Choline Dehydrogenase Contributes to Salt Tolerance in Dunaliella through Betaine Synthesis.

In Dunaliella tertiolecta, a microalga renowned for its extraordinary tolerance to high salinity levels up to 4.5 M NaCl, the mechanisms underlying its stress response have largely remained a mystery. In a groundbreaking discovery, this study identifies a choline dehydrogenase enzyme, termed DtCHDH, capable of converting choline to betaine aldehyde. Remarkably, this is the first identification of such an enzyme not just in D. tertiolecta but across the entire Chlorophyta. A 3D model of DtCHDH was constructed, and molecular docking with choline was performed, revealing a potential binding site for the substrate. The enzyme was heterologously expressed in E. coli Rosetta (DE3) and subsequently purified, achieving enzyme activity of 672.2 U/mg. To elucidate the role of DtCHDH in the salt tolerance of D. tertiolecta, RNAi was employed to knock down DtCHDH gene expression. The results indicated that the Ri-12 strain exhibited compromised growth under both high and low salt conditions, along with consistent levels of DtCHDH gene expression and betaine content. Additionally, fatty acid analysis indicated that DtCHDH might also be a FAPs enzyme, catalyzing reactions with decarboxylase activity. This study not only illuminates the role of choline metabolism in D. tertiolecta's adaptation to high salinity but also identifies a novel target for enhancing the NaCl tolerance of microalgae in biotechnological applications.

Effects of ZnWO4 nanoparticles on growth, photosynthesis, and biochemical parameters of the green microalga Raphidocelis subcapitata.

Nanoparticles have applications in many sectors in the society. ZnWO4 nanoparticles (ZnWO4-NPs) have potential in the fabrication of sensors, lasers, and batteries, and in environmental remediation. Thus, these NPs may reach aquatic ecosystems. However, we still do not know their effects on aquatic biota and, to our knowledge, this is the first study that evaluates the toxicity of ZnWO4-NPs in a eukaryotic organism. We evaluated the toxicity of ZnWO4-NPs on the green microalga Raphidocelis subcapitata for 96 h, in terms of growth, cell parameters, photosynthesis, and biochemical analysis. Results show that most of Zn was presented in its particulate form, with low amounts of Zn2+, resulting in toxicity at higher levels. The growth was affected from 8.4 mg L-1, with 96h-IC50 of 23.34 mg L-1. The chlorophyll a (Chl a) content increased at 30.2 mg L-1, while the fluorescence of Chl a (FL3-H) decreased at 15.2 mg L-1. We observed increased ROS levels at 44.4 mg L-1. Regarding photosynthesis, the NPs affected the oxygen evolving complex (OEC) and the efficiency of the photosystem II at 22.9 mg L-1. At 44.4 mg L-1 the qP decreased, indicating closure of reaction centers, probably affecting carbon assimilation, which explains the decay of carbohydrates. There was a decrease of qN (non-regulated energy dissipation, not used in photosynthesis), NPQ (regulated energy dissipation) and Y(NPQ) (regulated energy dissipation via heat), indicating damage to the photoprotection system; and an increase in Y(NO), which is the non-regulated energy dissipation via heat and fluorescence. The results showed that ZnWO4-NPs can affect the growth and physiological and biochemical parameters of the chlorophycean R. subcapitata. Microalgae are the base of aquatic food chains, the toxicity of emerging contaminants on microalgae can affect entire ecosystems. Therefore, our study can provide some help for better protection of aquatic ecosystems.

The efficient capture of polysaccharides in Tetradesmus obliquus of indole-3-acetic acid coupling sludge extraction.

