CsubMADS1, a lag phase transcription factor, controls development of polar eukaryotic microalga Coccomyxa subellipsoidea C-169.

MADS-box transcription factors (TFs) have not been functionally delineated in microalgae. In this study, the role of CsubMADS1 from microalga Coccomyxa subellipsoidea C-169 has been explored. Unlike Type II MADS-box proteins of seed plants with MADS, Intervening, K-box, and C domains, CsubMADS1 only has MADS and Intervening domains. It forms a group with MADS TFs from algae in the phylogenetic tree within the Type II MIKCC clade. CsubMADS1 is expressed strongly in the lag phase of growth. The CsubMADS1 monomer does not have a specific localization in the nucleus, and it forms homodimers to localize exclusively in the nucleus. The monomer has two nuclear localization signals (NLSs): an N-terminal NLS and an internal NLS. The internal NLS is functional, and the homodimer requires two NLSs for specific nuclear localization. Overexpression (OX) of CsubMADS1 slows down the growth of the culture and leads to the creation of giant polyploid multinucleate cells, resembling autospore mother cells. This implies that the release of autospores from autospore mother cells may be delayed. Thus, in wild-type (WT) cells, CsubMADS1 may play a crucial role in slowing down growth during the lag phase. Due to starvation in 2-month-old colonies on solid media, the WT colonies produce mucilage, whereas OX colonies produce significantly less mucilage. Thus, CsubMADS1 also negatively regulates stress-induced mucilage production and probably plays a role in stress tolerance during the lag phase. Taken together, our results reveal that CsubMADS1 is a key TF involved in the development and stress tolerance of this polar microalga.

Characterization of river biofilm responses to the exposure with heavy metals using a novel micro fluorometer biosensor.

River biofilms are a suitable indicator of toxic stress in aquatic ecosystems commonly exposed to various anthropogenic pollutants from industrial, domestic, and agricultural sources. Among these pollutants, heavy metals are of particular concern as they are known to interfere with various physiological processes of river biofilm, directly or indirectly related to photosynthetic performance. Nevertheless, only limited toxicological data are available on the mechanisms and toxicodynamics of heavy metals in biofilms. Pulse Amplitude Modulated (PAM) fluorometry is a rapid, non-disruptive, well-established technique to monitor toxic responses on photosynthetic performance, fluorescence-kinetics, and changes in yield in other non-photochemical processes. In this study, a new micro-PAM-sensor was tested to assess potential acute and chronic effects of heavy metals in river biofilm. Toxicity values across the three parameters considered in this study (photosynthetic yield YII, non-photochemical quenching NPQ, and basal fluorescence F0) were comparable, as determined EC50 were within one order of magnitude (EC50 ∼1-10 mg L-1). However, the stimulation of NPQ was more clearly associated with early acute effects, especially in illuminated samples, while depression of YII and F0 were more prevalent in chronic tests. These results have implications for the development of functional indicators for the biomonitoring of aquatic health, in particular for the use of river biofilm as a bioindicator of water quality. In conclusion, the approach proposed seems promising to characterize and monitor the exposure and impact of heavy metals on river periphyton communities. Furthermore, this study provides a fast, highly sensitive, inexpensive, and accurate laboratory method to test effects of pollutants on complex periphyton communities that can also give insights regarding the probable toxicological mechanisms of heavy metals on photosynthetic performance in the river biofilm.

Comparing Biochemical and Raman Microscopy Analyses of Starch, Lipids, Polyphosphate, and Guanine Pools during the Cell Cycle of Desmodesmus quadricauda.

