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The marine microalga Tisochrysis lutea is a potential and sustainable source of bioactive compounds such as carotenoids and omega-3 fatty acids. In the present work, the extraction of fucoxanthin and docosahexaenoic acid (DHA), the most abundant omega-3 fatty acid which constitutes polar lipids particularly in the brain, was studied using advanced extraction techniques with green and bio-based solvents compared to traditional extraction techniques with hazardous organic solvents. The experimental design to maximize the lipid extraction yield by ultrasound-assisted extraction (UAE) was developed, choosing as experimental factors the percentage of solvent (0, 50 and 100% of 2-methyl-tetrahydrofuran or 2-methyloxolane (2-me-THF) in ethanol), the extraction time (20, 30 and 40 min) and temperature (40, 50 and 60 ºC). The highest lipid extraction yields were obtained using ethanol as solvent. Nevertheless, the most interesting extracts based on their chemical composition were obtained when the presence of 2-me-THF in the extraction mixture was greater than that of ethanol. Through analytical techniques such as HPLC-ELSD/DAD and GC-MS as well as spectrophotometric techniques, the contents of polar lipids, fatty acids, total carotenoids and fucoxanthin were quantified. In addition, the antioxidant capacity of different selected extracts was studied, being once again the most interesting those extracted with different amounts of 2-me-THF due to its selectivity and enriched composition in high-added value bioactives, mainly fucoxanthin and DHA. Therefore, it is shown the importance of choosing an advanced extraction technique together with the use of green solvents not only to develop procedures that are in agreement with Green Chemistry but also to preserve its bioactivity. [ABSTRACT FROM AUTHOR]

Tisochrysis lutea is a highly nutritious marine microalga that has various applications in aquaculture and biotechnology. However, the effects of T. lutea extract (TLE) on osteoarthritis (OA) pathogenesis remain unexplored. In this study, we aimed to determine the effects of TLE on OA development. We found that TLE inhibits the expression of matrix metalloproteinases (MMPs) and reactive oxygen species (ROS) activity in an OA mouse model generated by the destabilization of the medial meniscus (DMM) surgery. In vivo assays of the OA model mice demonstrated that TLE has a protective effect against cartilage destruction by inhibiting MMP3 and MMP13 expression. To enable the medical use of TLE, the components of TLE were characterized using high-performance liquid chromatography (HPLC) analysis. Interestingly, we found that Fucoxanthin accounts for 41.2% of TLE and showed anti-catabolic and antioxidant effects under IL-1β-treated in vitro conditions. RNA sequencing analysis showed that fucoxanthin decreased p38, NF-κB, and JNK signaling pathway gene expression, all of which are activated by IL-1β. Furthermore, in vivo analysis showed that fucoxanthin inhibited the IL-1β-stimulated phosphorylation of p65, JNK, and p38. These results highlight new possibilities for the use of TLE as a source of fucoxanthin, an antioxidant, for OA treatment. [ABSTRACT FROM AUTHOR]

Oceanic phytoplankton species are generally composed of many strains, with intra-species diversity consisting of genetic and phenotypic variability. Despite its importance in ecological and biotechnological contexts, this intra-species diversity and variation among strains has been little studied. We investigated the intra-species diversity of the microalga Tisochrysis lutea, a haptophyte of the Isochrysidales order. Inter-strain diversity of T. lutea was studied because of the economic importance of the species as a feed in aquaculture and for antioxidant metabolite production, particularly fucoxanthin and other carotenoids, which have health benefits. We analysed Tara Ocean datasets which revealed that T. lutea was present in the Pacific, Atlantic and Indian Oceans but not in the Arctic or Austral Oceans. We next made phenotypic and genotypic comparisons of 11 strains of T. lutea from worldwide algal collections. All strains were cultivated in the same controlled conditions for one week, and several phenotypic traits were measured, notably antioxidant content. In parallel, the genomes of each strain were sequenced, and genetic variants identified. At the genetic and phenotypic levels, the strains were distinct from each other and our analysis revealed natural trait variations of interest in relation to further exploitation in domestication programmes. A large number of genetic variations were identified among the strains, but no major differences in genome size were observed. Moreover, limited genetic structure was observed among these strains, which could be a consequence of the complex life history of species within the Isochrysidales. Our study provides new knowledge on the intra-species diversity that should be considered in future environmental studies and breeding programmes. Tisochrysis lutea is found in many parts of the world's oceans. T. lutea has high inter-strain phenotypic and genomic variation. Genetic structure of strains from culture collections is limited. [ABSTRACT FROM AUTHOR]

Tisochrysis lutea is a highly nutritious marine microalga that has various applications in aquaculture and biotechnology. However, the effects of T. lutea extract (TLE) on osteoarthritis (OA) pathogenesis remain unexplored. In this study, we aimed to determine the effects of TLE on OA development. We found that TLE inhibits the expression of matrix metalloproteinases (MMPs) and reactive oxygen species (ROS) activity in an OA mouse model generated by the destabilization of the medial meniscus (DMM) surgery. In vivo assays of the OA model mice demonstrated that TLE has a protective effect against cartilage destruction by inhibiting MMP3 and MMP13 expression. To enable the medical use of TLE, the components of TLE were characterized using high-performance liquid chromatography (HPLC) analysis. Interestingly, we found that Fucoxanthin accounts for 41.2% of TLE and showed anti-catabolic and antioxidant effects under IL-1β-treated in vitro conditions. RNA sequencing analysis showed that fucoxanthin decreased p38, NF-κB, and JNK signaling pathway gene expression, all of which are activated by IL-1β. Furthermore, in vivo analysis showed that fucoxanthin inhibited the IL-1β-stimulated phosphorylation of p65, JNK, and p38. These results highlight new possibilities for the use of TLE as a source of fucoxanthin, an antioxidant, for OA treatment.

In our study we investigated the growth, biochemical composition, and ultrastructure of Tisochrysis lutea microalgae in enrichment culture during 30 days experiment. The number of T. lutea cells increased throughout the experiment. We noted an increase in the size and number of lipid droplets containing fatty acids and carotenoids, including fucoxanthin, in the exponential and stationary phases of their growth. It has been established that the total content of carotenoids reaches a maximum in the stationary phase and decreases in the dying phase. During the stationary phase, exocytosis was observed in cells with the release of lipid droplets. This study demonstrates the potential of the T. lutea clone MBRU_Tiso-08 from the Marine Biobank Bioresource Collection of the National Scientific Center of Marine Biology, Far Eastern Branch, Russian Academy of Sciences, as a raw material for domestic biotechnology aimed at the combined extraction of carotenoids (including fucoxanthin) and lipids (including docosahexaenoic and eicosapentaenoic fatty acids). [ABSTRACT FROM AUTHOR]

