Unique photosynthetic strategies employed by closely related Breviolum minutum strains under rapid short-term cumulative heat stress.

The thermal tolerance of symbiodiniacean photo-endosymbionts largely underpins the thermal bleaching resilience of their cnidarian hosts such as corals and the coral model Exaiptasia diaphana. While variation in thermal tolerance between species is well documented, variation between conspecific strains is understudied. We compared the thermal tolerance of three closely related strains of Breviolum minutum represented by two internal transcribed spacer region 2 profiles (one strain B1-B1o-B1g-B1p and the other two strains B1-B1a-B1b-B1g) and differences in photochemical and non-photochemical quenching, de-epoxidation state of photopigments, and accumulation of reactive oxygen species under rapid short-term cumulative temperature stress (26-40 °C). We found that B. minutum strains employ distinct photoprotective strategies, resulting in different upper thermal tolerances. We provide evidence for previously unknown interdependencies between thermal tolerance traits and photoprotective mechanisms that include a delicate balancing of excitation energy and its dissipation through fast relaxing and state transition components of non-photochemical quenching. The more thermally tolerant B. minutum strain (B1-B1o-B1g-B1p) exhibited an enhanced de-epoxidation that is strongly linked to the thylakoid membrane melting point and possibly membrane rigidification minimizing oxidative damage. This study provides an in-depth understanding of photoprotective mechanisms underpinning thermal tolerance in closely related strains of B. minutum.

Assessing the contribution of bacteria to the heat tolerance of experimentally evolved coral photosymbionts.

Coral reefs are extremely vulnerable to ocean warming, which triggers coral bleaching-the loss of endosymbiotic microalgae (Symbiodiniaceae) from coral tissues, often leading to death. To enhance coral climate resilience, the symbiont, Cladocopium proliferum was experimentally evolved for >10 years under elevated temperatures resulting in increased heat tolerance. Bacterial 16S rRNA gene metabarcoding showed the composition of intra- and extracellular bacterial communities of heat-evolved strains was significantly different from that of wild-type strains, suggesting bacteria responded to elevated temperatures, and may even play a role in C. proliferum thermal tolerance. To assess whether microbiome transplantation could enhance heat tolerance of the sensitive wild-type C. proliferum, we transplanted bacterial communities from heat-evolved to the wild-type strain and subjected it to acute heat stress. Microbiome transplantation resulted in the incorporation of only 30 low-abundance strains into the microbiome of wild-type cultures, while the relative abundance of 14 pre-existing strains doubled in inoculated versus uninoculated samples. Inoculation with either wild-type or heat-evolved bacterial communities boosted C. proliferum growth, although no difference in heat tolerance was observed between the two inoculation treatments. This study provides evidence that Symbiodiniaceae-associated bacterial communities respond to heat selection and may contribute to coral adaptation to climate change.

Profiling of grazed cultures of the chlorophyte alga Dunaliella tertiolecta using an untargeted LC-MS approach.

Extracellular signals are reported to mediate chemical cross-talk among pelagic microbes, including microalgal prey and predators. Water-soluble mediator compounds play a crucial role in extracellular communication which is vital for prey recognition, attraction, capture, and predator deterrence. A range of exo-metabolites including oxylipins and vitamins are released by prey in response to grazing stress. The temporal dynamics of such exo-metabolites largely remains unknown, especially in large-scale cultivation of microalgae such as closed or open ponds. In open ponds, infestation of predators is almost inevitable but highly undesirable due to the imminent threat of culture collapse. The early production of exo-metabolites emitted by microalgal prey in response to predator attack could be leveraged as diagnostic markers of possible culture collapse. This study uses an untargeted approach for temporal profiling of Dunaliella tertiolecta-specific exo-metabolites under grazing pressure from Oxyrrhis marina. We report 24 putatively identified metabolites, belonging to various classes such as short peptides, lipids, indole-derivatives, and free amino acids, as potential markers of grazing-mediated stress. In addition, this study outlines a clear methodology for screening of exo-metabolites in marine algal samples, the analysis of which is frequently hindered by high salt concentrations. In future, a chemistry-based targeted detection of these metabolites could enable a quick and on-site screening of predators in microalgal cultures.