Photosynthetic energy conversion and the resulting photoautotrophic growth of green algae can only occur in daylight, but DNA replication, nuclear and cellular divisions occur often during the night. With such a light/dark regime, an algal culture becomes synchronized. In this study, using synchronized cultures of the green alga Desmodesmus quadricauda, the dynamics of starch, lipid, polyphosphate, and guanine pools were investigated during the cell cycle by two independent methodologies; conventional biochemical analyzes of cell suspensions and confocal Raman microscopy of single algal cells. Raman microscopy reports not only on mean concentrations, but also on the distribution of pools within cells. This is more sensitive in detecting lipids than biochemical analysis, but both methods-as well as conventional fluorescence microscopy-were comparable in detecting polyphosphates. Discrepancies in the detection of starch by Raman microscopy are discussed. The power of Raman microscopy was proven to be particularly valuable in the detection of guanine, which was traceable by its unique vibrational signature. Guanine microcrystals occurred specifically at around the time of DNA replication and prior to nuclear division. Interestingly, guanine crystals co-localized with polyphosphates in the vicinity of nuclei around the time of nuclear division.

Revision of Coelastrella (Scenedesmaceae, Chlorophyta) and first register of this green coccoid microalga for continental Norway.

A terrestrial green microalga was isolated at Ås, in Akershus County, Norway. The strain corresponded to a coccoid chlorophyte. Morphological characteristics by light and electron microscopy, in conjunction with DNA amplification and sequencing of the 18 s rDNA gene and ITS sequences, were used to identify the microalgae. The characteristics agree with those of the genus Coelastrella defined by Chodat, and formed a sister group with the recently described C. thermophila var. globulina. Coelastrella is a relatively small numbered genus that has not been observed in continental Norway before; there are no previous cultures available in collections of Norwegian strains. Gas chromatography analyses of the FAME-derivatives showed a high percentage of polyunsaturated fatty acids (44-45%) especially linolenic acid (C18:3n3; 30-34%). After the stationary phase, the cultures were able to accumulate several carotenoids as neoxanthin, pheophytin a, astaxanthin, canthaxanthin, lutein, and violaxanthin. Due to the scarcity of visual characters suitable for diagnostic purposes and the lack of DNA sequence information, there is a high possibility that species of this genus have been neglected in local environmental studies, even though it showed interesting properties for algal biotechnology.

Pediludiella daitoensis gen. et sp. nov. (Scenedesmaceae, Chlorophyceae), a large coccoid green alga isolated from a Loxodes ciliate.

Freshwater protists often harbor unicellular green algae within their cells. In ciliates, possibly because of large host cell sizes and the small size of algal coccoids, a single host cell typically contains more than a hundred algal cells. While surveying such algae-bearing protists on Minami Daito Jima Island in Japan, we found a green Loxodes ciliate (Loxodida, Karyorelictea) that contained one or two dozens of very large coccoid algae. We isolated one of these algae and analyzed its characteristics in detail. A small subunit (SSU) rDNA phylogeny indicated Pseudodidymocystis species (Scenedesmaceae, Chlorophyceae) to be the taxon closest to the alga, although it was clearly separated from this by 39 or more different sites (inclusive of gaps). SSU rRNA structure analyses indicated that these displacements included eight compensatory base changes (CBCs) and seven hemi-CBCs. We therefore concluded that this alga belongs to a separate genus, and described it as Pediludiella daitoensis gen. et sp. nov. The shape of the isolated and cultured P. daitoensis was nearly spherical and reached up to 30 µm in diameter. Chloroplasts were arranged peripherally and often split and elongated. Cells were often vacuolated and possessed a net-like cytoplasm that resembled a football (soccer ball) in appearance, which was reflected in the genus name.

The potential growth and lipid accumulation in Coccomyxa subellipsoidea triggered by glucose combining with sodium acetate.

