Functional Analysis of the Channelrhodopsin Genes from the Green Algae of the White Sea Basin.

Optogenetics, the method of light-controlled regulation of cellular processes is based on the use of the channelrhodopsins that directly generate photoinduced currents. Most of the channelrhodopsin genes have been identified in the green microalgae Chlorophyta, and the demand for increasing the number of functionally characterized channelrhodopsins and the diversity of their photochemical parameters keeps growing. We performed the expression analysis of cation channelrhodopsin (CCR) genes in natural isolates of microalgae of the genera Haematococcus and Bracteacoccus from the unique Arctic Circle region. The identified full-length CCR transcript of H. lacustris is the product of alternative splicing and encodes the Hl98CCR2 protein with no photochemical activity. The 5'-partial fragment of the B. aggregatus CCR transcript encodes the Ba34CCR protein containing a conserved TM1-TM7 membrane domain and a short cytosolic fragment. Upon heterologous expression of the TM1-TM7 fragment in CHO-K1 cell culture, light-dependent current generation was observed with the parameters corresponding to those of the CCR. The first discovered functional channelrhodopsin of Bracteacoccus has no close CCR homologues and may be of interest as a candidate for optogenetics.

Interactive Effects of Ceftriaxone and Chitosan Immobilization on the Production of Arachidonic Acid by and the Microbiome of the Chlorophyte Lobosphaera sp. IPPAS C-2047.

Pharmaceuticals including antibiotics are among the hazardous micropollutants (HMP) of the environment. Incomplete degradation of the HMP leads to their persistence in water bodies causing a plethora of deleterious effects. Conventional wastewater treatment cannot remove HMP completely and a promising alternative comprises biotechnologies based on microalgae. The use of immobilized microalgae in environmental biotechnology is advantageous since immobilized cultures allow the recycling of the microalgal cells, support higher cell densities, and boost tolerance of microalgae to stresses including HMP. Here, we report on a comparative study of HMP (exemplified by the antibiotic ceftriaxone, CTA) removal by suspended and chitosan-immobilized cells of Lobosphaera sp. IPPAS C-2047 in flasks and in a column bioreactor. The removal of CTA added in the concentration of 20 mg/L was as high as 65% (in the flasks) or 85% (in the bioreactor). The adsorption on the carrier and abiotic oxidation were the main processes contributing 65-70% to the total CTA removal, while both suspended and immobilized cells took up 25-30% of CTA. Neither the immobilization nor CTA affected the accumulation of arachidonic acid (ARA) by Lobosphaera sp. during bioreactor tests but the subsequent nitrogen deprivation increased ARA accumulation 2.5 and 1.7 times in the suspended and chitosan-immobilized microalgae, respectively. The study of the Lobosphaera sp. microbiome revealed that the immobilization of chitosan rather than the CTA exposure was the main factor displacing the taxonomic composition of the microbiome. The possibility and limitations of the use of chitosan-immobilized Lobosphaera sp. IPPAS C-2047 for HMP removal coupled with the production of valuable long-chain polyunsaturated fatty acids is discussed.

Failure of Micractinium simplicissimum Phosphate Resilience upon Abrupt Re-Feeding of Its Phosphorus-Starved Cultures.

Microalgae are naturally adapted to the fluctuating availability of phosphorus (P) to opportunistically uptake large amounts of inorganic phosphate (Pi) and safely store it in the cell as polyphosphate. Hence, many microalgal species are remarkably resilient to high concentrations of external Pi. Here, we report on an exception from this pattern comprised by a failure of the high Pi-resilience in strain Micractinium simplicissimum IPPAS C-2056 normally coping with very high Pi concentrations. This phenomenon occurred after the abrupt re-supplementation of Pi to the M. simplicissimum culture pre-starved of P. This was the case even if Pi was re-supplemented in a concentration far below the level toxic to the P-sufficient culture. We hypothesize that this effect can be mediated by a rapid formation of the potentially toxic short-chain polyphosphate following the mass influx of Pi into the P-starved cell. A possible reason for this is that the preceding P starvation impairs the capacity of the cell to convert the newly absorbed Pi into a "safe" storage form of long-chain polyphosphate. We believe that the findings of this study can help to avoid sudden culture crashes, and they are also of potential significance for the development of algae-based technologies for the efficient bioremoval of P from P-rich waste streams.

