Physiological and Biochemical Responses in Microalgae Dunaliella salina, Cylindrotheca closterium and Phormidium versicolor NCC466 Exposed to High Salinity and Irradiation.

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.

Carbon Partitioning and Lipid Remodeling During Phosphorus and Nitrogen Starvation in the Marine Microalga Diacronema lutheri (Haptophyta).

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.

Modulation of lipid biosynthesis by stress in diatoms.

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'.

First evidence of mariner-like transposons in the genome of the marine microalga Amphora acutiuscula (Bacillariophyta).

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.

Relaxation of the non-photochemical chlorophyll fluorescence quenching in diatoms: kinetics, components and mechanisms.

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.

The potential of microalgae for the production of bioactive molecules of pharmaceutical interest.

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.

Zinc affects differently growth, photosynthesis, antioxidant enzyme activities and phytochelatin synthase expression of four marine diatoms.

Zinc-supplementation (20 µM) effects on growth, photosynthesis, antioxidant enzyme activities (superoxide dismutase, ascorbate peroxidase, catalase), and the expression of phytochelatin synthase gene were investigated in four marine diatoms (Amphora acutiuscula, Nitzschia palea, Amphora coffeaeformis and Entomoneis paludosa). Zn-supplementation reduced the maximum cell density. A linear relationship was found between the evolution of gross photosynthesis and total chlorophyll content. The Zn treatment decreased the electron transport rate except in A. coffeaeformis and in E. paludosa at high irradiance. A linear relationship was found between the efficiency of light to evolve oxygen and the size of the light-harvesting antenna. The external carbonic anhydrase activity was stimulated in Zn-supplemented E. paludosa but was not correlated with an increase of photosynthesis. The total activity of the antioxidant enzymes did not display any clear increase except in ascorbate peroxidase activity in N. palea. The phytochelatin synthase gene was identified in the four diatoms, but its expression was only revealed in N. palea, without a clear difference between control and Zn-supplemented cells. Among the four species, A. paludosa was the most sensitive and A. coffeaeformis, the most tolerant. A. acutiuscula seemed to be under metal starvation, whereas, to survive, only N. palea developed several stress responses.

Dynamics of short-term acclimation to UV radiation in marine diatoms.

In order to investigate the dynamics of the acclimation of marine diatoms to ultraviolet radiation (UVR), Amphora coffeaeformis, Odontella aurita and Skeletonema costatum were exposed for 5 h per day to a combination of UVA and UVB (UVBR/UVAR ratio 4.5%) with a total UVR daily dose of 110 kJ m(-2), which is equivalent to that observed in the natural environment. This treatment was applied in the middle of the photoperiod and was repeated on five successive days. During the UVR treatment, chlorophyll fluorescence parameters were monitored, damage and repair constants were calculated from effective quantum yield values (phi(PSII)), and rapid light curves (electron transport rate versus irradiance curves using short light steps of different intensity) were plotted to determine the maximum relative electron transport rate (rETR(max)) and maximum light use efficiency (alpha). In all species the growth rate was lower than control from day 1-3, but increased thereafter, except for S. costatum. The cellular chlorophyll a content increased significantly with repeated daily exposure to UVR for A. coffeaeformis only. In all species, the fluorescence parameters (F(m), the maximum fluorescence level measured in the dark, phi(PSII), rETR(max) and alpha) decreased during UVR exposure, in contrast to F(0) (the minimum fluorescence level measured in the dark). The response to UVR stress was species-specific. S. costatum was very sensitive, and failed to survive for more than three days, whereas A. coffeaeformis and O. aurita were able to acclimate to UVR stress. These two species used different strategies. In A. coffeaeformis, the repair constant was lower than the damage constant, but phi(PSII) values returned to baseline values at the beginning of each experimental day, indicating that an effective active recovery process occurred after stress. In O. aurita, the repair processes took place during the stress, and could account for the UVR tolerance of this species.

Osmotic adjustment, gas exchanges and chlorophyll fluorescence of a hexaploid triticale and its parental species under salt stress.

The effect of salt stress (NaCl 85.7 or 110 mmol/L) was investigated in the triticale T300 and its parental species, Triticum dicoccum farrum (Triticum df) and Secale cereale cv. Petkus. Triticum df and T300 were more salt-tolerant than the rye (110 mmol/L NaCl was the highest concentration allowing rye growth to the three-leaf stage). Na+, K+ and Cl- ions accounted for almost half of the osmotic adjustment in Triticum df and T300, and up to 90% in rye. Salinity decreased the net photosynthesis and transpiration rates of the three cereals as compared to control plants, but induced no significant change in chlorophyll a fluorescence parameters. Water-use efficiency (WUE) increased with salinity. In the presence of 110 mmol/L NaCl, the K+/Na+ ratio decreased markedly in rye as compared to the other two cereals. Proline concentration, which increased in Triticum df and T300, could have protected membrane selectivity in favour of K+. Proline content remained low in rye, and increasing soluble sugar content did not appear to prevent competition between Na+ and K+. The salt sensitivity of rye could be due to low K+ uptake in the presence of a high NaCl concentration.