Microalga Dunaliella Tertiolecta Research Articles

Cost-effective fibrinolytic enzyme production by microalga Dunaliella tertiolecta using medium supplemented with corn steep liquor.

A fibrinolytic enzyme from the microalga Dunaliella tertiolecta was produced under mixotrophic conditions using different corn steep liquor (CSL) concentrations ( 0 ≤ CLS ≤ 0.75%), purified using a combination of salting out and ion-exchange chromatography, and then biochemical characterized. Cultivation of this microalga using 0.5% CSL led to the highest maximum cell concentration (1.960±0.010 mg L-1) and cell productivity (0.140g L-1 day-1), besides a high fibrinolytic activity of the extract obtained by the homogenization method (102 ±1 U mL-1). The enzyme extracted from the microalgal biomass was 5-fold purified with a 20% yield and was found to have a specific activity of 670 U mg-1. The enzyme, whose molecular weight determined by fibrin zymography was 10 kDa, was shown to be stable at pH 3.0-9.0 and up to 70°C with optimal pH and temperature values of 8.0 and 50°C, respectively. When compared to other fibrinolytic enzymes, this protease stood out for its high fibrinolytic activity, which was enhanced by Fe2+, inhibited by Zn2+, Cu2+, Mg2+, and Ca2+, and strongly inhibited by phenylmethylsulfonyl fluoride, suggesting that it belongs to the serine metalloprotease family. Moreover, thanks to its thermal stability, the enzyme may be easily preserved and activated under high-temperature conditions.

Ultrastructure and stability of cellular nanoparticles isolated from Phaeodactylum tricornutum and Dunaliella tertiolecta conditioned media

Background: Cells in general secrete nanoparticles (NPs) which are believed to mediate intercellular communication. Recently, great efforts have been made to utilize them as delivery vectors. We aimed to harvest and identify NPs from liquid cultures of two marine microalgae Dunaliella tertiolecta and Phaeodactyum tricornutum. Methods: NPs were isolated from the culture conditioned media by differential ultracentrifugation by the protocol used for the isolation of extracellular vesicles. Microalgae and isolated NPs were examined by scanning electron microscopy (SEM) while isolated NPs were examined also by cryogenic transmission electron microscopy (cryo-TEM). The Triton X-100 detergent and temperature sensitivity of NPs was assessed by dynamic light scattering (DLS) through monitoring the intensity of the scattered light (I) and the distribution of hydrodynamic radii of NPs (Rh). Results: Two mechanisms of formation of NPs with average Rh 200 nm were observed in the D. tertiolecta culture: a disintegration of tubular protrusions, and cell decay. A part of the imaged D. tertiolecta NPs were membrane-enclosed vesicles, but the isolates also contained electron-dense NPs and nanofilaments. P. tricornutum NPs in the culture and in the isolate were homogeneous in size and shape. Their average Rh was 104 nm. The addition of surfactant to isolates resulted in a change in Rh distribution and a decrease of I in samples from both species, indicating decay of a part of NPs. Changes in the width of the I(Rh) peaks were observed at temperatures above 45 °C. Conclusions: A part of NPs found in isolates from microalgae D. tertiolecta and P. tricornutum were membrane-enclosed vesicles. However, the isolates obtained by a standard protocol for extracellular vesicle isolation by ultracentrifugation contained also a significant amount of other similar-sized nanoparticles. The isolates were partly susceptible to the addition of detergent and to temperature up to 80 degrees.

Hyposalinity induces significant polar lipid remodeling in the marine microalga Dunaliella tertiolecta (Chlorophyceae)