Photosynthetic and ultrastructural responses of the chlorophyte Lobosphaera to the stress caused by a high exogenic phosphate concentration.

Biotechnology of microalgae holds promise for sustainable using of phosphorus, a finite non-renewable resource. Responses of the green microalga Lobosphaera sp. IPPAS C-2047 to elevated inorganic phosphate (Pi) concentrations were studied. Polyphosphate (PolyP) accumulation and ultrastructural rearrangements were followed in Lobosphaera using light and electron microscopy and linked to the responses of the photosynthetic apparatus probed with chlorophyll fluorescence. High tolerance of Lobosphaera to ≤ 50 g L-1 Pi was accompanied by a retention of photosynthetic activity and specific induction of non-photochemical quenching (NPQ up to 4; Fv/Fm around 0.7). Acclimation of the Lobosphaera to the high Pi was accompanied by expansion of the thylakoid lumen and accumulation of the carbon-rich compounds. The toxic effect of the extremely high (100 g L-1) Pi inhibited the growth by ca. 60%, induced a decline in photosynthetic activity and NPQ along with contraction of the lumen, destruction of the thylakoids, and depletion of starch reserves. The Lobosphaera retained viability at the Pi in the range of 25-100 g L-1 showing moderate an increase of intracellular P content (to 4.6% cell dry weight). During the initial high Pi exposure, the vacuolar PolyP biosynthesis in Lobosphaera was impaired but recovered upon acclimation. Synthesis of abundant non-vacuolar PolyP inclusions was likely a manifestation of the emergency acclimation of the cells converting the Pi excess to less metabolically active PolyP. We conclude that the remarkable Pi tolerance of Lobosphaera IPPAS C-2047 is determined by several mechanisms including rapid conversion of the exogenic Pi into metabolically safe PolyP, the acclamatory changes in the cell population structure. Possible involvement of NPQ in the high Pi resilience of the Lobosphaera is discussed.

Identification of the Channelrhodopsin Genes in the Green and Cryptophytic Algae from the White and Black Seas.

Due to the unique capability of modulating cell membrane potential upon photoactivation, channelrhodopsins of green (Chlorophyta) and cryptophytic (Cryptophyta) algae are widely employed in optogenetics, a modern method of light-dependent regulation of biological processes. To enable the search for new genes perspective for optogenetics, we have developed the PCR tests for the presence of genes of the cation and anion channelrhodopsins. Six isolates of green algae Haematococcus and Bracteacoccus from the White Sea region and 2 specimens of Rhodomonas sp. (Cryptophyta) from the regions of White and Black Seas were analyzed. Using our PCR test we have demonstrated the known Haematococcus rhodopsin genes and have discovered novel rhodopsin genes in the genus of Bracteacoccus. Two distantly homologous genes of anion channelrhodopsins were also identified in the cryptophytic Rhodomonas sp. from the White and Black Seas. These results indicate that the developed PCR tests might be useful tool for a broad-range screening of the Chlorophyta and Cryptophyta algae to identify unique channelrhodopsin genes.

Chilling Upregulates Expression of the PsbS and LhcSR Genes in the Chloroplasts of the Green Microalga Lobosphaera incisa IPPAS C-2047.

Non-photochemical quenching (NPQ) of excited chlorophyll states is essential for protecting the photosynthetic apparatus (PSA) from the excessive light-induced damage in all groups of oxygenic photosynthetic organisms. The key component of the NPQ mechanism in green algae and some other groups of algae and mosses is the LhcSR protein of the light harvesting complex (LHC) protein superfamily. In vascular plants, LhcSR is replaced by PsbS, another member of the LHC superfamily and a subunit of photosystem II (PSII). PsbS also performs the photoprotective function in mosses. For a long time, PsbS had been believed to be nonfunctional in green algae, although the corresponding gene was discovered in the genome of these organisms. The first evidence of the PsbS accumulation in the model green alga Chlamydomonas reinhardtii in response to the increase in irradiance was obtained only six years ago. However, the observed increase in the PsbS content was short-termed (on an hour-timescale). Here, we report a significant (more than three orders of magnitude) and prolonged (four days) upregulation of PsbS expression in response to the chilling-induced high-light stress followed by a less significant (~ tenfold) increase in the PsbS expression for nine days. This is the first evidence for the long-term upregulation of the PsbS expression in green alga (Chlorophyta) in response to stress. Our data indicate that the role of PsbS in the PSA of Chlorophyta is not limited to the first-line defense against stress, as it was previously assumed, but includes full-scale participation in the photoprotection of PSA from the environmental stress factors.

