Photosynthetic Dinoflagellates Research Articles

Fatty Acids in Cnidaria: Distribution and Specific Functions

The phylum Cnidaria comprises five main classes—Hydrozoa, Scyphozoa, Hexacorallia, Octocorallia and Cubozoa—that include such widely distributed and well-known animals as hard and soft corals, sea anemones, sea pens, gorgonians, hydroids, and jellyfish. Cnidarians play a very important role in marine ecosystems. The composition of their fatty acids (FAs) depends on food (plankton and particulate organic matter), symbiotic photosynthetic dinoflagellates and bacteria, and de novo biosynthesis in host tissues. In cnidarian lipids, besides the common FA characteristics of marine organisms, numerous new and rare FAs are also found. All Octocorallia species and some Scyphozoa jellyfish contain polyunsaturated FAs (PUFAs) with 24 and 26 carbon atoms. The coral families can be distinguished by specific FA profiles: the presence of uncommon FAs or high/low levels of common fatty acids. Many of the families have characteristic FAs: Acroporidae are characterized by 18:3n6, eicosapentaenoic acid (EPA) 20:5n3, 22:4n6, and 22:5n3; Pocilloporidae by 20:3n6, 20:4n3, and docosahexaenoic acid 22:6n3 (DHA); and Poritidae by arachidonic acid (AA) and DHA. The species of Faviidae show elevated concentrations of 18:3n6 and 22:5n3 acids. Dendrophylliidae, being azooxanthellate corals, have such dominant acids as EPA and 22:5n3 and a low content of DHA, which is the major PUFA in hermatypic corals. The major and characteristic PUFAs for Milleporidae (class Hydrozoa) are DHA and 22:5n6, though in scleractinian corals, the latter acid is found only in trace amounts.

Defensive responses of most antioxidant genes in the freshwater dinoflagellate Palatinus apiculatus to cadmium stress and their implications

Photosynthetic dinoflagellates are one of the major microalgal taxa, playing essential roles in biogeochemical cycles and food webs in aquatic environments. Some freshwater dinoflagellates are known to be sensitive to environmental conditions, like water quality and contaminants; however, their molecular toxicological responses are insufficiently discovered. In the present study, we evaluated the physiological and transcriptomic responses of the freshwater dinoflagellate Palatinus apiculatus exposed to cadmium (Cd), focusing on stress-responsive genes. The cell number of P. apiculatus decreased significantly at Cd concentrations above 0.25 mg/L after 72 h, with an estimated EC50 value of 1.35 mg/L. In addition, we constructed 87,207 transcriptomic contigs from the P. apiculatus cells exposed to the Cd. Differential expression gene analysis showed that 21.0 % of total contigs were statistically significant, including 8647 up-regulated and 4195 down-regulated genes. Gene Ontology enrichment results revealed that genes responsive to stress and external stimuli were highly expressed in Cd-treated cells. Moreover, Cd significantly induced reactive oxygen species (ROS) production in P. apiculatus cells, and their patterns were similar to the expressions of certain antioxidant genes. Among the selected genes, GR expression levels were down-regulated, which may lead to the failure of cell defense against heavy metals. These results showed molecular defense pathways of the freshwater dinoflagellate P. apiculatus against the heavy metal that could be served as potential sensitive biomarkers for evaluating molecular toxicity in freshwater ecosystems.

Interpreting the complexities of the plastid genome in dinoflagellates: a mini-review of recent advances.