Marine microalgae often live in a fluctuating environment including a decrease in salinity caused by global warming induced sea ice melting and freshwater inflows. Under conditions of fluctuating salinity, microalgae have evolved a variety of survival mechanisms such as lipid accumulation and remodeling. The purpose of this study was to investigated the membrane lipid remodeling of the marine green microalga Dunaliella tertiolecta as a short-term acclimation mechanism in response to hyposalinity (20 and 3 PSU) with respect to growth at optimal salinity (38 PSU). We identified 34 lipid species belonging to seven polar lipid classes. Dunaliella tertiolecta accumulates cell lipids and remodels polar lipid classes and their fatty acids composition as response to hypoosmotic stress at 3 PSU. We found that the unsaturation of most polar lipids decreases overall, indicating decreased membrane fluidity and altered permeability, whereas shortening the length of fatty acids of polar lipids is not one of the strategies of D. tertiolecta to cope with the decrease in salinity. Increase in relative content (%) and unsaturation of monogalactosyldiacylglycerols (MGDG) and decrease in relative content (%) and unsaturation of phosphatidylglycerols (PG), suggesting changes in photosynthetic membranes of thylakoids at 20 and 3 PSU. At a very low salinity of 3 PSU, the relative content (%) of phosphatidylinositols (PI) increases, suggesting increased lipid trafficking and signaling in the cells. These changes are statistically significant and we hypothesize that D. tertiolecta is genetically adapted to withstand large salinity fluctuations through polar lipid composition.

Selenite as a Lipid Inductor in Marine Microalga Dunaliella tertiolecta: Comparison of One-Stage and Two-Stage Cultivation Strategies.

Microalgae have emerged as one of the most promising alternative sources of biofuels due to their high lipid accumulation ability. High lipid content is of pivotal importance for biodiesel production. In order to obtain high lipid content, modifications of culture conditions and development of an efficient lipid induction method are called for. In the present study, the possibility of using selenium in a form of sodium selenite as a lipid inductor in marine microalga Dunaliella tertiolecta was investigated during one- and two-stage cultivation modes. The effects of selenite on algal growth, pigment content, oxidative stress, and neutral lipid content were determined during both cultivation modes. The results revealed that the two-stage cultivation on 10.00-40.00mg L-1 of selenite resulted in up to twofold higher algal cell density compared to the one-stage cultivation. Selenite concentrations from 2.50 to 20.00mg L-1 increased lipid peroxidation during both cultivation modes, emphasizing the selenite-induced oxidative stress accompanied by the increased lipid accumulation in microalgae cells. During one- and two-stage cultivation on 20.00mg L-1 of selenite, lipid content increased 2.39- and 5.73-fold at days 9 and 14 of cultivation, respectively. Moreover, the highest obtained neutral lipid content during the two-stage cultivation was 5.40-fold higher than lipid content obtained during the one-stage cultivation. Collectively, these results suggest that the two-stage cultivation strategy, initiated with optimal culture conditions for biomass production and followed by the addition of selenite as a stress inductor, can be successfully deployed to enhance the lipid content in D. tertiolecta.

Lipid Productivity of Marine Microalgae Dunaliella tertiolecta in Marmara Seawater and Johnson’s Media with Different Salinities and Evaluation as A Raw Material Source for Biofuel Production

Microalgae are increasingly used in the production of various industrial chemicals. Lipids from major microalgae crops are similar to lipids obtained from oilseed plants and can be converted into biodiesel, moreover, microalgae are more advantageous than plants in oil production in many ways. Therefore, oil and biodiesel production technology from microalgae is a current and interesting research topic. The effect of salinity in the nutrient medium on the growth and lipid productivity of marine microalgae Dunaliella tertiolecta was investigated. The usability of Marmara Seawater (MSW) as a growth medium and its suita-bility for microalgae were also investigated. The highest cell concentration (1.53gdw / L), growth rate (μmax = 0.006h-1) and oil productivity (12.8 (g / L. Day) (10-3)) in nutrient medium (5g) were obtained. On the other hand, the cetane number was calculated according to the dominant arachidic acid (C20:0), it was determined as 74.9 and the average molecular weight of the oil was calculated as 849.1 g / mol. Ac-cording to the experimental results, it is predicted that microalgae lipids will be widely used as an alterna-tive to vegetable oils soon as raw materials.

