Dunaliella salina (Chlorophyceae), Phormidium versicolor (Cyanophyceae), and Cylindrotheca closterium (Bacillariophyceae) were isolated from three ponds in the solar saltern of Sfax (Tunisia). Growth, pigment contents, and photosynthetic and antioxidant enzyme activities were measured under controlled conditions of three light levels (300, 500, and 1000 µmol photons m−2 s−1) and three NaCl concentrations (40, 80, and 140 g L−1). The highest salinity reduced the growth of D. salina and P. versicolor NCC466 and strongly inhibited that of C. closterium. According to ΦPSII values, the photosynthetic apparatus of P. versicolor was stimulated by increasing salinity, whereas that of D. salina and C. closterium was decreased by irradiance rise. The production of carotenoids in D. salina and P. versicolor was stimulated when salinity and irradiance increased, whereas it decreased in the diatom. Catalase (CAT), Superoxide dismutase (SOD), and Ascorbate peroxidase (APX) activities were only detected when the three species were cultivated under E1000. The antioxidant activity of carotenoids could compensate for the low antioxidant enzyme activity measured in D. salina. Salinity and irradiation levels interact with the physiology of three species that have mechanisms of more or less effective stress resistance, hence different resistance to environmental stresses according to the species. Under these stress-controlled conditions, P. versicolor and C. closterium strains could provide promising sources of extremolyte for several purposes.

Dunaliella salina (Chlorophyceae), Phormidium versicolor (Cyanophyceae) and Cylindrotheca closterium (Bacillariophyceae) were isolated from three ponds in the solar saltern of Sfax (Tunisia) having an average salinity of 350, 100 and 90 respectively. Growth, pigment contents, photosynthetic and antioxidant enzyme activities were measured under controlled conditions: three light levels (300, 500 and 1000 µmol photons m-2 s-1) and three NaCl concentrations (40, 80 and 140 g L-1). The highest salinity reduced the growth of D. salina and P. versicolor, and strongly inhibited that of C. closterium. These results are in accordance with the species distibution in the salt marshes. Irradiance rise only induced a significant increase of net photosynthesis in C. closterium probably due to the efficient nonphotochemical quenching and antioxidative enzyme activities. According to □PSII values, the photosynthetic apparatus of P. versicolor was stimulated by increasing salinity whereas that of D. salina and C. closterium was decreased by irradiance rise. The production of carotenoids in D. salina and P. versicolor was stimulated when salinity and irradiance increased whereas it decreased in the diatom. Antioxidant activity of carotenoids could compensate the low antioxidant enzyme activity measured in D. salina.

The domesticated marine microalga Diacronema lutheri is of great interest for producing various highly valuable molecules like lipids, particularly long-chain polyunsaturated fatty acids (LC-PUFA). In this study, we investigated the impact of phosphorus (P) and nitrogen (N) starvation on growth, carbon fixation (photosynthetic activity) and partitioning, and membrane lipid remodeling in this alga during batch culture. Our results show that the photosynthetic machinery was similarly affected by P and N stress. Under N starvation, we observed a much lower photosynthetic rate and biomass productivity. The degradation and re-use of cellular N-containing compounds contributed to triacylglycerol (TAG) accumulation. On the other hand, P-starved cells maintained pigment content and a carbon partitioning pattern more similar to the control, ensuring a high biomass. Betaine lipids constitute the major compounds of non-plastidial membranes, which are rich in eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. Under P and N starvations, EPA was transferred from the recycling of membrane polar lipids, most likely contributing to TAG accumulation.

Diatoms are responsible for up to 40% of the carbon fixation in our oceans. The fixed carbon is moved through carbon metabolism towards the synthesis of organic molecules that are consumed through interlocking foodwebs, and this process is strongly impacted by the abiotic environment. However, it has become evident that diatoms can be used as ‘platform’ organisms for the production of high valuable bio-products such as lipids, pigments and carbohydrates where stress conditions can be used to direct carbon metabolism towards the commercial production of these compounds. In the first section of this review, some aspects of carbon metabolism in diatoms and how it is impacted by environmental factors are briefly described. The second section is focused on the biosynthesis of lipids and in particular omega-3 long-chain polyunsaturated fatty acids and how low temperature stress impacts on the production of these compounds. In a third section, we review the recent advances in bioengineering for lipid production. Finally, we discuss new perspectives for designing strains for the sustainable production of high-value lipids. This article is part of the themed issue ‘The peculiar carbon metabolism in diatoms’.