Photosynthetic dinoflagellates play crucial roles in global primary production and carbon fixation. Despite their success in filling various ecological niches, numerous mysteries about their plastid evolution and plastid genomes remain unsolved. The plastid genome of dinoflagellates presents one of the most complex lineages in the biological realm, mainly due to multiple endosymbiotic plastid events in their evolutionary history. Peridinin-containing dinoflagellates possess the most reduced and fragmented genome, with only a few genes located on multiple "minicircles", whereas replacement plastids in dinoflagellate lineages have undergone different degrees of endosymbiotic gene transfer. Recent advancements in high-throughput sequencing have improved our understanding of plastid genomes and plastid-encoded gene expression in many dinoflagellate species. Plastid transcripts of dinoflagellates exhibit two unconventional processing pathways: the addition of a 3' poly(U) tail and substitutional RNA editing. These pathways are widely employed across dinoflagellate lineages, which are possibly retained from the ancestral peridinin plastid. This mini-review summarizes the developments in the plastid genomes of dinoflagellates and pinpoints the research areas that necessitate further exploration, aiming to provide valuable insights into plastid evolution in these fascinating and important organisms.

Investigation of heterotrophs reveals new insights in dinoflagellate evolution

Dinoflagellates are diverse and ecologically important protists characterized by many morphological and molecular traits that set them apart from other eukaryotes. These features include, but are not limited to, massive genomes organized using bacterially-derived histone-like proteins (HLPs) and dinoflagellate viral nucleoproteins (DVNP) rather than histones, and a complex history of photobiology with many independent losses of photosynthesis, numerous cases of serial secondary and tertiary plastid gains, and the presence of horizontally acquired bacterial rhodopsins and type II RuBisCo. Elucidating how this all evolved depends on knowing the phylogenetic relationships between dinoflagellate lineages. Half of these species are heterotrophic, but existing molecular data is strongly biased toward the photosynthetic dinoflagellates due to their amenability to cultivation and prevalence in culture collections. These biases make it impossible to interpret the evolution of photosynthesis, but may also affect phylogenetic inferences that impact our understanding of character evolution. Here, we address this problem by isolating individual cells from the Salish Sea and using single cell, culture-free transcriptomics to expand molecular data for dinoflagellates to include 27 more heterotrophic taxa, resulting in a roughly balanced representation. Using these data, we performed a comprehensive search for proteins involved in chromatin packaging, plastid function, and photoactivity across all dinoflagellates. These searches reveal that 1) photosynthesis was lost at least 21 times, 2) two known types of HLP were horizontally acquired around the same time rather than sequentially as previously thought; 3) multiple rhodopsins are present across the dinoflagellates, acquired multiple times from different donors; 4) kleptoplastic species have nucleus-encoded genes for proteins targeted to their temporary plastids and they are derived from multiple lineages, and 5) warnowiids are the only heterotrophs that retain a whole photosystem, although some photosynthesis-related electron transport genes are widely retained in heterotrophs, likely as part of the iron-sulfur cluster pathway that persists in non-photosynthetic plastids.

Photosynthesis and other factors affecting the establishment and maintenance of cnidarian-dinoflagellate symbiosis.

Coral growth depends on the partnership between the animal hosts and their intracellular, photosynthetic dinoflagellate symbionts. In this study, we used the sea anemone Aiptasia, a laboratory model for coral biology, to investigate the poorly understood mechanisms that mediate symbiosis establishment and maintenance. We found that initial colonization of both adult polyps and larvae by a compatible algal strain was more effective when the algae were able to photosynthesize and that the long-term maintenance of the symbiosis also depended on photosynthesis. In the dark, algal cells were taken up into host gastrodermal cells and not rapidly expelled, but they seemed unable to reproduce and thus were gradually lost. When we used confocal microscopy to examine the interaction of larvae with two algal strains that cannot establish stable symbioses with Aiptasia, it appeared that both pre- and post-phagocytosis mechanisms were involved. With one strain, algae entered the gastric cavity but appeared to be completely excluded from the gastrodermal cells. With the other strain, small numbers of algae entered the gastrodermal cells but appeared unable to proliferate there and were slowly lost upon further incubation. We also asked if the exclusion of either incompatible strain could result simply from their cells' being too large for the host cells to accommodate. However, the size distributions of the compatible and incompatible strains overlapped extensively. Moreover, examination of macerates confirmed earlier reports that individual gastrodermal cells could expand to accommodate multiple algal cells. This article is part of the theme issue 'Sculpting the microbiome: how host factors determine and respond to microbial colonization'.

