Toxicity of cadmium selenide nanoparticles on the green microalgaChlorella vulgaris: inducing antioxidative defense response.

Green algae are dominant primary producers in aquatic environments. Thus, assessing the influences of pollutants such as nanoparticles on the algae is of high ecological significance. In the current study, cadmium selenide nanoparticles (CdSe NPs) were synthesized using the hydrothermal method and their characteristics were determined by X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and Fourier-transform infrared spectroscopy (FT-IR) techniques. Subsequently, the toxicity of synthesized nanoparticles on the green microalga Chlorella vulgaris was investigated. The observations by SEM confirmed that exposure to CdSe NPs had severe impacts on the algal morphology. Furthermore, the obtained results revealed the toxic effect of CdSe NPs by a decrease in the number of cells. Measurement of antioxidant enzymes activity showed an increase in the activity of catalase, and a decrease in the activity of superoxide dismutase (SOD) at high concentrations of CdSe NPs. The exposure of C. vulgaris to CdSe NPs resulted also in a change in algal pigments as well as total phenol content. Taken together, CdSe NPs appeared to have significant cytotoxic effects on C. vulgaris in the applied concentrations.

Aged microplastics polyvinyl chloride interact with copper and cause oxidative stress towards microalgae Chlorella vulgaris.

Microplastics (MPs) could pose potential risks to microalgae, the primary producer of marine ecosystems. Currently, few studies focus on the interaction of aged MPs with other pollutants and their toxic effects to microalgae. Therefore, the present study aimed to investigate i) the aging of microplastics polyvinyl chloride (mPVC) in simulated seawater and the changes in physical and chemical properties; ii) the effects of single mPVC (virgin and aged) and copper on microalgae Chlorella vulgaris; and iii) the interaction of aged mPVC and copper and the oxidative stress towards C. vulgaris. In this study, some wrinkles, rough and fractured surface textures can be observed on the aged mPVC, accompanying with increased hydroxyl groups and aromatic carbon-carbon double bond but decreased carbon hydrogen bond. It was found that single virgin or aged mPVC at low concentration (10 mg/L) had significant inhibition on the growth of C. vulgaris but no inhibition at higher concentration (100, 1,000 mg/L), which can be reasonably explained by the aggregation and precipitation of mPVC at high concentration. The aging of mPVC inhibited the growth of C. vulgaris with the maximum growth inhibition ratio (IR) of 35.26% as compared with that of virgin mPVC (IR = 28.5%). However, the single copper could significantly inhibit the growth of C. vulgaris and the inhibitory effects increased with concentration (0.2, 0.5, 1.0 mg/L). Furthermore, both the single aged mPVC (10 mg/L) and copper (0.5 mg/L) caused serious cell damage, although the concentration of superoxide dismutase (SOD) and the intracellular malonaldehyde (MDA) increased. In contrast to single treatment, the growth of C. vulgaris can be enhanced by the combined group with copper (0.5 mg/L) and aged mPVC (10 mg/L).

Combined effects of erythromycin and enrofloxacin on antioxidant enzymes and photosynthesis-related gene transcription in Chlorella vulgaris.

Multiple antibiotics are simultaneously detected in aquatic environment, so it is extremely important to study the combined effects of their mixtures. In this study, we investigated the toxic effects of erythromycin (ERY) and enrofloxacin (ENR), added individually or in combination, on Chlorella vulgaris and explored the toxic mechanisms. Results showed that the 96 h-EC50 values of ERY, ENR and ERY-ENR mixture to C. vulgaris were 85.7, 124.5 and 39.9 µg L-1 respectively, and combined toxicity assessment found that joint effect of the two antibiotics was synergism, which was proven by the chlorophyll content in algae. Antioxidant defense system and photosynthesis were involved in toxic mechanisms and the results revealed that both the activities of antioxidant enzymes, and the malondialdehyde (MDA) and glutathione (GSH) contents increased in antibiotic treatments. In addition, the increase was more significant in joint exposure treatment, which implied that the antioxidant defense system was synergistically affected. RT-PCR showed that ERY and ENR upregulated the transcript abundance of psaB, psbC and chlB at low concentrations and the transcription abundance was synergistically increased in combined treatment. Therefore, the risk of the toxicity of antibiotics to aquatic organisms in real environment both at organismal and molecular level increases as a result of their combined presence.