Abstract Marine microalgae have received much attention as a sustainable source of the two health beneficial omega-3-fatty acids docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5). However, photoautotrophic monocultures of microalgae can only produce either DHA or EPA enriched biomass. An alternative may be the photoautotrophic co-cultivation of Tisochrysis lutea as DHA-producer with Microchloropsis salina for simultaneous EPA production to obtain EPA- and DHA-rich microalgae biomass in a nutritionally balanced ratio. Photoautotrophic co-cultivation processes of T. lutea and M. salina were studied, applying scalable and fully controlled lab-scale gas-lift flat-plate photobioreactors with LED illumination for dynamic climate simulation of a repeated sunny summer day in Australia [day–night cycles of incident light (PAR) and temperature]. Monocultures of both marine microalgae were used as reference batch processes. Differences in the autofluorescence of both microalgae enabled the individual measurement, of cell distributions in co-culture, by flow cytometry. The co-cultivation of T. lutea and M. salina in artificial sea water with an inoculation ratio of 1:3 resulted in a balanced biomass production of both microalgae simultaneously with a DHA:EPA ratio of almost 1:1 (26 mgDHA gCDW −1, and 23 mgEPA gCDW −1, respectively) at harvest after depletion of the initially added fertilizer. Surprisingly, more microalgae biomass was produced within 8 days in co-cultivation with an increase in the cell dry weight (CDW) concentration by 31%, compared to the monocultures with the same amount of light and fertilizer. What is more, DHA-content of the microalgae biomass was enhanced by 33% in the co-culture, whereas EPA-content remained unchanged compared to the monocultures. Graphical Abstract

Pre-metabolic syndrome (pre-MetS) may represent the best transition phase to start treatments aimed at reducing cardiometabolic risk factors of MetS. In this study, we investigated the effects of the marine microalga Tisochrysis lutea F&M-M36 (T. lutea) on cardiometabolic components of pre-MetS and its underlying mechanisms. Rats were fed a standard (5% fat) or a high-fat diet (20% fat) supplemented or not with 5% of T. lutea or fenofibrate (100 mg/Kg) for 3 months. Like fenofibrate, T. lutea decreased blood triglycerides (p < 0.01) and glucose levels (p < 0.01), increased fecal lipid excretion (p < 0.05) and adiponectin (p < 0.001) without affecting weight gain. Unlike fenofibrate, T. lutea did not increase liver weight and steatosis, reduced renal fat (p < 0.05), diastolic (p < 0.05) and mean arterial pressure (p < 0.05). In visceral adipose tissue (VAT), T. lutea, but not fenofibrate, increased the β3-adrenergic receptor (β3ADR) (p < 0.05) and Uncoupling protein 1 (UCP-1) (p < 0.001) while both induced glucagon-like peptide-1 receptor (GLP1R) protein expression (p < 0.001) and decreased interleukin (IL)-6 and IL-1β gene expression (p < 0.05). Pathway analysis on VAT whole-gene expression profiles showed that T. lutea up-regulated energy-metabolism-related genes and down-regulated inflammatory and autophagy pathways. The multitarget activity of T. lutea suggests that this microalga could be useful in mitigating risk factors of MetS. [ABSTRACT FROM AUTHOR]

Microalgae, because of their high nutritional value and bioactive molecule content, are interesting candidates for functional foods, including fermented foods, in which the beneficial effects of probiotic bacteria combine with those of biomolecules lying in microalgal biomass. The aim of this work was to evaluate the potential of Tisochrysis lutea F&M-M36 as a substrate for Lactiplantibacillus plantarum ATCC 8014 and to verify fermentation effects on functionality. Bacterium selection among three lactobacilli was based on growth and resistance to in vitro digestion. Microalgal raw biomass and its digested residue were fermented in two matrixes, water and diluted organic medium, and analysed for biochemical composition and antioxidant activity along with their unfermented counterparts. Bacterial survivability to digestion and raw biomass digestibility after fermentation were also evaluated. Fucoxanthin was strongly reduced (>90%) in post-digestion residue, suggesting high bioavailability. Raw biomass in diluted organic medium gave the highest bacterial growth (8.5 logCFU mL−1) and organic acid production (5 mg L−1), while bacterial survivability to digestion (<3%) did not improve. After fermentation, the antioxidant activity of lipophilic extracts increased (>90%). Fermentation appears an interesting process to obtain T. lutea-based functional foods, although further investigations are needed to optimize bacterial growth and fully evaluate its effects on functionality and organoleptic features. [ABSTRACT FROM AUTHOR]

Given the potential of microalgae as new aquafeed ingredients, this study focuses on using a blend of microalgae, Tisochrysis lutea, Nannochloropsis gaditana, and Scenedesmus almeriensis, as a dietary ingredient for feeding Sparus aurata juveniles. The growth performance, carcass composition, tissue fatty acid profile, and intestinal microbiota were evaluated after a 30 day-feeding period. A microalgae-free diet was used as control, and three experimental diets were formulated containing 5%, 15%, and 25% of the microalgae blend (MB-5%, MB-15%, and MB-25%, respectively). After 7, 15, and 30 days of feeding experimental diets, biological samples were taken. Growth performance and nutrient utilization were not significantly modified at the end of the experiment. Microalgae inclusion tended to decrease body lipids and affected the fatty acid profile, especially MB-25 diet increased DHA levels. Diet MB-25 promoted appropriate microbial diversity, favoring the presence of probiotic bacteria, such as Lactobacillus, and significantly influencing the fatty acid composition and lipid metabolism in fish. In conclusion, using a short pulse of dietary administration of 25% microalgal blend in S. aurata modulates the intestinal microbiota and lipid composition while maintaining growth performance. [ABSTRACT FROM AUTHOR]

Marine microalgae have received much attention as a sustainable source of the two health beneficial omega-3-fatty acids docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5). However, photoautotrophic monocultures of microalgae can only produce either DHA or EPA enriched biomass. An alternative may be the photoautotrophic co-cultivation of Tisochrysis lutea as DHA-producer with Microchloropsis salina for simultaneous EPA production to obtain EPA- and DHA-rich microalgae biomass in a nutritionally balanced ratio. Photoautotrophic co-cultivation processes of T. lutea and M. salina were studied, applying scalable and fully controlled lab-scale gas-lift flat-plate photobioreactors with LED illumination for dynamic climate simulation of a repeated sunny summer day in Australia [day–night cycles of incident light (PAR) and temperature]. Monocultures of both marine microalgae were used as reference batch processes. Differences in the autofluorescence of both microalgae enabled the individual measurement, of cell distributions in co-culture, by flow cytometry. The co-cultivation of T. lutea and M. salina in artificial sea water with an inoculation ratio of 1:3 resulted in a balanced biomass production of both microalgae simultaneously with a DHA:EPA ratio of almost 1:1 (26 mgDHA gCDW−1, and 23 mgEPA gCDW−1, respectively) at harvest after depletion of the initially added fertilizer. Surprisingly, more microalgae biomass was produced within 8 days in co-cultivation with an increase in the cell dry weight (CDW) concentration by 31%, compared to the monocultures with the same amount of light and fertilizer. What is more, DHA-content of the microalgae biomass was enhanced by 33% in the co-culture, whereas EPA-content remained unchanged compared to the monocultures. [ABSTRACT FROM AUTHOR]

