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This study explores the use of chemical epigenetic modifiers to enhance the production of bioactive molecules in the dinoflagellate Amphidinium carterae. It was selected for its known ability to produce amphidinols with various pharmacological and agro-industrial applications. Growth inhibition, photosynthetic yield reduction, and stress-related variables were observed in response to different treatments using 5-azacitidine, hydroxamic suberoylanilide acid, metyrapone, tricyclazole, sodium butyrate, and jasplakinolide. Although no evidence of amphidinols overproduction was found, hydroxamic suberoylanilide acid and tricyclazole showed potential for stimulating the synthesis of antioxidant carotenoids such as peridinin and their derivatives. An increase of around 200% and 170% relative to control was observed at the highest non-lethal concentration. More research is required to elucidate the mechanisms underlying this response and to optimize production conditions. This pioneering research opens up new avenues for the biotechnological exploitation of dinoflagellate microalgae. [ABSTRACT FROM AUTHOR]

Background: In the search of a rapid and representative method for the approximate calculation of culture density and cell content of useful pigments, the study of absorption spectra of cultures of the dinoflagellate Amphidinium carterae, the haptophyte Isochrysis galbana, the chlorophyte Nephroselmis sp., and the filamentous cyanobacterium Anabaena sp. were selected as representative species of different taxa. Methods: The experimental cultures were established in small volumes by the discontinuous method under 20–21 °C, salinity of 30 or 40 ppt, and 2000 or 8000 lux illumination, and the absorbance spectra, density of the culture and concentration of the extracted pigments chlorophyll, total carotenoids, and phycocyanin were recorded during cultivation. Results: Algal density can be predicted sufficiently correctly because the regression equation of the correlation of the OD value of 750 nm from each absorption spectrum and the measured algal biomass was very strong. The same is true for the corresponding correlations between OD 750 nm and the detected pigments. Conclusions: Absorption spectra of microalgal cultures can be a simple, inexpensive, and non-invasive method for the growers to obtain the necessary information for predicting the right time to collect an ideal combination of maximum biomass and useful pigments, provided that the interpretation of the spectra is performed according to the method described herein.

Background: In the search of a rapid and representative method for the approximate calculation of culture density and cell content of useful pigments, the study of absorption spectra of cultures of the dinoflagellate Amphidinium carterae, the haptophyte Isochrysis galbana, the chlorophyte Nephroselmis sp., and the filamentous cyanobacterium Anabaena sp. were selected as representative species of different taxa. Methods: The experimental cultures were established in small volumes by the discontinuous method under 20-21 °C, salinity of 30 or 40 ppt, and 2000 or 8000 lux illumination, and the absorbance spectra, density of the culture and concentration of the extracted pigments chlorophyll, total carotenoids, and phycocyanin were recorded during cultivation. Results: Algal density can be predicted sufficiently correctly because the regression equation of the correlation of the OD value of 750 nm from each absorption spectrum and the measured algal biomass was very strong. The same is true for the corresponding correlations between OD 750 nm and the detected pigments. Conclusions: Absorption spectra of microalgal cultures can be a simple, inexpensive, and non-invasive method for the growers to obtain the necessary information for predicting the right time to collect an ideal combination of maximum biomass and useful pigments, provided that the interpretation of the spectra is performed according to the method described herein. [ABSTRACT FROM AUTHOR]