Pigments Production, Growth Kinetics, and Bioenergetic Patterns in Dunaliella tertiolecta (Chlorophyta) in Response to Different Culture Media

This work dealt with the study of growth parameters, pigments production, and bioenergetic aspects of the microalga Dunaliella tertiolecta in different culture media. For this purpose, cultures were carried out in Erlenmeyer flasks containing F/2 medium, Bold’s Basal medium, or an alternative medium made up of the same constituents of the Bold’s Basal medium dissolved in natural seawater instead of distilled water. D. tertiolecta reached the highest dry cell concentration (Xmax = 1223 mgDM·L−1), specific growth rate (µmax = 0.535 d−1), cell productivity (PX = 102 mgDM·L−1·d−1), and photosynthetic efficiency (PE = 14.54%) in the alternative medium, while the highest contents of carotenoids (52.0 mg·g−1) and chlorophyll (108.0 mg·g−1) in the biomass were obtained in Bold’s Basal medium. As for the bioenergetic parameters, the biomass yield on Gibbs energy dissipation was higher and comparable in both seawater-based media. However, the F/2 medium led to the highest values of moles of photons absorbed to produce 1 C-mol of biomass (nPh), total Gibbs energy absorbed by the photosynthesis (ΔGa) and released heat (Q), as well as the lowest cell concentration, thus proving to be the least suitable medium for D. tertiolecta growth. On the other hand, the highest values of molar development of O2 and consumption of H+ and H2O were obtained in the alternative medium, which also ensured the best kinetic parameters, thereby allowing for the best energy exploitation for cell growth. These results demonstrate that composition of culture medium for microalgae cultivation has different effects on pigments production, growth kinetics, and bioenergetics parameters, which should be taken into consideration for any use of biomass, including as raw material for biofuels production.

Rainbow trout farm effluent as a potential source of feed and medium for mass culture of Artemia parthenogenetica

This study was conducted to evaluate the effects of the trout farm effluent (TFE) co-feeding with microalgae Dunaliella tertiolecta (MA) on Artemia parthenogenetica biomass production. The feeding regimes were 95% TFE+5% MA, 85% TFE+15% MA, and 75% TFE+25% MA, with different Artemia densities of 250, 500, 1000 and 1500 individuals L−1 in 1-L tanks and 1000 individuals L−1 in 800-L tanks in three replicates in two distinct experiments, comprising 48 experimental units in 1-L tank stage (including four controls fed 0%TFE+100% MA in four densities) and 9 experimental units in 800-L stage. The growth performance and nutritional value of animals were measured on days 8, 11, 14, 17, 20 and 23. The highest growth and survival performance was observed in the treatment containing 75% TFE+25% MA at a density of 250 individuals L−1 (1-L tanks) and 85% TFE+15% MA at a density of 1000 individuals L−1 (800-L tanks). Furthermore, higher protein, carbohydrate, and lipid contents were obtained in the treatment feed 75% TFE+25% MA for both 1-L and 800-L tanks. In the same treatment, significantly higher contents of fatty acids were obtained too. However, the maximum amount for each fatty acids was different based on the different densities. Based on the present result, Artemia parthenogenetica can be cultured successfully using trout farm effluent even with high substitution ratio instead of microalgae as the main feed for Artemia biomass production.

Dietary Supplementation with Dunaliella Tertiolecta Prevents Whitening of Brown Fat and Controls Diet-Induced Obesity at Thermoneutrality in Mice.

We investigated the effect of evodiamine-containing microalga Dunaliella tertiolecta (DT) on the prevention of diet-induced obesity in a thermoneutral C57BL/6J male (30 °C). It attenuates the activity of brown adipose tissue (BAT), which accelerates diet-induced obesity. Nine-week-old mice were fed a high-fat diet supplemented with 10 g (Low group) or 25 g (High group) DT powder per kg food for 12 weeks. Compared to control mice without DT supplementation, body weight gain was significantly reduced in the High group with no difference in food intake. Tissue analyses indicated maintenance of multilocular morphology in BAT and reduced fat deposition in liver in DT-supplemented mice. Molecular analysis showed a significant decrease in mammalian target of rapamycin−ribosomal S6 protein kinase signaling pathway in white adipose tissue and upregulation in mRNA expression of brown fat-associated genes including fibroblast growth factor-21 (Fgf21) and uncoupling protein 1 (Ucp1) in BAT in the High group compared to the control. In the experiments using C3H10T1/2 adipocytes, DT extract upregulated mRNA expression of brown fat-associated genes in dose-dependent and time-dependent manners, accompanied by a significant increase in secreted FGF21 levels. Our data show the ability of DT as a nutraceutical to prevent brown fat attenuation and diet-induced obesity in vivo.

