Dinoflagellate Cells Research Articles

Heat stress in symbiotic dinoflagellates: Implications on oxidative stress and cellular changes

Global warming has been shown to harmfully affect symbiosis between Symbiodiniaceae and other marine invertebrates. When symbiotic dinoflagellates (the genus Breviolum) were in vitro exposed to acute heat stress of +7 °C for a period of 5 days, the results revealed the negative impact on all physiological and other cellular parameters measured. Elevated temperatures resulted in a severe reduction in algal density of up to 9.5-fold, as well as pigment concentrations, indicating the status of the physiological stress and early signs of photo-bleaching. Reactive oxygen species (ROS) were increased in all heated dinoflagellate cells, while the antioxidant-reduced glutathione levels initially dropped on day one but increased under prolonged temperature stress. The cell viability parameters were reduced by 97 % over the heating period, with an increased proportion of apoptotic and necrotic cells. Autofluorescence (AF) for Cy5-PE 660-20 was reduced from 1.7-fold at day 1 to up to 50-fold drop at the end of heating time, indicating that the AF changes were highly sensitive to heat stress and that it could be an extremely sensitive tool for assessing the functionality of algal photosynthetic machinery. The addition of the drug 5-AZA-2′-deoxycytidine (5-AZA), which inhibits DNA methylation processes, was assessed in parallel and contributed to some alterations in algal cellular stress response. The presence of drug 5-AZA combined with the temperature stress had an additional impact on Symbiodiniaceae density and cell complexity, including the AF levels. These variations in cellular stress response under heat stress and compromised DNA methylation conditions may indicate the importance of this epigenetic mechanism for symbiotic dinoflagellate thermal tolerance adaptability over a longer period, which needs further exploration. Consequently, the increased ROS levels and changes in AF signals reported during ongoing heat stress in dinoflagellate cells could be used as early stress biomarkers in these microalgae and potentially other photosynthetic species.

Comparison of methods for DMSP measurements in dinoflagellate cultures

AbstractA comparison of three analytical methods (the indirect GC‐FPD and MIMS, and direct LC‐MS/MS) for dimethylsulfoniopropionate (DMSP) measurements was conducted to assess their accuracy and reliability. The three methods showed a linear response but are distinguished by their linearity range, the largest being for MIMS. All three methods showed good precision on Alexandrium minutum samples (2–12%). The variability between the three methods when comparing analyses of A. minutum replicates was 11%, with the DMSP measurements by LC‐MS/MS being the highest. This result also confirms that indirect DMSP measurement after hydrolysis for GC or MIMS methods does not lead to an overestimation of DMSP values in A. minutum. A special focus was made on the more recent LC‐MS/MS method including further assays in sample preparation and storage from cultures of the dinoflagellate A. minutum. Dinoflagellate cells should be harvested by gentle filtration (< 5 cm Hg) or slow centrifugation (500 × g) to retrieve the largest DMSP pool. For the LC‐MS/MS method, MeOH used for cell extraction should be added prior to freezing (to prevent DMSP degradation). Samples will then be stable in frozen storage for at least 2 months. Finally, direct and indirect methods are complementary for identifying the exact DMSP fraction among dimethylsulfide‐producing compounds that compose total and particulate DMSP pools issued from newly screened organisms or environmental samples.

In vitro effects of the harmful benthic dinoflagellates Prorocentrum hoffmannianum and Ostreopsis cf. ovata on immune responses of the farmed oyster Crassostrea gasar

Oyster culture is a sustainable solution to food production. However, this activity can be severely impacted by the presence and proliferation of harmful microalgae such as the benthic dinoflagellates Prorocentrum hoffmannianum and Ostreopsis cf. ovata. This study aimed to evaluate the in vitro effects of P. hoffmannianum and O. cf. ovata on immune system cells (hemocytes) of the native cultured oyster Crassostrea gasar. The direct toxicity of both dinoflagellates was first evaluated assessing hemocyte viability exposed to eight concentrations of each HAB species. No reduction in hemocyte viability was found with the exposure to cell culture or the crude extract of P. hoffmannianum, but O. cf. ovata culture induced hemocyte death in a concentration-dependent manner. Ostreopsis cf. ovata concentration that promoted half of maximal reduction in hemocyte viability (EC50) was 779 cells mL−1. Posteriorly, hemocytes were exposed to both dinoflagellate cells and crude extracts to investigate their effects on hemocyte functional parameters. Despite no direct toxicity of the dinoflagellate cells, P. hoffmannianum extract caused a threefold increase in ROS production and decreased the phagocytosis rate by less than half. Ostreopsis cf. ovata cells and crude extracts also triggered an increase in ROS production (two-fold), but the phagocytosis rate was reduced (by half) only in response to the two lower cell concentrations. These results indicate a harmful potential of both dinoflagellates through a direct toxicity (only for O. cf. ovata) and functional impairment of hemocytes (both species) which could expose C. gasar oyster to opportunistic infections.