First record of the genus Togula (Gymnodiniales, Dinophyceae) and of the species T. jolla for the South-West Atlantic coastal waters

In the framework of a phytoplankton and biotoxin monitoring program implemented since 2008 in marine coastal waters of the Buenos Aires Province (Argentina), a gymnodinioid, photosynthetic dinoflagellate was isolated from a sample collected in Santa Teresita, establishing the strain LPCc007. The strain was identified as belonging to the psammophilic genus Togula by light microscopy based on the following distinctive characters: morphology of the cell dorso-ventrally flattened, and course of the cingulum, highly asymmetric, descending, with its ends displaced by about one to two thirds of the total length of the cell and joined by an oblique intercingular groove. The comparative morphological analysis between the motile vegetative cells of the isolated strain and the three species of the genus described to date, did not reveal significant differences that would allow it to be determined at a specific level. Instead, the pattern of development of longitudinal grooves on the hypocone of cells about to undergo mitosis observed in strain LPCc007, perfectly agrees with that described for T. jolla as a differential character. The morphological identification was consistent with LSU rDNA (D1-D2) based phylogeny that showed the sequence aggregation in the T. jolla clade. This is the first report of the genus Togula and the species T. jolla from Argentina and the South-West Atlantic Ocean coastal waters.

Structures and organizations of PSI–AcpPCI supercomplexes from red tidal and coral symbiotic photosynthetic dinoflagellates

Marine photosynthetic dinoflagellates are a group of successful phytoplankton that can form red tides in the ocean and also symbiosis with corals. These features are closely related to the photosynthetic properties of dinoflagellates. We report here three structures of photosystem I (PSI)-chlorophylls (Chls) a/c-peridinin protein complex (PSI-AcpPCI) from two species of dinoflagellates by single-particle cryoelectron microscopy. The crucial PsaA/B subunits of a red tidal dinoflagellate Amphidinium carterae are remarkably smaller and hence losing over 20 pigment-binding sites, whereas its PsaD/F/I/J/L/M/R subunits are larger and coordinate some additional pigment sites compared to other eukaryotic photosynthetic organisms, which may compensate for the smaller PsaA/B subunits. Similar modifications are observed in a coral symbiotic dinoflagellate Symbiodinium species, where two additional core proteins and fewer AcpPCIs are identified in the PSI-AcpPCI supercomplex. The antenna proteins AcpPCIs in dinoflagellates developed some loops and pigment sites as a result to accommodate the changed PSI core, therefore the structures of PSI-AcpPCI supercomplex of dinoflagellates reveal an unusual protein assembly pattern. A huge pigment network comprising Chls a and c and various carotenoids is revealed from the structural analysis, which provides the basis for our deeper understanding of the energy transfer and dissipation within the PSI-AcpPCI supercomplex, as well as the evolution of photosynthetic organisms.

Genomic Data Reveal Diverse Biological Characteristics of Scleractinian Corals and Promote Effective Coral Reef Conservation.

Reef-building corals (Scleractinia, Anthozoa, Cnidaria) are the keystone organisms of coral reefs, which constitute the most diverse marine ecosystems. Since the first decoded coral genome reported in 2011, about 40 reference genomes are registered as of 2023. Comparative genomic analyses of coral genomes have revealed genomic characters that may underlie unique biological characteristics and coral diversification. These include existence of genes for biosynthesis of mycosporine-like amino acids, loss of an enzyme necessary for cysteine biosynthesis in family Acroporidae, and lineage-specific gene expansions of DMSP lyase-like genes in the genus Acropora. While symbiosis with endosymbiotic photosynthetic dinoflagellates is a common biological feature among reef-building corals, genes associated with the intricate symbiotic relationship encompass not only those shared by many coral species, but also genes that were uniquely duplicated in each coral lineage, suggesting diversified molecular mechanisms of coral-algal symbiosis. Coral genomic data have also enabled detection of hidden, complex population structures of corals, indicating the need for species-specific, local-scale, carefully considered conservation policies for effective maintenance of corals. Consequently, accumulating coral genomic data from a wide range of taxa and from individuals of a species not only promotes deeper understanding of coral reef biodiversity, but also promotes appropriate and effective coral reef conservation. Considering the diverse biological traits of different coral species and accurately understanding population structure and genetic diversity revealed by coral genomic analyses during coral reef restoration planning could enable us to "archive" coral reef environments that are nearly identical to natural coral reefs.

