystous cyanobacterial strain, Fischerella muscicola PCC 73103, was identified using standard and inverse PCR. While in unicellular cyanobacterial strains isiA is organized in an operon with isiB (encoding flavodoxin), in Fischerella not an isiB gene but another chlorophyll-binding protein encoding gene was identified downstream of isiA, which shows significant similarities to Pcb-like protein encoding genes known from prochlorophytes. The expression of both genes was clearly activated under iron deficiency. Although isiA and pcbC were independently transcribed, the size of the pcbC transcript indicates a large iron-regulated operon. Beside a 10-fold increase of isiA transcript content iron-starved cells of Fischerella showed a blue-shift in the red chlorophyll a absorption peak. In addition, chlorophyll fluorescence at 77 K was dominated by an emission peak at 685 nm. These features are in accordance with the characteristics of IsiA accumulation in iron-starved unicellular cyanobacteria, suggesting identical IsiA function in heterocystous strains in spite of different genetic organization.

Amino Acid Sequence↗

Microscale vertical profiles of n(2) fixation, photosynthesis, o(2), chlorophyll a, and light in a cyanobacterial assemblage.

Profiles of N(2) fixation, O(2) production (gross photosynthesis), O(2) concentration, chlorophyll a concentration, and photon fluence rates were measured with 50-mum resolution in colonies of the heterocyst-forming cyanobacterium Nostoc parmelioides. Microelectrode measurements were made after 20 h of incubation under N(2) gas. Colonies were frozen, and 50-mum sections were prepared by using a freezing microtome and analyzed for N enrichment and chlorophyll a concentration. Colonies exhibited steep spatial gradients in rates of gross photosynthesis, O(2) concentration, and irradiance, with the highest values generally occurring at the surface. O(2) concentration, photosynthesis, and irradiance all showed positive correlations, but chlorophyll a concentrations varied independently of photosynthesis and irradiance. Forty-four percent of the variation in N incorporation was explained by gross photosynthesis (a positive correlation) when incorporation of N was expressed per unit of biomass (chlorophyll a).

Journal Article↗

Biogenesis of chlorophyll-binding proteins under iron stress in Synechocystis sp. PCC 6803.

The biogenesis of chlorophyll-binding proteins under iron stress has been investigated in vivo in a chlN deletion mutant of Synechocystis sp. PCC 6803. The chlN gene encodes one subunit of the light-independent protochlorophyllide reductase. The mutant is unable to synthesis chlorophyll in darkness, causing chlorophyll biosynthesis to become light dependent. When the mutant was propagated in darkness, essentially no chlorophyll and photosystems were detected. Upon return of the chlN deletion mutant to light, 77 K fluorescence emission spectra and oxygen evolution of greening cells under iron-sufficient or -deficient conditions were measured. The gradual blue shift of the photosystem I (PS I) peak upon greening under iron stress suggested the structural alteration of newly synthesized PS I. Furthermore, the rate of biogenesis of PS II was delayed under iron stress, which might be due to the presence of IsiA.

Bacterial Proteins↗

CHLOROPHYLL DEGRADATION.

Although the loss of green color in senescent leaves and ripening fruits is a spectacular natural phenomenon, research on chlorophyll breakdown has been largely neglected until recently. This review summarizes knowledge about the fate of chlorophyll in degreening tissues that has been gained during the past few years. Structures of end- and intermediary products of degradation as well as the biochemistry of the porphyrin-cleaving reaction have been elucidated. The intracellular localization of the catabolic pathway is particularly important in the regulation of chlorophyll breakdown. None of the genes encoding the related catabolic enzymes has so far been isolated, which makes chlorophyll degradation an area of opportunity for future research.

Journal Article↗

Remote sensing of oligotrophic waters: model divergence at low chlorophyll concentrations.

The performance of the OC2 Sea-viewing Wide Field-of-view Sensor (SeaWiFS) algorithm based on 490- and 555-nm water-leaving radiances at low chlorophyll contents is compared with those of semianalytical models and a Monte Carlo radiative transfer model. We introduce our model, which uses two particle phase functions and scattering coefficient parameterizations to achieve a backscattering ratio that varies with chlorophyll concentration. We discuss the various parameterizations and compare them with existent measurements. The SeaWiFS algorithm could be confirmed within an accuracy of 35% over a chlorophyll range from 0.1 to 1 mg m(-3), whereas for lower chlorophyll concentrations we found a significant overestimation of the OC2 algorithm.

Journal Article↗

[Interaction of metal complexes of bacterial chlorophyll with DNA].

