Effect of the exogenous anthocyanin extract on key metabolic pathways and antioxidant status of Brazilian elodea (Egeria densa (Planch.) Casp.) exposed to cadmium and manganese.

Present study deals with the effect of 24 h pre-incubation with exogenous anthocyanins (ANTH), extracted from red cabbage leaves, on key metabolic processes (photosynthesis and respiration) and pro-/antioxidant balance in the aquatic macrophyte Egeria densa (Planch.) Casp., Hydrocharitaceae family, treated with Cd and Mn (in sulfate form) at a concentration of 100 µmol. After five days of metal treatments, Cd was accumulated and the damage caused to metabolic processes was stronger than Mn. In Cd-treated leaves, the protein level, chlorophyll concentration and maximal photochemical efficiency of PS II decreased twofold, and net-photosynthesis was significantly inhibited, whereas lipid peroxidation and H2O2 production increased. In turn, protective responses developed, including an increase in the total soluble thiols, alternative respiratory pathway capacity and the activity of superoxide dismutase and peroxidases. Pre-incubation in the ANTH-enriched extract caused an increase in foliar ANTH content, enhanced Cd and reduced Mn uptake into the tissue. A decrease in the level of oxidative reactions, an increase in the protein and chlorophyll concentration compared to the control values and a partial improvement of the photosynthetic parameters confirmed the ability of ANTH to reduce Cd-induced damage effects and to mitigate ROS-driven stress reactions. Stimulation of catalase and ascorbate peroxidase activity, an alternative respiration capacity and non-enzymatic antioxidant (carotenoids, ascorbate and proline) synthesis by ANTH were also revealed. These data suggest that ANTH-enriched extract from red cabbage leaves has a protective action against metal toxicity in Egeria plants.

Study on chromium-binding capacity of Callitriche cophocarpa in an aquatic environment.

The aim of the present study was to investigate the binding strength of chromium (Cr) ions to aquatic macrophyte Callitriche cophocarpa. Shoots of the plants were incubated in a natural water solution containing Cr(III) or Cr(VI) at a concentration ranging from 0.5 to 4 mM under laboratory conditions. We found that C. cophocarpa has an extremely high capacity to bind Cr. The average level of accumulation reached 28,385 or 7,315 mg kg(-1) dry weight for plants incubated with Cr(III) or Cr(VI), respectively. Shoots incubated in a 0.5 mM concentration of Cr(III) for 5 days removed almost 100 % of the metal from solution. The major pool of the bound Cr(III) ions follows the strongest mechanism of metal-binding to an organic matter. In contrast, we found that only 25 % of Cr(VI) ions are bound into the metallo-organic compounds and 57 % of Cr(VI) exists in an easily remobilizable form. Activity of a photosynthetic electron transport (as F V/F M) was evaluated with respect to the Cr-binding mechanism. Our results contribute to the development of knowledge on processes controlling bioremediation of heavy-metallic compounds in aquatic systems.

Small effects of a large sediment contamination with heavy metals on aquatic organisms in the vicinity of an abandoned lead and zinc mine.

The effects of the long-term contamination of water reservoirs with mine effluents were investigated at an abandoned mine site in Upper Silesia, southern Poland. The studies covered metal content and mobility in bottom sediments as well as water chemistry in relation to the content of metals in selected macrophytes and their physiology and the composition of phyto- and zooplankton communities. Although it is 40 years since mining ceased, reservoir sediments are still heavily contaminated with cadmium, zinc and lead with concentrations (mg/kg), which vary roughly between 130­340, 10,000­50,000 and 4,000­12,000, respectively. About 50­80 % of these elements are associated with the reducible phase, and only a small percentage, <10%, is present in the most mobile exchangeable phase. Despite the high total metal concentration in sediments, their content in the submerged plants Myriophyllum spicatum and the emerged plants Phragmites australis was low. The observed effects of heavy metal contamination on photosynthetic activity in the leaves of P. australis were negligible, whereas those in M. spicatum show up only as a difference in the distribution of photosynthetic activity in leaves of different ages, which seems to be related to the very good water quality and to the generally small concentrations of metals in pond water. The physicochemical properties of water also seem to control the presence of planktonic species more than does sediment contamination. However, a shift toward groups of species known to be more resistant to heavy metals (diatoms, green algae and Rotifera) indicates some adaptative changes related to the longlasting contamination of ponds.

The role of antioxidant response and nonphotochemical quenching of chlorophyll fluorescence in long-term adaptation to Cu-induced stress in Chlamydomonas reinhardtii.