Carbon sources whether types or magnitudes were fateful in terms of stimulating growth and lipids accumulation in microalgae applied for biodiesel production. The set scenario of this work was to investigate the feasibilities of glucose (G) combining with sodium acetate (SA) carbon sources in enhancing biomass and lipid accumulation in Coccomyxa subellipsoidea. The results demonstrated that C. subellipsoidea subjected to the combination feeding of G (20 g/L) and SA (12 g/L) achieved the favorable biomass (5.22 g/L) and lipid content (52.16%). The resulting lipid productivity (388.96 mg/L/day) was 1.33- to 7.60-fold more than those of sole G or SA as well as other combinations of G and SA. Even though the total fatty acids of C. subellipsoidea cells treated with the optimal combination of G and SA showed no noticeable increment in comparison with sole G or SA, the proportion of monounsaturated C18:1 (over 48.69%) and the content of C18:3 (< 12%) were commendable in high-quality algal biodiesel production. Further, such fascinating lipid accumulation in C. subellipsoidea cells treated with G combining with SA might be attributed to that G promoted glycolysis as well as SA activated glyoxylate shunt and TCA cycle to synergistically provide sufficient acetyl-CoA precursors for lipid accumulation. These findings hinted the potential of the combination of carbon sources in enhancing the overall lipid productivity to offset alga-based biodiesel production cost and would guide other alga strains cultivation.

Growth and the cell cycle in green algae dividing by multiple fission.

Most cells divide into two daughter cells; however, some green algae can have different division patterns in which a single mother cell can sometimes give rise to up to thousands of daughter cells. Although such cell cycle patterns can be very complex, they are governed by the same general concepts as the most common binary fission. Moreover, cell cycle progression appears to be connected with size, since cells need to ensure that their size after division will not drop below the limit required for survival. Although the exact mechanism that lets cells measure cell size remains largely unknown, there have been several prominent hypotheses that try to explain it.

Embryogenesis of flattened colonies implies the innovation required for the evolution of spheroidal colonies in volvocine green algae.

BACKGROUND: Volvocine algae provide a suitable model for investigation of the evolution of multicellular organisms. Within this group, evolution of the body plan from flattened to spheroidal colonies is thought to have occurred independently in two different lineages, Volvocaceae and Astrephomene. Volvocacean species undergo inversion to form a spheroidal cell layer following successive cell divisions during embryogenesis. During inversion, the daughter protoplasts change their shape and develop acute chloroplast ends (opposite to basal bodies). By contrast, Astrephomene does not undergo inversion; rather, its daughter protoplasts rotate during successive cell divisions to form a spheroidal colony. However, the evolutionary pathways of these cellular events involved in the two tactics for formation of spheroidal colony are unclear, since the embryogenesis of extant volvocine genera with ancestral flattened colonies, such as Gonium and Tetrabaena, has not previously been investigated in detail. RESULTS: We conducted time-lapse imaging by light microscopy and indirect immunofluorescence microscopy with staining of basal bodies, nuclei, and microtubules to observe embryogenesis in G. pectorale and T. socialis, which form 16-celled or 4-celled flattened colonies, respectively. In G. pectorale, a cup-shaped cell layer of the 16-celled embryo underwent gradual expansion after successive cell divisions, with the apical ends (position of basal bodies) of the square embryo's peripheral protoplasts separated from each other. In T. socialis, on the other hand, there was no apparent expansion of the daughter protoplasts in 4-celled embryos after successive cell divisions, however the two pairs of diagonally opposed daughter protoplasts shifted slightly and flattened after hatching. Neither of these two species exhibited rotation of daughter protoplasts during successive cell divisions as in Astrephomene or the formation of acute chloroplast ends of daughter protoplasts as in volvocacean inversion. CONCLUSIONS: The present results indicate that the ancestor of Astrephomene might have newly acquired the rotation of daughter protoplasts after it diverged from the ancestor of Gonium, while the ancestor of Volvocaceae might have newly acquired the formation of acute chloroplast ends to complete inversion after divergence from the ancestor of Goniaceae (Gonium and Astrephomene).

Effect of algal surface area and species interactions in toxicity testing bioassays.