Polysaccharide is an important biomass of algae. The sludge extract is rich in organic substances, which can be used by algae for biomass growth and high-value biomass synthesis, but its organic toxicity has an inhibitory effect on algae. To overcome inhibition and improve polysaccharide enrichment, Tetradesmus obliquus was cultured with sludge extract with different indole-3-acetic acid (IAA) concentrations. Within 30 days of the culture cycle, T. obliquus showed in good condition at the IAA dosage content of 10-6 M, the maximum cell density and dry weight were respectively (106.78 ± 2.20) × 106 cell/mL and 2.941 ± 0.067 g/L while the contents of chlorophyll-a, chlorophyll-b, and carotenoid were 1.79, 1.91 and 2.80 times that of the blank group, respectively. The highest polysaccharide accumulation was obtained under this culture condition, reaching 533.15 ± 21.11 mg/L on the 30th day, which was 2.49 times that in the blank group. By FT-IR and NMR analysis, it was found that the polysaccharides of T. obliquus were sulfated polysaccharide with glucose and rhamnose as the main monosaccharides. Proteomic showed that the up-regulation of A0A383WL26 and A0A383WLM8 enhanced the light trapping ability, and A0A383WMJ2 enhanced the accumulation of NADPH. The up-regulation of A0A383WHD5 and A0A383WAY6 indicated that IAA culture could repair the damage caused by sludge toxicity, thus promoting the accumulation of biomass. The above findings provided new insights into the mechanism of sludge toxicity removal of T. obliquus and the enhancement of the polysaccharide accumulation effect under different concentrations of IAA.

Palmelloid-like phenotype in the alga Raphidocelis subcapitata exposed to pollutants: A generalized adaptive strategy to stress or a specific cellular response?

This work focuses on the formation of palmelloid-like phenotype in the freshwater alga Raphidocelis subcapitata (formerly known as Pseudokirchneriella subcapitata and Selenastrum capricornutum), when exposed to adverse conditions generated by the presence of organic [the antibiotic erythromycin (ERY) and the herbicide metolachlor (MET)] or inorganic [the heavy metals, cadmium (Cd) and zinc (Zn)] pollutants, at environmentally relevant concentrations. This alga in absence of stress or when exposed to ERY or Zn, up to 200 µg/L, essentially showed a single-nucleus state, although algal growth was reduced or stopped. R. subcapitata "switched" to a multinucleated state (palmelloid-like morphology) and accumulated energy-reserve compounds (neutral lipids) when stressed by 100-200 µg/L MET or 200 µg/L Cd; at these concentrations of pollutants, growth was arrested, however, the majority of the algal population (≥83 %) was alive. The formation of palmelloid-like phenotype, at sub-lethal concentrations of pollutants, was dependent on the pollutant, its concentration and exposure time. The multinucleated structure is a transitory phenotype since R. subcapitata population was able to revert to a single-nucleus state, with normal cell size, within 24-96 h (depending on the impact of the toxic in the alga), after being transferred to fresh OECD medium, without pollutants. The obtained results indicate that the formation of a palmelloid-like phenotype in R. subcapitata is dependent on the mode of action of toxics and their concentration, not constituting a generalized defense mechanism against stress. The observations here shown contribute to understanding the different strategies used by the unicellular alga R. subcapitata to cope with severe stress imposed by organic and inorganic pollutants.

Individual and Combined Effects of Manganese and Chromium on a Freshwater Chlorophyceae.

Manganese (Mn), an essential metal in trace amounts, and chromium (Cr), a nonessential metal to algae, are often found in effluent discharges and may co-occur in contaminated aquatic environments. Therefore, we investigated the effects of Mn and Cr, and their mixtures, on a freshwater Chlorophyceae, Raphidocelis subcapitata, using a multiple endpoint approach. Regarding the single exposure of metals, Mn was 4 times more toxic (median inhibitory concentration at 72 h [IC5072 h ] = 4.02 ± 0.45 µM Mn) than Cr (IC5072 h = 16.42 ± 4.94 µM Cr) for microalgae, considering the effects on the relative growth rate. Moreover, this species was the most sensitive to Mn, according to the species sensitivity distribution curve. Overall, the tested metals did not lead to significant changes in reactive oxygen species production, cellular complexity, and cell relative size but significantly decreased the algal growth and the mean cell chlorophyll a (Chl a) fluorescence at the highest concentrations (3.64-14.56 µM of Mn and 15.36-19.2 µM of Cr). The decreased mean cell Chl a fluorescence indicates an impact on pigment synthesis, which may be related to the observed growth inhibition. In the mixture tests, the reference models concentration addition and independent action were used to analyze the data, and the independent action model was the best fit to describe our results. Therefore, the Mn and Cr mixture was noninteractive, showing additive effects. This is the first study to address the combined toxicity of Mn and Cr regarding freshwater Chlorophyceae. Environ Toxicol Chem 2022;41:1004-1015. © 2022 SETAC.