Dinoflagellates make up the second largest marine group of marine unicellular eukaryotes in the world ocean and comprise both heterotrophic and autotrophic species, encompassing a wide genetic and chemical diversity. They produce a plethora of secondary metabolites that can be toxic to other species and are mainly used against predators and competing species. Dinoflagellates are indeed often responsible for harmful algal bloom, where their toxic secondary metabolites can accumulate along the food chain, leading to significant damages to the ecosystem and human health. Secondary metabolites from dinoflagellates have been widely investigated for potential biomedical applications and have revealed multiple antimicrobial, antifungal, and anticancer properties. Species from the genus Amphidinium seem to be particularly interesting for the production of medically relevant compounds. The present review aims at summarising current knowledge on the diversity and the pharmaceutical properties of secondary metabolites from the genus Amphidinium. Specifically, Amphidinium spp. produce a range of polyketides possessing cytotoxic activities such as amphidinolides, caribenolides, amphidinins, and amphidinols. Potent antimicrobial properties against antibiotic-resistant bacterial strains have been observed for several amphidinins. Amphidinols revealed instead strong activities against infectious fungi such as Candida albicans and Aspergillus fumigatus. Finally, compounds such as amphidinolides, isocaribenolide-I, and chlorohydrin 2 revealed potent cytotoxic activities against different cancer cell lines. Overall, the wide variety of antimicrobial, antifungal, and anticancer properties of secondary metabolites from Amphidinium spp. make this genus a highly suitable candidate for future medical applications, spanning from cancer drugs to antimicrobial products that are alternatives to currently available antibiotic and antimycotic products. [ABSTRACT FROM AUTHOR]

The genus Amphidinium is shown in many phylogenies to be basal to other peridinin-containing, photosynthetic dinoflagellates as one of the first photosynthetic genera to arise after the evolution of heterotrophic genera. As part of our continuing examination of the plastid-associated galactolipids, namely mono- and digalactosyldiacylglycerol (MGDG and DGDG, respectively), in dinoflagellates, we here examine the galactolipid composition of members of the genus Amphidinium. We show that this genus is characterized by an abundance of 20:5(n-3)/18:5(n-3) and 20:5(n-3)/18:4(n-3) forms of MGDG and DGDG (with sn-1/sn-2 regiochemical specificity of fatty acids), but also sometimes with generally lesser amounts of some polyunsaturated C18/C18 forms, thus placing the examined species within a previously identified cluster of C20/C18 MGDG- and DGDG-containing, peridinin-containing dinoflagellates. We also show that Testudodinium testudo, previously known as Amphidinium testudo, conversely falls within a previously identified C18/C18 cluster, indicating a distinct difference in galactolipid biosynthesis capability. While it is likely that further revision of the genus may occur in the future and/or more basal peridinin-containing, photosynthetic genera may be discovered, at the current time Amphidinium is the currently agreed-upon most basal dinoflagellate genus for which isolates are available for biochemical characterization such as what we describe in this paper. Thus, because of the presumed basal position of the genus Amphidinium, we present a hypothesis that its galactolipids currently represent those that are ancestral to other genera of peridinin-containing dinoflagellates, including those within the C18/C18 cluster. Amphidinium species' galactolipids reside within the C20/C18 peridinin dinoflagellate cluster. Conversely, Testudodinium testudo (formerly Amphidinium testudo) falls within the C18/C18 cluster. We hypothesize Amphidinium's galactolipids as basal to other peridinin dinoflagellates. [ABSTRACT FROM AUTHOR]

The dinoflagellates Amphidinium carterae and Amphidinium corpulentum have been previously characterized as having Δ8(14)‐nuclear unsaturated 4α‐methyl‐5α‐cholest‐8(14)‐en‐3β‐ol (C28:1) and 4α‐methyl‐5α‐ergosta‐8(14),24(28)‐dien‐3β‐ol (amphisterol; C29:2) as predominant sterols, where they comprise approximately 80% of the total sterol composition. These two sterols have hence been considered as possible major sterol biomarkers for the genus. Here, we have examined the sterols of four recently identified species of Amphidinium (Amphidinium fijiense, Amphidinium magnum, Amphidinium theodori, and Amphidinium tomasii) that are closely related to Amphidinium operculatum as part of what is termed the Operculatum Clade to show that each species has its sterol composition dominated by the common dinoflagellate sterol cholesterol (cholest‐5‐en‐3β‐ol; C27:1), which is found in many other dinoflagellate genera, rather than Δ8(14) sterols. While the Δ8(14) sterols 4α‐methyl‐5α‐cholest‐8(14)‐en‐3β‐ol and 4α,23,24‐trimethyl‐5α‐cholest‐8(14),22E‐dien‐3β‐ol (C30:2) were present as minor sterols along with another common dinoflagellate sterol, 4α,23,24‐trimethyl‐5α‐cholest‐22E‐en‐3β‐ol (dinosterol; C30:1), in some of these four species, amphisterol was not conclusively observed. From a chemotaxonomic perspective, while this does reinforce the genus Amphidinium's ability to produce Δ8(14) sterols, albeit here as minor sterols, these results demonstrate that caution should be used when considering Δ8(14) sterols, especially amphisterol, as Amphidinium‐specific biomarkers within these species where cholesterol is the predominant sterol. [ABSTRACT FROM AUTHOR]