Copper is an essential micronutrient, but at supraoptimal concentrations it is also highly toxic, inducing oxidative stress and disrupting photosynthesis. The aim of the present study was to analyze selected protective mechanisms in strains of Chlamydomonas reinhardtii adapted and not adapted for growth in the presence of elevated copper concentrations. Two algal lines (tolerant and non-tolerant to high Cu2+ concentrations) were used in experiments to study photosynthetic pigment content, peroxidase activity, and non-photochemical quenching. The content of prenyllipids was studied in four different algal lines (two of the same as above and two new ones). The copper-adapted strains contained about 2.6 times more α-tocopherol and plastoquinol and about 1.7 times more total plastoquinone than non-tolerant strains. Exposure to excess copper led to oxidation of the plastoquinone pool in non-tolerant strains, whereas this effect was less pronounced or did not occur in copper-tolerant strains. Peroxidase activity was approximately 1.75 times higher in the tolerant strain than in the non-tolerant one. The increase in peroxidase activity in the tolerant strain was less pronounced when the algae were grown in dim light. In the tolerant line nonphotochemical quenching was induced faster and was usually about 20-30% more efficient than in the non-tolerant line. The improvement of antioxidant defense and photoprotection may be important factors in the evolutionary processes leading to tolerance to heavy metals.

Arabidopsis BPG2: a phytochrome-regulated gene whose protein product binds to plastid ribosomal RNAs.

BPG2 (Brz-insensitive pale green 2) is a dark-repressible and light-inducible gene that is required for the greening process in Arabidopsis. Light pulse experiments suggested that light-regulated gene expression of BPG2 is mediated by phytochrome. The T-DNA insertion mutant bpg2-2 exhibited a reduced level of chlorophyll and carotenoid pigmentation in the plastids. Measurements of time resolved chlorophyll fluorescence and of fluorescence emission at 77 K indicated defective photosystem II and altered photosystem I functions in bpg2 mutants. Kinetic analysis of chlorophyll fluorescence induction suggested that the reduction of the primary acceptor (QA) is impaired in bpg2. The observed alterations resulted in reduced photosynthetic efficiency as measured by the electron transfer rate. BPG2 protein is localized in the plastid stroma fraction. Co-immunoprecipitation of a formaldehyde cross-linked RNA-protein complex indicated that BPG2 protein binds with specificity to chloroplast 16S and 23S ribosomal RNAs. The direct physical interaction with the plastid rRNAs supports an emerging model whereby BPG2 provides light-regulated ribosomal RNA processing functions, which are rate limiting for development of the plastid and its photosynthetic apparatus.

Cadmium inhibitory action leads to changes in structure of ferredoxin:NADP(+) oxidoreductase.

This study deals with the influence of cadmium on the structure and function of ferredoxin:NADP(+) oxidoreductase (FNR), one of the key photosynthetic enzymes. We describe changes in the secondary and tertiary structure of the enzyme upon the action of metal ions using circular dichroism measurements, Fourier transform infrared spectroscopy and fluorometry, both steady-state and time resolved. The decrease in FNR activity corresponds to a gentle unfolding of the protein, caused mostly by a nonspecific binding of metal ions to multiple sites all over the enzyme molecule. The final inhibition event is most probably related to a bond created between cadmium and cysteine in close proximity to the FNR active center. As a result, the flavin cofactor is released. The cadmium effect is compared to changes related to ionic strength and other ions known to interact with cysteine. The complete molecular mechanism of FNR inhibition by heavy metals is discussed.Electronic supplementary material The online version of this article (doi:10.1007/s10867-012-9262-z) contains supplementary material, which is available to authorized users.

Effect of cadmium on ferredoxin:NADP+ oxidoreductase activity.

Ferredoxin:NADP(+) oxidoreductase (FNR) was treated with cadmium and after that its diaphorase reaction in the presence of dibromothymoquinone (DBMIB) or ferricyanide (FeCy, K(3)Fe(CN)(6)) was examined. CdSO(4) (5 mM) caused 50% inhibition after half hour incubation. At least two components were distinguishable in the time-course inhibition, suggesting that more than one amino acid residues were engaged in reaction with the metal ion. The Lineweaver-Burk plots indicate that Cd(2+) is an uncompetitive inhibitor for DBMIB reduction but exerts non-competitive inhibition for the NADPH oxidation. The FeCy reduction did not follow Michaelis-Menten kinetics. Zn(2+) diminished inhibitory effect of Cd(2+) on the DBMIB reduction but enhanced inhibition of the FeCy reduction. Incubation with additional chelator (beta-mercaptoethanol, or histidine) abolished inhibitory effect of Cd(2+) on the FeCy reduction but not on the DBMIB reduction. The mode of Cd(2+) action on the diaphorase activity of FNR in the presence of DBMIB or FeCy is briefly discussed with the special reference to the implication of two distinct sites at the FNR molecule, which might be involved in the reduction of various non-physiological substrates.

Temperature effect on growth, and selected parameters of Phaeodactylum tricornutum in batch cultures.

The effect of optimal and stress temperatures on the growth kinetics of the Phaeodactylum tricornutum CCAP/1055/1 strain (a model diatom with a known genome sequence) in batch cultures was examined. The analysis of the obtained results showed two phases of culture growth. There were significant positive correlations between OD increase of chlorophyll a chlorophyll c and protein concentration at different temperatures. The Fv/Fm parameter achieved a maximum level on the 6(th) or 7(th) day and then decreased to the values registered on the first day of observation. Genetic material undergoes gradual degradation 10 days after inoculation. The size of the cells was invariable.