Biochemical and morphological characterization of freshwater microalga Tetradesmus obliquus (Chlorophyta: Chlorophyceae).

Tetradesmus is a microalgal genus with biotechnological potential due to its rapid production of biomass, which is plenty in proteins, carbohydrates, lipids, and bioactives. However, its morphology and physiology need to be determined to guide better research to optimize the species cultivation and biocompounds processing. Thus, this study describes the biochemistry and morphology of the strain Tetradesmus obliquus BR003, isolated from a sample of freshwater reservoirs in a Brazilian municipality. In the T. obliquus BR003 dry biomass, we identified 61.6% unsaturated fatty acids, and 3.4% saturated fatty acids. Regarding other compounds, 28.50 ± 1.47 g soluble proteins/100 g, 0.14 ± 0.009 g carotenoids/100 g, 0.76 ± 0.013 g chlorophyll a/100 g, and 0.42 ± 0.015 g chlorophyll b/100 g with a chlorophyll a/b ratio of 1.8 were detected. The main chemical elements found were S, Mg, and P. The cells of BR003 were elliptically curved at the ends and without appendages. Histochemical tests showed carbohydrates distributed in the cytoplasm and pyrenoids, some lipid droplets, and proteins. The cytoplasm is rich in vacuoles, rough endoplasmic reticulum, mitochondria, and chloroplasts. The nucleus has a predominance of decondensed chromatin, and the cell wall has three layers. Chloroplasts have many starch granules and may be associated with a spherical central pyrenoid. To the best of our knowledge, this was the first biochemical description combined with ultrastructural morphological characterization of the strain T. obliquus BR003, grown under standard conditions, to demonstrate specific characteristics of the species.

Cytotoxicity and genotoxicity of coated-gold nanoparticles on freshwater algae Pseudokirchneriella subcapitata.

Gold engineered nanoparticles (nAu) are increasingly detected in ecosystems, and this raises the need to establish their potential effects on aquatic organisms. Herein, cytotoxic and genotoxic effects of branched polyethylenimine (BPEI)- and citrate (cit)-coated nAu (5, 20, and 40 nm) on algae Pseudokirchneriella subcapitata were evaluated. The apical biological endpoints: growth inhibition and chlorophyll a (Chl a) content were investigated at 62.5-1000 µg/L over 168 h. In addition, the apurinic/apyrimidinic (AP) sites, randomly amplified polymorphic deoxyribonucleic acid (RAPD) profiles, and genomic template stability (GTS) were assessed to determine the genotoxic effects of nAu. The results show algal growth inhibition at 5 nm BPEI-nAu up to 96 h, and thereafter cell recovery except at the highest concentration of 1000 µg/L. Insignificant growth reduction for cit-nAu (all sizes), as well as 20 and 40 nm BPEI-nAu, was observed over 96 h, but growth promotion was apparent at all exposures thereafter except for 40 nm BPEI-nAu at 250 µg/L. A decrease in Chl a content following exposure to 5 nm BPEI-nAu at 1000 µg/L corresponded to significant algal growth reduction. In genotoxicity studies, a significant increase in AP sites content was observed relative to the control - an indication of nAu ability to induce genotoxic effects irrespective of their size and coating type. For 5 nm- and 20 nm-sized nAu for both coating types and exposure concentrations no differences in AP sites content were observed after 72 and 168 h. However, a significant reduction in AP sites was observed following algae exposure to 40 nm-sized nAu (irrespective of coating type and exposure concentration) at 168 h compared to 72 h. Thus, AP sites results at 40 nm-size suggest likely DNA damage recovery over a longer exposure period. The findings on AP sites content showed a good correlation with an increase in genome template stability and growth promotion observed after 168 h. In addition, RAPD profiles demonstrated that nAu can induce DNA damage and/or DNA mutation to P. subcapitata as evidenced by the appearance and/or disappearance of normal bands compared to the controls. Therefore, genotoxicity results revealed significant toxicity of nAu to algae at the molecular level although no apparent effects were detectable at the morphological level. Overall, findings herein indicate that long-term exposure of P. subcapitata to low concentrations of nAu may cause undesirable sub-lethal ecological effects.