Ocean acidification and food availability impacts on the metabolism and grazing in a cosmopolitan herbivorous protist Oxyrrhis marina

The heterotrophic dinoflagellate Oxyrrhis marina is an essential microzooplankton in coastal waters, linking the energy transfer from phytoplankton to higher trophic levels. It is of general significance to investigate how it responds and acclimates to ocean acidification (OA), especially under varied availabilities of food. Here, O. marina was exposed and acclimated to three pCO2 levels (LC: 415, MC:1000, HC:1500 μatm) for 60 days, and then was further grown under the CO2 levels with different levels of food (the microalgae Dunaliella salina) availability for about 8 generations. The OA treatments did not significantly hamper its growth and ingestion rates even under the reduced food availability and starvation (deprived of the microalgae), which significantly reduced its growth rate. While the impacts of OA on the growth and ingestion rates of O. marina were insignificant, the OA treatments appeared to have resulted in a faster decline of the heterotrophic dinoflagellate cells during the starvation period. Nevertheless, the acidic stress under the elevated pCO2 of 1000 or 1500 μatm decreased its respiration by about 53% or 59% with the high and by about 26% or 23% with the low food availability, respectively. Such OA-repressed respiration was also significant during the starvation period. On the other hand, the OA treatments and deprivation of the microalgae synergistically reduced the cellular quota of particulate organic C, N and P, resulting in a reduction of food value of the heterotrophic dinoflagellate as prey. In conclusion, our results show that O. marina is highly resilient to future ocean acidification by reducing its respiration and sustaining its ingestion of microalgae.

Functional analyses of bacterial genomes found in Symbiodiniaceae genome assemblies.

Bacterial-algal interactions strongly influence marine ecosystems. Bacterial communities in cultured dinoflagellates of the family Symbiodiniaceae have been characterized by metagenomics. However, little is known about whole-genome analysis of marine bacteria associated with these dinoflagellates. We performed in silico analysis of four bacterial genomes from cultures of four dinoflagellates of the genera Symbiodinium, Breviolum, Cladocopium and Durusdinium. Comparative analysis showed that the former three contain the alphaproteobacterial family Parvibaculaceae and that the Durusdinium culture includes the family Sphingomonadaceae. There were no large genomic reductions in the alphaproteobacteria with genome sizes of 2.9-3.9 Mb, implying they are not obligate intracellular bacteria. Genomic annotations of three Parvibaculaceae detected the gene for diacetylchitobiose deacetylase (Dac), which may be involved in the degradation of dinoflagellate cell surfaces. They also had metabolic genes for dissimilatory nitrate reduction to ammonium (DNRA) in the nitrogen (N) cycle and cobalamin (vitamin B12 ) biosynthetic genes in the salvage pathway. Those three characters were not found in the Sphingomonadaceae genome. Predicted biosynthetic gene clusters for secondary metabolites indicated that the Parvibaculaceae likely produce the same secondary metabolites. Our study suggests that the Parvibaculaceae is a major resident of Symbiodiniaceae cultures with antibiotics.

Two novel glutathione S-transferase (GST) genes in the toxic marine dinoflagellate Alexandrium pacificum and their transcriptional responses to environmental contaminants

The present study identified two novel glutathione S-transferase (GST) genes from the toxic dinoflagellate Alexandrium pacificum and examined their molecular characteristics and transcriptional responses to algicides and environmental contaminants. Bioinformatic analysis revealed that both ApGSTs are cytosolic, belonging to the chi-like class (ApGST1) and an undefined class (ApGST2). The overall expression of ApGSTs showed similar patterns depending on the exposed contaminants, while they were differently regulated by polychlorinated biphenyl (PCB). Copper treatments (CuCl2 and CuSO4) did not significantly induce the expression of ApGSTs. The highest up-regulations of ApGST1 and ApGST2 were under 6-h treatments of 0.10 and 0.50 mg L−1 NaOCl. Interestingly, only ApGST1 increased significantly after 0.10, 0.50, and 1.00 mg L−1 of PCB exposure (6 h). Intracellular reactive oxygen species (ROS) increased considerably under NaOCl; however, it was not significantly higher in the PCB-treated cells. GST activity was increased by NaOCl and PCB treatments, but only PCB caused apoptosis. These results suggest that GSTs are involved in the first line of phase II detoxification, protecting dinoflagellate cells against oxidative damage.