Combined Impact of Magnetic Force and Spaceflight Conditions on Escherichia coli Physiology.

Changes in bacterial physiology caused by the combined action of the magnetic force and microgravity were studied in Escherichia coli grown using a specially developed device aboard the International Space Station. The morphology and metabolism of E. coli grown under spaceflight (SF) or combined spaceflight and magnetic force (SF + MF) conditions were compared with ground cultivated bacteria grown under standard (control) or magnetic force (MF) conditions. SF, SF + MF, and MF conditions provided the up-regulation of Ag43 auto-transporter and cell auto-aggregation. The magnetic force caused visible clustering of non-sedimenting bacteria that formed matrix-containing aggregates under SF + MF and MF conditions. Cell auto-aggregation was accompanied by up-regulation of glyoxylate shunt enzymes and Vitamin B12 transporter BtuB. Under SF and SF + MF but not MF conditions nutrition and oxygen limitations were manifested by the down-regulation of glycolysis and TCA enzymes and the up-regulation of methylglyoxal bypass. Bacteria grown under combined SF + MF conditions demonstrated superior up-regulation of enzymes of the methylglyoxal bypass and down-regulation of glycolysis and TCA enzymes compared to SF conditions, suggesting that the magnetic force strengthened the effects of microgravity on the bacterial metabolism. This strengthening appeared to be due to magnetic force-dependent bacterial clustering within a small volume that reinforced the effects of the microgravity-driven absence of convectional flows.

Photosynthesis measurements on the upper and lower side of the thallus of the foliose lichen Nephroma arcticum (L.) Torss.

The measurements of chlorophyll fluorescence play an important role in studies of lichen physiology. Usually, for foliose lichens fluorescence kinetics is recorded from the upper thalline side often exhibiting green color reflecting the presence of photosynthetic pigments. The lower side of such lichens is grey, dark-brown or black. At the first time, we evaluated photosynthetic activity distribution by chlorophyll fluorescence analysis on both lower and upper thallus sides for the foliose lichen Nephroma arcticum. We have demonstrated that photosynthesis proceeds not only on the green-colored upper side, but also on the gray lower side of the curled growing edges of the thallus lobes. These sides were differed in terms of PSII photochemical quantum yield, activity of non-regulatory dissipation and non-photochemical quenching of excited chlorophyll states (NPQ). Upper side was characterized by higher maximal PSII efficiency, whereas the lower one of the curled edges was characterized by higher actual photochemical quantum yield during actinic light acclimation. NPQ was higher on the upper surface, whereas, on the lower side (of the curled edges) non-regulatory dissipation was predominant. In terms of photosynthetic activity measurements, these results show, that actinic and measuring light reached the layer of phycobiont despite its shielding by mycobiont hyphae. On the melanized lower side in the basal thalline zone attached to the substratum photosynthesis was not detected. Lower side demonstrated higher level of light scattering in the reflectance spectra. We believe that different photoprotective mechanisms against high light are crucial on the upper and lower sides: NPQ on the upper surface, and light scattering and shielding by mycobiont on the lower side. Possible biological role of photosynthesis on the lower side is discussed.

In vitro Biofilm Formation by Bioluminescent Bacteria Isolated from the Marine Fish Gut.