Chlamydiae as symbionts of photosynthetic dinoflagellates.

Chlamydiae are ubiquitous intracellular bacteria and infect a wide diversity of eukaryotes, including mammals. However, chlamydiae have never been reported to infect photosynthetic organisms. Here, we describe a novel chlamydial genus and species, Candidatus Algichlamydia australiensis, capable of infecting the photosynthetic dinoflagellate Cladocopium sp. (originally isolated from a scleractinian coral). Algichlamydia australiensis was confirmed to be intracellular by fluorescence in situ hybridization and confocal laser scanning microscopy and temporally stable at the population level by monitoring its relative abundance across four weeks of host growth. Using a combination of short- and long-read sequencing, we recovered a high-quality (completeness 91.73% and contamination 0.27%) metagenome-assembled genome of A. australiensis. Phylogenetic analyses show that this chlamydial taxon represents a new genus and species within the Simkaniaceae family. Algichlamydia australiensis possesses all the hallmark genes for chlamydiae-host interactions, including a complete type III secretion system. In addition, a type IV secretion system is encoded on a plasmid and has previously been observed for only three other chlamydial species. Twenty orthologous groups of genes are unique to A. australiensis, one of which is structurally similar to a protein known from Cyanobacteria and Archaeplastida involved in thylakoid biogenesis and maintenance, hinting at potential chlamydiae interactions with the chloroplasts of Cladocopium cells. Our study shows that chlamydiae infect dinoflagellate symbionts of cnidarians, the first photosynthetic organism reported to harbor chlamydiae, thereby expanding the breadth of chlamydial hosts and providing a new contribution to the discussion around the role of chlamydiae in the establishment of the primary plastid.

Introduction of the microbial symbiosis in marine organisms from the genus Tridacna and Kiwaidae and the insight in aquaculture gained from them

The global demand for essential nutrients, particularly protein, is steadily rising as the world's population increases. Aquatic animals contribute significantly to protein consumption, making up 15.3% of the world's protein intake. Aquaculture, the fastest-growing sector in agriculture over the past three decades, has emerged as a vital source of aquatic animal protein. However, the success of aquaculture is closely tied to environmental factors, such as water quality and nutrient availability. This paper explores the role of microbial symbiosis in nutrient acquisition by marine invertebrates and how these natural relationships can inspire innovative approaches in aquaculture. The first case study focuses on the giant clam, which forms a mutualistic relationship with photosynthetic dinoflagellates known as zooxanthellae. These clams, found in nutrient-poor waters, harness energy through photosynthesis, facilitated by these symbiotic microbes. The paper delves into the molecular mechanisms of this association and its potential applications in aquaculture. In the second case study, the yeti crab's relationship with chemoautotrophic bacteria is examined. These crabs actively cultivate epibiotic bacteria on their setae and rely on these microbes for sustenance. The paper discusses the microbial mechanisms involved in sulfur respiration and how the crab utilizes these bacteria. Drawing insights from these case studies, the paper proposes innovative approaches in the realm of aquaculture. These include the domestication of symbiotic species, the induction of symbiosis in aquacultural species, and the supplementation of aquacultural organisms with microbes as a potential food source. Various methods, such as mass selection, are suggested as potential means to carry out these experiments. It is important to note that while these ideas are theoretically compelling, they remain largely untested in practical applications, and the paper critically evaluates their feasibility in light of this nascent stage of research.

Unlocking the Complex Cell Biology of Coral-Dinoflagellate Symbiosis: A Model Systems Approach.