Single-species dinoflagellate cyst carbon isotope fractionation in core-top sediments: environmental controls, CO2 dependency and proxy potential

Abstract. Sedimentary bulk organic matter and various molecular organic components exhibit strong CO2-dependent carbon isotope fractionation relative to dissolved inorganic carbon sources. This fractionation (εp) has been employed as a proxy for paleo-pCO2. Yet, culture experiments indicate that CO2-dependent εp is highly specific at genus and even species level, potentially hampering the use of bulk organic matter and non-species-specific organic compounds. In recent years, significant progress has been made towards a CO2 proxy using controlled growth experiments with dinoflagellate species, also showing highly species-specific εp values. These values were, however, based on motile specimens, and it remains unknown whether these relations also hold for the organic-walled resting cysts (dinocysts) produced by these dinoflagellate species in their natural environment. We here analyze dinocysts isolated from core tops from the Atlantic Ocean and Mediterranean Sea, representing several species (Spiniferites elongatus, S. (cf.) ramosus, S. mirabilis, Operculodinium centrocarpum sensu Wall and Dale (1966) (hereafter referred to as O. centrocarpum) and Impagidinium aculeatum) using laser ablation–nano-combustion–gas-chromatography–isotope ratio mass spectrometry (LA/nC/GC-IRMS). We find that the dinocysts produced in the natural environment are all appreciably more 13C-depleted compared to the cultured motile dinoflagellate cells, implying higher overall εp values, and, moreover, exhibit large isotope variability. Where several species could be analyzed from a single location, we often record significant differences in isotopic variance and offsets in mean δ13C values between species, highlighting the importance of single-species carbon isotope analyses. The most geographically expanded dataset, based on O. centrocarpum, shows that εp correlates significantly with various environmental parameters. Importantly, O. centrocarpum shows a CO2-dependent εp above ∼ 240 µatm pCO2. Similar to other marine autotrophs, relative insensitivity at low pCO2 is in line with active carbon-concentrating mechanisms at low pCO2, although we here cannot fully exclude that we partly underestimated εp sensitivity at low pCO2 values due to the relatively sparse sampling in that range. Finally, we use the relation between εp and pCO2 in O. centrocarpum to propose a first pCO2 proxy based on a single dinocyst species.

PACMan: A software package for automated single-cell chlorophyll fluorometry.

Microalgae, small photosynthetic unicells, are of great interest to ecology, ecotoxicology and biotechnology and there is a growing need to investigate the ability of cells to photosynthesize under variable conditions. Current strategies involve hand-operated pulse-amplitude-modulated (PAM) chlorophyll fluorimeters, which can provide detailed insights into the photophysiology of entire populations- or individual cells of microalgae but are typically limited in their throughput. To increase the throughput of a commercially available MICROSCOPY-PAM system, we present the PAM Automation Control Manager ('PACMan'), an open-source Python software package that automates image acquisition, microscopy stage control and the triggering of external hardware components. PACMan comes with a user-friendly graphical user interface and is released together with a stand-alone tool (PAMalysis) for the automated calculation of per-cell maximum quantum efficiencies (= Fv /Fm ). Using these two software packages, we successfully tracked the photophysiology of >1000 individual cells of green algae (Chlamydomonas reinhardtii) and dinoflagellates (genus Symbiodiniaceae) within custom-made microfluidic devices. Compared to the manual operation of MICROSCOPY-PAM systems, this represents a 10-fold increase in throughput. During experiments, PACMan coordinated the movement of the microscope stage and triggered the MICROSCOPY-PAM system to repeatedly capture high-quality image data across multiple positions. Finally, we analyzed single-cell Fv /Fm with the manufacturer-supplied software and PAMalysis, demonstrating a median difference <0.5% between both methods. We foresee that PACMan, and its auxiliary software package will help increase the experimental throughput in a range of microalgae studies currently relying on hand-operated MICROSCOPY-PAM technologies.

