Unicellular Green Alga Scenedesmus Obliquus Research Articles

Dissolved organic matter and aluminum oxide nanoparticles synergistically cause cellular responses in freshwater microalgae

ABSTRACTThis study investigated the impact of dissolved organic matters (DOM) on the ecological toxicity of aluminum oxide nanoparticles (Al2O3NPs) at a relatively low exposure concentration (1 mg L−1). The unicellular green alga Scenedesmus obliquus was exposed to Al2O3NP suspensions in the presence of DOM (fulvic acid) at various concentrations (1, 10, and 40 mg L−1). The results show that the presence of DOM elevated the growth inhibition toxicity of Al2O3NPs towards S. obliquus in a dose-dependent manner. Moreover, the combination of DOM at 40 mg L−1 and Al2O3NPs resulted in a synergistic effect. The relative contribution of Al-ions released from Al2O3NPs to toxicity was lower than 5%, indicating that the presence of the particles instead of the dissolved ions in the suspensions was the major toxicity sources, regardless of the presence of DOM. Furthermore, DOM at 10 and 40 mg L−1 and Al2O3NPs synergistically induced the upregulation of intercellular reactive oxygen species levels and superoxide dismutase activities. Analysis of the plasma malondialdehyde concentrations and the observation of superficial structures of S. obliquus indicated that the mixtures of DOM and Al2O3NPs showed no significant effect on membrane lipid peroxidation damage. In addition, the presence of both DOM and Al2O3NPs contributed to an enhancement in both the mitochondrial membrane potential and the cell membrane permeability (CMP) in S. obliquus. In particular, Al2O3NPs in the presence of 10 and 40 mg L−1 DOM caused a greater increase in CMP compared to Al2O3NPs and DOM alone treatments. In conclusion, these findings suggest that DOM at high concentrations and Al2O3NPs synergistically interrupted cell membrane functions and triggered subsequent growth inhibition toxicity.

Bioenergetic reprogramming plasticity under nitrogen depletion by the unicellular green alga Scenedesmus obliquus.

Simultaneous nitrogen depletion and 3,4-dichlorophenol addition induce a bioenergetic microalgal reprogramming, through strong Cyt b 6 f synthesis, that quench excess electrons from dichlorophenol's biodegradation to an overactivated photosynthetic electron flow and H 2 -productivity. Cellular energy management includes "rational" planning and operation of energy production and energy consumption units. Microalgae seem to have the ability to calculate their energy reserves and select the most profitable bioenergetic pathways. Under oxygenic mixotrophic conditions, microalgae invest the exogenously supplied carbon source (glucose) to biomass increase. If 3,4-dichlorophenol is added in the culture medium, then glucose is invested more to biodegradation rather than to growth. The biodegradation yield is enhanced in nitrogen-depleted conditions, because of an increase in the starch accumulation and a delay in the establishment of oxygen-depleted conditions in a closed system. In nitrogen-depleted conditions, starch cannot be invested in PSII-dependent and PSII-independent pathways for H2-production, mainly because of a strong decrease of the cytochrome b 6 f complex of the photosynthetic electron flow. For this reason, it seems more profitable for the microalga under these conditions to direct the metabolism to the synthesis of lipids as cellular energy reserves. Nitrogen-depleted conditions with exogenously supplied 3,4-dichlorophenol induce reprogramming of the microalgal bioenergetic strategy. Cytochrome b 6 f is strongly synthesized (mainly through catabolism of polyamines) to manage the electron bypass from the dichlorophenol biodegradation procedure to the photosynthetic electron flow (at the level of PQ pool) and consequently through cytochrome b 6 f and PSI to hydrogenase and H2-production. All the above showed that the selection of the appropriate cultivation conditions is the key for the manipulation of microalgal bioenergetic strategy that leads to different metabolic products and paves the way for a future microalgal "smart" biotechnology.

Co-exposure of Freshwater Microalgae to Tetrabromobisphenol A and Sulfadiazine: Oxidative Stress Biomarker Responses and Joint Toxicity Prediction.