Parameter Estimation for Dynamic Resource Allocation in Microorganisms: A Bi-level Optimization Problem

Given their key roles in almost all ecosystems and in several industries, understanding and predicting microorganism growth is of utmost importance. In compliance with evolutionary principles, coarse-grained or genome-scale models of microbial growth can be used to determine optimal resource allocation scheme under dynamic environmental conditions. Resource allocation approaches have given important qualitative results, but it still remains a gap towards quantitiative predictions. The first step in this direction is parameter calibration with experimental data. But fitting these models results in a bi-level optimization problem, whose numerical resolution involves complex optimization issues. As a case study, we present here a coarse-grained model describing how microalgae acclimate to a change in light intensity. We first determine using the Pontryagin maximum principle and numerical simulations the optimal strategy, corresponding to a turnpike with a chattering arc. Then, a bi-level optimization problem is proposed to calibrate the model with experimental data. To solve it, a classical parameter identification routine is used, calling at each iteration the bocop solver to solve the optimal control problem (by a direct method). The calibrated model is able to represent the photoacclimation dynamics of the microalga Dunaliella tertiolecta facing a down-shift of light intensity.

Gene Expression Analysis of Zeaxanthin Epoxidase from the Marine Microalga Dunaliella tertiolecta in Response to Light/Dark Cycle and Salinity.

Zeaxanthin is an important pigment in the photo-protection mechanism of microalgae. However, zeaxanthin epoxidase, an enzyme involved in the accumulation and conversion of zeaxanthin, has not been extensively studied in microalgae. In this work, we report the expression pattern of zeaxanthin epoxidase in Dunaliella tertiolecta (DtZEP) at different light and diverse salinity conditions. To confirm the responsiveness to light conditions, the ZEP expression pattern was investigated in photoperiodic (16 h of light and 8 h of dark) and continuous (24 h of light and 0 h of dark) light conditions. mRNA expression levels in photoperiodic conditions fluctuated along with the light/dark cycle, whereas those in continuous light remained unchanged. In varying salinity conditions, the highest mRNA and protein levels were detected in cells cultured in 1.5 M NaCl, and ZEP expression levels in cells shifted from 0.6 M NaCl to 1.5 M NaCl increased gradually. These results show that mRNA expression of DtZEP responds rapidly to the light/dark cycle or increased salinity, whereas changes in protein synthesis do not occur within a short period. Taken together, we show that DtZEP gene expression responds rapidly to light irradiation and hyperosmotic stress. In addition, ZEP expression patterns in light or salinity conditions are similar to those of higher plants, even though the habitat of D. tertiolecta is different.

Extremum-Seeking for micro-algae biomass productivity maximization: an experimental validation

This paper presents experimental results of a lab-scale implementation of an extremum seeking control strategy for maximizing the biomass productivity of cultures of the micro-algae Dunaliella tertiolecta in a flat-panel photobioreactor operated in continuous mode. The real-time optimization is based on a recursive least squares adaptation where the input (Dilution rate)/output (Biomass concentration) relation is approximated by a linear Hammerstein regression from which a productivity gradient estimate can be inferred. Lab-scale instrumentation and operating conditions are described and the results of two experiments are presented. They demonstrate the fast convergence of the extremum seeking scheme and practical considerations related to stability are discussed.

Study on biocharacteristics and puruvate production of moderately halophilic bacteria isolated from <i> dunalliella tertiolecta’s </i> culture medium

A moderately halophilic bacteria designed strain D34 was isolated from the culture medium of green microalga Dunaliella tertiolecta. The isolate was Gram-negative, aerobic, rod-shaped, approximately 0.45–0.60 mm wide and 1.25–5.10 mm long, occuring singly, non-motile, and flagellum- less. Colonies on solid media are cream, circular, and smooth. This strain was able to produce exopolysaccharide, poly hydroxybutyrate, oxidase and catalase positive. Growth occurred in a temparature range of 20–40°C, a salts concentration of 0.1–25% (w/v), and pH range 6–12. The major fatty acids were C16:0 (35.59%), C16:1w-7 (20.54%), C18: 1w-7 (30.14%), and C12:0 (10.03% of total fatty acids). Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain D34 belonged to the genus Halomonas. The highest levels of 16S rRNA gene sequence similarity were found between the strain D34 and H. aquamarina (sequence similarity 98.6 %).Pyruvate, a central intermediate in metabolism processes in all organisms, is widely used for the synthesis of various chemicals and polymers as well as ingredient or additive in food, cosmetics, and pharmaceuticals. In this study, pyruvate production by strain D34 following changes in culture medium, glucose and nitrate concentrations and culture temperature were also studied. In 84 hours of batch- cultivation, pyruvate production by wild-type Halomonas sp. D34 reached 37.24 g/L at 37°C with 20% glucose and 30 g/L sodium nitrate adding to SOT medium. These data provided evidences for pyruvate production using novel wild-type strains.