AIM: Understanding the modes and activities of metal bacterial chlorophylls as PHD sensitizers with DNA. METHODS: The modes and activities of the interaction of DNA and metal complexes of bacterial chlorophyll, which have been prepared by extraction and synthesis reaction, have been discussed according to the ultraviolet-visual spectrum and nucleic acid electrophoresis. RESULTS: It indicates that the system of DNA and metal complexes have enchanced the interaction by the ultraviolet-visual spectrum. At the same time, it also indicates that metal complexes of bacterial chlorophyll and DNA have different combining way and have strong cutting effect in illumination by the nucleic acid electrophoresis. CONCLUSION: This paper proved that metal bacterial chlorophylls as PHD sensitizers have great advantage.

Bacteriochlorophylls↗

Protective properties of chlorophylls against the covalent binding of heterocyclic amines to DNA in vitro and in vivo.

The protective properties of chlorophylls have been studied using inhibition of carcinogen-DNA binding as an end point. Rats were co-administered sodium/copper chlorophyllin (CHL) and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) by single oral gavage. Eight hours after dosing, CHL inhibited IQ-DNA binding in liver by approximately 60% compared with rats given IQ alone. CHL treatment reduced the total radiolabel eliminated in urine and bile but caused increased excretion via the feces, including enhanced elimination of unmetabolized IQ. In a second study, CHL produced dose-related inhibition of IQ-DNA binding in the liver, large intestine and small intestine, and blocked IQ absorption from ligated sections of the intestine in situ. In in vitro studies CHL competitively inhibited microsome-mediated IQ-DNA binding, and interacted with liver microsomes to produce a difference spectrum with a peak at approximately 420 nm and a trough at 383 nm. CHL also interacted directly with heterocyclic amines to form 2:1 (IQ-type) or 1:1 complexes (non-IQ-type) with dissociation constants in the range 0.333 to 0.083 mM. Finally, in hepatocytes chlorophyll a inhibited the metabolism of 3-amino-1-methyl-5H-pyrido [4,3-b]indole (Trp-P-2) and produced dose-related inhibition of Trp-P-2-DNA binding. Collectively, these studies suggest that chlorophylls operate as interceptor molecules and limit carcinogen bioavailability. As a contributing mechanism, chlorophylls may interfere with cytochrome P450-mediated activation of chemical carcinogens.

Administration, Oral↗

Excitation of micelle-solubilized chlorophyll during the peroxidase-catalyzed aerobic oxidation of isonicotinic acid hydrazide.

Addition of micelle (hexadecyl-trimethylammoniumbromide)-solubilized chlorophyll alpha to the isoniazid/peroxidase/Mn2+/O2 system promotes light emission, identified as chlorophyll fluorescence. Based on O2 consumption, the quantum yield of chlorophyll excitation to the S1 state exceeds 6 X 10(-6). At least part of the excitation has its origin in the conversion of an intermediate--presumably a diazene--to pyridine-4-carboxaldehyde. On the basis of the present and earlier results [K. Zinner, C. C. C. Vidigal, N. Durán, and G. Cilento (1977) Arch. Biochem. Biophys. 180, 452-458], it is inferred that isoniazid, an important chemotherapeutic and also a carcinogenic agent, can lead to a substantial generation of electronically excited states.

Cetrimonium↗

A clover mutant lacking the chlorophyll a- and b-containing protein antenna complexes.

A chlorophyll deficient mutant of clover has been examined by SDS-PAGE, spectrophotometric and electron microscopic methods. By a comparison of the absorption and first derivative spectra of acetone extracts from the mutant and normal biotypes, we observed a deficiency in chlorophyll b for the mutant biotype. The calculated chlorophyll a/b ratios, using the method of Arnon (Plant Physiol. 24, 1-15, 1949), approached infinity for the mutant whereas it ranged from 3.0-3.3 in the wild-type. The low temperature (77 degrees K) fluorescence emission bands in the 685-695 nm region could not be differentiated for the biotypes; however, the long wavelength emission band (near 740 nm in the wild-type) was shifted to shorter wavelengths (ca 720-725 nm) in the mutant indicating loss of photosystem I antenna. The SDS-PAGE profile of the mutant biotype showed a dramatic decline in the Coomassie stained polypeptides of apparent molecular weights similar to those of LHC II. Transmission electron micrographs of the mutant and normal tissue exhibited similar extents of grana-stacking, indicating that a component(s) other than the LHC II may be responsible for membrane adhesion in this mutant.