Combined toxicity and oxidative stress biomarker responses were determined for tetrabromobisphenol A (TBBPA) and sulfadiazine (SDZ) to the unicellular green alga Scenedesmus obliquus. Concentration-response analyses were performed for single toxicants and for mixtures containing TBBPA and SDZ with two different mixture ratios. The effect concentrations and the observed effects of the mixtures were compared to the predictions of the joint toxicity by the concentration addition (CA) model and independent action (IA) model. Results showed that the observed joint toxicity was within the scope of the highest (TBBPA) and lowest (SDZ) toxicity observed for the individual components. Furthermore, co-exposure of S. obliquus to TBBPA and SDZ provided preliminary evidence that the mixtures induced oxidative stress leading to cell damage. The CA and IA models proved to be valid for the prediction of the joint toxicity of TBBPA and SDZ. This study highlights a combined environmental risk assessment for two emerging pollutants.

Production and characterization of biodiesel from the unicellular green alga Scenedesmus obliquus

ABSTRACTRegardless of whether biodiesel is represented in the form of oil from grains, agricultural crops, or microorganisms, it is a clean fuel depends mainly on biomass conversion. The present work indicates that the green microalga Scenedesmus obliquus showed higher biomass productivity of 0.15 g (CDW) L−1 d−1. The increase in NaCl concentration up to +100% caused an increase in growth, esterified fatty acid (EFA) content, and EFA productivity of S. obliquus. The decrease or increase of pH values reduced the growth, EFA content, and productivity. Biomass productivity increased from 0.06 g L−1d−1 under a light intensity of 80 μmol m−2s−1 to 0.1 g L−1d−1 at 160 μmol m−2 s−1. Biodiesel characterization showed that the iodine value of biodiesel of S. obliquus was 70 g iodine/100 g oil. Except viscosity, all biodiesel characteristics are in accordance with the European standards for fuel diesel (EN 590:1999). These results nominate S. obliquus as a promising candidate for biodiesel production by outdoor cultivation.

Impacts of low-molecular-weight organic acids on aquatic behavior of graphene nanoplatelets and their induced algal toxicity and antioxidant capacity.

Knowledge of the interaction between graphene-based materials and low-molecular-weight organic acids (LOAs) is essential to understand fate and effects of graphene-based materials in the aquatic environment, but this interaction remains poorly elucidated. In this study, the effects of LOAs on the physicochemical properties of graphene nanoplatelets (GNPs) in an aqueous medium and on the GNP toxicity to algae were studied. The unicellular green alga Scenedesmus obliquus was exposed to GNP suspensions in the presence of benzoic acid or gallic acid at various concentrations. The GNPs had smaller hydrodynamic sizes and the GNP suspensions were more stable and had higher or lower surface zeta potentials in the presence of LOAs than when LOAs were not present. The toxic effects in S. obliquus cultures incubated with GNP suspensions containing LOAs were related to the LOA concentration, and the presence of LOAs caused three effects: stimulation, alleviation, and synergistic inhibition. The intensities of the effects mainly correlated with the LOA concentration, the extent of agglomeration, and particle-induced oxidative stress. The results indicate that the environmental fates and toxicities of GNPs are strongly affected by the binding of GNPs to LOAs.

Bioenergetic Strategy for the Biodegradation of p-Cresol by the Unicellular Green Alga Scenedesmus obliquus

Cultures from the unicellular green alga Scenedesmus obliquus biodegrade the toxic p-cresol (4-methylphenol) and use it as alternative carbon/energy source. The biodegradation procedure of p-cresol seems to be a two-step process. HPLC analyses indicate that the split of the methyl group (first step) that is possibly converted to methanol (increased methanol concentration in the growth medium), leading, according to our previous work, to changes in the molecular structure and function of the photosynthetic apparatus and therefore to microalgal biomass increase. The second step is the fission of the intermediately produced phenol. A higher p-cresol concentration results in a higher p-cresol biodegradation rate and a lower total p-cresol biodegradability. The first biodegradation step seems to be the most decisive for the effectiveness of the process, because methanol offers energy for the further biodegradation reactions. The absence of LHCII from the Scenedesmus mutant wt-lhc stopped the methanol effect and significantly reduced the p-cresol biodegradation (only 9%). The present contribution deals with an energy distribution between microalgal growth and p-cresol biodegradation, activated by p-cresol concentration. The simultaneous biomass increase with the detoxification of a toxic phenolic compound (p-cresol) could be a significant biotechnological aspect for further applications.