Physiological and enzymatic responses of Chlorella vulgaris exposed to produced water and its potential for bioremediation.

In South America, Colombia is known as an important oil-producing country. However, the environmental impact of crude oil industry has not been studied deeply and few studies have been carried out for evaluating responses of algae and its adaptation under specific conditions. Enzymatic and physiological effects in Chlorella vulgaris and its potential for bioremediation after exposure to produced water (PW) were assessed using different PW concentrations (0, 25, 50, 75 and 100%) and crude oil. Variables such as cell density, growth rate (µ), percentage of growth inhibition (% I), chlorophyll a and b and cell diameter were evaluated during 5 days. Furthermore, enzymatic biomarkers such as superoxide dismutase (SOD) and catalase (CAT) were also measured. Results showed that the treatment with 100% PW had the highest cell density and µ; similarly, 25% PW treatment had a similar behaviour, being these two treatments with the highest growth. A dose-dependent response was seen for chlorophyll a and b and cell diameter, showing significant differences between treatments and the control. Different levels of SOD and CAT were observed in algae exposed to PW. At 24 h, an increase in SOD and CAT activity was observed, probably due to effects caused by xenobiotics. After 72 h, a decrease in the activity of both enzymes was observed. The results evidenced that C. vulgaris can adapt easily to PW, showing an increase on its growth and stabilisation in its antioxidant activity. Additionally, cell diameter results and decrease of hydrocarbons and phenols show the potential of these algae to degrade xenobiotics from PW.

Novel combination of feed enzymes to improve the degradation of Chlorella vulgaris recalcitrant cell wall.

In this study, a rational combination of 200 pre-selected Carbohydrate-Active enzymes (CAZymes) and sulfatases were tested, individually or combined, according to their ability to degrade Chlorella vulgaris cell wall to access its valuable nutritional compounds. The disruption of microalgae cell walls by a four-enzyme mixture (Mix) in comparison with the control, enabled to release up to 1.21 g/L of reducing sugars (p < 0.001), led to an eight-fold increase in oligosaccharides release (p < 0.001), and reduced the fluorescence intensity by 47% after staining with Calcofluor White (p < 0.001). The Mix treatment was successful in releasing proteins (p < 0.001), some MUFA (p < 0.05), and the beneficial 18:3n-3 fatty acid (p < 0.05). Even if no variation was detected for chlorophylls (p > 0.05), total carotenoids were increased in the supernatant (p < 0.05) from the Mix treatment, relative to the control. Taken together, these results indicate that this four-enzyme Mix displays an effective capacity to degrade C. vulgaris cell wall. Thus, these enzymes may constitute a good approach to improve the bioavailability of C. vulgaris nutrients for monogastric diets, in particular, and to facilitate the cost-effective use of microalgae by the feed industry, in general.

Effect of metformin exposure on growth and photosynthetic performance in the unicellular freshwater chlorophyte, Chlorella vulgaris.