Symbiotic interactions occur in all domains of life, providing organisms with resources to adapt to new habitats. A prime example is the endosymbiosis between corals and photosynthetic dinoflagellates. Eukaryotic dinoflagellate symbionts reside inside coral cells and transfer essential nutrients to their hosts, driving the productivity of the most biodiverse marine ecosystem. Recent advances in molecular and genomic characterization have revealed symbiosis-specific genes and mechanisms shared among symbiotic cnidarians. In this review, we focus on the cellular and molecular processes that underpin the interaction between symbiont and host. We discuss symbiont acquisition via phagocytosis, modulation of host innate immunity, symbiont integration into host cell metabolism, and nutrient exchange as a fundamental aspect of stable symbiotic associations. We emphasize the importance of using model systems to dissect the cellular complexity of endosymbiosis, which ultimately serves as the basis for understanding its ecology and capacity to adapt in the face of climate change.

Characterizing the Influence of a Heterotrophic Bicosoecid Flagellate Pseudobodo sp. on the Dinoflagellate Gambierdiscus balechii.

Microbial interactions including competition, mutualism, commensalism, parasitism, and predation, which can be triggered by nutrient acquisition and chemical communication, are universal phenomena in the marine ecosystem. The interactions may influence the microbial population density, metabolism, and even their environmental functions. Herein, we investigated the interaction between a heterotrophic bicosoecid flagellate, Pseudobodo sp. (Bicoecea), and a dinoflagellate, Gambierdiscus balechii (Dinophyceae), which is a well-known ciguatera food poisoning (CFP) culprit. The presence of Pseudobodo sp. inhibited the algal proliferation and decreased the cardiotoxicity of zebrafish in the algal extract exposure experiment. Moreover, a significant difference in microbiome abundance was observed in algal cultures with and without Pseudobodo sp. Chemical analysis targeting toxins was performed by using liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with molecular networking (MN), showing a significant alteration in the cellular production of gambierone analogs and some super-carbon chain compounds. Taken together, our results demonstrated the impact of heterotrophic flagellate on the photosynthetic dinoflagellates, revealing the complex dynamics of algal toxin production and the ecological relationships related to dinoflagellates in the marine environment.

Establishment of oxidative stress biomarkers in oocytes from healthy and bleached scleractinian corals

Corals have a symbiotic relationship with photosynthetic dinoflagellates associated with the host's tissue, those responsible for most of their daily energy gain. Warming ocean water breaks down symbiosis, leading to a phenomenon known as bleaching. Without their primary nutritional source, corals depend on heterotrophy to survive, which can stagnate the reproductive cycle. There are corals, however, that manage to maintain gametogenesis even when bleached. However, the physiology of these gametes in such a situation is unknown. Our study is the first to evaluate the effects of bleaching on the oocytes of a scleractinian coral, as well as the strategies of these cells to maintain the balance of the antioxidant system and cellular homeostasis. We evaluated and quantified markers of oxidative balance in oocytes released from colonies of Mussismilia harttii with different physiological conditions: bleached and healthy. Healthy and bleached coral oocytes were collected considering a post-spawning time curve (0, 5 and 10 h) and frozen to evaluate the relationship between oxidative balance markers and the physiological conditions of coral colonies. Total protein levels, the activity of the antioxidants superoxide dismutase (SOD) and catalase (CAT), and the levels of lipoperoxidation (TBARS), a marker of lipid damage, were measured. The oocytes presented a significant difference between 0 h and 5 h after spawning for all the parameters regardless of the colony's health. Health status modulated SOD activity and TBARS levels, with oocytes from the bleached colony suffering the most lipid damage. These organisms seem to preserve the quality of female gametic cells even 10 h after spawning in both colonies, suggesting a robust antioxidant system capable of prolonging their lifespan and, possibly, their fertilization capability. This response may be related to an intensification of heterotrophy, ensuring nutritional support and thus reproductive effort and quality of gametes even in bleached corals.