Modeling the dynamic modulation of light energy in photosynthetic algae

An integrated cell-based dynamic mathematical model that take into account the role of the photon absorbing process, the partition of excitation energy, and the photoinactivation and repair of photosynthetic units, under variable light and dissolved inorganic carbon (DIC) availability is proposed. The modeling of the photon energy absorption and the energy dissipation is based on the photoadaptive changes of the underlying mechanisms. The partition of the excitation energy is based on the relative availability of light and DIC to the cell. The modeling of the photoinactivation process is based on the common aspect that it occurs under any light intensity and the modeling of the repair process is based on the evidence that it is controlled by chloroplast and nuclear-encoded enzymes. The present model links the absorption of photons and the partitioning of excitation energy to the linear electron flow and other quenchers with chlorophyll fluorescence emission parameters, and the number of the functional photosynthetic units with the photosynthetic oxygen production rate. The energy allocation to the LEF increases as DIC availability increases and/or light intensity decreases. The rate of rejected energy increases with light intensity and with DIC availability. The resulting rate coefficient of photoinactivation increases as light intensity and/or as DIC concentration increases. We test the model against chlorophyll fluorescence induction and photosynthetic oxygen production rate measurements, obtained from cultures of the unicellular green alga Scenedesmus obliquus, and find a very close quantitative and qualitative correspondence between predictions and data.

Fast and reversible response of thylakoid-associated polyamines during and after UV-B stress: a comparative study of the wild type and a mutant lacking chlorophyll b of unicellular green alga Scenedesmus obliquus

The functional and biochemical aspects of the photosynthetic apparatus in response to UV-B radiation were examined in unicellular oxygenic algae Scenedesmus obliquus. The wild type (Wt) and a chlorophyll b-less mutant (Wt-lhc) were used as a specific tool for the understanding of antenna role. Photosynthesis was monitored during and after UV-B stress by time resolved fluorescence spectroscopy and polarography. Carotenoids, such as neoxanthin, loroxanthin, lutein, violaxanthin, antheraxanthin, zeaxanthin, alpha- and beta-carotene, cellular and thylakoid-associated putrescine, spermidine, spermine and subcomplexes of light-harvesting complex (LHCII) of photosystem II (PSII) were investigated to assess their possible involvement in response to UV-B. Oxygen evolution depression by UV-B was higher in the Wt-lhc mutant than in the Wt. Photosynthesis recovery occurred in the Wt, but not in the mutant. The dissipation of excess excitation energy during UV-B stress was accompanied by changes in the thylakoid-associated polyamines which were much higher than changes in xanthophylls. We conclude that, at least in the unicellular green alga S. obliquus, mutants lacking chlorophyll b have significant lower capacity for recovery after UV-B stress. In addition, the comparison of xanthophylls and thylakoid-associated polyamines reveals that the latter are more responsive to UV-B stress and in a reversible manner.

Changes in the LHCII-mediated energy utilization and dissipation adjust the methanol-induced biomass increase

Considerably low methanol concentrations of 0.5% (v/v), induce an immense increase in biomass production in cultures of the unicellular green alga Scenedesmus obliquus compared to controls without additional methanol. The effect is light-regulated and it mimics high-CO 2 induced changes of the molecular structure and function of the photosynthetic apparatus. There is evidence that methanol enhances under high light conditions by molecular changes in the LHCII – a decrease of the functional antenna-size per active reaction center – the photochemical effectiveness of the absorbed energy. This means that the non-photochemical quenching (NPQ) is minimized and thereby the overall dissipation energy. Experiments with mutants of Scenedesmus Wt produced evidence that the LHCII is the locus of the mechanism which regulates the methanol effect. The employed mutants were Wt-LHC, lacking a functioning LHCII, the light-dependent greening mutant C-2A′, and the double mutant C-2A′-LHC, combining both mutations.