Many pharmaceuticals have negative effects on biota when released into the environment. For example, recent work has shown that the commonly prescribed antidiabetic drug, metformin (N,N-dimethylbiguanide), has endocrine disrupting effects on fish. However, effects of metformin on aquatic primary producers are poorly known. We exposed cultured isolates of a freshwater chlorophyte, Chlorella vulgaris, to a range of metformin concentrations (0-767.9 mg L-1) to test the hypothesis that exposure negatively affects photosynthesis and growth. A cessation of growth, increase in non-photochemical quenching (NPQ, NPQmax), and reduced electron transport rate (ETR) were observed 24 h after exposure to a metformin concentration of 767.8 mg L-1 (4.6 mM). By 48 h, photosynthetic efficiency of photosystem II (Fv/Fm), α, the initial slope of the ETR-irradiance curve, and Ek (minimum irradiance required to saturate photosynthesis) were reduced. At a lower concentration (76.8 mg L-1), negative effects on photosynthesis (increase in NPQ, decrease in ETR) were delayed, occurring between 72 and 96 h. No negative effects on photosynthesis were observed at an exposure concentration of 1.5 mg L-1. It is likely that metformin impairs photosynthesis either through downstream effects from inhibition of complex I of the electron transport chain or via activation of the enzyme, SnRK1 (sucrose non-fermenting-related kinase 1), which acts as a cellular energy regulator in plants and algae and is an ortholog of the mammalian target of metformin, AMPK (5' adenosine monophosphate-activated protein kinase).

Phylogeny and comparative modeling of phytochelatin synthase from Chlorella sp. as an efficient bioagent for detoxification of heavy metals.

Phytochelatins (PCs) found extensively in algae and plants are important for detoxification of heavy metals from soil and wastewater, and their synthesis is mediated by an enzyme phytochelatin synthase (PCS). In this study, a phylogram was generated to study evolutionary relationships of PCS from various organisms. It was revealed that PCS from green algae and plants are orthologs as both have evolved from a common ancestor. PCS from cyanobacteria appeared in two different clades showing that they have followed different lineages during evolution. Structural modeling was also carried out by building a 3D model of PCS from Chlorella variabilis using software Modeller v9.16. The predicted structure will be helpful for protein engineering strategies and to understand its interactions with other proteins. The biological biosorption capacity of Chlorella vulgaris (a green alga) was determined to remove Cd, Cu and Pb from industrial effluents. The biosorption of three heavy metals from industrial waste water was investigated under various conditions like pH, biomass concentration, contact time and temperature. Bio-removal of heavy metals was carried out by exposing culture of C. vulgaris to water samples of different heavy metal concentrations. The decrease in Cd, Cu and Pb quantities after 1 to 7 days of incubation period were 83%, 84% and 82.5%, respectively. In view of this, Chlorella spp. could be used on a large scale to detoxify heavy metals and clean up contaminated environments.

In vitro thrombolytic activity of a purified fibrinolytic enzyme from Chlorella vulgaris.

A fibrinolytic enzyme was produced by microalga Chlorella vulgaris cultivated in autotrophic and mixotrophic conditions added corn steep liquor, purified by a single chromatographic step, then biochemical characterization and in vitro thrombolytic activity was performed. Maximum cell concentration (1637.45 ±â€¯15 mg L-1) and productivity (181.93 mg L-1 day-1) was obtained in mixotrophic culture using 1% corn steep liquor. Enzyme-extracted microalgal biomass was purified by acetone precipitation and DEAE Sephadex anion exchange chromatography up to 2 fold with recovery of 4.0%. After purification, fibrinolytic activity was 1834.6 U mg-1 and 226.86 mm2 by spectrophotometry and fibrin plate assays, respectively. SDS-PAGE results exhibited a protein band of about 45 kDa and fibrinolytic band was detected by fibrin zymography. Enzyme activity was enhanced in the presence of Fe2+ and inhibited by phenylmethane sulfonyl fluoride (PMSF) and ethylenediamine tetracetic acid (EDTA), which suggest it to be a metal-dependent serine protease. The extract also showed a red blood cell lysis <4% and in vitro thrombolytic activity of 25.6% in 90 min of reaction. These results indicate that the fibrinolytic enzyme from C. vulgaris may have potential applications in the prevention and treatment of thrombosis.

Effects of sodium pentaborate pentahydrate exposure on Chlorella vulgaris growth, chlorophyll content, and enzyme activities.

