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.

In this study, we investigated the responses of Phaeodactylum tricornutum cells acclimated to 300 µmol m−2 s−1 photon flux density to an increase (1000 µmol m−2 s−1) or decrease (30 µmol m−2 s−1) in photon flux densities. The light shift occurred abruptly after 5 days of growth and the acclimation to new conditions was followed during the next 6 days at the physiological and molecular levels. The molecular data reflect a rearrangement of carbon metabolism towards the production of phosphoenolpyruvic acid (PEP) and/or pyruvate. These intermediates were used differently by the cell as a function of the photon flux density: under low light, photosynthesis was depressed while respiration was increased. Under high light, lipids and proteins accumulated. Of great interest, under high light, the genes coding for the synthesis of aromatic amino acids and phenolic compounds were upregulated suggesting that the shikimate pathway was activated. This article is part of the themed issue ‘The peculiar carbon metabolism in diatoms’.

This study assessed the responses of molecular biomarkers and heavy metal levels in Cerastoderma glaucum exposed for 1 week to two industrial effluents (1%) discharged into the Tunisian coastal area, F1 and F2, produced by different units of production of a phosphate treatment plant. A significant uptake of metals (Cd, Cu, Zn, and Ni) was observed in exposed cockles compared to controls, with an uptake higher for F1 than for F2. A decrease in LT50 (stress on stress test) was also observed after an exposure to the effluent F1. Treatments resulted in different patterns of messenger RNA (mRNA) expression of the different genes tested in this report. Gene transcription monitoring performed on seven genes potentially involved in the tolerance to metal exposure showed that for both exposures, mechanisms are rapidly and synchronically settled down to prevent damage to cellular components, by (1) handling and exporting out metal ions through the up-regulation of ATP-binding cassette xenobiotic transporter (ABCB1) and metallothionein (MT), (2) increasing the mRNA expression of antioxidant enzymes (catalase (CAT), superoxide dismutases, CuZnSOD and MnSOD), (3) protecting and/or repairing proteins through the expression of heat shock protein 70 (HSP70) mRNAs, and (4) increasing ATP production (through the up-regulation of cytochrome c oxidase 1 (CO1)) to provide energy for cells to tolerate stress exposure. The tools developed may be useful both for future control strategies and for the use of the cockle C. glaucum as a sentinel species.

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 particular gene complement in diatoms, inherited from various types of organisms, has contributed to the development of metabolic networks that contrast with those found in other photosynthetic organisms. To understand these networks and how they are linked, transcriptomic, proteomic and metabolomic approaches have been used over the last decade. Understanding how these networks developed and interact remains a major goal for physiologists. Metabolic compartmentalization and fluxes between compartments are still poorly known, requiring: (1) the localization of proteins and biological activities, as well as potential protein isoforms and (2) relating metabolite measurements to pathway fluxes. Moreover, when considering metabolism, the identification of transcription factors, which are largely unknown for diatoms, is necessary. Integration of the results from these different approaches will complete our understanding of cell functioning and how differences impact metabolic reorientation.

This study investigates cadmium effects on key messenger RNA (mRNA) expression (MT, MnSOD, CuZnSOD, CAT, ABCB1, HSP70, and CO1) by qPCR in the cockle Cerastoderma glaucum after chronic exposure to two high but environmentally relevant concentrations of CdCl2 (50 μg/L and 5 mg/L) for 12 h to 18 days. Cd accumulation measured in cockles' tissues is significantly higher in both treatment conditions compared to controls and in a dose-dependent manner. Stress on stress tests performed at different times of the experiment clearly demonstrated that exposure to both concentrations of Cd significantly affects cockle survival time in air. Important changes in gene transcription were also highlighted. In particular, MT, HSP70, CAT, and CuZnSOD seem to be relevant biomarkers of Cd exposure because (1) their mRNA levels increase upon exposure and (2) they are highly correlated to Cd accumulation in tissues. Results may be useful for control strategies and for the use of cockles as sentinel organisms.

Hydrothermal vent conditions can alter DNA and hydrothermal organisms may develop detoxification mechanisms and/or genetic adaptations. Hydrothermal vent animals notably synthesize a high quantity of metallothioneins (MT). Recent studies have revealed that the levels of MT within hydrothermal crustacean tissues are higher than those found in other vent animals. To improve our understanding of the environmental impacts exerted on the vent organisms, we characterized the metallothioneins (cDNA and Mt genes) of several members of the Bythograeidae (Bythograea thermydron, Cyanagraea praedator and Segonzacia mesatlantica) which is the only endemic hydrothermal crab family. In comparison, the isolation of metallothionein cDNA was also carried out in several coastal crab families. The results showed that the hydrothermal crabs possess Mt composed of three exons and two introns presenting conserved splicing signals. The cDNA sequences isolated from distinct crabs showed multiple substitutions. In spite of the unique environmental conditions, the protein sequence analysis revealed no specific amino acid residue for the MT of the three hydrothermal crabs. However, gene expression analysis performed by real-time PCR based on S. mesatlantica (hydrothermal crab) compared to Pachygrapsus marmoratus (coastal crab) confirmed the higher metallothionein induction in hydrothermal crabs suggested by others authors.

Crabs of the Bythograeidae family (Crustacea: Brachyura: Bythogreoidea) are the only endemic crab family living in hydrothermal fields. The hydrothermal environment is characterized by unique ecological parameters, such as the high temperature gradient around the hydrothermal chimney (2-350 degrees C), a fluid environment containing high levels of metals and numerous gases. The 70-kDa Heat Shock Protein (HSP70) group is the most-studied HSP, because it is ubiquitous, and a strong positive correlation has been found between the amounts of HSP70 produced in response to stress, and the ability of the organism to withstand stressful conditions. The 70-kDa heat shock protein genes from Bythograeids (species analyzed: Bythograea thermydron, Cyanagraea praedator and Segonzacia mesatlantica) were characterized. Our results revealed that Bythograeidae possess genes which are similar with those present in Xanthids (coastal crabs). The deduced protein sequences displayed motifs distinct from those in the other crustacean HSC70/HSP70s available in the databases. Phylogenetic analysis showed that these members of HSP70 family identified in Bythograeidae and Xanthidae constitute a new subgroup within this family. (c) 2007 Elsevier B.V. All rights reserved.