The succession of epiphytic microalgae conditions fungal community composition: how chytrids respond to blooms of dinoflagellates

This study aims to investigate the temporal dynamics of the epiphytic protist community on macroalgae, during the summer months, with a specific focus on fungi, and the interactions between zoosporic chytrid parasites and the proliferation of the dinoflagellates. We employed a combination of environmental sequencing techniques, incubation of natural samples, isolation of target organisms and laboratory experiments. Metabarcoding sequencing revealed changes in the dominant members of the epiphytic fungal community. Initially, fungi comprised < 1% of the protist community, mostly accounted for by Basidiomycota and Ascomycota, but with the emergence of Chytridiomycota during the mature phase of the biofilm, the fungal contribution increased to almost 30%. Chytridiomycota became dominant in parallel with an increase in the relative abundance of dinoflagellates in the community. Microscopy observations showed a general presence of chytrids following the peak proliferation of the dinoflagellate Ostreopsis sp., with the parasite, D. arenysensis as the dominant chytrid. The maximum infection prevalence was 2% indicating host-parasite coexistence. To further understand the in-situ prevalence of chytrids, we characterised the dynamics of the host abundance and prevalence of chytrids through co-culture. These laboratory experiments revealed intraspecific variability of D. arenysensis in its interaction with Ostreopsis, exhibiting a range from stable coexistence to the near-extinction of the host population. Moreover, while chytrids preferentially parasitized dinoflagellate cells, one of the strains examined displayed the ability to utilize pollen as a resource to maintain its viability, thus illustrating a facultative parasitic lifestyle. Our findings not only enrich our understanding of the diversity, ecology, and progression of epiphytic microalgal and fungal communities on Mediterranean coastal macroalgae, but they also shed light on the presence of zoosporic parasites in less-explored benthic habitats.

The enigmatic nucleus of the marine dinoflagellate Prorocentrum cordatum.

The marine, bloom-forming dinoflagellate Prorocentrum cordatum CCMP 1329 (formerly P. minimum) has a genome atypical of eukaryotes, with a large size of ~4.15 Gbp, organized in plentiful, highly condensed chromosomes and packed in a dinoflagellate-specific nucleus (dinokaryon). Here, we apply microscopic and proteogenomic approaches to obtain new insights into this enigmatic nucleus of axenic P. cordatum. High-resolution focused ion beam/scanning electron microscopy analysis of the flattened nucleus revealed highest density of nuclear pores in the vicinity of the nucleolus, a total of 62 tightly packed chromosomes (~0.4-6.7 µm3), and interaction of several chromosomes with the nucleolus and other nuclear structures. A specific procedure for enriching intact nuclei was developed to enable proteomic analyses of soluble and membrane protein-enriched fractions. These were analyzed with geLC and shotgun approaches employing ion-trap and timsTOF (trapped-ion-mobility-spectrometry time-of-flight) mass spectrometers, respectively. This allowed identification of 4,052 proteins (39% of unknown function), out of which 418 were predicted to serve specific nuclear functions; additional 531 proteins of unknown function could be allocated to the nucleus. Compaction of DNA despite very low histone abundance could be accomplished by highly abundant major basic nuclear proteins (HCc2-like). Several nuclear processes including DNA replication/repair and RNA processing/splicing can be fairly well explained on the proteogenomic level. By contrast, transcription and composition of the nuclear pore complex remain largely elusive. One may speculate that the large group of potential nuclear proteins with currently unknown functions may serve yet to be explored functions in nuclear processes differing from those of typical eukaryotic cells. IMPORTANCE Dinoflagellates form a highly diverse group of unicellular microalgae. They provide keystone species for the marine ecosystem and stand out among others by their very large, unusually organized genomes embedded in the nuclei markedly different from other eukaryotic cells. Functional insights into nuclear and other cell biological structures and processes of dinoflagellates have long been hampered by the paucity of available genomic sequences. The here studied cosmopolitan P. cordatum belongs to the harmful algal bloom-forming, marine dinoflagellates and has a recently de novo assembled genome. We present a detailed 3D reconstruction of the P. cordatum nucleus together with comprehensive proteogenomic insights into the protein equipment mastering the broad spectrum of nuclear processes. This study significantly advances our understanding of mechanisms and evolution of the conspicuous dinoflagellate cell biology.