The internal surface of the animal gastrointestinal tract is covered by microbial biofilms. They play an important role in the development and functioning of the host organism and protect it against pathogens. Microbial communities of gastrointestinal biofilms are less elucidated than luminal microbiota. Therefore, the studies of biofilm formation by gastrointestinal microorganisms are a topical issue. For the first time, we report the formation of a biofilm in vitro by the strains of bioluminescent bacteria isolated from the intestines of marine fish. These bacteria exhibit co-aggregation and tend to attach to solid surfaces. The attachment of cells is accompanied by appearance of the pili. Then, we observed the formation of microcolonies and the production of extracellular polymer substances (EPSs) connecting bacterial cells into an integrated system. The presence of acidic polysaccharides is shown in the EPS when using the ruthenium red staining. Acidic polysaccharides in this matrix is a biochemical evidence of microbial biofilms. On the fibers of the polymer matrix, these bacteria form the "mushroom body"-type structures. Matured biofilms exhibit a specific three-dimensional architecture with pores and channels formed by cells and EPS. We also demonstrated the formation of a biofilm by binary culture of the luminous enterobacterium Kosakonia cowanii and a Gram-positive Macrococcus sp. The data obtained help to understand the role of these bacteria in the intestines of fish. They lead to a new study in the field of investigation of the intestinal microbiome of fish.

The microalga Haematococcus lacustris (Chlorophyceae) forms natural biofilms in supralittoral White Sea coastal rock ponds.

MAIN CONCLUSION: Haematococcus lacustris inhabits supralittoral rock ponds and forms, under natural conditions, biofilms including layered cyanobacterial and fermentative microbial mats. Dry mats, formed under extremely stressful conditions, contained only haematocysts. Under favorable growth conditions, modeled for dry biofilms in vitro, microalgal free-living stages were detected. Haematococcus lacustris is the microalga known for its high potential to survive under a wide range of unfavorable conditions, particularly in the supralittoral temporal rock ponds of the White Sea. Previously, we described microbial communities containing H. lacustris in this region. In many cases, they were organized into systems exhibiting complex three-dimensional structure similar to that of natural biofilms. In this study, for the first time, we clarify structural description and provide microscopic evidence that these communities of H. lacustris and bacteria are assembled into the true biofilms. There are (1) simple single layer biofilms on the surface of rocks and macrophytic algae, (2) floccules (or flocs) not attached to a surface, (3) as well as stratified (layered) biofilms, wet, and dehydrated in nature. Being involved into primary organic production, H. lacustris and cyanobacteria are located exclusively in the upper layers of stratified biofilms, where they are capable to absorb sufficient for photosynthesis amount of light. The presence of acidic polysaccharides in the extracellular matrix revealed by specific staining with ruthenium red in the H. lacustris-containing microbial communities is a biochemical evidence of biofilm formation. Meanwhile, the presence of bacterial L-form is an ultrastructural confirmation of that fact. Under favorable conditions, modeled in vitro, H. lacustris from the dry microbial mats moves to the free-living states represented by vegetative palmelloid cells and motile zoospores. Owing to the fact that inside biofilms cells of microorganisms exist under stable conditions, we consider the biofilm formation as an additional mechanism that contributes to the survival of H. lacustris in the supralittoral zone of the White Sea.

Natural Communities of Carotenogenic Chlorophyte Haematococcus lacustris and Bacteria from the White Sea Coastal Rock Ponds.

Haematococcus lacustris is a biotechnologically important green unicellular alga producing widely used keta-karotenoid astaxanthin. In natural habitats, it exists in the form of algal-bacterial community, and under laboratory conditions, it is also accompanied by bacteria. The issue of the bacterial composition of industrial algal cultures is widely recognized as important. However, there is a dearth of information about bacterial composition of H. lacustris communities. In current work, we analyze the composition of natural H. lacustris communities from the White Sea coastal temporal rock ponds. For the first time, a 16S rRNA gene-based metagenome of natural H. lacustris bacterial communities has been generated. Main results of its analysis are as follow. Bacterial families Comamonadaceae, Cytophagaceae, Xanthomonadaceae, Acetobacteraceae, Rhodobacteraceae, and Rhodocyclaceae were observed in all studied H. lacustris natural communities. They also contained genera Hydrogenophaga and Cytophaga. Bacteria from the Hydrogenophaga genus were present in H. lacustris cultures after their isolation under the conditions of laboratory cultivation. Similar to other planktonic microalgae, H. lacustris forms a phycosphere around the cells. In this zone, bacteria attached to the algal surface. The contact between H. lacustris and bacteria is maintained even after sample drying. The study provides information about possible members of H. lacustris core microbiome, which can be presented in the industrial and laboratory cultures of the microalga.