Thylakoid‐associated Polyamines Adjust the UV‐B Sensitivity of the Photosynthetic Apparatus by Means of Light‐harvesting Complex II Changes¶

AbstractThe sensitivity of the photosynthetic apparatus to ultraviolet‐B (UV‐B) irradiation was studied in cultures of unicellular green alga Scenedesmus obliquus incubated in low light (low photosynthetically active radiation intensity [LL]) and high light (high photosynthetically active radiation intensity [HL]) conditions, treated or not with exogenous polyamines. Biochemical and physicochemical measurement showed that UV‐B radiation induces a decrease in the thylakoid‐associated putrescine (Put) and an increase in spermine (Spm), so that the reduction of Put/Spm ratio leads to the increase of light‐harvesting complex II (LHCII) size per active reaction center and, consequently, the amplification of UV‐B effects of the photosynthetic apparatus. The separation of oligomeric and monomeric froms of LHCII from isolated thylakoids showed that UV‐B induces an increase in the oligomeric forms of LHCII, which was more intense in LL than in HL. By manipulating the LHCII size with exogenous polyamines, the sensitivity degree of the photosynthetic apparatus to UV‐B changed significantly. Specifically, the addition of Put decreased highly the sensitivity of LL culture to UV‐B because of the inhibitory effect of Put on the LHCII size increasing, wheres the addition of Spm enhanced the UV‐B injury induced in HL culture because of the increasing of LHCII size. The ability of the photosynthetic apparatus to recover the UV‐B induced changes was also investigated.

The impact of high CO 2 concentrations on the structure and function of the photosynthetic apparatus and the role of polyamines

Here we examined the influence of high CO2 concentrations on the structure and functioning of the photosynthetic apparatus in the unicellular green alga Scenedesmus obliquus. Presented in this work are: chlorophyll (Chl) a fluorescence induction kinetics, measurements of photosynthetic and respiration rates, estimation of Chl a/Chl b ratios, isolation and quantitative assessment of the photosynthetic subcomplexes, quantitative analyses of thylakoid bound polyamines, and experiments with exogenously supplied polyamines with cultures grown in low- and high-CO2 concentrations. Together, they indicated that high-CO2 concentrations affect polyamines and, more specifically, increase the thylakoid bound putrescine (PUT) level that leads to an increase of the active reaction center density combined with a decrease in the LHCII-size and the ratio of LHCII-oligomeres/LHCII-monomeres. This reorganization of the photosynthetic apparatus leads to enhanced photosynthetic rates, which in combination with the high-CO2 concentrations, leads to an immense increase of biomass (800%). Further incubation for longer time periods under the same conditions produces, due to an increase in cell density, a self-shading effect and photoadaptation of the photosynthetic apparatus to low light conditions and therefore also results in reduction of the high-CO2 effect. The photoadaptation of the photosynthetic apparatus to high-light conditions (Kotzabasis et al. 1999) and the acclimation to high-CO2 concentrations (present work) lead to the same changes in the structure and function of the photosynthetic apparatus. These changes could be induced or inhibited through the manipulation of intracellular polyamines, especially through the putrescine/spermine ratio. The possibility that polyamines influence the photoadaptation of the photosynthetic apparatus and its acclimation to high-CO2 concentrations through a common mechanism is discussed.