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.

Chronological age-related metabolome responses in the dinoflagellate Karenia mikimotoi, can predict future bloom demise

Karenia mikimotoi is a common harmful algal bloom (HAB)-forming dinoflagellate and has caused severe financial loss in aquaculture. There are limited metabolomic studies on dinoflagellate biology. Here, we examined alterations in metabolic profiles over the growth curve of K. mikimotoi under nitrogen or phosphorus deficiency and further explored a key criterion for the diagnosis of late stationary phase to identify when the dinoflagellate cells will enter bloom demise. The results demonstrate the differential expression of metabolites for coping with chronological aging or nutrient deprivation. Furthermore, an increase in the glucose to glycine ratio in the late stationary phase was indicative of dinoflagellate cells entering bloom demise; this was also detected in the cultured diatom, Chaetoceros tenuissimus, indicating that this may be the general criterion for phytoplankton species. Our findings provide insights regarding chronological aging and the criterion for the prediction of phytoplankton bloom demise.

Dinoflagellate cell density limits explored using Scrippsiella lachrymosa cultured in flow-through cages

Dinoflagellates constitute one of the most important groups of primary producers and micro-zooplankton on earth, common in both marine and freshwater environments. Despite their prominent position among phytoplankton, they are difficult to grow into dense cultures in the laboratory. This discrepancy between field and laboratory indicates serious limitations caused by the laboratory culturing conditions. A difficult to study but important factor is the constraints of enclosure in a limited volume of water. We conducted an experiment wherein the dinoflagellate Scrippsiella lachrymosa was grown in “flow cells” – 100 cm3 cylindrical cages constructed from plankton net, inserted in larger volumes of growth medium, allowing an exchange of medium without dilution of the culture. Cell numbers far exceeding the normal for culturing of this species and dinoflagellates in general were attained, even though the experiment was terminated before cultures reached stationary phase. A cell number ten times higher than under regular batch culturing was achieved (up to 340,000 cells mL−1). Pattern formation was distinct in cultures when cells were plentiful and water movements caused cell accumulation, not dispersion. High cell density concurrent with access to new growth medium promoted induction of the sexual cell cycle. The results indicate serious limitations to growth set by enclosure in a limited water volume in laboratory experiments; thus, maximum growth rates of dinoflagellates in favourable field conditions may be vastly underestimated. Cell accumulation behavior of dinoflagellates during the sexual life cycle may together with physical transport by larger forces in nature explain sudden bloom occurrences.

Oleaginous Heterotrophic Dinoflagellates—Crypthecodiniaceae

The heterotrophic Crypthecodinium cohnii is a major model for dinoflagellate cell biology, and a major industrial producer of docosahexaenoic acid, a key nutraceutical and added pharmaceutical compound. Despite these factors, the family Crypthecodiniaceae is not fully described, which is partly attributable to their degenerative thecal plates, as well as the lack of ribotype-referred morphological description in many taxons. We report here significant genetic distances and phylogenetic cladding that support inter-specific variations within the Crypthecodiniaceae. We describe Crypthecodinium croucheri sp. nov. Kwok, Law and Wong, that have different genome sizes, ribotypes, and amplification fragment length polymorphism profiles when compared to the C. cohnii. The interspecific ribotypes were supported by distinctive truncation-insertion at the ITS regions that were conserved at intraspecific level. The long genetic distances between Crypthecodiniaceae and other dinoflagellate orders support the separation of the group, which includes related taxons with high oil content and degenerative thecal plates, to be ratified to the order level. The current study provides the basis for future specific demarcation-differentiation, which is an important facet in food safety, biosecurity, sustainable agriculture feeds, and biotechnology licensing of new oleaginous models.