Cationic penetrating antioxidants switch off Mn cluster of photosystem II in situ.

Mitochondria-targeted antioxidants (also known as 'Skulachev Ions' electrophoretically accumulated by mitochondria) exert anti-ageing and ROS-protecting effects well documented in animal and human cells. However, their effects on chloroplast in photosynthetic cells and corresponding mechanisms are scarcely known. For the first time, we describe a dramatic quenching effect of (10-(6-plastoquinonyl)decyl triphenylphosphonium (SkQ1) on chlorophyll fluorescence, apparently mediated by redox interaction of SkQ1 with Mn cluster in Photosystem II (PSII) of chlorophyte microalga Chlorella vulgaris and disabling the oxygen-evolving complex (OEC). Microalgal cells displayed a vigorous uptake of SkQ1 which internal concentration built up to a very high level. Using optical and EPR spectroscopy, as well as electron donors and in silico molecular simulation techniques, we found that SkQ1 molecule can interact with Mn atoms of the OEC in PSII. This stops water splitting giving rise to potent quencher(s), e.g. oxidized reaction centre of PSII. Other components of the photosynthetic apparatus proved to be mostly intact. This effect of the Skulachev ions might help to develop in vivo models of photosynthetic cells with impaired OEC function but essentially intact otherwise. The observed phenomenon suggests that SkQ1 can be applied to study stress-induced damages to OEC in photosynthetic organisms.

Eukaryotic Sequences in the 16SrRNA Metagenomic Dataset of Algal-bacterial Consortia of the White Sea Coastal Zone.

The libraries of bacterial 16SrRNA gene fragment from algal-bacterial consortia of the White Sea coastal zone are analyzed. Up to 3% of the reads have revealed to correspond to eukaryotic rRNA. They related to following main eukaryotic clades: Discoba, Stramenopiles, Ciliata, Amoebozoa, and Nucletmycea. Amoebae, especially Vermamoeba, were present in all samples. In one sample, heterolobose amoeba Paravahlkampfia was detected. These microorganisms are parasites of microalgae, which can induce significant damage to industrial cultures. However, the data on their physiology and distribution are scarce. This study provides new evidence about the diversity of herbivorous eukaryotic microorganisms in natural algal-containing consortia.

Spatial organization of the three-component lichen Peltigera aphthosa in functional terms.

The cephalolichen Peltigera aphthosa (L.) Willd. is characterized by lateral heterogeneity, which manifests itself in the presence of three thallus zones, referred to as the apical, basal and medial zone. These zones differ in terms of interaction between lichen bionts and their physiological activity. The apical thallus zone is more efficient in establishing a contact with cyanobacteria, because of a higher lectin content and a larger overall thallus surface area due to the presence of numerous mycobiont hyphae. Cephalodia are formed in this zone. The interaction between the mycobiont and cyanobiont is more intense in the medial zone. However, the establishment of the contact with cyanobacteria in this zone less probable. The spatial distribution of lectins in the thallus was determined. To reveal the differences in photosynthetic activity in three thallus zones, transient analysis of chlorophyll a fluorescence and the assessment of non-photochemical quenching of excited chlorophyll states were performed. Assimilation of absorbed light energy was more effective in the medial zone. The basal zone was characterized by decreased photosynthetic activity, lichen dissociation and thallus death.

Coordinated rearrangements of assimilatory and storage cell compartments in a nitrogen-starving symbiotic chlorophyte cultivated under high light.

A quantitative micromorphometric study of the cell compartment rearrangements was performed in a symbiotic chlorophyte Desmodesmus sp. 3Dp86E-1 grown on nitrogen (N) replete or N-free medium under 480 µmol PAR quanta m(-2) s(-1). The changes in the chloroplast, intraplastidial, and cytoplasmic inclusions induced by high light (HL) and N starvation were similar to those characteristic of free-living chlorophytes. The N-sufficient culture responded to HL by a transient swelling of the thylakoid lumen and a decline in photosynthetic efficiency followed by its recovery. In the N-starving cells, a more rapid expansion and thylakoid swelling occurred along with the irreversible decline in the photosynthetic efficiency. Differential induction of starch grains, oil bodies, and cell wall polysaccharides depending on the stress exposure and type was recorded. Tight relationships between the changes in the assimilatory and storage compartments in the stressed Desmodesmus sp. cells were revealed.

