Crosstalk between sterol and neutral lipid metabolism in the alga Haematococcus pluvialis exposed to light stress.

The presence, biosynthesis and functional role of sterols in the green microalga Haematococcus pluvialis remain poorly understood. In this work we studied the effect of high-light (HL) stress on sterol synthesis in H. pluvialis UTEX 2505 cells. HL stress induced the synthesis of sterols in parallel with that of triacylglycerides (TAG), giving rise to the synthesis of cholesterol over that of phytosterols. Blockage of the carotenogenic 1-deoxy-D-xylulose 5-phosphate (MEP) pathway is shown to be involved in HL-induced sterol synthesis. In addition, high irradiance exposure induced MEP- and fatty acid (FA)-biosynthetic transcripts. The pharmacological inhibition of these pathways suggests a possible feedback regulation of sterol and FA homeostasis. Finally, both lipid classes proved crucial to the adequate photosynthetic performance of H. pluvialis grown under HL intensity stress. Our findings reveal new insights into H. pluvialis lipid metabolism that contribute to the development of value-added bioproducts from microalgae.

Neochloris oleoabundans grown in enriched natural seawater for biodiesel feedstock: evaluation of its growth and biochemical composition.

The freshwater microalga Neochloris oleoabundans was used to study algal lipid production in enriched natural seawater, in order to assess its suitability as biodiesel feedstock. Optimal and nitrogen-stress (N-stress) conditions were analyzed. Under optimal conditions, the strain's growth rate was 0.73 div day(-1) and the biomass concentration was 1.5 g L(-1), while it had a maximum lipid yield under N-stress conditions (lipid content: 26% of dry weigh and lipid productivity: 56 mg L(-1) day(-1)). Lipid accumulation was mainly due to a significant increase of triacylglycerol content. Neutral lipids were characterized by a dominance of monounsaturated fatty acids and displayed a fatty acid profile that is suitable for biodiesel. This work offers an interesting alternative for sustainable microalgal oil synthesis for biodiesel production without using freshwater resources. However, further studies are necessary in order to optimize the lipid productivities required for commercial biodiesel production.

Lipid analysis in Haematococcuspluvialis to assess its potential use as a biodiesel feedstock.

The lipid content and composition of Haematococcuspluvialis exposed to stress conditions were analyzed to assess the potential of this microalga as a biodiesel feedstock. The total lipid content of control cells was 15.61% dw, whereas that of cells exposed to continuous high light intensity with nitrogen-sufficient medium (A-stress condition) or under continuous high light intensity with nitrogen-deprivation medium (B-stress condition) was 34.85% dw and 32.99% dw, respectively. The fatty acid profile was similar under all conditions and indicated that the main components were palmitic, stearic, oleic, linoleic, linolenic and linolelaidic acids. The neutral lipid fraction increased about 2-fold under both stress conditions. The percentage of saturated fatty acids in the neutral lipid fraction was 30.36% and 29.62% in cultures grown under A-stress and B-stress, respectively, and 27.81% under control conditions. The monounsaturated fatty acid content was not significantly different in control and A-stress cultures (20.07% and 19.91%, respectively), but was 18.96% under B-stress. The content of polyunsaturated fatty acids was 47.23% under B-stress and 43.15% under A-stress. Growth-rate was higher under A-stress compared to B-stress. This is the first study of H.pluvialis that provides a detailed characterization of its lipid content in relation to bioenergy. The results indicate the potential of this microalga as a biodiesel feedstock; however, culture conditions still have to be improved in order to achieve an adequate energy balance in mass culture.

CELL WALL CARBOHYDRATE EPITOPES IN THE GREEN ALGA OEDOGONIUM BHARUCHAE F. MINOR (OEDOGONIALES, CHLOROPHYTA)(1).