Molecular Mechanism of Lipid Accumulation and Metabolism of Oleaginous Chlorococcum sphacosum GD from Soil under Salt Stress.

The oleaginous microalgae species Chlorococcum sphacosum GD is a promising feedstock for biodiesel production from soil. However, its metabolic mechanism of lipid production remains unclear. In this study, the lipid accumulation and metabolism mechanisms of Chlorococcum sphacosum GD were analyzed under salt stress based on transcriptome sequencing. The biomass and lipid content of the alga strain were determined under different NaCl concentrations, and total RNA from fresh cells were isolated and sequenced by HiSeq 2000 high throughput sequencing technology. As the salt concentration increased in culture medium, the algal lipid content increased but the biomass decreased. Following transcriptome sequencing by assembly and splicing, 24,128 unigenes were annotated, with read lengths mostly distributed in the 200-300 bp interval. Statistically significant differentially expressed unigenes were observed in different experimental groups, with 2051 up-regulated genes and 1835 down-regulated genes. The lipid metabolism pathway analysis showed that, under salt stress, gene-related fatty acid biosynthesis (ACCase, KASII, KAR, HAD, FATA) was significantly up-regulated, but some gene-related fatty acid degradation was significantly down-regulated. The comprehensive results showed that salt concentration can affect the lipid accumulation and metabolism of C. sphacosum GD, and the lipid accumulation is closely related to the fatty acid synthesis pathway.

Using multiple endpoints to assess the toxicity of cadmium and cobalt for chlorophycean Raphidocelis subcapitata.

Metals may cause damage to the biota of contaminated environments. Moreover, using multiple endpoints in ecotoxicological studies is useful to better elucidate the mechanisms of toxicity of these compounds. Therefore, this study aimed to evaluate the effects of cadmium (Cd) and cobalt (Co) on growth, biochemical and photosynthetic parameters of the microalgae Raphidocelis subcapitata, through quantification of lipid classes composition, chlorophyll a (Chl a) content, maximum (ΦM) and effective (Φ'M) quantum yields and efficiency of the oxygen-evolving complex (OEC). Both metals affected the algal population growth, with an IC50-96h of 0.67 and 1.53 µM of Cd and Co, respectively. Moreover, the metals led to an increase in the total lipid content and reduced efficiency of OEC and ΦM. Cell density was the most sensitive endpoint to detect Cd toxicity after 96 h of treatment. Regarding Co, the photosynthetic parameters were the most affected and the total lipid content was the most sensitive endpoint as it was altered by the exposure to this metal in all concentrations. Cd led to increased contents of the lipid class wax esters (0.89 µM) and phospholipids (PL - at 0.89 and 1.11 µM) and decreased values of triglycerides (at 0.22 µM) and acetone-mobile polar lipids (AMPL - at 0.44 and 1.11 µM). The percentage of free fatty acids (FFA) and PL of microalgae exposed to Co increased, whereas AMPL decreased in all concentrations tested. We were able to detect differences between the toxicity mechanisms of each metal, especially how Co interferes in the microalgae at a biochemical level. Furthermore, to the best of our knowledge, this is the first study reporting Co effects in lipid classes of a freshwater Chlorophyceae. The damage caused by Cd and Co may reach higher trophic levels, causing potential damage to the aquatic communities as microalgae are primary producers and the base of the food chain.