Nitrogen (N) and phosphorus (P) limitations induce triacylglycerol (TAG) accumulation and membrane lipid remodelling in the marine diatom Phaeodactylum tricornutum. However, a clear understanding of the metabolic reorientation is still lacking. Carbon partitioning is of great interest because this microalga produces various highly valuable molecules such as lipids and polyunsaturated fatty acids. This study compared growth, photosynthetic activity, biochemical and transcriptional responses of P. tricornutum throughout batch culture under N or P limitation. The integrated results show that the photosynthetic intensity was greatly reduced under N or P limitation. Under N limitation, the degradation and re-use of cellular N-containing compounds contributed to TAG accumulation, whilst P limitation favoured TAG accumulation due to the efficiency of carbon fixation, without massive degradation of essential compounds at cellular level. There was no difference in the partitioning of carbon to neutral lipids between N and P limitation. Substitution of phospholipids with betaine lipids appeared to be a P-specific acclimation strategy in P. tricornutum, which was largely regulated at the gene expression level. Betaine lipid synthesis was induced by P limitation. The lipid remodelling began once the medium became deficient in P. While the phospholipid biosynthesis pathway was not completely inhibited, a shift of lipid classes occurred immediately after their synthesis via phospholipid-recycling mechanisms.

Nitrogen (N) and phosphorus (P) limitations induce neutral lipid accumulation and membrane lipid remodeling in the domesticated oleaginous microalga Tisochrysis lutea. This study compared growth, photosynthetic activity, biochemical and transcriptional responses of T. lutea throughout batch cultures under N or P limitation compared with non-limiting nutrient condition (NP). The results show that, under N limitation, the breakdown and re-use of cellular N-containing compounds contributed to carbohydrates and further triacylglycerol (TAG) accumulation, where the accumulation of docosahexaenoic acid (DHA) in neutral lipids might mainly be derived from the recycling of membrane polar lipids. Conversely, P limitation did not hinder the photosynthetic capacity; a higher efficiency of carbon fixation fueled the allocation of carbon fluxes to the reserves of carbohydrates and neutral lipids. These latter accumulated without massive degradation of essential cellular compounds. Betaine lipids constitute the major compounds of non-plastidial membranes in T. lutea. Given an extremely low constitutive phospholipid level, phospholipids might not be involved in the regulation of P storage. However, transfer of P from non-plastidial to chloroplast membranes might occur, keeping a stable lipid composition of thylakoid membranes and maintaining a relatively high photosynthetic capacity under P deprivation.

Colour is an often overlooked sensory character that certainly influences flavour perception. Pigments colouring food are generally unstable and are modified during processing. To maintain or restore product colour uniformity, colouring agents, considered worldwide as food additives, are intentionally added to food products. The natural food additives market has been growing extensively since the last century due to the potential hazards of artificial food additives and the potential benefits of biologically active compounds. In this chapter, a fairly compressed overview of the most important colours of natural origin as well as information about less common or/and promising colouring molecules are provided.

Transposable elements (TEs) are mobile DNA sequences thought to be involved in the genomic response to environmental stresses of organisms. The thermal stress behaviour of two retrotransposons (Surcouf and Blackbeard) was investigated in the genome of two strains of the marine diatom Phaeodactylum tricornutum. Using quantitative reverse transcriptase polymerase chain reaction, the retrotransposon expression pattern was investigated after thermal stresses and compared with that of the small heat-shock protein (shsp). While the expression of Blackbeard was not affected by temperature changes, the retroelement Surcouf was markedly overexpressed in response to high temperature in correlation with shsp overexpression. An in silico analysis investigated the presence of regulatory heat-inducible motifs such as heat-shock elements, stress response elements and CCAAT boxes within the promoters of shsp and retrotransposons in the genome of P. tricornutum. The results of mRNA quantification are supported by the presence of heat-shock response motifs found in the promoters of shsp and the retrotransposon Surcouf.

This study investigated the major environmental factors structuring, for a year, phytoplankton assemblages in the Sfax saltern (Tunisia): salinity and nutrients. A STATICO analysis based on 11 environmental variables and the abundances of 64 phytoplanktonic species was conducted. STATICO is used to analyze the stable part of the relationships between the environment and species, and then to determine how these relationships change over time. The analysis confirmed that the salinity gradient had a considerable influence on the composition of the phytoplanktonic communities. Bacillariophyceae and Dinophyceae dominated in the least salty ponds, whereas Chlorophyceae and Cyanophyceae dominated in the saltiest ponds, in accordance with the halotolerance level estimated for each species by calculating the optimum salinity and salt tolerance. Nitrogen (N) and phosphorus (P) ions could have a secondary influence on the phytoplankton distribution and its dynamics. Dinophyceae seem to be favored by high ammonium (NH4 +) concentrations, whereas diatoms seem to be favored by high orthophosphates (PO4 3−) and nitrates (NO3 −) values. The Chlorophyceae Dunalliela salina thrived in the saltiest ponds when the NO3 − concentrations increased.