The biochemical nature and composition of fatty acids (FA's) in microalgae influence the nutritional quality and efficiency of this food source in bivalve larviculture. This research analyses the effect FA's content of Chaetoceros gracilis and Tisochrysis lutea on the FA's composition, growth and survival of Striostrea prismatica larvae. We analysed the FA's profiles of the microalgae, and the tissue of the larvae during a S. prismatica larviculture at day 1 ("D" larvae), day 8 (umbonate larvae) and day 22 (pediveliger) post fertilization (PF). Larvae growth and survival were evaluated during two phases: phase I from day 1 to day 8 PF and phase II from day 9 to day 22 PF. In phase I, we recorded a daily growth rate of 4.76 μm.day−1, while in phase II we recorded increases of 13.24 μm.day−1. In phase I, survival was 36.6%, and decreased further to 9.3% at the end of phase II. The sharp decrease in survival and low growth during phase I could possibly be explained by a specific nutritional deficiency of the microalgae diet (absence of FA's 15:0, 20:4n‐3, 20:4n‐6 and 22:5n‐ 3), especially if we consider the starting FA's profile of the "D" larvae. The larvae showed a high nutritional requirement for palmitic, stearic and oleic FA's and long‐chain PUFA's, especially 20:5n‐3 and 22:6n‐3. The absence of these FA's in the diet and their presence in the larval profile suggest that S. prismatica could have the ability to synthesize these FA's; however, more research is needed to corroborate this hypothesis. [ABSTRACT FROM AUTHOR]

We propose metabolic models for the haptophyte microalgae Tisochrysis lutea with different possible organic carbon excretion mechanisms. These models--based on the DRUM(Dynamic Reduction of UnbalancedMetabolism) methodology--are calibrated with an experiment of nitrogen starvation under day/night cycles, and then validated with nitrogen-limited chemostat culture under continuous light. We show that models including exopolysaccharide excretion offer a better prediction capability. It also gives an alternative mechanistic interpretation to the Droop model for nitrogen limitation, which can be understood as an accumulation of carbon storage during nitrogen stress, rather than the common belief of a nitrogen pool driving growth. Excretion of organic carbon limits its accumulation, which leads to a maximal C/N ratio (corresponding to the minimum Droop N/C quota). Although others phenomena--including metabolic regulations and dissipation of energy--are possibly at stake, excretion appears as a key component in our metabolic model, that we propose to include in the Droop model. [ABSTRACT FROM AUTHOR]

Haptophyte microalgae are key contributors to microbial communities in many environments. It has been proposed recently that members of this group would be virtually all dependent on vitamin B12 (cobalamin), an enzymatic cofactor produced only by some bacteria and archaea. Here, we examined the processes of vitamin B12 acquisition by haptophytes. We tested whether co-cultivating the model species Tisochrysis lutea with B12-producing bacteria in vitamin-deprived conditions would allow the microalga to overcome B12 deprivation. While T. lutea can grow by scavenging vitamin B12 from bacterial extracts, co-culture experiments showed that the algae did not receive B12 from its associated bacteria, despite bacteria/algae ratios supposedly being sufficient to allow enough vitamin production. Since other studies reported mutualistic algae–bacteria interactions for cobalamin, these results question the specificity of such associations. Finally, cultivating T. lutea with a complex bacterial consortium in the absence of the vitamin partially rescued its growth, highlighting the importance of microbial interactions and diversity. This work suggests that direct sharing of vitamin B12 is specific to each species pair and that algae in complex natural communities can acquire it indirectly by other mechanisms (e.g., after bacterial lysis). [ABSTRACT FROM AUTHOR]

A statistical experimental design using response surface methodology (RSM) was carried out to optimize the mixotrophic growth using glutamic acid, tri-sodium citrate, succinic acid, sodium aspartate, and sodium pyruvate to improve the production of fucoxanthin. First, a fractional factorial design as a screening test was done with the above-mentioned intermediates. Then, glutamic acid, sodium aspartate, and sodium pyruvate were chosen according to their statistically significant (P<0.05) and positive effects on fucoxanthin production. The three selected metabolic intermediates were optimized employing RSM to obtain a high level of fucoxanthin. The data were adjusted with a second-order polynomial equation to determine the relationship between fucoxanthin production and three optimized metabolic intermediates. Using multiple regression techniques and to attain the high fucoxanthin productivity (22.4 mg L−1 day−1), the optimum amount of the metabolic intermediates were found as follows: sodium aspartate, 7.5 mM; sodium pyruvate, 7.5 mM; glutamic acid, 3.29 mM. [ABSTRACT FROM AUTHOR]

We propose metabolic models for the haptophyte microalgae Tisochrysis lutea with different possible organic carbon excretion mechanisms. These models—based on the DRUM (Dynamic Reduction of Unbalanced Metabolism) methodology—are calibrated with an experiment of nitrogen starvation under day/night cycles, and then validated with nitrogen-limited chemostat culture under continuous light. We show that models including exopolysaccharide excretion offer a better prediction capability. It also gives an alternative mechanistic interpretation to the Droop model for nitrogen limitation, which can be understood as an accumulation of carbon storage during nitrogen stress, rather than the common belief of a nitrogen pool driving growth. Excretion of organic carbon limits its accumulation, which leads to a maximal C/N ratio (corresponding to the minimum Droop N/C quota). Although others phenomena—including metabolic regulations and dissipation of energy—are possibly at stake, excretion appears as a key component in our metabolic model, that we propose to include in the Droop model.

Pre-metabolic syndrome (pre-MetS) may represent the best transition phase to start treatments aimed at reducing cardiometabolic risk factors of MetS. In this study, we investigated the effects of the marine microalga Tisochrysis lutea F&M-M36 (T. lutea) on cardiometabolic components of pre-MetS and its underlying mechanisms. Rats were fed a standard (5% fat) or a high-fat diet (20% fat) supplemented or not with 5% of T. lutea or fenofibrate (100 mg/Kg) for 3 months. Like fenofibrate, T. lutea decreased blood triglycerides (p < 0.01) and glucose levels (p < 0.01), increased fecal lipid excretion (p < 0.05) and adiponectin (p < 0.001) without affecting weight gain. Unlike fenofibrate, T. lutea did not increase liver weight and steatosis, reduced renal fat (p < 0.05), diastolic (p < 0.05) and mean arterial pressure (p < 0.05). In visceral adipose tissue (VAT), T. lutea, but not fenofibrate, increased the β3-adrenergic receptor (β3ADR) (p < 0.05) and Uncoupling protein 1 (UCP-1) (p < 0.001) while both induced glucagon-like peptide-1 receptor (GLP1R) protein expression (p < 0.001) and decreased interleukin (IL)-6 and IL-1β gene expression (p < 0.05). Pathway analysis on VAT whole-gene expression profiles showed that T. lutea up-regulated energy-metabolism-related genes and down-regulated inflammatory and autophagy pathways. The multitarget activity of T. lutea suggests that this microalga could be useful in mitigating risk factors of MetS.