Sodium pentaborate pentahydrate (SPP) is a rare mineral. In this study, SPP was synthesized from boric acid and borax through low-temperature crystallization, and its effects on the growth of the alga, Chlorella vulgaris (C. vulgaris) were assessed. The newly synthesized SPP was characterized by chemical analysis, X-ray diffraction, Fourier-transform infrared spectroscopy, Raman spectroscopy, thermogravimetric analysis, and differential thermal analysis. The changes in C. vulgaris growth, chlorophyll content, and enzyme activities upon exposure to SPP for 168h were evaluated. Results showed that SPP treatment was detrimental to C. vulgaris growth during the first 24-120h of exposure. The harmful effects, however, diminished over time (168h), even at an effective medium concentration of 226.37mg BL(-1) (the concentration of boron applied per liter of culture medium). A similar trend was observed for chlorophyll content (chlorophyll a and b) and indicated that the photosynthesis of C. vulgaris was not affected and that high levels of SPP may even promote chlorophyll synthesis. Superoxide dismutase and catalase activities of C. vulgaris increased during 24-120h exposure to SPP, but these activities gradually decreased as culture time progressed. In other words, the initial detrimental effects of synthetic SPP on C. vulgaris were temporary and reversible. This research provides a scientific basis for applications of SPP in the environment.

Selective extraction of intracellular components from the microalga Chlorella vulgaris by combined pulsed electric field-temperature treatment.

The synergistic effect of temperature (25-65 °C) and total specific energy input (0.55-1.11 kWh kgDW(-1)) by pulsed electric field (PEF) on the release of intracellular components from the microalgae Chlorella vulgaris was studied. The combination of PEF with temperatures from 25 to 55 °C resulted in a conductivity increase of 75% as a result of cell membrane permeabilization. In this range of temperatures, 25-39% carbohydrates and 3-5% proteins release occurred and only for carbohydrate release a synergistic effect was observed at 55 °C. Above 55 °C spontaneous cell lysis occurred without PEF. Combined PEF-temperature treatment does not sufficiently disintegrate the algal cells to release both carbohydrates and proteins at yields comparable to the benchmark bead milling (40-45% protein, 48-58% carbohydrates).

Crystal Structure of the Homing Endonuclease I-CvuI Provides a New Template for Genome Modification.

Homing endonucleases recognize and generate a DNA double-strand break, which has been used to promote gene targeting. These enzymes recognize long DNA stretches; they are highly sequence-specific enzymes and display a very low frequency of cleavage even in complete genomes. Although a large number of homing endonucleases have been identified, the landscape of possible target sequences is still very limited to cover the complexity of the whole eukaryotic genome. Therefore, the finding and molecular analysis of homing endonucleases identified but not yet characterized may widen the landscape of possible target sequences. The previous characterization of protein-DNA interaction before the engineering of new homing endonucleases is essential for further enzyme modification. Here we report the crystal structure of I-CvuI in complex with its target DNA and with the target DNA of I-CreI, a homologue enzyme widely used in genome engineering. To characterize the enzyme cleavage mechanism, we have solved the I-CvuI DNA structures in the presence of non-catalytic (Ca(2+)) and catalytic ions (Mg(2+)). We have also analyzed the metal dependence of DNA cleavage using Mg(2+) ions at different concentrations ranging from non-cleavable to cleavable concentrations obtained from in vitro cleavage experiments. The structure of I-CvuI homing endonuclease expands the current repertoire for engineering custom specificities, both by itself as a new scaffold alone and in hybrid constructs with other related homing endonucleases or other DNA-binding protein templates.

Azoxystrobin-induced excessive reactive oxygen species (ROS) production and inhibition of photosynthesis in the unicellular green algae Chlorella vulgaris.