Thylakoid-associated Polyamines Adjust the UV-B Sensitivity of the Photosynthetic Apparatus by Means of Light-harvesting Complex II Changes¶

The sensitivity of the photosynthetic apparatus to ultraviolet-B (UV-B) irradiation was studied in cultures of unicellular green alga Scenedesmus obliquus incubated in low light (low photosynthetically active radiation intensity [LL]) and high light (high photosynthetically active radiation intensity [HL]) conditions, treated or not with exogenous polyamines. Biochemical and physicochemical measurements showed that UV-B radiation induces a decrease in the thylakoid-associated putrescine (Put) and an increase in spermine (Spm), so that the reduction of Put/Spm ratio leads to the increase of light-harvesting complex II (LHCII) size per active reaction center and, consequently, the amplification of UV-B effects on the photosynthetic apparatus. The separation of oligomeric and monomeric forms of LHCII from isolated thylakoids showed that UV-B induces an increase in the oligomeric forms of LHCII, which was more intense in LL than in HL. By manipulating the LHCII size with exogenous polyamines, the sensitivity degree of the photosynthetic apparatus to UV-B changed significantly. Specifically, the addition of Put decreased highly the sensitivity of LL culture to UV-B because of the inhibitory effect of Put on the LHCII size increasing, whereas the addition of Spm enhanced the UV-B injury induced in HL culture because of the increasing of LHCII size. The ability of the photosynthetic apparatus to recover the UV-B induced changes was also investigated.

Pigments as biomarkers of exposure to the vineyard herbicide flazasulfuron in freshwater algae

Weed control in Champagne vineyards has long relied on the use of diuron and substituted triazines; these compounds are now being replaced by flazasulfuron, a sulfonylurea that is used at a much lower dosage. The vineyards of Champagne are planted on steep slopes and runoff is important, and even though low doses of these herbicides are used, they may present some potential risk for freshwater ecosystems. Therefore, the effects of the sulfonylurea herbicide, flazasulfuron (formulated as Katana) was investigated on the unicellular green alga Scenedesmus obliquus. The pigment content of the algal suspensions was followed as a biomarker of exposure to the herbicide. The results demonstrate that flazasulfuron induced a reduction in chlorophyll content at concentrations of 10 μg/L, while the increase of pigment content in the culture was reduced with the lowest concentration tested (0.1 μg/L). Among the three pigments tested, chlorophyll a appeared to be the most sensitive biomarker. In the algal medium, flazasulfuron was slowly degraded (DT 50 approximately 8 days) in a compound that was tentatively identified. The toxicity of this herbicide for the algae was comparable to that of older herbicides which are used at a much higher rate. Therefore, we may speculate that even if flazasulfuron comes into contact with freshwater ecosystems, its effects on algae will be less deleterious than that of traditional herbicides.

Light-dependent induction of strongly increased microalgal growth by methanol

Low methanol concentrations (about 0.5% v/v) induce biomass production in cultures of the unicellular green alga Scenedesmus obliquus by more than 300%, compared to controls without this solvent. This effect on the microalgal growth was found to be dependent on the solvent concentration, the packed cell volume (PCV), light intensity and light quality. It could be shown that methanol addition leads to a decrease in size of the light harvesting complex (LHC) on the basis of chlorophylls and proteins, and thus to changes in structure and functioning of the photosynthetic apparatus. These alterations lead to enhanced photosynthesis and respiration rates. The action of methanol on the photosynthetic apparatus is thus comparable to the effect of enhanced CO 2 concentrations. These findings support the previously proposed pathway for methanol metabolization with CO 2 as the final product. We conclude that the subsequent assimilation of the increased CO 2 amounts by the Calvin–Benson cycle is a possible explanation for the methanol-mediated increase in biomass production in terms of PCV. The methanol effect is observed only in the light and in the presence of a functioning photosynthetic apparatus. Preliminary action spectra suggest that the primary photoreceptor is a chlorophyll–protein complex with two absorption maxima at 680 and 430 nm, which may possibly be attributed to the reaction center of photosystem II (PSII).