[Display omitted] • Ultrasonic probe method (5 min, ethanol) for extracting microalgal carotenoids. • Ultrasound as a greener and shorter alternative to the 24 h reference method. • AGREEprep™ software demonstrated the greenness of the ultrasonic method. • The application of ultrasound to defatted biomass shows potential for microalgal valorization. • Carotenoid and chlorophyll profiles varied depending on the species, biomass, and method. Carotenoids, versatile natural pigments with numerous health benefits, face environmental concerns associated with conventional petrochemical-based extraction methods and limitations of their synthetic equivalents. In this context, this study aims to introduce eco-friendly approaches using ultrasound-based strategies (probe and bath) for the extraction of carotenoids from microalgae, initially focusing on Microchloropsis gaditana and subsequently evaluating the versatility of the method by applying it to other microalgae species of interest (Tisochrysis lutea , Porphyridium cruentum , and Phaeodactylum tricornutum) and defatted microalgal residues. Among the approaches evaluated, the 5-min ultrasonic probe system with ethanol showed comparable carotenoid recovery efficiency to the reference method (agitation, 24 h, acetone) (9.4 ± 2.5 and 9.6 ± 3.2 mg g−1 carotenoids per dry biomass, for the green and the reference method, respectively). Moreover, the method's sustainability was demonstrated using the AGREEprep™ software (scored 0.62 out of 1), compared to the traditional method (0.22 out of 1). The developed method yielded high carotenoid contents across species with diverse cell wall compositions (3.1 ± 0.2, 2.1 ± 0.3, and 4.1 ± 0.1 mg g−1 carotenoid per dry biomass for T. lutea , P. cruentum , and P. tricornutum , respectively). Moreover, the application of the method to defatted biomass showed potential for microalgal valorization with carotenoid recovery rates of 41 %, 60 %, 61 %, and 100 % for M. gaditana , P. tricornutum , T. lutea , and P. cruentum , compared to the original biomass, respectively. Furthermore, by using high-performance liquid chromatography with a diode array detector (HPLC-DAD) and high-resolution mass spectrometry (HRMS), we reported the carotenoid and chlorophyll profiles of the different microalgae and evaluated the impact of the eco-friendly methods. The carotenoid and chlorophyll profiles varied depending on the species, biomass, and method used. In summary, this study advances a green extraction method with improved environmental sustainability and shorter extraction time, underscoring the potential of this approach as a valuable alternative for the extraction of microalgal pigments. [ABSTRACT FROM AUTHOR]

Microalgae, because of their high nutritional value and bioactive molecule content, are interesting candidates for functional foods, including fermented foods, in which the beneficial effects of probiotic bacteria combine with those of biomolecules lying in microalgal biomass. The aim of this work was to evaluate the potential of Tisochrysis lutea F&M-M36 as a substrate for Lactiplantibacillus plantarum ATCC 8014 and to verify fermentation effects on functionality. Bacterium selection among three lactobacilli was based on growth and resistance to in vitro digestion. Microalgal raw biomass and its digested residue were fermented in two matrixes, water and diluted organic medium, and analysed for biochemical composition and antioxidant activity along with their unfermented counterparts. Bacterial survivability to digestion and raw biomass digestibility after fermentation were also evaluated. Fucoxanthin was strongly reduced (>90%) in post-digestion residue, suggesting high bioavailability. Raw biomass in diluted organic medium gave the highest bacterial growth (8.5 logCFU mL−1) and organic acid production (5 mg L−1), while bacterial survivability to digestion (90%). Fermentation appears an interesting process to obtain T. lutea-based functional foods, although further investigations are needed to optimize bacterial growth and fully evaluate its effects on functionality and organoleptic features.

Fucoxanthin is a brown-colored pigment from algae, with great potential as a bioactive molecule due to its numerous properties. This review aims to present current knowledge on this high added-value pigment. An accurate analysis of the biological function of fucoxanthin explains its wide photon absorption capacities in golden-brown algae. The specific chemical structure of this pigment also leads to many functional activities in human health. They are outlined in this work and are supported by the latest studies in the literature. The scientific and industrial interest in fucoxanthin is correlated with great improvements in the development of algae cultures and downstream processes. The best fucoxanthin producing algae and their associated culture parameters are described. The light intensity is a major influencing factor, as it has to enable both a high biomass growth and a high fucoxanthin content. This review also insists on the most eco-friendly and innovative extraction methods and their perspective within the next years. The use of bio-based solvents, aqueous two-phase systems and the centrifugal partition chromatography are the most promising processes. The analysis of the global market and multiple applications of fucoxanthin revealed that Asian companies are major actors in the market with macroalgae. In addition, fucoxanthin from microalgae are currently produced in Israel and France, and are mostly authorized in the USA. [ABSTRACT FROM AUTHOR]

Fucoxanthin is one of the most vital pigments during photosynthesis and is extracted from golden-brown micro-algae such as Tisochrysis lutea. The present study investigates the constant volumetric mass transfer coefficient (k L a), for the first time as the scale-up strategy to change the scale from 500 mL to 2-L cultivation flasks, and 5-L bubble column photobioreactor for fucoxanthin production in T. lutea. The cell density and fucoxanthin content were improved because of through fine aeration, nutrients, and light availability by successful laboratory scale up. Fucoxanthin productivity obtained 21.20, 22.99, and 24.96 mg L–1day–1 for 500 mL, 2-L bottle, and 5-L bubble column photobioreactor, respectively. In addition, the biomass productivity enhanced from 267.5 to 275 and 284 mg L–1day–1 in three mentioned scales, respectively. Eventually, the scale up process for the production of fucoxanthin was succeeded from 500 mL bottle to 5-L photobioreactor using constant (k L a) under laboratory conditions. • Fucoxanthin is one of the most important carotenoids with several health benefits. • The production of this valuable metabolite from Tisochrysis lutea was investigated in a bubble column photobioreactor. • The constant volumetric mass transfer coefficient, (kLa) used as the scale-up strategy for the production of fucoxanthin from 500 mL to 5-L bubble column photobioreactor. • Fucoxanthin productivity was enhanced up to 24.96 mg L–1 day–1 in f/2 enriched medium by starch. [ABSTRACT FROM AUTHOR]

We observed differences in lhc classification in Chromista. We proposed a classification of the lhcf family with two groups specific to haptophytes, one specific to diatoms, and one specific to seaweeds. Identification and characterization of the Fucoxanthin and Chlorophyll a/c -binding Protein (FCP) of the haptophyte microalgae Tisochrysis lutea were performed by similarity analysis. The FCP family contains 52 lhc genes in T. lutea. FCP pigment binding site candidates were characterized on Lhcf protein monomers of T. lutea , which possesses at least nine chlorophylls and five fucoxanthin molecules, on average, per monomer. The expression of T. lutea lhc genes was assessed during turbidostat and chemostat experiments, one with constant light (CL) and changing nitrogen phases, the second with a 12 h:12 h sinusoidal photoperiod and changing nitrogen phases. RNA-seq analysis revealed a dynamic decrease in the expression of lhc genes with nitrogen depletion. We observed that T. lutea lhcx2 was only expressed at night, suggesting that its role is to protect \cells from return of light after prolonged darkness exposure. [ABSTRACT FROM AUTHOR]