Accumulation of astaxanthin by a new Haematococcus pluvialis strain BM1 from the white sea coastal rocks (Russia).

We report on a novel arctic strain BM1 of a carotenogenic chlorophyte from a coastal habitat with harsh environmental conditions (wide variations in solar irradiance, temperature, salinity and nutrient availability) identified as Haematococcus pluvialis Flotow. Increased (25‰) salinity exerted no adverse effect on the growth of the green BM1 cells. Under stressful conditions (high light, nitrogen and phosphorus deprivation), green vegetative cells of H. pluvialis BM1 grown in BG11 medium formed non-motile palmelloid cells and, eventually, hematocysts capable of a massive accumulation of the keto-carotenoid astaxanthin with a high nutraceutical and therapeutic potential. Routinely, astaxanthin was accumulated at the level of 4% of the cell dry weight (DW), reaching, under prolonged stress, 5.5% DW. Astaxanthin was predominantly accumulated in the form of mono- and diesters of fatty acids from C16 and C18 families. The palmelloids and hematocysts were characterized by the formation of red-colored cytoplasmic lipid droplets, increasingly large in size and number. The lipid droplets tended to merge and occupied almost the entire volume of the cell at the advanced stages of stress-induced carotenogenesis. The potential application of the new strain for the production of astaxanthin is discussed in comparison with the H. pluvialis strains currently employed in microalgal biotechnology.

Multimodal non-invasive probing of stress-induced carotenogenesis in the cells of microalga Bracteacoccus aggregatus.

Microalgae are the richest source of natural carotenoids-accessory photosynthetic pigments used as natural antioxidants, safe colorants, and nutraceuticals. Microalga Bracteacoccus aggregatus IPPAS C-2045 responds to stresses, including high light, with carotenogenesis-gross accumulation of secondary carotenoids (the carotenoids structurally and energetically uncoupled from photosynthesis). Precise mechanisms of cytoplasmic transport and subcellular distribution of the secondary carotenoids under stress are still unknown. Using multimodal imaging combining micro-Raman imaging (MRI), fluorescent lifetime (τ) imaging (FLIM), and transmission electron microscopy (TEM), we monitored ultrastructural and biochemical rearrangements of B. aggregatus cells during the stress-induced carotenogenesis. MRI revealed a decline in the diversity of molecular surrounding of the carotenoids in the cells compatible with the relocation of the bulk of the carotenoids in the cell from functionally and structurally heterogeneous photosynthetic apparatus to the more homogenous lipid matrix of the oleosomes. Two-photon FLIM highlighted the pigment transformation in the cell during the stress-induced carotenogenesis. The structures co-localized with the carotenoids with shorter τ (mainly chloroplast) shrunk, whereas the structures harboring secondary carotenoids with longer τ (mainly oleosomes) expanded. These changes were in line with the ultrastructural data (TEM). Fluorescence of B. aggregatus carotenoids, either in situ or in acetone extracts, possessed a surprisingly long lifetime. We hypothesize that the extension of τ of the carotenoids is due to their aggregation and/or association with lipids and proteins. The propagation of the carotenoids with prolonged τ is considered to be a manifestation of the secondary carotenogenesis suitable for its non-invasive monitoring with multimodal imaging.

A local or a stranger? Comparison of autochthonous vs. allochthonous microalgae potential for bioremediation of coal mine drainage water.