Genomic architecture constrains macromolecular allocation in dinoflagellates

Dinoflagellate genomes have a unique architecture that may constrain their physiological and biochemical responsiveness to environmental stressors. Here we quantified how nitrogen (N) starvation influenced macromolecular allocation and C:N:P of three photosynthetic marine dinoflagellates, representing different taxonomic classes and genome sizes. Dinoflagellates respond to nitrogen starvation by decreasing cellular nitrogen, protein and RNA content, but unlike many other eukaryotic phytoplankton examined RNA:protein is invariant. Additionally, 2 of the 3 species exhibit increases in cellular phosphorus and very little change in cellular carbon with N-starvation. As a consequence, N starvation induces moderate increases in C:N, but extreme decreases in N:P and C:P, relative to diatoms. Dinoflagellate DNA content relative to total C, N and P is much higher than similar sized diatoms, but similar to very small photosynthetic picoeukaryotes such as Ostreococcus. In aggregate these results indicate the accumulation of phosphate stores may be an important strategy employed by dinoflagellates to meet P requirements associated with the maintenance and replication of their large genomes.

Host nutrient sensing is mediated by mTOR signaling in cnidarian-dinoflagellate symbiosis

To survive in the nutrient-poor waters of the tropics, reef-building corals rely on intracellular, photosynthetic dinoflagellate symbionts. Photosynthates produced by the symbiont are translocated to the host, and this enables corals to form the structural foundation of the most biodiverse of all marine ecosystems. Although the regulation of nutrient exchange between partners is critical for ecosystem stability and health, the mechanisms governing how nutrients are sensed, transferred, and integrated into host cell processes are largely unknown. Ubiquitous among eukaryotes, the mechanistic target of the rapamycin (mTOR) signaling pathway integrates intracellular and extracellular stimuli to influence cell growth and cell-cycle progression and to balance metabolic processes. A functional role of mTOR in the integration of host and symbiont was demonstrated in various nutritional symbioses, and a similar role of mTOR was proposed for coral-algal symbioses. Using the endosymbiosis model Aiptasia, we examined the role of mTOR signaling in both larvae and adult polyps across various stages of symbiosis. We found that symbiosis enhances cell proliferation, and using an Aiptasia-specific antibody, we localized mTOR to symbiosome membranes. We found that mTOR signaling is activated by symbiosis, while inhibition of mTOR signaling disrupts intracellular niche establishment and symbiosis altogether. Additionally, we observed that dysbiosis was a conserved response to mTOR inhibition in the larvae of a reef-building coral species. Our data confim that mTOR signaling plays a pivotal role in integrating symbiont-derived nutrients into host metabolism and symbiosis stability, ultimately allowing symbiotic cnidarians to thrive in challenging environments.

Composition of galactolipids, betaine lipids and triglyceride‐associated fatty acids of the symbiotic dinoflagellate Zooxanthella (Brandtodinium) nutricula: A glimpse into polyunsaturated fatty acids available to its polycystine radiolarian host