Loss of Function in Zeaxanthin Epoxidase of Dunaliella tertiolecta Caused by a Single Amino Acid Mutation within the Substrate-Binding Site.

The zea1 mutant of marine microalga Dunaliella tertiolecta accumulates zeaxanthin under normal growth conditions, and its phenotype has been speculated to be related to zeaxanthin epoxidase (ZEP). In this study, we isolated the ZEP gene from both wild-type D. tertiolecta and the mutant. We found that the zea1 mutant has a point mutation of the 1337th nucleotide of the ZEP sequence (a change from guanine to adenine), resulting in a change of glycine to aspartate in a highly conserved region in the catalytic domain. Similar expression levels of ZEP mRNA and protein in both wild-type and zea1 were confirmed by using qRT-PCR and western blot analysis, respectively. Additionally, the enzyme activity analysis of ZEPs in the presence of cofactors showed that the inactivation of ZEP in zea1 was not caused by deficiency in the levels of cofactors. From the predicted three-dimensional ZEP structure of zea1, we observed a conformational change on the substrate-binding site in the ZEP. A comparative analysis of the ZEP structures suggested that the conformational change induced by a single amino acid mutation might impact the interaction between the substrate and substrate-binding site, resulting in loss of zeaxanthin epoxidase function.

Growth bottlenecks of microalga Dunaliella tertiolecta in response to an up-shift in light intensity

ABSTRACTMicroalgae are a potential source of valuable products including drugs, chemicals, food supplements, biofuels, animal feed and fertilizers. In outdoor culture, biomass productivity is dependent on how well the microalgae respond to fluctuations in light during the day. Dunaliella tertiolecta is a halotolerant, photoautotrophic microalga. Our objective is to identify rate-limiting steps in growth as D. tertiolecta acclimates to changing light intensity. We studied the physiological, metabolic and transcriptomic changes of D. tertiolecta in a low dilution rate chemostat culture to determine rate-limiting processes in cell growth in response to changes in light intensity. Upon a 10-fold increase in light intensity from 40 to 400 µmol photons m–2 s–1 (HL), the photosynthetic efficiency and maximum photosynthetic rate (PR) per cell, or per chlorophyll, did not change significantly during lag (2 h) and exponential growth (8 h) phases, which suggests that the photosynthetic system was not the growth rate limiting step in response to higher light intensity. The expression of most genes in 2 h and 8 h samples was similar. Some genes involved in synthesis of cellular structures, protein processing and the cell cycle might be bottlenecks in the HL response, because they were expressed only at 8 h. It might not be practical to increase biomass by genetic modification of D. tertiolecta because of the numerous pathways and genes that are potential bottlenecks. Instead, future research can be directed to improving carbon sequestration by channelling it to storage compounds or excretion as extracellular glycerol.

Ecotoxicological effects of polystyrene microbeads in a battery of marine organisms belonging to different trophic levels

The aim of this study was to detect ecotoxicological effects of 0.1 μm polystyrene microbeads in marine organisms belonging to different trophic levels. MP build up, lethal and sub-lethal responses were investigated in the bacterium Vibrio anguillarum (culturability), in the green microalga Dunaliella tertiolecta (growth inhibition), in the rotifer Brachionus plicatilis (mortality and swimming speed alteration) and in the sea urchin Paracentrotus lividus (immobility and swimming speed alteration) exposed to a wide range of microplastic (MP) concentrations (from 0.001 to 10 mg L−1). Survival was not affected in all organisms up to 10 mg L−1, while algal growth inhibition, rotifer and sea urchin larvae swimming behaviour alterations were observed after exposure to MPs. Ingestion was only observed in rotifers and it was directly correlated with sub-lethal effects.These results account for the ecotoxicological risk associated to the polystyrene microbeads, which are able to affect different endpoints in primary producers and consumers (rotifers and sea urchins) since no effects were observed in decomposers.This study points out the importance of using a battery of marine organisms belonging to different trophic levels by studying acute toxicity of MPs at low and high contamination levels, and investigating sub-lethal responses. Further investigations aimed at studying the transfer of these materials through the web are particularly recommended.