Chlorophyll↗

A micellar model system for the role of zeaxanthin in the non-photochemical quenching process of photosynthesis--chlorophyll fluorescence quenching by the xanthophylls.

To get an insight to the mechanism of the zeaxanthin-dependent non-photochemical quenching in photosystem II of photosynthesis, we probed the interaction of some xanthophylls with excited chlorophyll-a by trapping both pigments in micelles of triton X-100. Optimal distribution of pigments among micelles was obtained by proper control of the micelle concentration, using formamide in the reaction mixture, which varies the micellar aggregation number over three orders of magnitude. The optimal reaction mixture was obtained around 40% (v/v) formamide in 0.2-0.4% (v/v) triton X-100 in water. Zeaxanthin in the micellar solution exhibited initially absorption and circular dichroism spectral features corresponding to a J-type aggregate. The spectrum was transformed over time (half-time values vary-an average characteristic figure is roughly 20 min) to give features representing an H-type aggregate. The isosbestic point in the series of spectral curves favors the supposition of a rather simple reaction between two pure J and H-types dimeric species. Violaxanthin exhibited immediately stable spectral features corresponding to a mixture of J-type and more predominately H-type dimers. Lutein, neoxanthin and beta-carotene did not show any aggregated spectral forms in micelles. The spectral features in micelles were compared to spectra in aqueous acetone, where the assignment to various aggregated types was established previously. The specific tendency of zeaxanthin to form the J-type dimer (or aggregate) could be important for its function in photosynthesis. The abilities of five carotenoids (zeaxanthin, violaxanthin, lutein, neoxanthin and beta-carotene) to quench chlorophyll-a fluorescence were compared. Zeaxanthin, in its two micellar dimeric forms, and beta-carotene were comparable good quenchers of chlorophyll-a fluorescence. Violaxanthin was a much weaker quencher, if at all. Lutein and neoxanthin rather enhanced the fluorescence. The implications to non-photochemical quenching process in photosynthesis are discussed.

Carotenoids↗

Association of chlorophyll a/b-binding Pcb proteins with photosystems I and II in Prochlorothrix hollandica.

Action spectra for photosystem II (PSII)-driven oxygen evolution and of photosystem I (PSI)-mediated H(2) photoproduction and photoinhibition of respiration were used to determine the participation of chlorophyll (Chl) a/b-binding Pcb proteins in the functions of pigment apparatus of Prochlorothrix hollandica. Comparison of the in situ action spectra with absorption spectra of PSII and PSI complexes isolated from the cyanobacterium Synechocystis 6803 revealed a shoulder at 650 nm that indicated presence of Chl b in the both photosystems of P. hollandica. Fitting of two action spectra to absorption spectrum of the cells showed a chlorophyll ratio of 4:1 in favor of PSI. Effective antenna sizes estimated from photochemical cross-sections of the relevant photoreactions were found to be 192+/-28 and 139+/-15 chlorophyll molecules for the competent PSI and PSII reaction centers, respectively. The value for PSI is in a quite good agreement with previous electron microscopy data for isolated Pcb-PSI supercomplexes from P. hollandica that show a trimeric PSI core surrounded by a ring of 18 Pcb subunits. The antenna size of PSII implies that the PSII core dimers are associated with approximately 14 Pcb light-harvesting proteins, and form the largest known Pcb-PSII supercomplexes.

Bacterial Proteins↗

The long wavelength forms of chlorophyll a.

When Euglena gracilis is cultured with light of low intensity (ca. 250 ft-c), an absorption band at 695 mmu is formed in an amount equal to about 20 per cent of the total chlorophyll absorption in this red region. An equally large proportion of C(a)695 is observed in Ochromonas danica, irrespective of light intensity. Other algae tested appear to contain approximately 3 to 5 per cent of their chlorophyll as C(a)695; this proportion does not increase as strikingly with lowering of the light intensity as it does in Euglena. C(a)695 bleaches more readily than the other chlorophyll forms both reversibly, in whole cells, and irreversibly, in homogenates. Cells containing a large proportion of C(a)695 have a fluorescence maximum at 708 mmu, as contrasted to the 687 mmu maximum in other algae. Occasionally, old cultures of Euglena contain cells with an absorption band at approximately 710 mmu. This absorption band is quite stable in aqueous extracts; when the pigment is transferred to ether an equivalent amount of pheophytin a is found to be present. Conditions leading to the formation of the 710 mmu absorption band are not yet known.