[Display omitted] • Winter strain of Tisochrysis lutea obtained from >180-day cold adaptation and two rounds of FACS selection. • Fx content increased from 1.11 to 4.24 mg g−1 during strain improvement. • TFA polar lipid fraction increased from 21 % to 55 % during strain improvement. • Fx productivity in winter strain at 15 °C was identical to original one at 30 °C. Tisochrysis lutea is an important microalgal species for fucoxanthin and docosahexaenoic acid (DHA) production with an optimum cultivation temperature of approximately 30 °C. The aim of the present work was to develop a winter strain with high productivity at 15 °C. The response of the original strain to a decrease in temperature from 30 °C to 15 °C was investigated in continuous turbidostat experiments. This was followed by adaptation for >180 days at 15 °C and 2 rounds of sorting for cells with high chlorophyll fluorescence (top 5%) using fluorescence-activated cell sorting (FACS). For the original strain the productivity of biomass, fucoxanthin, and DHA decreased by 92 %, 98 % and 85 % respectively when decreasing the temperature from 30 °C to 15 °C. In the sorted cold-adapted 'winter strain', biomass, fucoxanthin, and DHA productivities were similar to those at 30 °C. In addition, the fucoxanthin concentration increased from 1.11 to 4.24 mg g−1 dry weight and the polar lipid fraction in total fatty acids increased from 21 % to 55 %. The winter strain showed a robust and stable phenotype after one year of cultivation, expanding the outdoor fucoxanthin and lipid production seasons for this species. [ABSTRACT FROM AUTHOR]

We observed differences in lhc classification in Chromista. We proposed a classification of the lhcf family with two groups specific to haptophytes, one specific to diatoms, and one specific to seaweeds. Identification and characterization of the Fucoxanthin and Chlorophyll a/c-binding Protein (FCP) of the haptophyte microalgae Tisochrysis lutea were performed by similarity analysis. The FCP family contains 52 lhc genes in T. lutea. FCP pigment binding site candidates were characterized on Lhcf protein monomers of T. lutea, which possesses at least nine chlorophylls and five fucoxanthin molecules, on average, per monomer. The expression of T. lutea lhc genes was assessed during turbidostat and chemostat experiments, one with constant light (CL) and changing nitrogen phases, the second with a 12 h:12 h sinusoidal photoperiod and changing nitrogen phases. RNA-seq analysis revealed a dynamic decrease in the expression of lhc genes with nitrogen depletion. We observed that T. lutea lhcx2 was only expressed at night, suggesting that its role is to protect \cells from return of light after prolonged darkness exposure.

Given the potential of microalgae as new aquafeed ingredients, this study focuses on using a blend of microalgae, Tisochrysis lutea, Nannochloropsis gaditana, and Scenedesmus almeriensis, as a dietary ingredient for feeding Sparus aurata juveniles. The growth performance, carcass composition, tissue fatty acid profile, and intestinal microbiota were evaluated after a 30 day-feeding period. A microalgae-free diet was used as control, and three experimental diets were formulated containing 5%, 15%, and 25% of the microalgae blend (MB-5%, MB-15%, and MB-25%, respectively). After 7, 15, and 30 days of feeding experimental diets, biological samples were taken. Growth performance and nutrient utilization were not significantly modified at the end of the experiment. Microalgae inclusion tended to decrease body lipids and affected the fatty acid profile, especially MB-25 diet increased DHA levels. Diet MB-25 promoted appropriate microbial diversity, favoring the presence of probiotic bacteria, such as Lactobacillus, and significantly influencing the fatty acid composition and lipid metabolism in fish. In conclusion, using a short pulse of dietary administration of 25% microalgal blend in S. aurata modulates the intestinal microbiota and lipid composition while maintaining growth performance.

The biotechnological potential of microalgae has gained considerable importance in many applied fields: biomass production for food and feed, cosmeceutical and pharmaceutical products, energy and phytoremediation. The driving force that inspires the progress in microalgae production is the need for new cultivation systems to obtain simultaneously the maximum yield, reduction of water and nutrients use, and production of economically interesting molecules, such as pigments, fatty acids and polysaccharides. We aim to test, for the first time, the co-cultivation in saline medium of Tisochrysis lutea (Haptophyta) and Nannochloropsis oculata (Ochrophyta) to obtain valuable compounds, i.e. pigments and lipids characteristic of each species, using a single culture process. Mono-cultures of each strain were used as controls. The two strains showed an increase in the concentration of chlorophylls and carotenoids in co-culture. At the end of the experiment, the fatty acid profile was analysed by gas chromatography–mass spectrometry. The lipids in the co-cultivated cell extracts were mainly attributable to N. oculata, which represented 97% of the total cells (ca. 83% of the total biomass) at the end of the experiment. Nevertheless, the ω-3 characteristic of T. lutea (DHA and SDA, absent in N. oculata) was also detectable. Although the co-cultivation of these two phylogenetically different species of microalgae did not show positive effects on the growth and on the total lipid production, however, this process resulted in a reduction of the production costs and a lower consumption of water and nutrients. [ABSTRACT FROM AUTHOR]

Haptophyte microalgae are key contributors to microbial communities in many environments. It has been proposed recently that members of this group would be virtually all dependent on vitamin B12 (cobalamin), an enzymatic cofactor produced only by some bacteria and archaea. Here, we examined the processes of vitamin B12 acquisition by haptophytes. We tested whether co-cultivating the model species Tisochrysis lutea with B12-producing bacteria in vitamin-deprived conditions would allow the microalga to overcome B12 deprivation. While T. lutea can grow by scavenging vitamin B12 from bacterial extracts, co-culture experiments showed that the algae did not receive B12 from its associated bacteria, despite bacteria/algae ratios supposedly being sufficient to allow enough vitamin production. Since other studies reported mutualistic algae–bacteria interactions for cobalamin, these results question the specificity of such associations. Finally, cultivating T. lutea with a complex bacterial consortium in the absence of the vitamin partially rescued its growth, highlighting the importance of microbial interactions and diversity. This work suggests that direct sharing of vitamin B12 is specific to each species pair and that algae in complex natural communities can acquire it indirectly by other mechanisms (e.g., after bacterial lysis).

The effect of light on biomass and fucoxanthin (Fx) productivities was studied in two microalgae, Tisochrysis lutea and Phaeodactylum tricornutum. High and low biomass concentrations (1.1 and 0.4 g L−1) were tested in outdoor pilot‐scale flat‐panel photobioreactors at semi‐continuous cultivation mode. Fluorescence spectroscopy coupled with chemometric modeling was used to develop prediction models for Fx content and for biomass concentration to be applied for both microalgae species. Prediction models showed high R2 for cell concentration (.93) and Fx content (.77). Biomass productivity was lower for high biomass concentration than low biomass concentration, for both microalgae (1.1 g L−1: 75.66 and 98.14 mg L−1 d−1, for T. lutea and P. tricornutum, respectively; 0.4 g L−1: 129.9 and 158.47 mg L−1 d−1, T. lutea and P. tricornutum). The same trend was observed in Fx productivity (1.1 g L−1: 1.14 and 1.41 mg L−1 d−1, T. lutea and P. tricornutum; 0.4 g L−1: 2.09 and 1.73 mg L−1 d−1, T. lutea and P. tricornutum). These results show that biomass and Fx productivities can be set by controlling biomass concentration under outdoor conditions and can be predicted using fluorescence spectroscopy. This monitoring tool opens new possibilities for online process control and optimization. [ABSTRACT FROM AUTHOR]

Fucoxanthin is a brown-colored pigment from algae, with great potential as a bioactive molecule due to its numerous properties. This review aims to present current knowledge on this high added-value pigment. An accurate analysis of the biological function of fucoxanthin explains its wide photon absorption capacities in golden-brown algae. The specific chemical structure of this pigment also leads to many functional activities in human health. They are outlined in this work and are supported by the latest studies in the literature. The scientific and industrial interest in fucoxanthin is correlated with great improvements in the development of algae cultures and downstream processes. The best fucoxanthin producing algae and their associated culture parameters are described. The light intensity is a major influencing factor, as it has to enable both a high biomass growth and a high fucoxanthin content. This review also insists on the most eco-friendly and innovative extraction methods and their perspective within the next years. The use of bio-based solvents, aqueous two-phase systems and the centrifugal partition chromatography are the most promising processes. The analysis of the global market and multiple applications of fucoxanthin revealed that Asian companies are major actors in the market with macroalgae. In addition, fucoxanthin from microalgae are currently produced in Israel and France, and are mostly authorized in the USA.