Amino acid sequence, crystallization and structure determination of reduced and oxidized cytochrome c6 from the green alga Scenedesmus obliquus

Cytochrome c 6 from the unicellular green alga Scenedesmus obliquus was sequenced, crystallized in its reduced and oxidized state and the three-dimensional structure of the protein in both redox states was determined by X-ray crystallography. Reduced cytochrome c 6 crystallized as a monomer in the space group P2 12 12, whereas the oxidized protein crystallized as a dimer in the space group P3 121. The structures were solved by molecular replacement and refined to 1.9 and 2.0 Å, respectively. Comparison of the structures of both redox states revealed only slight differences on the protein surface, whereas a distortion along the axis between the heme iron and its coordinating Met61 residue was observed. No redox-dependent movement of internal water molecules could be detected. The high degree of similarity of the surfaces and charge distributions of both redox states, as well as the dimerization of cytochrome c 6 as observed in the oxidized crystal, is discussed with respect to its biological relevance and its implications for the reaction mechanisms between cytochrome c 6 and its redox partners. The dimer of oxidized cytochrome c 6 may represent a molecular structure occurring in a binary complex with cytochrome b 6 f. This assembly might be required for the correct orientation of cytochrome c 6 with respect to its redox partner cytochrome b 6 f, facilitating the electron transfer within the complex. If the dimerization is not redox-dependent in vivo, the almost identical surfaces of both redox states do not support a long range differentiation between reduced and oxidized cyt c 6, i.e. a random collision model for the formation of an electron transfer complex must be assumed.

The regulatory role of polyamines in structure and functioning of the photosynthetic apparatus during photoadaptation

In this contribution we describe the changes in structure and functioning of the photosynthetic apparatus of the unicellular green alga Scenedesmus obliquus induced by inhibition or induction of polyamine biosynthesis. The synthesis or inhibition is controlled by white light of low and high intensities, as well as by blue and red irradiation. We observe that a decrease of the intracellular putrescine level and, vice versa, an increase of spermine, indicated by a raised ratio of spermine to putrescine, simulate a low-light photoadapted photosynthetic apparatus. Action spectra of the spermine/putrescine ratio compared with action spectra on characteristics of photoadaptation, e.g., chlorophyll biosynthesis, or the rate of primary photochemistry and electron transport, show that the photoreceptors for both the adaptation of the photosynthetic apparatus to low-light conditions and for the regulation of the spermine/putrescine ratio are, at least, very similar. We show that the photoreceptor is primarily a blue-light receptor with a superimposed red-light receptor that absorbs around 680 nm.

Differential Changes in the Photosynthetic Pigments and Polyamine Content during Photoadaptation and Photoinhibition in the Unicellular Green Alga Scenedesmus obliquus

In the unicellular green alga Scenedesmus obliquus the level of photoinhibition and the recovery of the cells after reversal to the initial light conditions in relation to the pre-photoadaptation of the culture to low, medium and high light intensity was determined. The changes in the photosynthetic pigment content and in the intracellular polyamine concentration allowed to distinguish between photoadaptation and photoinhibition. In particular, the level of chlorophylls, xanthophylls and carotenoids decreased inversely proportional to the light intensity applied during photoadaptation, whereas their concentrations remained constant during photoinhibition. The violaxanthin/zeaxanthin and the loroxanthin/lutein cycle work only under photoinhibitory conditions, but not under photoadaptive premises. Changes in the level of these carotenoids in relation to the changes in the photosynthetic apparatus during photoadaptation are discussed. In addition, it was found that the intracellular polyamine level increased only under stress conditions, i. e. during photoinhibition, and decreased during recovery of the cells after reversal to the initial light conditions. The increase of the putrescine level during photoinhibition is inversely proportional to the light intensity used for pre-adaptation. This rise of the polyamine level in the cells photoadapted to high light conditions is an additional indication for the finding that photoadaptation and photoinhibition are different phenomena which are clearly distinguishable from each other. Finally, the changes of the chlorophyll, violaxanthin, zeaxanthin, loroxanthin, lutein and polyamine levels under photoadaptation in high light intensity (50 W m -2) in relation to the range of photoadaptation in Scenedesmus obliquus are discussed.