Acute growth inhibition & toxicity analysis of nano-polystyrene spheres on Raphidocelis subcapitata.

Micro/nano-plastics (MNPs) have been found within many environments and organisms including humans, making them a significant and growing concern. Initial research into the potential detrimental effects these MNPs both from acute and chronic exposure has been ongoing but still requires substantially more data to clarify. This research presents the response of nano-polystyrene (NPS) on Raphidocelis subcapitata, a freshwater alga, under an existing acute toxicity test along with additional analytical techniques to try identifying possible sources of toxicity. R. subcapitata cells were exposed for 72 h to a concentration range of 0-100 mg/l NPS. Growth Inhibition (GI) testing showed the R. subcapitata demonstrated statistically distinct reductions in growth over 72 h at all NPS exposure concentrations while not suffering culture collapse. By the 100 mg/l NPS exposure the R. subcapitata has suffered almost a 33.7% reduction in cell concentration after 72 h compared to control samples. Confocal imaging showed the NPS wasn't permeating into the algal plasma membrane or individual organelles but agglomerated onto the algal cell wall. The agglomeration was irregular but increased in total surface area covered as NPS concentration increases. UV-Vis fluorimetry testing produced a linear response of emission intensities to algae exposed to the 0-100 mg/l range of NPS. However, comparisons of emission intensity values of algae exposed to NPS to emission intensities of pure NPS at identical concentrations showed consistent intensity reduction. This response further indicated NPS agglomerating within the media and onto the alga cells seen from confocal imaging. Finally, Raman spectroscopy on R. subcapitata attempted to distinguish the key 1001 cm-1 peak or other crucial identifier peaks of polystyrene from overall Raman spectra. This was not successful as emissions from algal component (e.g. phenylaniline) completely suppressed the signal region.

Exposure of the alga Pseudokirchneriella subcapitata to environmentally relevant concentrations of the herbicide metolachlor: Impact on the redox homeostasis.

This study investigated the effect of the herbicide metolachlor (MET) on the redox homeostasis of the freshwater green alga Pseudokirchneriella subcapitata. At low MET concentrations (≤40 µg L-1), no effects on algal cells were detected. The exposure of P. subcapitata to 45-235 µg L-1 MET induced a significant increase of reactive oxygen species (ROS). The intracellular levels of ROS were particularly increased at high (115 and 235 µg L-1) but environmentally relevant MET concentrations. The exposure of algal cells to 115 and 235 µg L-1 MET originated a decrease in the levels of antioxidants molecules (reduced glutathione and carotenoids) as well as a reduction of the activity of scavenging enzymes (superoxide dismutase and catalase). These results suggest that antioxidant (non-enzymatic and enzymatic) defenses were affected by the excess of MET. As consequence of this imbalance (ROS overproduction and decline of the antioxidant system), ROS inflicted oxidative injury with lipid peroxidation and damage of cell membrane integrity. The results provide further insights about the toxic modes of action of MET on a non-target organism and emphasize the relevance of toxicological studies in the assessment of the impact of herbicides in freshwater environments.

Ameliorating effects of dietary Haematococcus pluvialis on arsenic-induced oxidative stress in rainbow trout (Oncorhynchus mykiss) fillet.