Adaptive laboratory evolution is a powerful tool for microorganism improvement likely to produce enhanced microalgae better tailored to their industrial uses. In this work, 12 wild-type strains of Tisochrysis lutea were co-cultivated under increasing thermal stress for 6 months. Indeed, temperature was oscillating daily between a high and a low temperature, with increasing amplitude along the experiment. The goal was to enhance the polyunsaturated fatty acid content of the polar lipids. Samples were taken throughout the evolution experiment and cultivated in standardized conditions to analyze the evolution of the lipid profile. Genomic analysis of the final population shows that two strains survived. The lipid content doubled, impacting all lipid classes. The fatty acid analyses show a decrease in SFAs correlated with an increase in monounsaturated fatty acids (MUFAs), while changes in polyunsaturated fatty acid (PUFAs) vary between both photobioreactors. Hence, the proportion of C18-MUFAs (18:1 n-9) and most C18-PUFAs (18:2 n-6, 18:3 n-3, and 18:4 n-3) increased, suggesting their potential role in adjusting membrane fluidity to temperature shifts. Of particular interest, DHA in polar lipids tripled in the final population while the growth rate was not affected. Key points: • Adaptive laboratory evolution on a mix of 12 T. lutea strains led to survival of 2 • Thermal stress impacted cell size, total lipid cell content, and all lipid classes • DHA cell content partitioned to polar lipids tripled throughout the experiment [ABSTRACT FROM AUTHOR]

Cobalamin (vitamin B12) is a cobalt-containing enzymatic cofactor involved in methionine synthesis. Provided only by select bacteria and archaea in marine systems, this vitamin is known to limit primary production in different oceanic areas. Understanding the consequences of cobalamin limitation on phytoplankton physiology is of great interest, notably for cobalamin-dependent haptophytes that significantly contribute to oceanic carbon fixation and sulfur cycle through dimethyl sulfonio propionate (DMSP) synthesis. Here, the effect of cobalamin limitation was compared to nitrogen limitation on the model haptophyte Tisochrysis lutea grown in chemostats, combining comparative proteomics with the analysis of major macromolecules and specific osmolytes. Our results highlight the interconnection of carbon and DMSP metabolisms through the cobalamin-dependent methionine synthesis by showing that cobalamin scarcity impacts the mechanisms of carbon allocation and reduces DMSP quota. Conversely, proline production seemed to anticorrelate with cobalamin availability. In a boarder context, analysis of transcriptomes or genomes of main DMSP producers from different phytoplankton lineages suggests that most of them are cobalamin-dependent, which means that prokaryotic cobalamin synthesis exerts an important control on phytoplankton DMSP production in some regions of the world ocean.

In recent years, a large number of sunscreens have emerged to protect our skin. Most of them are made up of simple or compound aromatic structures, which can pose a threat to marine ecosystems. In order to understand their effects on the marine environment, different ecotoxicological bioassays were carried out using planktonic organisms from three phyla and two different trophic levels: larvae of the sea urchin Paracentrotus lividus, the copepod Acartia tonsa, and the microalga Tisochrysis lutea. The aim of these tests was to expose these organisms to leachates from eight sunscreen formulations. All of them showed a great variability in toxicity on the different plankton organisms. The highest toxicity level was found for cream number 4 when tested on sea urchin, exhibiting an EC50 = 122.4 mg/L. The toxicity of the UV filter 2-phenyl-5-benzimidazolesulfonic acid, exclusively present in that cream, was evaluated in sea urchin, where an EC10 = 699.6 mg/L was obtained under light exposure. According to our results, all tested creams become nontoxic to plankton upon 30,000-fold dilution in seawater; thus, only local effects are expected. This study highlights the need to understand the toxic effects generated by solar protection products, as well as their ingredients, on marine organisms.

Abstract In this study, Tisochrysis lutea was cultivated in mixotrophic and heterotrophic cultures with glycerol as a carbon source and with glucose and acetate for comparison; autotrophic cultivation was the control group without a carbon source. It was found that T. lutea used glycerol and did not use glucose and acetate under mixotrophy. Mixotrophy slightly elevated the docosahexaenoic acid (DHA) and total fatty acids (TFA) content in the dry-weight and enhanced the DHA and TFA production in medium (41.3 and 31.9% respectively) at the end of a 16-day cultivation, while heterotrophy reduced the DHA content and TFA production. Under the mixotrophy, the glycerol contribution to the DHA production (16.19 mg/L) and the TFA production (97.8 mg/L) was not very high and the DHA yield [2.63% chemical oxygen demand (COD)] and TFA yield (13.1% COD) were also very low. Furthermore, T. lutea using glycerol had a period of adaptation, indicating that T. lutea was not an ideal microalga for organic carbon utilization.

Tisochrysis lutea is a haptophyte microalga important in aquaculture. The 105,837-bp chloroplast genome encodes 113 proteins, 27 different transfer-RNAs and ribosomal-RNAs. Phylogenetic analysis on currently available chloroplast sequences showed T. lutea was most closely related to Emiliania huxleyi. Unlike E. huxleyi chloroplast DNA, T. lutea contains directly-repeated ribosomal repeats following multiple genome rearrangements during evolution. The non-identical 4.9-kb ribosomal direct-repeats are distinguished by 42 nucleotide polymorphisms and three single- or double-nucleotide INDELs. This is unlike land plant and green algal chloroplast genomes in which efficient copy correction by homologous recombination ensures identical long inverted repeats containing the rRNA operons. Imperfect rDNA repeats are also found in the chloroplast genomes of cryptophytes, rhodophytes and other haptophytes. Reduced copy correction rates in these algal lineages raise questions on the mechanism(s) by which genetic uniformity of multi-copy chloroplast genomes is maintained and has implications for developing chloroplast transformation in these species.