Ferredoxin:NADP+ oxidoreductase as a target of Cd2+ inhibitory action--biochemical studies.

The ferredoxin:NADP+ oxidoreductase (FNR) catalyses the ferredoxin-dependent reduction of NADP+ to NADPH in linear photosynthetic electron transport. The enzyme also transfers electrons from reduced ferredoxin (Fd) or NADPH to the cytochrome b(6)f complex in cyclic electron transport. In vitro, the enzyme catalyses the NADPH-dependent reduction of various substrates, including ferredoxin, the analogue of its redox centre - ferricyanide, and the analogue of quinones, which is dibromothymoquinone. This paper presents results on the cadmium-induced inhibition of FNR. The K(i) value calculated for research condition was 1.72 mM. FNR molecule can bind a large number of cadmium ions, as shown by the application of cadmium-selective electrode, but just one ion remains bound after dialysis. The effect of cadmium binding is significant disturbance in the electron transfer process from flavin adenine dinucleotide (FAD) to dibromothymoqinone, but less interference with the reduction of ferricyanide. However, it caused a strong inhibition of Fd reduction, indicating that Cd-induced changes in the FNR structure disrupt Fd binding. Additionally, the protonation of the thiol groups is shown to be of great importance in the inhibition process. A mechanism for cadmium-caused inhibition is proposed and discussed with respect to the in vitro and in vivo situation.

Photoprotective function of chloroplast avoidance movement: in vivo chlorophyll fluorescence study.

Light-induced chloroplast avoidance movement has long been considered to be a photoprotective mechanism. Here, we present an experimental model in which this function can be shown for wild type Arabidopsis thaliana. We used blue light of different fluence rates for chloroplast positioning, and strong red light inactive in chloroplast positioning as a stressing light. The performance of photosystem II was measured by means of chlorophyll fluorescence. After stressing light treatment, a smaller decrease in photosystem II quantum yield was observed for leaves with chloroplasts in profile position as compared with leaves with chloroplasts in face position. Three Arabidopsis mutants, phot2 (no avoidance response), npq1 (impaired zeaxanhtin accumulation) and stn7 (no state transition), were examined for their chloroplast positioning and chlorophyll fluorescence parameters under identical experimental conditions. The results obtained for these mutants revealed additional stressing effects of blue light as compared with red light.

Chromium(VI) bioremediation by aquatic macrophyte Callitriche cophocarpa Sendtn.

Callitriche cophocarpa (water-starwort)--aquatic widespread macrophyte--was found to be an excellent chromium accumulator. The plants were exposed to various chromium(VI) concentration ranging from 50 to 700 microM in a hydroponic culture up to ca. 3 weeks. Physiological conditions of shoots were monitored via measuring potential photosynthesis quantum efficiency (F(v)/F(m)) and photosynthetic pigment contents. Additionally, the structure of leaves was analyzed using optical and atomic force microscopy (AFM). It has been shown that plants grown in 50 microM Cr(VI) solution exhibited photosynthetic activity and shoot and leaf morphology similar to control plants. Moreover, at the same time the average Cr concentration in their shoots reached about 470 mg kg(-1)d.w. after 10d and up to 1000 mg kg(-1)d.w. after 3 weeks of culture while in control plants did not exceed a few mgkg(-1)d.w. Our results point to Callitriche cophocarpa as a very promising species to be used in the investigation of chromium(VI) phytoremediation mechanisms as well as a good candidate for wastewaters remediation purpose.

Dioxygen uptake by isolated thylakoids from lettuce (Lactuca sativa L.): simultaneous measurements of dioxygen uptake, pH change of the medium and chlorophyll fluorescence parameters.

The setup has been elaborated for the simultaneous measurements of dioxygen uptake, pH changes, and chlorophyll a fluorescence parameters of an isolated thylakoid suspension. Using this equipment we have found at least three kinetically distinguishable components in the response of dioxygen uptake and pH increase to light intensity in the range of 0-1600 microE m(-2) s(-1). The pH changes were not observed in the presence of uncouplers (2 microM valinomycin plus 2 microM nigericin) while O(2) uptake increased by about 10% and F (v)/F (m) ratio appeared to be unaffected by this treatment. Treatment with DNP-INT, an inhibitor of plastoquinol oxidation, led to a significant reduction of pH increase and O(2) consumption whereas F (v) /F (m) was impaired only to 71% of the control. Incubation with catalase (580 U/ml) caused a total inhibition of oxygen uptake, while the pH increased and the F (v) /F (m) ratio decreased to about 60% and 85% of the control, respectively. The addition of catalase after the irradiation period led to an evolution of the same amount of dioxygen as was consumed during the light period. These results show that hydrogen peroxide was formed in the investigated system and accumulated during illumination. On the basis of the obtained data, three sites of dioxygen reduction within isolated thylakoid membranes and the dependence of dioxygen uptake on the photosystem activities were discussed.