Symbiodiniaceae in and ex hospite have differential physiological responses under different heat stress scenarios

ABSTRACT Extreme seawater temperatures are linked to increasingly frequent cnidarian bleaching, a breakdown of the symbiosis between host and endosymbiotic dinoflagellates. This can be driven by several anthropogenic stressors, of which seawater warming is a major and frequent one. It remains unclear whether this is caused by intrinsic properties of the host or exclusively by the symbiont. This study investigates the response to elevated temperatures of dinoflagellate symbionts in and ex hospite, particularly to find out whether thermal tolerance differs between endosymbionts and free-living micro-algae. Maximum quantum yield of photosystem II (Fv/Fm ) and symbiont cell density were measured in seven different cnidarian species and five cultures of isolated endosymbionts subjected to temperatures of 26°C (control), 30°C and 34°C for 21 days. Isolated dinoflagellate cells showed a susceptibility to elevated temperatures of 30°C, evidenced by a decrease in photochemical efficiency and cell density of the cultures, as well as a progressive disintegration of cellular structures and loss of pigmentation of all but two cultures during the first week of exposure. Bleaching of coral holobionts at 30°C could be explained by reduced density of algae cells in host tissue. This effect was particularly evident in soft corals. Exposure to 34°C resulted in drastic bleaching of stony coral species, anemones and jellyfish, and even the death of soft corals. Data from this study show different responses to elevated thermal conditions across several species and different genera of symbionts and emphasize the important role of dinoflagellate symbionts in the holobiont response to elevated seawater temperatures.

A candidate transporter allowing symbiotic dinoflagellates to feed their coral hosts

The symbiotic partnership between corals and dinoflagellate algae is crucial to coral reefs. Corals provide their algal symbionts with shelter, carbon dioxide and nitrogen. In exchange, the symbiotic algae supply their animal hosts with fixed carbon in the form of glucose. But how glucose is transferred from the algal symbiont to the animal host is unknown. We reasoned that a transporter resident in the dinoflagellate cell membrane would facilitate outward transfer of glucose to the surrounding host animal tissue. We identified a candidate transporter in the cnidarian symbiont dinoflagellate Breviolum minutum that belongs to the ubiquitous family of facilitative sugar uniporters known as SWEETs (sugars will eventually be exported transporters). Previous gene expression analyses had shown that BmSWEET1 is upregulated when the algae are living symbiotically in a cnidarian host by comparison to the free-living state [1, 2]. We used immunofluorescence microscopy to localise BmSWEET1 in the dinoflagellate cell membrane. Substrate preference assays in a yeast surrogate transport system showed that BmSWEET1 transports glucose. Quantitative microscopy showed that symbiotic B. minutum cells have significantly more BmSWEET1 protein than free-living cells of the same strain, consistent with export during symbiosis but not during the free-living, planktonic phase. Thus, BmSWEET1 is in the right place, at the right time, and has the right substrate to be the transporter with which symbiotic dinoflagellate algae feed their animal hosts to power coral reefs.

First Record of The Thecate Marine Dinoflagellate Protoperidinium abei (Peridiniales, Dinophyceae) in Indonesia

Two identical dinoflagellate cell specimens were found in a seawater sample collected from Tanjung Pasir Beach (TPB; 6.01 S, 106.8 N), Banten, Indonesia and subjected to morphological characterization light microscopy (LM). The cells measured 71.97–72.86 µm long and 52.55–52.76 µm wide, fusiform or biconical-shaped, with tapered apical and antapical horns. A sharp and pointy spine was present at the antapex. The Kofoidean thecal plates arrangement pattern of the specimens was formulated as Po, 4', 2a, 7'', 3(?)c, 4s, 5''', 2''''. Based on this thecal plate tabulation formula combined with the cell morphologies, the specimens conformed to the original description of, and therefore identified as, Protoperidinium abei (Paulsen) Balech. This study constitutes the first record of P. abei in Indonesia and the first to illustrate and describe its detailed morphology using LM.

Deciphering interactions between the marine dinoflagellate Prorocentrum lima and the fungus Aspergillus pseudoglaucus.

The comprehension of microbial interactions is one of the key challenges in marine microbial ecology. This study focused on exploring chemical interactions between the toxic dinoflagellate Prorocentrum lima and a filamentous fungal species, Aspergillus pseudoglaucus, which has been isolated from the microalgal culture. Such interspecies interactions are expected to occur even though they were rarely studied. Here, a co-culture system was designed in a dedicated microscale marine-like condition. This system allowed to explore microalgal-fungal physical and metabolic interactions in presence and absence of the bacterial consortium. Microscopic observation showed an unusual physical contact between the fungal mycelium and dinoflagellate cells. To delineate specialized metabolome alterations during microalgal-fungal co-culture metabolomes were monitored by high-performance liquid chromatography coupled to high-resolution mass spectrometry. In-depth multivariate statistical analysis using dedicated approaches highlighted (1) the metabolic alterations associated with microalgal-fungal co-culture, and (2) the impact of associated bacteria in microalgal metabolome response to fungal interaction. Unfortunately, only a very low number of highlighted features were fully characterized. However, an up-regulation of the dinoflagellate toxins okadaic acid and dinophysistoxin 1 was observed during co-culture in supernatants. Such results highlight the importance to consider microalgal-fungal interactions in the study of parameters regulating toxin production.