Tisochrysis lutea is a haptophyte microalga commonly used as a commercial feed for juvenile fish and shellfish larvae. Genetic modification is of considerable importance for developing the potential economic value of T. lutea. However, the genetic transformation system of T. lutea has not yet been established, which limits functional genomic studies and strain improvement of this important microalgal species. In the current study, a chloroplast transformation vector harboring the phosphinothricin resistance gene (bar) as a selectable marker was established, and two short peptide-encoding genes (ant1 and ant2) driven by the endogenous psbA and rbcL promoters were cloned in this vector. The recombinant plasmid was transformed using a biolistic method into the trnI/trnA locus of the chloroplast genome via homologous recombination. After continuous selection on phosphinothricin, the integration of foreign genes and the expression of specific products in the transformants were detected using polymerase chain reaction (PCR), Southern blotting, and western blot analysis. This is the first report of establishment of a stable transformation system in T. lutea, which is a prerequisite for functional genomics and applied research on this species. [ABSTRACT FROM AUTHOR]

The marine microalga Tisochrysis lutea, a Haptophyta with a thin cell wall and currently used mainly in aquaculture is a potential source of several bioactive compounds of interest such as carotenoids. In the present study, the simultaneous extraction and purification of fucoxanthin, the main carotenoid from T. lutea, was optimized using pressurized fluid extraction followed by in‐cell purification. An experimental design was employed to maximize carotenoids' extraction; the experimental factors chosen were: (i) percentage of ethanol/ethyl acetate (0–100 %), (ii) temperature (40–150°C), and (iii) number of static extraction cycles (1–3). The maximum carotenoids' recovery, mainly fucoxanthin, was obtained with pure ethyl acetate at 40°C using one extraction cycle, achieving values of 132.8 mg of carotenoids per gram of extract. Once the optimum extraction conditions were confirmed, in‐cell purification strategies using different adsorbents were developed to obtain fucoxanthin‐enriched extracts. Activated charcoal showed potential retention of chlorophylls allowing an effective purification of fucoxanthin in the obtained extracts. Chemical characterization of extracts was carried out by reversed‐phase high‐performance liquid chromatography with diode array detection. Therefore, a selective fractionation of high value compounds was achieved using the proposed green downstream platform based on the use of compressed fluids. [ABSTRACT FROM AUTHOR]

By answering the need for increasing sustainability in aquaculture, the present study aimed to compare growth, gene expression involved in appetite regulation, physical characteristics, and chemical composition of Sparus aurata fed alternative protein sources. Fish were fed ten iso-proteic, iso-lipidic, and isoenergetic diets: a vegetable-based (CV) and a marine ingredient-rich (CF) diet were set as control diets. The others were prepared by replacing graded levels (10, 20 or 40%) of the vegetable proteins in the CV with proteins from a commercial defatted Hermetia illucens pupae meal (H), poultry by-product meal (PBM) singly (H10, H20, H40, P20, P40) or in combination (H10P30), red swamp crayfish meal (RC10) and from a blend (2:1, w:w) of Tisochrysis lutea and Tetraselmis suecica (MA10) dried biomasses. The increase in ghre gene expression observed in MA10 fed fish matched with increased feed intake and increased feed conversion ratio. Besides, the MA10 diet conferred a lighter aspect to the fish skin (p < 0.05) than the others. Overall, no detrimental effects of H, PBM, and RC meal included in the diets were observed, and fish fatty acid profile resulted as comparable among these groups and CV, thus demonstrating the possibility to introduce H, PBM, and RC in partial replacement of vegetable proteins in the diet for Sparus aurata.

In this study, we compared the effects of a Tisochrysis lutea (T. lutea) F&M-M36 methanolic extract with those of fucoxanthin (FX) at equivalent concentration, on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The T. lutea F&M-M36 methanolic extract contained 4.7 mg of FX and 6.22 mg of gallic acid equivalents of phenols per gram. HPLC analysis revealed the presence of simple phenolic acid derivatives. The T. lutea F&M-M36 extract exhibited a potent and concentration-dependent inhibitory activity against COX-2 dependent PGE2 production compared to FX alone. Compared to LPS, T. lutea F&M-M36 extract and FX reduced the expression of IL-6 and of Arg1 and enhanced that of IL-10 and of HO-1; T. lutea F&M-M36 extract also significantly abated the expression of NLRP3, enhanced mir-223 expression and reduced that of mir-146b, compared to LPS (p < 0.05). These findings indicate that T. lutea F&M-M36 methanolic extract has a peculiar anti-inflammatory activity against COX-2/PGE2 and NLRP3/mir-223 that might be attributable to the known anti-inflammatory effects of simple phenolic compounds found in the extract that may synergize with FX. Our data suggest that T. lutea F&M-M36 may serve as a source of anti-inflammatory compounds to be further evaluated in in vivo models of inflammation.

Abstract This study investigated the effect of nitrogen (N) and phosphorous (P) stress on the production of DHA or EPA and total fatty acids (TFAs) in the marine microalga Tisochrysis lutea and the freshwater microalga Monodus subterraneus. Five N or P starvation/limitation conditions (N sufficient and P limited, N sufficient and P starved, N starved and P sufficient, N starved and P limited, and N and P starved) and one N and P sufficient condition (control) were studied. The results demonstrated that the proportion of DHA or EPA among TFAs and production in the microalgae suspensions decreased (57%, 73% for N stress and 18%, 51% for P stress, respectively) under N or P stress in both microalgae compared with the N and P sufficient group. Differently, DHA dry weight content of T. lutea decreased significantly, and EPA dry weight content of M. subterraneus decreased slightly under N starved conditions. Clear differences in TFA content/production and the relationship between TFA and DHA or EPA production/content and CO 2 fixation were observed between the two microalgae. These results give a new sight on the difference between marine microalgae and freshwater microalgae. Meanwhile, it gave a potential application to produce DHA or EPA and TFA combining with CO 2 fixation by these microalgae. Graphical abstract Unlabelled Image Highlights • DHA or EPA proportion and production decreased under N or P stress for two microalgae. • TFA content/production and CO 2 fixation of two microalgae were obviously different. • Potential to mitigate CO 2 emission with DHA or EPA production by the two microalgae. [ABSTRACT FROM AUTHOR]

Tisochrysis lutea is a marine haptophyte rich in omega-3 polyunsaturated fatty acids (e.g., docosahexaenoic acid (DHA)) and carotenoids (e.g., fucoxanthin). Because of the nutraceutical applications of these compounds, this microalga is being used in aquaculture to feed oyster and shrimp larvae. In our earlier report, T. lutea organic crude extracts exhibited in vitro cytotoxic activity against human hepatocarcinoma (HepG2) cells. However, so far, the compound(s) accountable for the observed bioactivity have not been identified. Therefore, the aim of this study was to isolate and identify the chemical component(s) responsible for the bioactivity observed. Bioassay-guided fractionation through a combination of silica-gel column chromatography, followed by preparative thin layer chromatography (PTLC), led to the isolation of two diastereomers of a monoterpenoid lactone, namely, loliolide (1) and epi-loliolide (2), isolated for the first time in this species. The structural elucidation of both compounds was carried out by GC-MS and 1D (1H and 13C APT) and 2D (COSY, HMBC, HSQC-ed, and NOESY) NMR analysis. Both compounds significantly reduced the viability of HepG2 cells and were considerably less toxic towards a non-tumoral murine stromal (S17) cell line, although epi-loliolide was found to be more active than loliolide.