This study investigated the short-term toxicity of azoxystrobin (AZ), one of strobilurins used as an effective fungicidal agent to control the Asian soybean rust, on aquatic unicellular algae Chlorella vulgaris. The median percentile inhibition concentration (IC50) of AZ for C. vulgaris was found to be 510 µg L(-1). We showed that the algal cells were obviously depressed or shrunk in 300 and 600 µg L(-1) AZ treatments by using the electron microscopy. Furthermore, 19, 75, and 300 µg L(-1) AZ treatments decreased the soluble protein content and chlorophyll concentrations in C. vulgaris and altered the energy-photosynthesis-related mRNA expression levels in 48- and 96-h exposure periods. Simultaneously, our results showed that AZ could increase the total antioxidant capacity (T-AOC) level and compromise superoxide dismutase (SOD), peroxidase (POD), glutathione S transferase (GST), glutathione peroxidase (GPx) activities, and glutathione (GSH) content. These situations might render C. vulgaris more vulnerable to oxidative damage. Overall, the present study indicated that AZ might be toxic to the growth of C. vulgaris, affect energy-photosynthesis-related mRNA expressions, and induce reactive oxygen species (ROS) overproduction in C. vulgaris.

Accumulation of fatty acids in Chlorella vulgaris under heterotrophic conditions in relation to activity of acetyl-CoAcarboxylase, temperature, and co-immobilization with Azospirillum brasilense [corrected].

The relation between fatty acid accumulation, activity of acetyl-CoA carboxylase (ACC), and consequently lipid accumulation was studied in the microalgae Chlorella vulgaris co-immobilized with the plant growth-promoting bacterium Azospirillum brasilense under dark heterotrophic conditions with Na acetate as a carbon source. In C. vulgaris immobilized alone, cultivation experiments for 6 days showed that ACC activity is directly related to fatty acid accumulation, especially in the last 3 days. In co-immobilization experiments, A. brasilense exerted a significant positive effect over ACC activity, increased the quantity in all nine main fatty acids, increased total lipid accumulation in C. vulgaris, and mitigated negative effects of nonoptimal temperature for growth. No correlation between ACC activity and lipid accumulation in the cells was established for three different temperatures. This study demonstrated that the interaction between A. brasilense and C. vulgaris has a significant effect on fatty acid and lipid accumulation in the microalgae.

Crystallization and preliminary X-ray diffraction analysis of the homing endonuclease I-CvuI from Chlorella vulgaris in complex with its target DNA.

Homing endonucleases are highly specific DNA-cleaving enzymes that recognize long stretches of DNA. The engineering of these enzymes provides novel instruments for genome modification in a wide range of fields, including gene targeting, by inducing specific double-strand breaks. I-CvuI is a homing endonuclease from the green alga Chlorella vulgaris. This enzyme was purified after overexpression in Escherichia coli. Crystallization experiments of I-CvuI in complex with its DNA target in the presence of Mg(2+) yielded crystals suitable for X-ray diffraction analysis. The crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 62.83, b = 83.56, c = 94.40 Å. The self-rotation function and the Matthews coefficient suggested the presence of one protein-DNA complex per asymmetric unit. The crystals diffracted to a resolution limit of 1.9 Å using synchrotron radiation.

Alleviating CTAC and Flu combined pollution damage in Chlorella vulgaris by exogenous nitric oxide.

This study investigates the effect of sodium nitroprussiate (SNP), an exogenous NO-donor, on the joint toxicity of binary mixtures of cetyltrimethylammonium chloride (CTAC) and fluoranthene (Flu) (CTAC/Flu), which are representatives for surfactants and polycyclic aromatic hydrocarbons (PAHs) respectively, in a unicellular green alga Chlorella vulgaris (C. vulgaris). The results showed that the addition of low SNP (20µM) alleviated the CTAC/Flu combined pollution damage in C. vulgaris. Supplement of low SNP significantly increased the algae biomass, chlorophyll content, soluble protein content and the activities of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) as compared to CTAC/Flu treatment alone. SNP also reduced the content of malondialdehyde (MDA) and the reactive oxygen species (ROS), as compared with CTAC/Flu treated alone. On the contrary, the above phenomena were reversed when high concentration of SNP (100µM) was added. Our study indicated that the damage of the joint action of surfactants and PAHs on hydrobios can be alleviated through protecting against oxidant substances and increasing the activity of antioxidant enzymes with an exogenous supply of NO in certain concentration range.