In silico Analyses of Transcriptomes of the Marine Green Microalga Dunaliella tertiolecta: Identification of Sequences Encoding P-type ATPases

De novo assembled transcriptomes of the marine microalga Dunaliella tertiolecta (Chlorophyta) were analyzed. Transcriptome assemblies were performed using short-read RNA-seq data deposited in the SRA database (DNA and RNA Sequence Read Archive, NCBI). A merged transcriptome was assembled using a pooled RNA-seq data set. The goal of the study was in silico identification of nucleotide sequences encoding P-type ATPases in D. tertiolecta transcriptomes. P-type ATPases play a considerable role in the adaptation of an organism to a variable environment, and this problem is particularly significant for microalgae inhabiting an environment with an unstable ionic composition. Particular emphasis was given to searching for a sequence coding Na^(+)-ATPase. This enzyme is expected to function in the plasma membrane of D. tertiolecta like in some marine algae, in particular, in the closely related alga Dunaliella maritima. An ensemble of 12 P-type ATPases consisting of members belonging to the five main subfamilies of the P-type ATPase family was revealed in the assembled transcriptomes. The genes of the following P-type ATPases were found: (1) heavy metal ATPases (subfamily PIB); (2) Ca^(2+)-ATPases of SERCA type (subfamily P2A); (3) H^(+)-ATPases (subfamily P3); (4) phospholipid-transporting ATPases (flippases) (subfamily P4); (5) cation-transporting ATPases of uncertain specificities (subfamily P5). The presence of functional Na^(+)-ATPases in marine algae is presently undoubted. However, contrary to expectations, we failed to find a nucleotide sequence encoding a protein that could unequivocally be considered a Na^(+)-ATPase. Further study is necessary to elucidate the roles of in silico revealed D. tertiolecta ATPases in Na^(+) transport.

Long-term toxicity of surface-charged polystyrene nanoplastics to marine planktonic species Dunaliella tertiolecta and Artemia franciscana

Plastic pollution has been globally recognized as a critical issue for marine ecosystems and nanoplastics constitute one of the last unexplored areas to understand the magnitude of this threat. However, current difficulties in sampling and identifying nano-sized debris make hard to assess their occurrence in marine environment. Polystyrene nanoparticles (PS NPs) are largely used as nanoplastics in ecotoxicological studies and although acute exposures have been already investigated, long-term toxicity on marine organisms is unknown. Our study aims at evaluating the effects of 40nm PS anionic carboxylated (PS-COOH) and 50nm cationic amino-modified (PS-NH2) NPs in two planktonic species, the green microalga Dunaliella tertiolecta and the brine shrimp Artemia franciscana, respectively prey and predator. PS NP behaviour in exposure media was determined through DLS, while their toxicity to microalgae and brine shrimps evaluated through 72h growth inhibition test and 14 d long-term toxicity test respectively. Moreover, the expression of target genes (i.e. clap and cstb), having a role in brine shrimp larval growth and molting, was measured in 48h brine shrimp larvae. A different behaviour of the two PS NPs in exposure media as well as diverse toxicity to the two planktonic species was observed. PS-COOH formed micro-scale aggregates (Z-Average>1μm) and did not affect the growth of microalgae up to 50μg/ml or that of brine shrimps up to 10μg/ml. However, these negatively charged NPs were adsorbed on microalgae and accumulated (and excreted) in brine shrimps, suggesting a potential trophic transfer from prey to predator. On the opposite, PS-NH2-formed nano-scale aggregates (Z-Average<200nm), caused inhibition of algal growth (EC50=12.97μg/ml) and mortality in brine shrimps at 14 d (LC50=0.83μg/ml). Moreover, 1μg/ml PS-NH2 significantly induced clap and cstb genes, explaining the physiological alterations (e.g. increase in molting) previously observed in 48h larvae, but also suggesting an apoptotic pathway triggered by cathepsin L-like protease in brine shrimps upon PS-NH2 exposure. These findings provide a first insight into long-term toxicity of nanoplastics to marine plankton, underlining the role of the surface chemistry in determining the behaviour and effects of PS NPs, in terms of adsorption, growth inhibition, accumulation, gene modulation and mortality. The use of long-term end-point has been identified as valuable tool for assessing the impact of nanoplastics on marine planktonic species, being more predictable of real exposure scenarios for risk assessment purposes.