Single and multispecies algal bioassays were assessed using copper toxicity with three green algae (Scenedesmus subspicatus, Scenedesmus quadricauda and Ankistrodesmus angustus) and one blue-green algae species (Oscillatoria prolifera). Single and multispecies toxicity tests were conducted based on cell density as per standard toxicity testing, and on equivalent surface area. A higher copper sulfate toxicity was registered for O. prolifera, followed by S. subspicatus, S. quadricauda, and A. angustus in single-species toxicity tests based on cell density. Single species toxicity tests based on surface area showed increased copper toxicity with increasing algal surface area except for A. angustus. In multispecies control bioassays, the growth of A. angustus was inhibited in the presence of other species in surface area-based tests. As compared to single species bioassays, O. prolifera, and S. quadricauda showed a decreased sensitivity to copper sulfate in both cell density and surface area based multispecies tests. However, for the algae species with the smallest surface area, S. subspicatus, 96h-EC50 value decreased in multispecies bioassays based on surface area as compared to the single species test, while it increased in multispecies bioassays based on cell density. The difference in S. subspicatus sensitivity to copper between tests based on cell density and surface area supports the need to adopt multispecies toxicity testing based on surface area to avoid the confounding effect on copper toxicity of increased biomass for metal binding. 96h-EC50 values for all species combined in the multispecies test based on cell density and on surface area were significantly different from 96h-EC50 values obtained in single species bioassays. These results demonstrate that single-species bioassays may over- or underestimate metal toxicity in natural waters.

Diversity of photosynthetic picoeukaryotes in eutrophic shallow lakes as assessed by combining flow cytometry cell-sorting and high throughput sequencing.

Photosynthetic picoeukaryotes (PPE) are key components of primary production in marine and freshwater ecosystems. In contrast with those of marine environments, freshwater PPE groups have received little attention. In this work, we used flow cytometry cell sorting, microscopy and metabarcoding to investigate the composition of small photosynthetic eukaryote communities from six eutrophic shallow lakes in South America, Argentina. We compared the total molecular diversity obtained from PPE sorted populations as well as from filtered total plankton samples (FTP). Most reads obtained from sorted populations belonged to the classes: Trebouxiophyceae, Chlorophyceae and Bacillariophyceae. We retrieved sequences from non-photosynthetic groups, such as Chytridiomycetes and Ichthyosporea which contain a number of described parasites, indicating that these organisms were probably in association with the autotrophic cells sorted. Dominant groups among sorted PPEs were poorly represented in FTP and their richness was on average lower than in the sorted samples. A significant number of operational taxonomic units (OTUs) were exclusively found in sorting samples, emphasizing that sequences from FTP underestimate the diversity of PPE. Moreover, 22% of the OTUs found among the dominant groups had a low similarity (<95%) with reported sequences in public databases, demonstrating a high potential for novel diversity in these lakes.

Morphoanatomical characterization of filamentous green algae of district Lahore based on classical and modern microscopic technique.

INTRODUCTION: For sample collection, four sites were selected, sites where fresh water were present, that is, Ravi syphon, BRB Lahore, Shahdara, and Head Baloki. The latitude and altitude of the locations were recorded at the time of collection. RESULTS: A total of 21 species of algae belonging to eight genera and four families were examined in the current studies. The collected samples were mainly filamentous and they were primarily green in color as the photosynthetic pigment found to be dominated was Chlorophyll. It was observed that all the genera varied mainly on the basis of cell shape, size, number of pyramids and on the arrangement of cells. The site mainly selected was those where fresh water was mainly found and at the time of collection latitude and altitude was also recorded. CONCLUSIONS: The classical microscopic technique (Light Microscopy) and Modern microscopic technique (Staining Electron Microscopy) of some samples were done that, played a lively part in the correct identification of species as a slight difference was found among species that were only evident when the Modern microscopic technique was done.

Differences between Motile and Nonmotile Cells of Haematococcus pluvialis in the Production of Astaxanthin at Different Light Intensities.