Testudodinium testudo is a peridinin‐containing dinoflagellate recently renamed from Amphidinium testudo. While T. testudo has been shown via phylogenetic analysis of small subunit ribosomal RNA genes to reside in a clade separate from the genus Amphidinium, it does possess morphological features similar to Amphidinium sensu stricto. Previous studies of Amphidinium carterae and Amphidinium corpulentum have found the sterols to be enriched in Δ8(14) sterols, such as 4α‐methyl‐5α‐ergosta‐8(14),24(28)‐dien‐3β‐ol (amphisterol), uncommon to most other dinoflagellate taxa and thus considered possible biomarkers for the genus Amphidinium. Here, we provide an examination of the sterols of T. testudo and show they are dominated not by amphisterol, but rather by a different Δ8(14) sterol, (24R)‐4α‐methyl‐5α‐ergosta‐8(14),22‐dien‐3β‐ol (gymnodinosterol), previously thought to be a major sterol only within the Kareniaceae genera Karenia, Karlodinium, and Takayama. Also found to be present at low levels were 4α‐methyl‐5α‐ergosta‐8,14,22‐trien‐3β‐ol, a sterol previously observed in Karenia brevis to be an intermediate in the production of gymnodinosterol, and cholesterol, a sterol common to many other dinoflagellates. The presence of gymnodinosterol in T. testudo is the first report of this sterol as the sole major sterol in a dinoflagellate outside of the Kareniaceae. The implication of this chemotaxonomic relationship to the Kareniaceae is discussed. [ABSTRACT FROM AUTHOR]

While a longstanding method for identifying dinoflagellates, morphology alone can be inaccurate because of convergent evolution of truly different species toward a particular cellular shape. The dinoflagellate genus Togula is a case in point in that its type species Togula britannica was previously assigned to the genus Amphidinium based on morphology but was recently recognized as an independent genus following phylogenetic characterization. Chemotaxonomy can be a useful tool to complement the characterization of dinoflagellates by phylogeny. To this point, some, but not all, species of Amphidinium, such as Amphidinium carterae, have been observed to produce the rare 4α-methyl-substituted, Δ 8(14) -nuclear-unsaturated major sterol 4α-methyl-5α-ergosta-8(14),24(28)-dien-3β-ol (amphisterol) that has historically been considered a potential chemotaxonomic biomarker for the genus as it is found in very few other dinoflagellate genera. To date, no isolates within the genus Togula have had their sterols characterized; our objective was thus to provide a first examination of the sterols of an isolate of Togula to compare to the sterols of Amphidinium. To this end, we have characterized the sterols of Togula jolla, one of the few members of Togula available for study, to demonstrate the production of amphisterol, among other Δ 8(14) -nuclear unsaturated sterols, as its major sterol., (© 2025 International Society of Protistologists.)

The dinoflagellate Amphidinium carterae is a promising source of unique bioactive compounds with potential applications in diverse industries. Abiotic strategies for increasing bioactive cell quota have been preferred for cost-effectiveness, controllability, and existing knowledge. The supernatants of Heterosigma akashiwo and Pavlova sp. enhanced haemolytic activity of A. carterae by 2.3 times and improved its final biomass by 10 % when added to the growth medium. Prymnesium parvum supernatant increased the total lipid content of A. carterae by 50 %, while Pleurochrysis roscoffensis supernatant boosted its total carotenoid content by 40 %. This study demonstrated a rapid screening strategy for evaluating growth responses and highlights the potential of using allelopathic strategies to enhance bioactive metabolite production. Utilizing supernatants for the formulation of growth media fosters a circular economy by repurposing resources, minimizing waste, and promoting sustainability in the microalgal biotechnological industry. [Display omitted] • 38 microalgal supernatants were evaluated for stimulating A. carterae. • Quorum sensing allowed increasing total lipid and carotenoid content. • Evidence of amphidinols overproduction was found for many species. • Repurposing supernatants as a biostimulant enhances circularity. [ABSTRACT FROM AUTHOR]