The current study was performed to investigate the protective effects of dietary Haematococcus pluvialis (H. pluvialis) on the oxidative stress induced by arsenic in rainbow trout (Oncorhynchus mykiss). The fish (20.70 ± 0.09 g) were fed with H. pluvialis at the levels of 0.28, 0.56, and 1.12 g 100 g-1 diet for 60 days. Then, each group was divided into two subgroups. In one of the subgroups, fish were exposed to arsenic challenge at a level of 9.1 mg/L. The other subset was used as the negative control. After the 96 h of toxicity test, protein and lipid oxidative levels, antioxidant-relevant gene expression as well as several chemical factors, including pH and peroxide value and moisture content, were evaluated in the fillet samples. Results showed that feeding with H. pluvialis decreased the levels of pH, peroxide value, as well as protein and lipid oxidation levels in treatment groups. Besides, the expression of antioxidant genes was significantly increased in the groups administrated with H. pluvialis. Based on the results of this study, feeding H. pluvialis attenuated the oxidative stress induced by arsenic in rainbow trout fillet through improving the antioxidant defense system.

Detachment of Dunaliella tertiolecta Microalgae from a Glass Surface by a Near-Infrared Optical Trap.

We report on the observation of the detachment in situ and in vivo of Dunaliella tertiolecta microalgae cells from a glass surface using a 1064 nm wavelength trapping laser beam. The principal bends of both flagella of Dunaliella were seen self-adhered to either the top or bottom coverslip surfaces of a 50 µm thick chamber. When a selected attached Dunaliella was placed in the trapping site, it photoresponded to the laser beam by moving its body and flagellar tips, which eventually resulted in its detachment. The dependence of the time required for detachment on the trapping power was measured. No significant difference was found in the detachment time for cells detached from the top or bottom coverslip, indicating that the induced detachment was not due solely to the optical forces applied to the cells. After detachment, the cells remained within the optical trap. Dunaliella detached from the bottom were seen rotating about their long axis in a counterclockwise direction, while those detached from the top did not rotate. The rotation frequency and the minimal force required to escape from the trap were also measured. The average rotation frequency was found to be independent of the trapping power, and the swimming force of a cell escaping the laser trap ranged from 4 to 10 picoNewtons. Our observations provide insight into the photostimulus produced when a near-infrared trapping beam encounters a Dunaliella. The microalgae frequently absorb more light than they can actually use in photosynthesis, which could cause genetic and molecular changes. Our findings may open new research directions into the study of photomovement in species of Dunaliella and other swimming microorganisms that could eventually help to solve technological problems currently confronting biomass production. In future work, studies of the response to excess light may uncover unrecognized mechanisms of photoprotection and photoacclimation.

A non-photosynthetic green alga illuminates the reductive evolution of plastid electron transport systems.

BACKGROUND: Plastid electron transport systems are essential not only for photosynthesis but also for dissipating excess reducing power and sinking excess electrons generated by various redox reactions. Although numerous organisms with plastids have lost their photoautotrophic lifestyles, there is a spectrum of known functions of remnant plastids in non-photosynthetic algal/plant lineages; some of non-photosynthetic plastids still retain diverse metabolic pathways involving redox reactions while others, such as apicoplasts of apicomplexan parasites, possess highly reduced sets of functions. However, little is known about underlying mechanisms for redox homeostasis in functionally versatile non-photosynthetic plastids and thus about the reductive evolution of plastid electron transport systems. RESULTS: Here we demonstrated that the central component for plastid electron transport systems, plastoquinone/plastoquinol pool, is still retained in a novel strain of an obligate heterotrophic green alga lacking the photosynthesis-related thylakoid membrane complexes. Microscopic and genome analyses revealed that the Volvocales green alga, chlamydomonad sp. strain NrCl902, has non-photosynthetic plastids and a plastid DNA that carries no genes for the photosynthetic electron transport system. Transcriptome-based in silico prediction of the metabolic map followed by liquid chromatography analyses demonstrated carotenoid and plastoquinol synthesis, but no trace of chlorophyll pigments in the non-photosynthetic green alga. Transient RNA interference knockdown leads to suppression of plastoquinone/plastoquinol synthesis. The alga appears to possess genes for an electron sink system mediated by plastid terminal oxidase, plastoquinone/plastoquinol, and type II NADH dehydrogenase. Other non-photosynthetic algae/land plants also possess key genes for this system, suggesting a broad distribution of an electron sink system in non-photosynthetic plastids. CONCLUSION: The plastoquinone/plastoquinol pool and thus the involved electron transport systems reported herein might be retained for redox homeostasis and might represent an intermediate step towards a more reduced set of the electron transport system in many non-photosynthetic plastids. Our findings illuminate a broadly distributed but previously hidden step of reductive evolution of plastid electron transport systems after the loss of photosynthesis.