Haematococcus pluvialis, as the best natural resource of astaxanthin, is widely used in nutraceuticals, aquaculture, and cosmetic industries. The purpose of this work was to compare the differences in astaxanthin accumulation between motile and nonmotile cells of H. pluvialis and to determine the relationship between the two cells and astaxanthin production. The experiment design was achieved by two different types of H. pluvialis cell and three different light intensities for an eight day induction period. The astaxanthin concentrations in nonmotile cell cultures were significantly increased compared to motile cell cultures. The increase of astaxanthin was closely associated with the enlargement of cell size, and the nonmotile cells were more conducive to the formation of large astaxanthin-rich cysts than motile cells. The cyst enlargement and astaxanthin accumulation of H. pluvialis were both affected by light intensity, and a general trend was that the higher the light intensity, the larger the cysts formed, and the larger the quantity of astaxanthin accumulated. In addition, the relatively low cell mortality rate in the nonmotile cell cultures indicated that the nonmotile cells have a stronger tolerance to photooxidative stress. We suggest that applying nonmotile cells as the major cell type of H. pluvialis to the induction period may help to enhance the content of astaxanthin and the stability of astaxanthin production.

Growth and secretome analysis of possible synergistic interaction between green algae and cyanobacteria.

Synergistic coexistence of nitrogen fixing cyanobacteria such as Anabaena variabilis, Nostoc muscorum and Westiellopsis prolifica with green algae namely Scenedesmus obliquus, Chlorella vulgaris and Botryococcus braunii was studied under nitrogen deficient conditions. The effect of these interactions was investigated on growth, fixed nitrogen content, lipid content and their secretomes in individual cultures and cocultures. Based on the cocultivation studies, it was found that out of the nine interactions studied, B. braunii-N. muscorum synergism was best established. This interaction resulted in a maximum of 50% enhancement in nitrogen fixation in B. braunii-N. muscorum co-culture leading to 27% enhancement in lipid content (membrane and neutral lipid). In general, B. braunii co-cultures showed an enhancement in biomass content of up to 38%. Secretome analysis showed presence of new and modified secondary metabolites having roles in quorum sensing/quenching, interspecies signaling, N-fixation, carbon metabolism, lipid metabolism, antimicrobial activity. Compounds such as trichloroacetic acid and hexadecane were identified that are known to have roles in nitrogen assimilation and carbon metabolism, respectively, were present in some of the co-culture secretomes. The combination of B. braunii-N. muscorum led to the formation of new compounds such as triacontanol which have role in improvement of glucose-lipid metabolism and 9-octadecenamide that is known to be a phytohormone.

Effects of reaction conditions on light-dependent silver nanoparticle biosynthesis mediated by cell extract of green alga Neochloris oleoabundans.

Silver nanoparticles (AgNPs) were synthesized by incubating the mixture of AgNO3 solution and whole-cell aqueous extracts (WCAEs) of Neochloris oleoabundans under light conditions. By conducting single-factor and multi-factor optimization, the effects of parameters including AgNO3 concentration, pH, and extraction time were quantitatively evaluated. The optimal conditions in terms of AgNP yield were found to be 0.8 mM AgNO3, pH 5, and 9-h extraction. The AgNPs thus synthesized were quasi-spherical with a mean particle diameter of 16.63 nm and exhibited decent uniformity as well as antibacterial activities, which may facilitate AgNP biosynthesis's application in the near future.

Dielectrophoretic separation of microalgae cells in ballast water in a microfluidic chip.

The composition of the ship's ballast water is complex and contains a large number of microalgae cells, bacteria, microplastics, and other microparticles. To increase the accuracy and efficiency of detection of the microalgae cells in ballast water, a new microfluidic chip for continuous separation of microalgae cells based on alternating current dielectrophoresis was proposed. In this microfluidic chip, one piece of 3-dimensional electrode is embedded on one side and eight discrete electrodes are arranged on the other side of the microchannel. An insulated triangular structure between electrodes is designed for increasing the inhomogeneity of the electric field distribution and enhancing the dielectrophoresis (DEP) force. A sheath flow is designed to focus the microparticles near the electrode, so as to increase the suffered DEP force and improve separation efficiency. To demonstrate the performance of the microfluidic separation chip, we developed two species of microalgae cells (Platymonas and Closterium) and a kind of microplastics to be used as test samples. Analyses of the related parameters and separation experiments by our designed microfluidic chip were then conducted. The results show that the presented method can separate the microalgae cells from the mixture efficiently, and this is the first time to separate two or more species of microalgae cells in a microfluidic chip by using negative and positive DEP force simultaneously, and moreover it has some advantages including simple operation, high efficiency, low cost, and small size and has great potential in on-site pretreatment of ballast water.