Responses of dinoflagellate cells to ultraviolet-C irradiation.

Dinoflagellates are important aquatic microbes and major harmful algal bloom (HAB) agents that form invasive species through ship ballast transfer. UV-C installations are recommended for ballast treatments and HAB controls, but there is a lack of knowledge in dinoflagellate responses to UV-C. We report here dose-dependent cell cycle delay and viability loss of dinoflagellate cells irradiated with UV-C, with significant proliferative reduction at 800 Jm-2 doses or higher, but immediate LD50 was in the range of 2400-3200 Jm-2 . At higher dosages, some dinoflagellate cells surprisingly survived after days of recovery incubation, and continued viability loss, with samples exhibiting DNA fragmentations per proliferative resumption. Sequential cell cycle postponements, suggesting DNA damages were repaired over one cell cycle, were revealed with flow cytometric analysis and transcriptomic analysis. Over a sustained level of other DNA damage repair pathways, transcript elevation was observed only for several components of base pair repair and mismatch repair. Cumulatively, our findings demonstrated special DNA damage responses in dinoflagellate cells, which we discussed in relation to their unique chromo-genomic characters, as well as indicating resilience of dinoflagellate cells to UV-C.

Deciphering the potential of Ba/Ca, Mo/Ca and Li/Ca profiles in the bivalve shell Pecten maximus as proxies for the reconstruction of phytoplankton dynamics

Shells of the fast-growing bivalve Pecten maximus serve as a geochemical archive for the reconstruction of past phytoplankton dynamics. Specifically, high-resolution, temporally accurately aligned molar barium-to-calcium (Ba/Cashell), molybdenum-to-calcium (Mo/Cashell) and lithium-to-calcium ratios (Li/Cashell) of the shell calcite revealed distinct peaks which are closely linked to phytoplankton dynamics. Yet, the development and applicability of these geochemical proxies is still at an early stage and needs further calibration. In this study, we examined the relationship between the timing and magnitude of Ba/Cashell, Mo/Cashell and Li/Cashell peaks of P. maximus and the occurrence of various phytoplankton species (diatoms and dinoflagellates) from a statistical perspective. Studied shell samples (three specimens per calendar year) as well as detailed phytoplankton observation data were derived from the well-studied costal ecosystem of the Bay of Brest (France) over three years (2011, 2012 and 2019). An algorithm-based pseudo-random sampling simulation technique was established that analyzed the complex phytoplankton datasets with respect to the profiles of Ba/Cashell, Mo/Cashell and Li/Cashell to identify potential patterns between phytoplankton and trace element time-series. The simulation results indicate that the timing and magnitude of Ba/Cashell, Mo/Cashell and Li/Cashell peaks agreed best with the occurrence of specific phytoplankton blooms that developed ca. one to two weeks earlier. The data suggest that the formation of transient Ba/Cashell peaks had a plurispecific origin, i.e., potentially linked to blooms of ingestible diatom, dinoflagellate and flagellate species enriched in Ba that occurred 8 to 12 days earlier. Observed peaks in Mo/Cashell profiles demonstrably followed the timing and intensity of blooms of the dominant dinoflagellate genus Gymnodinium spp. after a short time lag of around 8 days, potentially linked to an enhanced enzyme activity of nitrate reductase that requires the presence of Mo in the dinoflagellate cells. In addition, Mo/Cashell peaks agreed with periods of diatom aggregate formation which were hypothesized to induce the formation of Mo/Cashell peaks in scallop shells. Li/Cashell profiles revealed similar patterns as large blooms of the diatom Chaetoceros spp. as well as to neurotoxin producing diatoms of the genus Pseudo-nitzschia when considering a short time lag of 8 to 12 days. These findings highlight the great potential of using Ba/Cashell, Mo/Cashell and Li/Cashell chronologies in P. maximus shells as proxies of past phytoplankton dynamics.