Monogalactosyldiacylglycerol synthase isoforms play diverse roles inside and outside the diatom plastid.

Diatoms derive from a secondary endosymbiosis event, which occurred when a eukaryotic host cell engulfed a red alga. This led to the formation of a complex plastid enclosed by four membranes: two innermost membranes originating from the red alga chloroplast envelope, and two additional peri- and epiplastidial membranes (PPM, EpM). The EpM is linked to the endoplasmic reticulum (ER). The most abundant membrane lipid in diatoms is monogalactosyldiacylglycerol (MGDG), synthesized by galactosyltransferases called MGDG synthases (MGDs), conserved in photosynthetic eukaryotes and considered to be specific to chloroplast membranes. Similar to angiosperms, a multigenic family of MGDs has evolved in diatoms, but through an independent process. We characterized MGDα, MGDß and MGDγ in Phaeodactylum tricornutum, combining molecular analyses, heterologous expression in Saccharomyces cerevisiae, and studying overexpressing and CRISPR-Cas9-edited lines. MGDα localizes mainly to thylakoids, MGDß to the PPM, and MGDγ to the ER and EpM. MGDs have distinct specificities for diacylglycerol, consistent with their localization. Results suggest that MGDα is required for thylakoid expansion under optimal conditions, while MGDß and MGDγ play roles in plastid and non-plastid membranes and in response to environmental stress. Functional compensation among MGDs likely contributes to diatom resilience under adverse conditions and to their ecological success.

Lipid Droplets in Unicellular Photosynthetic Stramenopiles.

The Heterokonta or Stramenopile phylum comprises clades of unicellular photosynthetic species, which are promising for a broad range of biotechnological applications, based on their capacity to capture atmospheric CO2 via photosynthesis and produce biomolecules of interest. These molecules include triacylglycerol (TAG) loaded inside specific cytosolic bodies, called the lipid droplets (LDs). Understanding TAG production and LD biogenesis and function in photosynthetic stramenopiles is therefore essential, and is mostly based on the study of a few emerging models, such as the pennate diatom Phaeodactylum tricornutum and eustigmatophytes, such as Nannochloropsis and Microchloropsis species. The biogenesis of cytosolic LD usually occurs at the level of the endoplasmic reticulum. However, stramenopile cells contain a complex plastid deriving from a secondary endosymbiosis, limited by four membranes, the outermost one being connected to the endomembrane system. Recent cell imaging and proteomic studies suggest that at least some cytosolic LDs might be associated to the surface of the complex plastid, via still uncharacterized contact sites. The carbon length and number of double bonds of the acyl groups contained in the TAG molecules depend on their origin. De novo synthesis produces long-chain saturated or monounsaturated fatty acids (SFA, MUFA), whereas subsequent maturation processes lead to very long-chain polyunsaturated FA (VLC-PUFA). TAG composition in SFA, MUFA, and VLC-PUFA reflects therefore the metabolic context that gave rise to the formation of the LD, either via an early partitioning of carbon following FA de novo synthesis and/or a recycling of FA from membrane lipids, e.g., plastid galactolipids or endomembrane phosphor- or betaine lipids. In this review, we address the relationship between cytosolic LDs and the complex membrane compartmentalization within stramenopile cells, the metabolic routes leading to TAG accumulation, and the physiological conditions that trigger LD production, in response to various environmental factors.

Dynamics of the Metabolome of Aliinostoc sp. PMC 882.14 in Response to Light and Temperature Variations.

Cyanobacteria are microorganisms able to adapt to a wide variety of environmental conditions and abiotic stresses. They produce a large number of metabolites that can participate in the dynamic adaptation of cyanobacteria to a range of different light, temperature, and nutrient conditions. Studying the metabolite profile is one way to understand how the physiological status of cells is related to their adaptive response. In this study, we sought to understand how the diversity and dynamics of the whole metabolome depended on the growth phase and various abiotic factors such as light intensity and temperature. The cyanobacterium, Aliinostoc sp. PMC 882.14, was selected for its large number of biosynthetic gene clusters. One group of cells was grown under normal conditions as a control, while other groups were grown under higher light or temperature. Metabolomes were analyzed by mass spectrometry (qTOF-MS/MS) combined with untargeted analysis to investigate metabolite dynamics, and significant variation was found between exponential and stationary phases, regardless of culture conditions. In the higher light group, the synthesis of several metabolites, including shinorine, was induced while other metabolites, such as microviridins, were synthesized under higher temperature conditions. Among highly regulated metabolites, we observed the presence of mycosporine-like amino acids (MAAs) and variants of somamides, microginins, and microviridins. This study demonstrated the importance of considering the physiological state of cyanobacteria for comparative global metabolomics and studies of the regulatory processes involved in production of specific metabolites. Our results also open up new perspectives on the use of organisms such as cyanobacteria for the targeted production of bioactive metabolites.