Spectral microscopic imaging of heterocysts and vegetative cells in two filamentous cyanobacteria based on spontaneous Raman scattering and photoluminescence by 976 nm excitation.

Photosynthetic pigment-protein complexes are highly concentrated in thylakoid membranes of chloroplasts and cyanobacteria that emit strong autofluorescence (mainly 600-800 nm). In Raman scattering microscopy that enables imaging of pigment concentrations of thylakoid membranes, near infrared laser excitation at 1064 nm or visible laser excitation at 488-532 nm has been often employed in order to avoid the autofluorescence. Here we explored a new approach to Raman imaging of thylakoid membranes by using excitation wavelength of 976 nm. Two types of differentiated cells, heterocysts and vegetative cells, in two diazotrophic filamentous cyanobacteria, Anabaena variabilis, and Rivularia M-261, were characterized. Relative Raman scattering intensities of phycobilisomes of the heterocyst in comparison with the nearest vegetative cells of Rivularia remained at a significantly higher level than those of A. variabilis. It was also found that the 976 nm excitation induces photoluminescence around 1017-1175 nm from the two cyanobacteria, green alga (Parachlorella kessleri) and plant (Arabidopsis thaliana). We propose that this photoluminescence can be used as an index of concentration of chlorophyll a that has relatively small Raman scattering cross-sections. The Rivularia heterocysts that we analyzed were clearly classified into at least two subgroups based on the Chla-associated photoluminescence and carotenoid Raman bands, indicating two physiologically distinct states in the development or aging of the terminal heterocyst.

Behavior of the extremophile green alga Coccomyxa melkonianii SCCA 048 in terms of lipids production and morphology at different pH values.

The extremophile green alga Coccomyxa melkonianii SCCA 048 was investigated to evaluate its ability to grow in culture media with different pH. Specifically, Coccomyxa melkonianii was sampled in the Rio Irvi river (Sardinia, Italy) which is severely polluted by heavy metals as a result of abandoned mining activities. In this study, the strain was cultivated in growth media where the pH was kept fixed at the values of 4.0, 6.8 and 8.0, respectively. During the investigation, a significant phenotypic plasticity of this strain was observed. The strain grew well in the pH range 4.0-8.0, while the optimal value for its growth was 6.8. Furthermore, maximum lipid contents of about 24 and 22 %wt were achieved at the end of cultivation when using pH 4.0 and 8.0, respectively. Finally, the analysis of fatty acid methyl esters (FAMEs) highlights the presence of suitable amounts of compounds which can be profitably exploited in the food, nutraceutical, and cosmetic industry. This aspect, coupled with the possibility of cultivating Coccomyxa melkonianii under extreme pH conditions in economic open ponds, makes this strain an interesting candidate for several biotechnological applications.

Effects of cyclin-dependent kinase activity on the coordination of growth and the cell cycle in green algae at different temperatures.