Changes in the polyamine content of plastidal membranes in light- and dark-grown wildtype and pigment mutants of the unicellular green alga Scenedesmus obliquus and their possible role in chloroplast photodevelopment

Polyamines are involved in a great variety of developmental processes in plant cells, however, in many cases the mechanism and the place of action are unknown. In the present paper attempts are made to elucidate, in part, the physiological role of putrescine, spermidine, spermine and norspermidine in chloroplast development. It is shown that cultures of the unicellular green alga Scenedesmus obliquus and its pigment mutants C-2A' and C-6D, grown heterotrophically in darkness, exhibit, when exposed to light, an immediate and continuing decrease of the intracellular putrescine level, whereas the polyamines spermidine, spermine and norspermidine present only slight changes. Additionally, a considerable redistribution of the bound polyamines in the isolated plastid membranes from various chloroplast developmental stages is observed. The fluctuation of the bound polyamines in the plastid membranes during darkness and illumination was determined and correlated to different ways of chloroplast development represented by the three types of Scenedesmus cultures (wildtype, mutants C-2A' and C-6D). A possible role of polyamines in the assembly of the various photosynthetic complexes during the formation of the photosynthetic apparatus is discussed.

The relation between the photochemical yield and variable fluorescence of photosystem II in the green alga Scenedesmus obliquus

The redox state of the primary quinone acceptor (Q A) of photosystem II (PS II) and specific non-photochemical effects determine the relation between PS II photochemistry and chlorophyll fluorescence. For the unicellular green alga Scenedesmus obliquus, the fluorescence yield ( F), the fluorescence yield in the presence of reduced Q A ( F M′) and the yield of respiration-corrected oxygen evolution (ф ox, evolution rate divided by light intensity) were determined during exposure to actinic light of various intensities (0.1 – 1600 μE m −2 s −1). Irrespective of the growth conditions, the parameter ф p = (F M ′ − F)/F M ′ was found to be proportional to ф ox for each algal culture. This finding is in full agreement with non-photochemical fluorescence quenching by increased thermal deactivation of excited antenna states, but is difficult to reconcile with non-photochemical quenching due to modified PS II electron transfer reactions. In particular, no indications of cyclic PS II electron flow were found.

Charging of Both, Plastidal tRNAgln and tRNAglu with Glutamate and Subsequent Amidation of the Misacylated tRNAgln by a Glutamyl-tRNA Amidotransferase in the Unicellular Green Alga Scenedesmus obliquus, Mutant C-2A'

Abstract , 1995 C5-Pathway, Glutamyl-tRNAglu-Synthetase (E.C.6.1.1.17), Misacylation of tR N A glu, Amidotransferase, Scenedesmus obliquus In a previous paper we described the purification of a glutamyl-tRNA synthetase from the unicellular green alga Scenedesmus obliquus, m utant C-2A'. We now dem onstrate that, firstly, this enzyme is capable of mischarging plastidal tR N A gln from barley with glutamate, as well as it regularly charges the plastidal tR N A glu from Scenedesmus. Secondly, we show that the mischarged glutamyl-tRNAgln is subsequently am idated by a glutamyl-tRNA am idotransfer­ ase to form the glutaminyl-tRNAglri required for plastidal protein biosynthesis. This phenom ­ enon could already be dem onstrated for higher plant chloroplasts, mitochondria, cyanobact­ eria and gram-positive bacteria, as far as investigated. As recently shown the applied glutamyl-tRNA synthetase from Scenedesmus is a plastidal enzyme. In this paper we prove by treatm ent with m onobromobim ane and cyanogen bromide that the "regular" substrate of the enzyme, tR N A glu from Scenedesmus, is a plastidal tRNA with the plastid-specific sulfur modification in the anticodon. In the case of cyanogen bromide treatm ent, a total inactivation of the tRNA was achieved, revealing the presence of a sulfur modification in the plastid-tRNA glu anticodon.