SUMMARYZooxanthella nutricula is a photosynthetic dinoflagellate symbiont of polycystine radiolarians. As such, it is hypothesized to provide fixed organic carbon, including in the form of acylglycerolipids and sterols, to its non‐photosynthetic host. We have previously characterized the sterols of Z. nutricula that may be transferred to its host and, in the present study, have turned our attention to three classes of fatty acid‐containing lipids, chloroplast‐associated galactolipids, betaine lipids, which are non‐phosphorylated phospholipid analogs present in many eukaryotes, and triglycerides. Zooxanthella nutricula was observed using positive‐ion electrospray/mass spectrometry (ESI/MS) and ESI/MS/MS to produce the galactolipids mono‐ and digalactosyldiacylglycerol (MGDG and DGDG, respectively) enriched in octadecapentaenoic (18:5(n‐3)) and octadecatetraenoic (18:4(n‐3)) acid to place it within a group of peridinin‐containing dinoflagellates in a C18/C18 (sn‐1/sn‐2 fatty acid regiochemistry) cluster, as opposed to another cluster with C20/C18 MGDG and DGDG, where the C20 fatty acid is eicosapentaenoic acid (20:5(n‐3)) and the C18 fatty acid is either 18:5(n‐3) or 18:4(n‐3). Zooxanthella nutricula was also observed to produce 38:10 (total number of fatty acid carbons:total number of double bonds), 38:6, and 44:7 diacylglycerylcarboxyhydroxymethylcholine (DGCC) as the sole type of betaine lipid. Although it is more difficult to determine which fatty acids are present in the sn‐1 and sn‐2 positions on the glycerol backbone of DGCC using ESI/MS/MS, gas chromatography/mass spectrometry (GC/MS)‐based examination indicated the putatively DGCC‐associated polyunsaturated fatty acid (PUFA) docosahexaenoic acid (22:6(n‐3)). Coupled with the C18 PUFAs of MGDG and DGDG, and fatty acids associated with triglycerides (also examined via GC/MS), Z. nutricula could serve as a rich source of PUFAs for its radiolarian host. These data demonstrate that Z. nutricula produces a similar set of PUFA‐containing lipids as Symbiodinium microadriaticum, a photosynthetic dinoflagellate symbiont of cnidarians, indicating a metabolic commonality in these phylogenetically discrete dinoflagellate symbionts with unrelated host organisms.

Targeted volume correlative light and electron microscopy of an environmental marine microorganism.

Photosynthetic microalgae are responsible for an important fraction of CO2 fixation and O2 production on Earth. Three-dimensional (3D) ultrastructural characterization of these organisms in their natural environment can contribute to a deeper understanding of their cell biology. However, the low throughput of volume electron microscopy (vEM) methods along with the complexity and heterogeneity of environmental samples pose great technical challenges. In the present study, we used a workflow based on a specific electron microscopy sample preparation method compatible with both light and vEM imaging in order to target one cell among a complex natural community. This method revealed the 3D subcellular landscape of a photosynthetic dinoflagellate, which we identified as Ensiculifera tyrrhenica, with quantitative characterization of multiple organelles. We show that this cell contains a single convoluted chloroplast and show the arrangement of the flagellar apparatus with its associated photosensitive elements. Moreover, we observed partial chromatin unfolding, potentially associated with transcription activity in these organisms, in which chromosomes are permanently condensed. Together with providing insights in dinoflagellate biology, this proof-of-principle study illustrates an efficient tool for the targeted ultrastructural analysis of environmental microorganisms in heterogeneous mixes.

Dinoflagellates as sustainable cellulose source: Cultivation, extraction, and characterization

The global demand for manufacturing and consuming biodegradable materials from natural sources has created a great interest in microalgae such as dinoflagellates. Photosynthetic dinoflagellates are a sustainable source of natural materials such as cellulose as they grow using only sunlight and CO2 at near-neutral pH without any fertilizers. In this paper, the cultivation of two species of dinoflagellates (Peridinium sp. and Prorocentrum micans) is established under lab conditions (up to 20 l), cellulose extraction is optimized, and the resulting material is thoroughly characterized. Dinoflagellate cellulose was extracted at room temperature by sequential treatment with highly concentrated 30 % NaOH and 6 M HCl, followed by bleaching with 10 % H2O2. The overall yield of cellulose is around 73 % (w/w), and roughly 85 % of the original dinoflagellate cellulosic morphology remains intact. Chemical purity, morphology, and porosity of the dinoflagellate-derived cellulose are analysed by different characterization techniques (ICP-OES, SEM, XRD, ATR-FTIR, Raman, ssNMR, TGA, BET, and GPC). XRD characterization of the extracted cellulose shows no characteristic reflexes corresponding to a cellulose II allomorph which is mainly amorphous. This result is further supported by ATR-FTIR, Raman, and ssNMR spectroscopy. Overall, these results show that the extracted cellulose is a highly porous, lignin-free material that is thermally stable up to 260 °C. Its high degree of purity and porosity make dinoflagellate-derived cellulose a promising, sustainable candidate for the development of functional hybrid materials for biomedical applications.