Cell wall changes in vegetative and suffultory cells (SCs) and in oogonial structures from Oedogonium bharuchae N. D. Kamat f. minor Vélez were characterized using monoclonal antibodies against several carbohydrate epitopes. Vegetative cells and SCs develop only a primary cell wall (PCW), whereas mature oogonial cells secrete a second wall, the oogonium cell wall (OCW). Based on histochemical and immunolabeling results, (1→4)-ß-glucans in the form of crystalline cellulose together with a variable degree of Me-esterified homogalacturonans (HGs) and hydroxyproline-rich glycoprotein (HRGP) epitopes were detected in the PCW. The OCW showed arabinosides of the extensin type and low levels of arabinogalactan-protein (AGP) glycans but lacked cellulose, at least in its crystalline form. Surprisingly, strong colabeling in the cytoplasm of mature oogonia cells with three different antibodies (LM-5, LM-6, and CCRC-M2) was found, suggesting the presence of rhamnogalacturonan I (RG-I)-like structures. Our results are discussed relating the possible functions of these cell wall epitopes with polysaccharides and O-glycoproteins during oogonium differentiation. This study represents the first attempt to characterize these two types of cell walls in O. bharuchae, comparing their similarities and differences with those from other green algae and land plants. This work represents a contribution to the understanding of how cell walls have evolved from simple few-celled to complex multicelled organisms.

Cytochemical localization of acid phosphatase in Stigeoclonium tenue (Chaetophorales, Chlorophyceae).

Nonspecific acid phosphatases are a group of enzymes whose activity increases the availability of exogenous and endogenous orthophosphate either through extra- or intracellular hydrolysis of phosphate compounds. Our study demonstrates the activity of acid phosphatases in the filamentous freshwater alga Stigeoclonium tenue. These enzymes were detected following a cerium-based method in which cerium was used as an orthophosphate-capture reagent. In thalli from S. tenue from the natural environment, acid phosphatases were found in the longitudinal cell wall, plasmalemma, and vacuole. In thalli from Bold's Basal Medium culture, these enzymes were found mainly in the plasmalemma; they were scarce in the cell wall. In the thalli grown in phosphate-enriched culture medium, enzymes were found only in the plasmalemma. The low availability of orthophosphate in the medium seems to induce the transport of these enzymes to the cell wall. Its abundance, on the contrary, seems to attenuate this response without affecting the localization of acid phosphatases in the plasmalemma.

FINE STRUCTURE AND TAXONOMY OF MONOMORPHINA AENIGMATICA COMB. NOV. (EUGLENOPHYTA)(1).

The euglenoid genus Monomorphina was defined by Mereschowsky in 1877 to include rigid euglenoids that were pyriform in lateral view, had a hyaline spine at the posterior end, and one to few parietal chloroplasts typically without pyrenoids. The genus included taxa previously assigned to Phacus Dujardin or Euglena Ehrenberg. The general structure of Monomorphina aenigmatica comb. nov. is described on the basis of light microscopy and scanning and transmission electron microscopy. Cells were pear-shaped in lateral view, rounded at the anterior end and narrowed posteriorly, tapering into a long twisted tail. The pellicle had helically arranged strips spiralled in a counter-clockwise fashion. A distinctive feature of M. aenigmatica was the presence of a single chloroplast bearing a pyrenoid, capped with a paramylon plate. The large parietal chloroplast extended along most of the cell with three prominent cup-shaped paramylon caps on the external face. In transverse section, the chloroplast appeared C-shaped. Because of the ambiguity surrounding the original descriptions used to diagnose this taxon, we designated an epitype for Monomorphina aenigmatica. Morphological features of this species were compared to other members of the genus.

FINE STRUCTURE OF BIFLAGELLATE ZOOSPORES OF ASTEROCOCCUS SUPERBUS (TETRASPORALES, CHLOROPHYCEAE), INCLUDING THE ABSOLUTE CONFIGURATION OF THE FLAGELLAR APPARATUS.

The ultrastructure of zoospores of Asterococcus superbus (Cienk.) Scherffel was studied to provide ultrastructural data relevant to the systematic position of the genus. Our results demonstrated that the motile cells of A. superbus were similar to those of the tetrasporalean algae, such as Tetraspora sp. and Tetrasporidium javanicum Moebius. The flagellar apparatus of A. superbus had the same clock-wise orientation of basal bodies and the V-shaped alignment of basal bodies as Tetraspora cylindrica (Wahlb.) Ag. and T. lubrica (Roth) Ag., but differed by having rhizoplasts. The motile cells of A. superbus displayed chlamydomonadal ultrastructure, similar to Chlamydomonas reinhardtii Dangeard, including the absolute configuration of the flagellar apparatus. The pyrenoid matrix in A. superbus, however, showed a large lateral invagination occupied by chloroplast stroma, a characteristic that has never been observed in Chlorophyta.