Transposable elements (TEs) are mobile DNA sequences thought to be involved in the genomic response to environmental stresses of organisms. The thermal stress behaviour of two retrotransposons (Surcouf and Blackbeard ) was investigated in the genome of two strains of the marine diatom Phaeodactylum tricornutum. Using quantitative reverse transcriptase polymerase chain reaction, the retrotransposon expression pattern was investigated after thermal stresses and compared with that of the small heat-shock protein (shsp). While the expression of Blackbeard was not affected by temperature changes, the retroelement Surcouf was markedly overexpressed in response to high temperature in correlation with shsp overexpression. An in silico analysis investigated the presence of regulatory heat-inducible motifs such as heat-shock elements, stress response elements and CCAAT boxes within the promoters of shsp and retrotransposons in the genome of P. tricornutum. The results of mRNA quantification are supported by the presence of heat-shock response motifs found in the promoters of shsp and the retrotransposon Surcouf.

The diatom Odontella aurita has now been industrially cultured and commercialized as a dietary supplement rich in omega-3 fatty acids for several years. In this study, we investigated the effect of three temperatures (8, 16, and 24 °C) on the growth and fatty acid composition of cells harvested during the exponential and stationary growth phases. These temperatures were selected on the basis of photosynthesis responses previously obtained at different temperatures using a modulated fluorometer. Our results confirm that both growth and lipid composition were sensitive to culture temperature. Growth was reduced when O. aurita was cultured at low temperature (8 °C) compared to when it was cultured at high temperatures (16 and 24 °C), but the proportion of polyunsaturated fatty acids (PUFAs, 20:5 n-3 and 22:6 n-3) increased while the level of saturated fatty acids (SFAs, 14:0 and 16:0) decreased in the cells harvested during both the exponential and stationary growth phases. On the other hand, the cells grown at 24 °C displayed a marked decrease in PUFA and an increase in SFA levels. Harvesting time is also a critical parameter in achieving optimum n-3 PUFA productivity during batch cultivation. Indeed, changes in fatty acid composition with growth phase seem to be dependent on the culture temperature, with the most marked effects being observed at 24 °C. PUFA levels (i.e., levels of 20:5 n-3 and 22:6 n-3) increased during the stationary growth phase, while the proportion of SFAs and monounsaturated fatty acids (MUFAs) fell with time. As this species is currently grown in outdoor ponds with seasonal temperature variations (minimal and maximal average temperatures in winter and summer, from 3 to 9 °C and from 13 to 26 °C, respectively), this factor can be expected to have a strong influence on the fatty acid content and composition of the algal biomass harvested and commercialized.

Diatoms are especially important microorganisms because they constitute the larger group of microalgae. To survive the constant variations of the light environment, diatoms have developed mechanisms aiming at the dissipation of excess energy, such as the xanthophyll cycle and the non-photochemical chlorophyll (Chl) fluorescence quenching. This contribution is dedicated to the relaxation of the latter process when the adverse conditions cease. An original nonlinear regression analysis of the relaxation of non-photochemical Chl fluorescence quenching, qN, in diatoms is presented. It was used to obtain experimental evidence for the existence of three time-resolved components in the diatom Phaeodactylum tricornutum: qNf, qNi and qNs. qNf (s time-scale) and qNs (h time-scale) are exponential in shape. By contrast, qNi (min time-scale) is of sigmoidal nature and is dominant among the three components. The application of metabolic inhibitors (dithiothreitol, ammonium chloride, cadmium and diphenyleneiodonium chloride) allowed the identification of the mechanisms on which each component mostly relies. qNi is linked to the relaxation of the ΔpH gradient and the reversal of the xanthophyll cycle. qNs quantifies the stage of photoinhibition caused by the high light exposure, qNf seems to reflect fast conformational changes within thylakoid membranes in the vicinity of the photosystem II complexes.

Mariner-like elements (MLEs) are transposable elements able to move in the host genomes by a "cut and paste" mechanism. They have been found in numerous organisms. We succeeded in amplifying complete and truncated MLEs in the marine diatom Amphora acutiuscula. Full-length MLEs of 2,100bp delimited by imperfect Terminal Inverted Repeats revealed an intact Open Reading Frame, suggesting that the MLEs could be active. The DNA binding domain of the corresponding putative transposase could have two Helix-Turn-Helix and a Nuclear Location Site motifs, and its catalytic domain includes a particular triad of aspartic acids DD43D not previously reported. The number of copies was estimated to be 38, including approximately 20 full-length elements. Phylogenetic analysis shows that these peculiar MLEs differ from plant and other stramenopile MLEs and that they could constitute a new sub-family of Tc1-mariner elements.