Chromate tolerance and accumulation in Chlorella vulgaris L.: role of antioxidant enzymes and biochemical changes in detoxification of metals.

A concentration-dependent increase in activity of antioxidant enzymes (catalase, ascorbate peroxidase, glutathione, superoxide dismutase) and carotenoid, MDA level have been observed in the green alga Chlorella vulgaris following chromium exposure at different concentrations (0.01-100 µg ml(-1)). Simultaneously, decrease in growth rate, chlorophyll and protein contents was observed. In case of ascorbate peroxidase, glutathione peroxidase and superoxide dismutase a bell shaped dose response was evident, however, lipid peroxidation followed a linear relationship along with catalase activity, which could be used as biomarker of Cr toxicity and played important role in providing tolerance and subsequently, high accumulation potential of chromium in C. vulgaris. In present investigation, the green alga C. vulgaris respond better under chromium stress in terms of tolerance, growth and metal accumulating potential at higher concentration of Cr (VI) which could be employed in decontamination of chromium for environmental cleanup.

Gene transfer of Chlorella vulgaris n-3 fatty acid desaturase optimizes the fatty acid composition of human breast cancer cells

Chlorella vulgaris has the gene of n-3 fatty acid desaturase (CvFad3), which can synthesize the precursor of n-3 polyunsaturated fatty acids (PUFAs) or convert n-6 to n-3 PUFAs. The objective of the present study was to examine whether the CvFad3 gene from C. vulgaris can be functionally and efficiently expressed in human breast cancer cells and whether its expression can exert a significant effect on cell fatty acid composition. We inserted the CvFad3 gene into the plasmid pEGFP-C3 to construct the eukaryotic expression vector pEGFP-C3-n-3 and to express the n-3 Fad gene in human breast cancer cells (MCF-7 cells). Transfection of MCF-7 cells with the recombinant vector resulted in a high expression of n-3 fatty acid desaturase. Lipid analysis indicated that the ratio of n-6/n-3 PUFAs was decreased from 6:1 in the control cells to about 1:1 in the cells expressing the n-3 fatty acid desaturase. Accordingly, the CvFad3 gene significantly decreased the ratio of n-6/n-3 PUFAs of the MCF-7 cell membrane. The expression of the CvFad3 gene can decrease cell proliferation and promote cell apoptosis. This study demonstrates that the CvFad3 gene can dramatically balance the ratio of n-6/n-3 PUFAs and may provide an effective approach to the modification of the fatty acid composition of mammalian cells, also providing a basis for potential applications of its transfer in experimental and clinical settings.

Chloroplast NADPH-dependent thioredoxin reductase from Chlorella vulgaris alleviates environmental stresses in yeast together with 2-Cys peroxiredoxin.

Chloroplast NADPH-dependent thioredoxin reductase (NTRC) catalyzes the reduction of 2-Cys peroxiredoxin (2-Cys Prx) and, thus, probably functions as an antioxidant system. The functions of the enzyme in oxidative and salt stresses have been reported previously. We have previously identified and characterized NTRC in Chlorella vulgaris. In the present study, we isolated a full-length cDNA clone encoding 2-Cys Prx from C. vulgaris and investigated the involvement of Chlorella NTRC/2-Cys Prx system in several environmental stress tolerances by using yeast as a eukaryotic model. Deduced Chlorella 2-Cys Prx was homologous to those of chloroplast 2-Cys Prxs from plants, and two conserved cysteine residues were found in the deduced sequence. Enzyme assay showed that recombinant mature C. vulgaris NTRC (mCvNTRC) transferred electrons from NADPH to recombinant mature C. vulgaris 2-Cys Prx (mCvPrx), and mCvPrx decomposed hydrogen peroxide, tert-butyl hydroperoxide, and peroxynitrite by cooperating with mCvNTRC. Based on the results, the mCvNTRC/mCvPrx antioxidant system was identified in Chlorella. The antioxidant system genes were expressed in yeast separately or coordinately. Stress tolerances of yeast against freezing, heat, and menadione-induced oxidative stresses were significantly improved by expression of mCvNTRC, and the elevated tolerances were more significant when both mCvNTRC and mCvPrx were co-expressed. Our results reveal a novel feature of NTRC: it functions as an antioxidant system with 2-Cys Prx in freezing and heat stress tolerances.