The microbiome of the endosymbiotic Symbiodiniaceae in corals exposed to thermal stress

The coral reef crisis has influenced research for over two decades, during which time the capacity of corals to withstand and respond to environmental stress has been documented from the cellular to ecosystem level. Over the past decade, research is increasingly working towards uncovering the extent of coral–bacterial interactions, finding that diverse and stable microbial interactions can be indicative of the health of the coral host. However, we have yet to determine at which level of organismal organisation these interactions occur, in particular those with the coral’s photosynthetic dinoflagellate symbionts. This information is critical if we are to understand the impact of stress on meta-organism functioning. Using 16S gene amplicon sequencing, we investigated the bacterial microbiome of endosymbiotic Symbiodiniaceae from thermally stressed Acropora aspera, under 3 ecologically relevant temperature trajectories (defined as protective, repetitive and single) that are expected under a changing climate. We show that endosymbiotic Symbiodiniaceae host a distinct and diverse bacterial assemblage when compared with the A. aspera host. Alphaproteobacteria (mainly Rhodobacteraceae and Bradyrhizobiaceae), from the Rhizobiales order dominated the Symbiodiniaceae microbiome, while Gammaproteobacteria (mainly Endozoicomonadaceae) dominated the coral microbiome. The Symbiodiniaceae core microbiome also reflected the distinct microbiomes of the two partners, specifically, Rhizobiales were not present in the A. aspera core, while Endozoicomonadaceae were not present in the Symbiodiniaceae core. We show the Symbiodiniaceae-associated microbiome was highly responsive to increases in temperature, and the microbial consortium was significantly altered in the Symbiodiniaceae retained in the host exposed to different temperature. Most notably, Myxococcolaes were up to 25-fold higher relative abundance in dinoflagellate partner microbiomes under the single temperature trajectory, compared with the repetitive and control treatments. The distinct composition of bacteria associated with Symbiodiniaceae suggests a previously unrecognised, yet important functional role of these associations to overall coral health, which is increasingly important as reefs decline worldwide. Our study provides the first characterisation of Symbiodiniaceae-associated microbes from a coral host under a range of temperature trajectories occurring on the Great Barrier Reef.

Uptake and Effects of Nanoplastics on the Dinoflagellate Gymnodinium corollarium.

Plastic nanoparticles (NPs) are the final state of plastic degradation in the environment before they disintegrate into low-molecular-weight organic compounds. Unicellular organisms are highly sensitive to the toxic effects of nanoplastics, because they are often capable of phagotrophy but are unable to consume a foreign material such as synthetic plastic. We studied the effect of polystyrene, poly(vinyl chloride), poly(methyl acrylate), and poly(methyl methacrylate) NPs on the photosynthetic dinoflagellate Gymnodinium corollarium Sundström, Kremp et Daugbjerg. Fluorescent tagged particles were used to visualize plastic capture by dinoflagellate cells. We found that these dinoflagellates are capable of phagotrophic nutrition and thus should be regarded as mixotrophic species. This causes their susceptibility to the toxic effects of plastic NPs. Living cells ingest plastic NPs and accumulate in the cytoplasm as micrometer-level aggregates, probably in food vacuoles. The action of nanoplastics leads to a dose-dependent increase in the level of reactive oxygen species in dinoflagellate cells, indicating plastic degradation in the cells. The introduction of a methyl group into the main chain in the α-position in the case of poly(methyl methacrylate) causes a drastic reduction in toxicity. We expect that such NPs can be a tool for testing unicellular organisms in terms of heterotrophic feeding ability. We suggest a dual role of dinoflagellates in the ecological fate of plastic waste: the involvement of nanoplastics in the food chain and its biochemical destruction. Environ Toxicol Chem 2023;42:1124-1133. © 2023 SETAC.