Effects of Ocean Warming on the Underexplored Members of the Coral Microbiome.

The climate crisis is one of the most significant threats to marine ecosystems. It is leading to severe increases in sea surface temperatures and in the frequency and magnitude of marine heatwaves. These changing conditions are directly impacting coral reef ecosystems, which are among the most biodiverse ecosystems on Earth. Coral-associated symbionts are particularly affected because summer heatwaves cause coral bleaching-the loss of endosymbiotic microalgae (Symbiodiniaceae) from coral tissues, leading to coral starvation and death. Coral-associated Symbiodiniaceae and bacteria have been extensively studied in the context of climate change, especially in terms of community diversity and dynamics. However, data on other microorganisms and their response to climate change are scarce. Here, we review current knowledge on how increasing temperatures affect understudied coral-associated microorganisms such as archaea, fungi, viruses, and protists other than Symbiodiniaceae, as well as microbe-microbe interactions. We show that the coral-microbe symbiosis equilibrium is at risk under current and predicted future climate change and argue that coral reef conservation initiatives should include microbe-focused approaches.

Temporal acclimation of Microchloropsis gaditana CCMP526 in response to hypersalinity.

Evaporation from culture ponds and raceways can subject algae to hypersalinity stress, and this is exacerbated by global warming. We investigated the effect of salinity on a marine microalga, Microchloropsis gaditana, which is of industrial significance because of its high lipid-accumulating capability. Both short-term (hours) and medium-term (days) effects of salinity were studied across various salinities (37.5, 55, 70 and 100 PSU). Salinity above 55 PSU suppressed cell growth and specific growth rate was significantly reduced at 100 PSU. Photosynthesis (Fv/Fm, rETRmax and Ik) was severely affected at high salinity conditions. Total carbohydrate per cell increased ∼1.7-fold after 24 h, which is consistent with previous findings that salinity induces osmolyte production to counter osmotic shock. In addition, accumulation of lipid increased by ∼4.6-fold in response to salinity. Our findings indicate a possible mechanism of acclimation to salinity, opening up new frontiers for osmolytes in pharmacological and cosmetics applications.

Time for Multiple Extraction Methods in Proteomics? A Comparison of Three Protein Extraction Methods in the Eustigmatophyte Alga Microchloropsis gaditana CCMP526.

Proteomics is a crucial postgenomic biotechnology for functional and systems scale analyses in cell and integrative biology, not to mention clinical and precision medicine research. However, a fundamental requirement for an accurate examination of the protein complement of cells is an efficient method for extracting the proteins. This study reports on the evaluation of three protein extraction methods: trichloroacetic acid (TCA)-acetone, phenol, and TRIzol, in the eustigmatophyte alga Microchloropsis gaditana CCMP526 for proteomic analysis. M. gaditana is a potential candidate for algal-based biofuels. This industrially important strain is also rich in dietary oil and pigments and is used as feed in the aquaculture industry. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomic analysis was performed for proteins obtained using the three extraction methods and their effects were examined by the abundance ratio. Protein yield was higher using the TCA-acetone and phenol methods than with the TRIzol method. The TCA method was superior than other methods examined here in terms of protein coverage and abundance. Subcellular localization of the identified proteins revealed no significant difference among the extraction methods. Importantly, each method revealed a unique set of proteins. To the best of our knowledge, this is the first report on evaluation of protein extraction methods for the proteomic analysis of M. gaditana CCMP526. These observations underscore the importance of using multiple protein extraction methods for comprehensive proteome coverage, as the field of proteomics edges toward diverse applications in biofuels, aquaculture industry, marine biology, and agriculture.