Gene transfer of Chlorella vulgaris n-3 fatty acid desaturase optimizes the fatty acid composition of human breast cancer cells.

Chlorella vulgaris has the gene of n-3 fatty acid desaturase (CvFad3), which can synthesize the precursor of n-3 polyunsaturated fatty acids (PUFAs) or convert n-6 to n-3 PUFAs. The objective of the present study was to examine whether the CvFad3 gene from C. vulgaris can be functionally and efficiently expressed in human breast cancer cells and whether its expression can exert a significant effect on cell fatty acid composition. We inserted the CvFad3 gene into the plasmid pEGFP-C3 to construct the eukaryotic expression vector pEGFP-C3-n-3 and to express the n-3 Fad gene in human breast cancer cells (MCF-7 cells). Transfection of MCF-7 cells with the recombinant vector resulted in a high expression of n-3 fatty acid desaturase. Lipid analysis indicated that the ratio of n-6/n-3 PUFAs was decreased from 6:1 in the control cells to about 1:1 in the cells expressing the n-3 fatty acid desaturase. Accordingly, the CvFad3 gene significantly decreased the ratio of n-6/n-3 PUFAs of the MCF-7 cell membrane. The expression of the CvFad3 gene can decrease cell proliferation and promote cell apoptosis. This study demonstrates that the CvFad3 gene can dramatically balance the ratio of n-6/n-3 PUFAs and may provide an effective approach to the modification of the fatty acid composition of mammalian cells, also providing a basis for potential applications of its transfer in experimental and clinical settings.

Phytohormones as regulators of heavy metal biosorption and toxicity in green alga Chlorella vulgaris (Chlorophyceae).

The present study was undertaken to test the influence of exogenously applied phytohormones: auxins (IAA, IBA, NAA, PAA), cytokinins (BA, CPPU, DPU, 2iP, Kin, TDZ, Z), gibberellin (GA(3)), jasmonic acid (JA) as well as polyamine - spermidine (Spd) upon the growth and metabolism of green microalga Chlorella vulgaris (Chlorophyceae) exposed to heavy metal (Cd, Cu, Pb) stress. The inhibitory effect of heavy metals on algal growth, metabolite accumulation and enzymatic as well as non-enzymatic antioxidant system was arranged in the following order: Cd > Pb > Cu. Exogenously applied phytohormones modify the phytotoxicity of heavy metals. Auxins, cytokinins, gibberellin and spermidine (Spd) can alleviate stress symptoms by inhibiting heavy metal biosorption, restoring algal growth and primary metabolite level. Moreover, these phytohormones and polyamine stimulate antioxidant enzymes' (superoxide dismutase, ascorbate peroxidase, catalase) activities and ascorbate as well as glutathione accumulation by producing increased antioxidant capacity in cells growing under abiotic stress. Increased activity of antioxidant enzymes reduced oxidative stress expressed by lipid peroxidation and hydrogen peroxide level. In contrast JA enhanced heavy metal toxicity leading to increase in metal biosorption and ROS generation. The decrease in cell number, chlorophylls, carotenoids, monosaccharides, soluble proteins, ascorbate and glutathione content as well as antioxidant enzyme activity was also obtained in response to JA and heavy metals. Determining the stress markers (lipid peroxidation, hydrogen peroxide) and antioxidants' level as well as antioxidant enzyme activity in cells is important for understanding the metal-specific mechanisms of toxicity and that these associated novel endpoints may be useful metrics for accurately predicting toxicity. The data suggest that phytohormones and polyamine play an important role in the C. vulgaris responding to abiotic stressor and algal adaptation ability to metal contamination of aquatic environment.