Coal mining endangers the environment by contaminating of soil, surface, and ground water with coal mine drainage water (CMW) polluted by heavy metals. Microalgal cultures, hyper-accumulators of heavy metals, represent a promising solution for CMW biotreatment. A bottleneck of this approach is the availability of microalgal strains that combine a large capacity for heavy metal biocapture with a high resilience to their toxic effects. Biotopes contaminated with heavy metals are frequently inhabited by microalgae evolved to be resilient to heavy metal toxicity. Therefore, the autochthonous (locally isolated) microalgal strains are a priori considered to be superior for biotreatment of heavy metal-polluted waste streams. Still, strains from biocollections combine a high pollutant resilience with other biotechnologically important traits such as high productivity, high CO2 sequestration rate etc. Moreover, the strains available "off-the-shelf" would enable rapid development of bioprocesses. Here, we compared the efficiency of CMW biotreatment with autochthonous (isolated from the coal mine drainage sump) and allochthonous microalgae (from a geographically distant phosphate-polluted site). Both autochthonous strains and allochthonous strains turned to be interchangeable under our experimental conditions. Still, the autochthonous strains showed a higher capacity for sequestration of iron, zinc, and manganese, the specific pollutants of the studied CMW. It can be important when the duration of unattended exploitation of the CMW treatment facility is a priority or spikes of the heavy metal concentration in CMW are expected. Therefore, the "off-the-shelf" strains can be a plausible solution for rapid development of CMW treatment technologies from scratch (although screening for acute toxicity of CMW is imperative). On the other hand, locally isolated strains can offer distinct advantages and should be always considered if sufficient time and other resources are available for the development of microalgae-based process for CMW treatment.

Assessment of the Tolerance of a Chlorophyte Desmodesmus to CuO-NP for Evaluation of the Nanopollution Bioremediation Potential of This Microalga.

Broad application of CuO nanoparticles (CuO-NP) for industrial and household purposes leads to a continuous increase in their discharge to, and, hence, ever-increasing environmental hazards for aquatic ecosystems. Microalgae-based technologies hold promise for bioremediation of diverse hazardous micropollutants (HMP), including NP, from wastewater. In this study, we tested the ability of the green microalga Desmodesmus sp. to accumulate CuO-NP or their components. We also assessed the tolerance of this microalga to the environmentally relevant concentrations of CuO-NP. Using scanning electron microscopy, we demonstrated that the average size of CuO-NP was 50-100 nm, and their purity was confirmed with elemental composition analysis. Tests of the colloidal suspensions of CuO-NP showed that the hydrodynamic diameter of CuO-NP and their aggregates was below 100 nm. Flow cytometry analysis showed that CuO-NP at a concentration of 100 µg L-1 slightly inhibited the viability of microalgae cells and led to an increase in their oxidative stress. The assessment of the condition of photosystem II showed that CuO-NP exert a multifaceted effect on the photosynthetic apparatus of Desmodesmus sp., depending on the concentration of and the exposure to the CuO-NP. Desmodesmus sp. turned to be relatively tolerant to CuO-NP. In addition, the ICP-MS method revealed increased bioaccumulation of copper by microalgae cells in the experimental groups. The outcomes of this study indicate that the Desmodesmus sp. has a significant potential for bioremoval of the copper-based nanostructured HMP from an aquatic environment.

Chlorophyll fluorescence as a valuable multitool for microalgal biotechnology.

Variable fluorescence of chlorophyll (CF) of the photosynthetic apparatus is an ample source of valuable information on physiological condition of photosynthetic organisms. Currently, the most widespread CF-based technique is represented by recording pulse-amplitude modulated (PAM) induction of CF by saturating light. The CF-based monitoring techniques are increasingly employed for characterization of performance and stress resilience of microalgae in microalgal biotechnology. Analysis of CF induction curves reveals the fate of light energy absorbed by photosynthetic apparatus, the proportions of the energy that have been utilized for photochemistry (culture growth), and heat dissipated by photoprotective mechanisms. Hence CF and its derived parameters are an accurate proxy of the metabolic activity of the photosynthetic cell and the engagement of photoprotective mechanisms. This information is a solid foundation for making decisions on the microalgal culture management during the lab-scale and industrial-scale cultivation. Applications of CF and PAM include the monitoring of stressor (high light, nutrient deprivation, extreme temperatures, etc.) effects for assessment of the culture robustness. It also serves as a non-invasive express test for gauging the effect of assorted toxicants in microalgae. This approach is becoming widespread in ecological toxicology and environmental biotechnology, particularly for bioprospecting strains capable of the destruction of dangerous pollutants such as pharmaceuticals. In the review, we discuss the advantages and drawbacks of using CF-based methods for assessment of the culture conditions. Special attention is paid to the potential caveats and applicability of different variations of CF and PAM measurements for solving problems of microalgal biotechnology.