The progression of the cell cycle in green algae dividing by multiple fission is, under otherwise unlimited conditions, affected by the growth rate, set by a combination of light intensity and temperature. In this study, we compared the cell cycle characteristics of Desmodesmus quadricauda at 20 °C or 30 °C and upon shifts between these two temperatures. The duration of the cell cycle in cells grown under continuous illumination at 20 °C was more than double that at 30 °C, suggesting that it was set directly by the growth rate. Similarly, the amounts of DNA, RNA, and bulk protein content per cell at 20 °C were approximately double those of cells grown at the higher temperature. For the shift experiments, cells grown at either 20 °C or 30 °C were transferred to darkness to prevent further growth, and then cultivated at the same or the other temperature. Upon transfer to the lower temperature, fewer nuclei and daughter cells were produced, and not all cells were able to finish the cell cycle by division, remaining multinuclear. Correspondingly, cells placed in the dark at the higher temperature divided faster into more daughter cells than the control cells. These differences correlated with shifts in the preceding cyclin-dependent kinase activity, suggesting that cell cycle progression was not related to growth rate or cell biomass but correlated with cyclin-dependent kinase activity.

Responses of freshwater algal cell density to hydrochemical variables in an urban aquatic ecosystem, northern China.

In this paper, the algal cell density of cyanobacteria, green algae, and diatoms and their responses to the hydrochemical factors were analyzed to reveal the structural characteristics of water quality in an urban river. A total of nine sampling sites from upstream to downstream was explored in our study. At each site, the density of algae was identified every week during the wet season (June-October) from 2012 to 2017, and in situ detection was used for the relative 11 hydrochemical variables. The temporal and spatial characteristics of 14 variables were analyzed using a heatmap coupled with the cluster analysis method. The trend of each parameter was analyzed using the smoothing method with locally weighted regression. The nonmetric multidimensional scaling method was employed to detect the temporal and spatial similarities among algae along hydrochemical gradients. The responses of algal density to hydrochemical variables were analyzed using a redundancy analysis. The results showed that the water temperature (Wtemp), pH, dissolved oxygen (DO), cyanobacteria, and diatoms exhibited significant declining trends, and significant increasing trends were shown in the permanganate index, chemical oxygen demand, total nitrogen, ammonia nitrogen, and total phosphorus; the cyanobacteria exhibited certain differences with green algae and diatoms in summer and the downstream areas of the river. The temporal-spatial homogeneity of algal to hydrochemical variables showed the key influencing factors of Wtemp for cyanobacteria density, chlorophyll for green algae density, DO, and pH for diatoms. The results presented here are valuable for deepening our understanding of river ecosystem evaluations and effective environmental management, as well as an important reference for the sustainable development of aquatic biological resources.

Exogenously applied auxins and cytokinins ameliorate lead toxicity by inducing antioxidant defence system in green alga Acutodesmus obliquus.

The effects of auxins (IAA, IBA, PAA) and cytokinins (tZ, Kin, DPU) on the growth, oxidative damage, level of antioxidants and the activity of antioxidant enzymes as well as the contents of proteins and photosynthetic pigments in green alga Acutodesmus obliquus were investigated under 100 µM lead (Pb) stress. Heavy metal induced oxidative damage as evidenced by a decrease in cell number and reduction in the contents of proteins and chlorophylls as a consequence of an increase in reactive oxygen species (ROS) formation and lipid peroxidation. The application of exogenous auxins and cytokinins modulated biosorption of Pb by algal cells significantly alleviated the growth inhibition and stimulated the accumulation of proteins, chlorophylls and carotenes. Phytohormones also activated the xanthophyll cycle which is extensively involved in the protection of the photosynthetic apparatus in adverse environmental conditions. The reduction in oxidative stress caused by the presence of toxic Pb was observed in algal cultures treated with phytohormones. Cytokinins were more effective in lowering hydrogen peroxide and lipid peroxidation levels in comparison with auxins. This improving effect of cytokinins seems to be mediated by a decrease in Pb accumulation by algal cells, whereas auxins promoted metal uptake. Importantly, auxins and cytokinins enhanced the redox status of algal cells inducing the increase in the content of antioxidants (ascorbate, glutathione, and proline) and in the activity of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase) involved in ROS scavenging. The results of the present study strongly suggest that exogenous auxins and cytokinins enhanced the resistance of microalga A. obliquus against Pb toxicity through the activation of the antioxidant defence system.