Exposure to environmental concentrations of fipronil and 2,4-D mixtures causes physiological, morphological and biochemical changes in Raphidocelis subcapitata.

The occurrence of pesticides and their mixtures in the environment can alter the ecological relationships between aquatic food chains. Since fipronil and 2,4-dichlorophenoxyacetic acid (2,4-D) are commonly found together in Brazilian water bodies, the present study aimed to investigate through an integrative approach the toxicity mechanisms of environmentally relevant concentrations of pesticides Regent® 800 WG (active ingredient - a.i. fipronil), DMA® 806 BR (a.i. 2,4-D) isolated and in mixtures on the green alga Raphidocelis subcapitata using multiple parameters: physiological (growth rate and chlorophyll a fluorescence), morphological (cell complexity and size), biochemical (composition of lipid classes) and related to the photosynthetic activity (variable fluorescence, the maximum quantum yield of the photosystem II - PSII - and the efficiency of the oxygen evolving complex - OEC - of PSII). The results indicated that fipronil significantly inhibited algal population growth, increased the chlorophyll a content (observed by fluorescence), cell size and lipid class content of triacylglycerol (TAG), free fatty acid (FFA) and acetone mobile polar lipid (AMPL) and, on the other hand, decreased variable fluorescence of algae. The tested concentrations of 2,4-D increased the chlorophyll a fluorescence, the cell size and the lipid classes TAG and FFA. The pesticide mixtures have had more effects on algae than isolated compounds, causing alterations in all parameters analyzed, including photosynthetic activity (maximum quantum yield and efficiency of the oxygen evolving complex of the PSII), in which no alterations were observed for the toxicity of the single pesticides. The results suggest that these analyses are important to evaluate pesticide toxicity mechanisms in ecological risk assessments of tropical regions. Thus, here we demonstrate the importance of using multiple parameters in ecotoxicological studies to obtain a better understanding of the toxicity of these compounds for phytoplankton.

Mechanisms protect airborne green microalgae during long distance dispersal.

Viable microalgae occur in the air. Whether they can survive the stresses such as UV, desiccation and freezing temperatures at high altitudes during long distance dispersal is rarely studied. If yes, what mechanisms confer the tolerance? Four freshwater airborne green microalgae were isolated from Dongsha Atoll in the South China Sea, classified as Scenedesmus sp. DSA1, Coelastrella sp. DSA2, Coelastrella sp. DSA3 and Desmodesmus sp. DSA6 based on their morphologies and ITS sequences. Their survival rates under UV stress were tightly correlated with their cell wall thickness. All the four airborne green microalgae survived the air-dry stress on benchtop followed by - 20 °C freeze-desiccation stress for 4 weeks, but not the two waterborne green microalgae Desmodesmus sp. F5 and Neodesmus sp. UTEX 2219-4 used as controls. Three of the four airborne microalgae survived the lyophilization treatment, excluding Desmodesmus sp. DSA6 and the two waterborne microalgae. The four airborne microalgae produced carotenoids under prolonged stress conditions, which might help detoxify the reactive oxygen species generated under environmental stresses and shield from the high-light stress in the air. Characterization of these airborne microalgae may help answer how the descendants of green algae survived on the land about 450 MYA.

Shared up-regulation and contrasting down-regulation of gene expression distinguish desiccation-tolerant from intolerant green algae.