Chlorophyll↗

Chlorophyll a and carotenoid triplet states in light-harvesting complex II of higher plants.

Laser-flash-induced transient absorption measurements were performed on trimeric light-harvesting complex II to study carotenoid (Car) and chlorophyll (Chl) triplet states as a function of temperature. In these complexes efficient transfer of triplets from Chl to Car occurs as a protection mechanism against singlet oxygen formation. It appears that at room temperature all triplets are being transferred from Chl to Car; at lower temperatures (77 K and below) the transfer is less efficient and chlorophyll triplets can be observed. In the presence of oxygen at room temperature the Car triplets are partly quenched by oxygen and two different Car triplet spectral species can be distinguished because of a difference in quenching rate. One of these spectral species is replaced by another one upon cooling to 4 Ki demonstrating that at least three carotenoids are in close contact with chlorophylls. The triplet minus singlet absorption (T-S) spectra show maxima at 504-506 nm and 517-523 nm, respectively. In the Chl Qy region absorption changes can be observed that are caused by Car triplets. The T-S spectra in the Chl region show an interesting temperature dependence which indicates that various Car's are in contact with different Chl a molecules. The results are discussed in terms of the crystal structure of light-harvesting complex II.

Aerobiosis↗

The effects of variability in the inherent optical properties on estimations of chlorophyll a by remote sensing in Swedish freshwaters.

During late summer of 1997 measurements were made of the inherent optical properties in Lake Mälaren, Sweden. These included measurements of the total particulate absorption, phytoplankton absorption, yellow substance absorption and total backscattering. Measurements were also made of concentrations known to affect the inherent optical properties; chlorophyll a and suspended particulate organic and inorganic matter. At the same time measurements of underwater radiance reflectance were made. The above measurements were used to better define the relationships between the inherent optical properties and the concentrations affecting them. Correlation analyses were also made in order to study covariation of the concentrations of suspended and dissolved substances in the data set. These relationships were then used to better parameterize a semi-analytical model predicting remote sensing reflectance. Multivariate sensitivity analyses showed that large variations on the Rr(700-710)/Rr(678-685) ratio vs. chlorophyll concentrations could be expected in a heterogeneous environment such as Lake Mälaren. This could lead to large errors in subsequent estimates of chlorophyll concentrations.

Adsorption↗

Spectroscopic study of trypsin, heat and triton X-100-induced denaturation of the chlorophyll-binding protein CP43.

Trypsin-, heat- and Triton X-100-induced denaturation of CP43, the core antenna complex of photosystem II purified from spinach, has been investigated using absorption, fluorescence and circular dichroism spectroscopy. Triton X-100 was found to bring about considerable dissolution of pigments from the protein to the monomeric state in solution and destruction of the interactions among the chlorophyll, carotene and protein. Heat induced significant unfolding of the protein secondary structure and loss of excitonic interactions of the pigments, but no apparent dissolution of the pigments from CP43. Trypsin caused structural changes in the extrinsic part of the protein but no change of the native state of the pigments. Trypsin, heat and Triton X-100 treatments increased the light sensitivity of chlorophyll in CP43 to different extents. The results suggest that the protein and beta-carotene can protect the chlorophyll from light-induced destruction in CP43.

Binding Sites↗

Investigation of genotoxic and antigenotoxic activities of chlorophylls and chlorophyllin in cultured V79 cells.

Chlorophyll and its derivatives are examples of plant compounds (purified and/or extracted) which appear to protect DNA from damage caused by chemical or physical agents, although some studies have identified clastogenic activity of these compounds. This study was carried out to assess the genotoxic activity of chlorophyll-a (Chl-a), -b (Chl-b) and chlorophyllin (Chl) and their antigenotoxic activity against the DNA damage induced by methyl methanesulphonate (MMS) under conditions of simultaneous, pre-, post-treatment, and simultaneous treatment after pre-incubation of the chemical with MMS. The micronucleus (MN) test was used in binucleated cells (induced by cytochalasin-B) of a mammalian cell line (V79). The three concentrations of Chl-a, Chl-b or Chl (0.1375, 0.275, 0.55microM) were not genotoxic and the genotoxic action of MMS (400microM) decreased (74-117%) under all treatment conditions. The results showed that there was no significant difference among the treatment types, the concentration or the nature of chlorophyll used. The data obtained suggest that Chl-a, Chl-b and Chl when associated with the DNA damaging agent, MMS, may protect the DNA by desgenotoxic action and/or by bio-antigenotoxic mechanisms, with the similar efficiency.

Animals↗