Transposable elements (TEs) are DNA sequences that are able to move and replicate within the genomes of virtually all organisms, ranging from animals to plants, and also including protozoans, bacteria and diatoms. Initially considered to be ‘junk DNA’ with no function, TEs are now thought to play a pivotal role in the evolution of genomes and organisms. This review is intended to provide non-specialists with an introduction to the world of TEs by summarizing the information available about these elements, ranging from their impact on genome structure to their potential involvement in the evolution of species. In this review, we propose to bring together what is known about the TEs and provide an overview of recent advances concerning TEs in diatoms.

The effects of continuous light exposure (24L:0D) and a 12 h:12 h light/dark regime (12L:12D) were compared on the growth and carotenoid, protein, sugar, lipid, and fatty acid contents in Chlorella vulgaris, Nannochloropsis sp., Isochrysis galbana, and Dunaliella salina cultured in a batchwise facility. These microalgae were grown axenically under a low photon flux density (PFD) of 27 μmol photons m−2 s−1. C. vulgaris, Nannochloropsis sp., and I. galbana exhibited the highest cell densities when cultured under 24L:0D, whereas D. salina grew better under the alternating light/dark regime. I. galbana accumulated high levels of proteins, sugars, and lipids and exhibited the highest carotenoid content under 24L:0D. Protein production was enhanced in C. vulgaris under 24L:0D. The highest total lipid content was recorded for D. salina, reaching 74.6 % of total proteins, sugars, and lipids in cells at the stationary phase when grown under 12L:12D. The light/dark regime at low PFD was sufficient to stimulate the accumulation of monounsaturated and polyunsaturated fatty acids in all four algae. Their levels, like those of saturated fatty acids, did not differ significantly under the two light regimes. D. salina was an important source of tetradecenoic acid 14:1(n-5). Nannochloropsis sp. produced a large amount of the essential eicosapentaenoic acid, which reached 20 % of total fatty acids under 12L:12D, while I. galbana exhibited the highest level of docosahexaenoic acid, which reached 21 % under both light regimes. This study demonstrated the feasibility of culturing microalgae under low PFD in order to produce large quantities of valuable metabolites, especially various lipids with neutraceutical value.

Diatom development depends on several environmental factors, including the availability in metals. When micronutrients are present of adequate amount, cells exhibit a strong fitness and develop at their maximum growth rate. In many circumstances, the optimal metal amount in the cell environment is disrupted and cells experience starvation or excess for one or more elements. The metals in excess interfere with biochemical and cellular processes triggering a dysfunctioning that reduces growth and may ultimately lead to cell death. The ability of diatoms to adapt/resist to environmental changes has ecological consequences in term of biodiversity. To survive, diatoms activate defence mechanisms, such as the production of antioxidants or/and metal chelators. In this contribution, the diatom requirements for cadmium, copper, zinc and sodium are briefly reviewed. Then the impacts of an excess or a deprivation in one of these elements on diatom physiology is discussed from the molecular and biochemical point of views. The defence mechanisms enabling diatoms to overcome the metal stress are presented. At the end of this contribution, an assay on the integration of the defence mechanisms is presented.

Through the photosynthetic activity, microalgae process more than 25% of annual inorganic carbon dissolved in oceans into carbohydrates that ultimately, serve to feed the other levels of the trophic networks. Besides, microalgae synthesize bioactive molecules such as pigments and lipids that exhibit health properties. In addition, abiotic stresses, such as high irradiance, nutrient starvation, UV irradiation, trigger metabolic reorientations ending with the production of other bioactive compounds such as ω-3 fatty acids or carotenoids. Traditionally, these compounds are acquired through the dietary alimentation. The increasing, and often unsatisfied, demand for compounds from natural sources, combined with the decrease of the halieutic resources, forces the search for alternative resources for these bioactive components. Microalgae possess this strong potential. For instance, the diatom Odontella aurita is already commercialized as dietary complement and compete with fish oil for human nutrition. In this contribution, the microalga world is briefly presented. Then, the different types of biologically active molecules identified in microalgae are presented together with their potential use. Due to space limitation, only the biological activities of lipids and pigments are described in details. The contribution ends with a description of the possibilities to play with the environmental constrains to increase the productivity of biologically active molecules by microalgae and by a description of the progresses made in the field of alga culturing.