Hydrothermal liquefaction phase behavior of microalgae & model compounds in fused silica capillary reactor

ABSTRACTA fused silica capillary reactor combined with a heating/cooling stage, a microscope and a digital camera were used to investigate phase behavior during the hydrothermal liquefaction of microalgae (Dunaliella tertiolecta) and model compounds, including soya protein and glycine, starch, glucose and xylose, stearic acid and palmitic acid. Bubbles were generated at 246°C and disappeared at 360°C upon heating when Dunaliella tertiolecta used as feedstocks. Moreover, liquid products were generated at 300°C upon heating and oily liquid products began to separate out at 250°C upon cooling. The phase behavior of soya protein was similar to that of the Dunaliella tertiolecta. Meanwhile, there only observed the bubbles generation during hydrothermal liquefaction of glycine. Heating the starch, glucose and xylose above 350°C generated black solids from carbonization. Stearic acid and palmitic acid only had the process of melting, dissolution, dispersion and precipitation.

Effects of Salt Concentrations and Nitrogen and Phosphorus Starvations on Neutral Lipid Contents in the Green Microalga Dunaliella tertiolecta.

Dunaliella tertiolecta, a halotolerant alga, can accumulate large amounts of neutral lipid, which makes it a potential biodiesel feedstock. In this study, neutral lipids of D.tertiolecta induced by different salinities or N or P starvation were analyzed by thin-layer chromatography (TLC), flow cytometry (FCM), and confocal laser scanning microscopy (CLSM). High salinities or N or P starvation resulted in a decrease in cell growth and chlorophyll contents of D.tertiolecta. Neutral lipid contents increased markedly after 3-7 days of N starvation or at low NaCl concentrations (0.5-2.0 M). N starvation had a more dramatic effect on the neutral lipid contents of D.tertiolecta than P starvation. Four putative ME isozymes in different conditions can be detected by using isozyme electrophoresis. Two alternative acetyl-CoA producers, ACL and ACS genes, were up-regulated under low salinities and N starvation. It was suggested that low salinities and N starvation are considered efficient ways to stimulate lipid accumulation in D.tertiolecta.

Exploring the transcriptome of non-model oleaginous microalga Dunaliella tertiolecta through high-throughput sequencing and high performance computing

BackgroundRNA-Seq technology has received a lot of attention in recent years for microalgal global transcriptomic profiling. It is widely used in transcriptome-wide analysis of gene expression., particularly for microalgal strains with potential as biofuel sources. However, insufficient genomic or transcriptomic information of non-model microalgae has limited the understanding of their regulatory mechanisms and hampered genetic manipulation to enhance biofuel production. As such, an optimal microalgal transcriptomic database construction is a subject of urgent investigation.ResultsDunaliella tertiolecta, a non-model oleaginous microalgal species, was sequenced via Illumina MISEQ and HISEQ 4000 in RNA-Seq studies. The high quality high-throughout sequencing data were explored using high performance computing (HPC) in a petascale data center and subjected to de novo assembly and parallelized mpiBLASTX search with multiple species. As a result, a transcriptome database of 17,845 was constructed (~95% completeness). This enlarged database constructed fueled the RNA-Seq data analysis, which was validated by a nitrogen deprivation (ND) study that induces triacylglycerol (TAG) production.ConclusionsThe new paralleled assembly and annotation method under HPC presented here allows the solution of large-scale data processing problems in acceptable computation time. There is significant increase in the number of transcriptomic data achieved and observable heterogeneity in the performance to identify differentially expressed genes in the ND treatment paradigm. The results provide new insights as to how response to ND treatment in microalgae is regulated. ND analyses highlight the advantages of this database generated in this study that could also serve as a useful resource for future gene manipulation and transcriptome-wide analysis. We thus demonstrate the usefulness of exploring the transcriptome as an informative platform for functional studies and genetic manipulations in similar species.