Among green plants, desiccation tolerance is common in seeds and spores but rare in leaves and other vegetative green tissues. Over the last two decades, genes have been identified whose expression is induced by desiccation in diverse, desiccation-tolerant (DT) taxa, including, e.g., late embryogenesis abundant proteins (LEA) and reactive oxygen species scavengers. This up-regulation is observed in DT resurrection plants, mosses, and green algae most closely related to these Embryophytes. Here we test whether this same suite of protective genes is up-regulated during desiccation in even more distantly related DT green algae, and, importantly, whether that up-regulation is unique to DT algae or also occurs in a desiccation-intolerant relative. We used three closely related aquatic and desert-derived green microalgae in the family Scenedesmaceae and capitalized on extraordinary desiccation tolerance in two of the species, contrasting with desiccation intolerance in the third. We found that during desiccation, all three species increased expression of common protective genes. The feature distinguishing gene expression in DT algae, however, was extensive down-regulation of gene expression associated with diverse metabolic processes during the desiccation time course, suggesting a switch from active growth to energy-saving metabolism. This widespread downshift did not occur in the desiccation-intolerant taxon. These results show that desiccation-induced up-regulation of expression of protective genes may be necessary but is not sufficient to confer desiccation tolerance. The data also suggest that desiccation tolerance may require induced protective mechanisms operating in concert with massive down-regulation of gene expression controlling numerous other aspects of metabolism.

Genome analyses provide insights into the evolution and adaptation of the eukaryotic Picophytoplankton Mychonastes homosphaera.

BACKGROUND: Picophytoplankton are abundant and can contribute greatly to primary production in eutrophic lakes. Mychonastes species are among the common eukaryotic picophytoplankton in eutrophic lakes. We used third-generation sequencing technology to sequence the whole genome of Mychonastes homosphaera isolated from Lake Chaohu, a eutrophic freshwater lake in China. RESULT: The 24.23 Mbp nuclear genome of M.homosphaera, harboring 6649 protein-coding genes, is more compact than the genomes of the closely related Sphaeropleales species. This genome streamlining may be caused by a reduction in gene family number, intergenic size and introns. The genome sequence of M.homosphaera reveals the strategies adopted by this organism for environmental adaptation in the eutrophic lake. Analysis of cultures and the protein complement highlight the metabolic flexibility of M.homosphaera, the genome of which encodes genes involved in light harvesting, carbohydrate metabolism, and nitrogen and microelement metabolism, many of which form functional gene clusters. Reconstruction of the bioenergetic metabolic pathways of M.homosphaera, such as the lipid, starch and isoprenoid pathways, reveals characteristics that make this species suitable for biofuel production. CONCLUSION: The analysis of the whole genome of M. homosphaera provides insights into the genome streamlining, the high lipid yield, the environmental adaptation and phytoplankton evolution.

Agro-industrial by-product in photoheterotrophic and mixotrophic culture of Tetradesmus obliquus: Production of ω3 and ω6 essential fatty acids with biotechnological importance.

In recent years, researchers have highlighted the role of low cost-efficient agro-industrial by-products used as supplements in algal culture media. The aim of the study was to identify and characterize the basic metabolic pathways in Tetradesmus obliquus cells induced by supplementation with beet molasses in photoheterotrophic and mixotrophic culture conditions. To assess the impact of the nutritional strategy in unicellular algae, growth curves were plotted and lipid, carbohydrate, and protein levels were determined. Fourier Transform Infrared Spectroscopy was applied to measure the Tetradesmus obliquus cell composition. Additionally, the C16-C18 fatty acid profile of Tetradesmus obliquus was determined by gas chromatograph/mass spectrometry. The switch from autotrophy to photoheterotrophy and mixotrophy contributes to shortening of the adaptation growth phase. The highest protein content was obtained in the mixotrophic growth. This study has demonstrated high content of 18:1, cisΔ9, 18:2, cisΔ9,12, ω6, and 18:3, cisΔ9,12,15, ω3 in photoheterotrophic and mixotrophic culture conditions. High levels of proteins and essential fatty acids make Tetradesmus obliquus cell biomass important for human and animals health.