Iodine metabolism is essential for the antioxidant defense of marine algae and in the biogeochemical cycle of iodine. Moreover, some microalgae can synthetize thyroid hormone-like compounds that are essential to sustain food webs. However, knowledge regarding iodine-related molecular processes in microalgae is still scarce. In this study, a de novo transcriptome of Tisochrysis lutea cultured under high iodide concentrations (5 mM) was assembled using both long and short reads. A database termed IsochrysisDB was established to host all genomic information. Gene expression analyses during microalgal growth showed that most of the antioxidant- (aryl, ccp, perox, sod1, sod2, sod3, apx3, ahp1) and iodide-specific deiodinase (dio) genes increased their mRNA abundance progressively until the stationary phase to cope with oxidative stress. Moreover, the increase of dio mRNA abundance in aging cultures indicated that this enzyme was also involved in senescence. Cell treatments with iodide modified the expression of perox whereas treatments with iodate changed the transcript levels of gpx1 and ccp. To test the dependence of perox on iodide, microalgae cells were treated with hydrogen peroxide (H2O2) either in presence or absence of iodide observing that several genes related to reactive oxygen species (ROS) deactivation (perox, gpx1, apx2, apx3, ahp1, ahp2, sod1, sod3, and aryl) were transcriptionally activated although with some temporal differences. However, only the expression of perox was dependent on iodide levels indicating this enzyme, acquired by horizontal gene transfer (HGT), could act as a haloperoxidase. All these data indicate that T. lutea activates coordinately the expression of antioxidant genes to cope with oxidative stress. The identification of a phase-regulated deiodinase and a novel haloperoxidase provide new clues about the origin and evolution of thyroid signaling and the antioxidant role of iodine in the marine environment.

The lipid class and the fatty acid compositions of microalgae highly influence bivalve larval and post-larval development. Light is an essential environmental factor for microalgal culture, and quantity and quality of light may induce changes in the biochemical composition of the algae. The objective of this study was to investigate the effect of light spectrum (blue vs. white light) on lipid class and fatty acid compositions of Tisochrysis lutea cultured in a chemostat. Two different dilution rates (D) were assayed for each light spectrum: 0.2 and 0.7 day−1. Triacylglycerol (TAG), sterol, and hydrocarbon (HC) content increased sharply at low D. The proportion of alkenones was significantly reduced under blue light. Polyunsaturated fatty acids (PUFA), and particularly n-3 PUFA, content in phospholipids (PL) increased under blue light compared to white light at low D. Thus, blue light raised 22:6(n-3) levels in total lipids of T. lutea at low D. The cultivation of T. lutea in a chemostat at low D under blue light may improve nutritional value as feed for bivalve larvae by modifying the PUFA profile, especially increasing 22:6(n-3). [ABSTRACT FROM AUTHOR]

Background: Transposable elements (TEs) are mobile DNA sequences known as drivers of genome evolution. Their impacts have been widely studied in animals, plants and insects, but little is known about them in microalgae. In a previous study, we compared the genetic polymorphisms between strains of the haptophyte microalga Tisochrysis lutea and suggested the involvement of active autonomous TEs in their genome evolution. Results: To identify potentially autonomous TEs, we designed a pipeline named PiRATE (Pipeline to Retrieve and Annotate Transposable Elements, download: 10.17882/51795), and conducted an accurate TE annotation on a new genome assembly of T. lutea. PiRATE is composed of detection, classification and annotation steps. Its detection step combines multiple, existing analysis packages representing all major approaches for TE detection and its classification step was optimized for microalgal genomes. The efficiency of the detection and classification steps was evaluated with data on the model species Arabidopsis thaliana. PiRATE detected 81% of the TE families of A. thaliana and correctly classified 75% of them. We applied PiRATE to T. lutea genomic data and established that its genome contains 15.89% Class I and 4.95% Class II TEs. In these, 3.79 and 17.05% correspond to potentially autonomous and non-autonomous TEs, respectively. Annotation data was combined with transcriptomic and proteomic data to identify potentially active autonomous TEs. We identified 17 expressed TE families and, among these, a TIR/Mariner and a TIR/hAT family were able to synthesize their transposase. Both these TE families were among the three highest expressed genes in a previous transcriptomic study and are composed of highly similar copies throughout the genome of T. lutea. This sum of evidence reveals that both these TE families could be capable of transposing or triggering the transposition of potential related MITE elements. Conclusion: This manuscript provides an example of a de novo transposable element annotation of a non-model organism characterized by a fragmented genome assembly and belonging to a poorly studied phylum at genomic level. Integration of multi-omics data enabled the discovery of potential mobile TEs and opens the way for new discoveries on the role of these repeated elements in genomic evolution of microalgae. [ABSTRACT FROM AUTHOR]

Microalgae are an outstanding source of high value products with applications in food, feed or biofuel production. Among these promising organisms, the haptophyte Tisochrysis lutea is commonly used as a feed for shellfish and shrimps in aquaculture. Its capacity to produce high amounts of docosahexanoic acid and storage lipids is also of economic interest for nutrition and energy production. Consequently, understanding its lipid accumulation under nitrogen deprivation is of major interest. Here, we aimed to identify Transcription Factors (TFs) involved in the establishment of the mutant phenotype of the 2Xc1 strain of T. lutea , which accumulates double the quantity of storage lipids under nitrogen deprivation than the wild type strain (WTc1). Strains were grown in chemostats and subjected to different nitrogen availability (limitation, repletion and depletion). Using RNA-seq data, the differentially expressed genes (DEGs) between strains were identified and summarized as a co-expression network to pinpoint putative major TFs in mutant phenotypes. This analysis was followed by a complementary Weighted Gene Correlation Network Analysis in order to classify genes based on their relative importance to mutant phenotype features, regardless of annotation biases due to the lack of functional annotation of the Tisochrysis lutea draft genome. This network-like strategy allowed the identification of seven TF candidates related to key functions in the mutant strain compared with WTc1. In particular, MYB-2R_14 and NF-YB_2 TFs are related to photosynthesis, oxidative stress response and triacylglycerol synthesis. GATA_2, MYB-rel_11 and MYB-2R_20 TFs are likely to be related to nitrogen uptake or carbon and nitrogen recycling, feeding carbohydrate synthesis in the form of chrysolaminarin. Finally, a q-RT-PCR approach further characterized the role of MYB-rel_11 and MYB-2R_20, revealing an expression pattern dependent on nitrogen availability. [ABSTRACT FROM AUTHOR]

Tisochrysis lutea is a microalga species currently used in aquaculture as a feed for shellfish, oysters and shrimps. It also has many other potential industrial applications, such as the production of neutral lipids for biofuels or the production of ω-3 fatty acids for nutraceuticals (human food complements). To efficiently exploit the potential of this microalga, however, higher lipid productivities are needed. To this end, improvement programs need to be developed and optimized. The diversity of strains available in microalgae has not yet been exploited in such improvement programs. In this study, the intra-strain diversity was observed and exploited to increase neutral lipid productivity. New clonal strains with higher neutral lipid productivity were successfully selected. The best clonal strain selected accumulated 520% more triacylglycerols, with a similar growth rate to the wild-type strain in continuous light and nitrogen starvation conditions. In a photoperiod culture condition, this clonal stain also accumulated 84% more storage lipids and 30% less carbohydrates, compared to the wild-type strain. This clonal strain thus had a higher productivity which is of great interest for feed or biofuel applications. This study also focused on identifying the genomic mechanisms responsible for the improvements in these clonal strains. With this objective, the genome of Tisochrysis lutea was sequenced for the first time. It is the third genome of a Haptophyte microalga sequenced so far. Different genetic polymorphisms were identified between the sequenced genomes of the wild-type strain and clonal strains. Activity of transposable elements seems to have been involved in the genome reshuffling obtained through the improvement program. The contribution of transposable elements to the adaptive capacity of microalgae remains to be demonstrated. [ABSTRACT FROM AUTHOR]