In the quest of finding local strains of marine ciliates that can be easily cultured under a broad range of salinity and fed with microalgae, Fabrea salina Henneguy, 1890 and Condylostoma sp. Bory de St. Vincent, 1826 were cultured for 22 days in small volumes at a temperature of 16–18 °C and fed with flagellated microalgae. F. salina presented a clear preference for the salinity of 40 ppt and Condylostoma. sp. for 20 ppt. Rhodomonas salina Hill and Wetherbee, 1989 were the most efficient feeds, resulting in 30 ind./mL in F. salina and 73 ind./mL in Condylostoma. Dunaliella salina Teodoresco, 1905 and Nephroselmis sp. F. Stein, 1878 also resulted in considerable ciliate densities while Isochrysis galbana Parke, 1949 came last with the highest density in Condylostoma. The strain of Tetraselmis sp. F. Stein, 1878 (var. red pappas) which is transformed in immobilized palmelloid cells and the dinoflagellate Amphidinium carterae Hulburt, 1957, which is suspected of toxin production, were inappropriate for both ciliates. These ciliates can be easily cultured and can serve as useful organisms in bioassays and probably as live food in marine fish hatcheries. [ABSTRACT FROM AUTHOR]

Abstract Background Some dinoflagellates cause harmful algal blooms, releasing toxic secondary metabolites, to the detriment of marine ecosystems and human health. Our understanding of dinoflagellate toxin biosynthesis has been hampered by their unusually large genomes. To overcome this challenge, for the first time, we sequenced the genome, microRNAs, and mRNA isoforms of a basal dinoflagellate, Amphidinium gibbosum, and employed an integrated omics approach to understand its secondary metabolite biosynthesis. Results We assembled the ~ 6.4-Gb A. gibbosum genome, and by probing decoded dinoflagellate genomes and transcriptomes, we identified the non-ribosomal peptide synthetase adenylation domain as essential for generation of specialized metabolites. Upon starving the cells of phosphate and nitrogen, we observed pronounced shifts in metabolite biosynthesis, suggestive of post-transcriptional regulation by microRNAs. Using Iso-Seq and RNA-seq data, we found that alternative splicing and polycistronic expression generate different transcripts for secondary metabolism. Conclusions Our genomic findings suggest intricate integration of various metabolic enzymes that function iteratively to synthesize metabolites, providing mechanistic insights into how dinoflagellates synthesize secondary metabolites, depending upon nutrient availability. This study provides insights into toxin production associated with dinoflagellate blooms. The genome of this basal dinoflagellate provides important clues about dinoflagellate evolution and overcomes the large genome size, which has been a challenge previously.

Biologically rich rock-pools (RPs) are prominent component of rocky intertidal habitat and this study elucidates the factors affecting the phytoplankton distribution from high-tide (HT), mid-tide (MT) and low-tide (LT) RPs (Anjuna rocky shores, Goa, India), during the summer season. Results revealed that the RP location determines the nature of phytoplankton assemblage. MT-RPs and LT-RPs supported rich microalgal diversity (species number/morphological shapes) than HT-RPs. However, the cell abundance was higher in latter than former RPs. In frequently flushed MT-RPs and LT-RPs, pennate diatoms (Pseudonitzschia, Nitzschia, Thalasionema, Navicula, and Licmophora) representing elongated shapes dominated, whereas in stagnant HT-RPs spherical/combined shaped dinoflagellates (Amphidinium carterae sensu stricto and Bysmatrum gregarium) dominated. Interestingly, even though the taxonomic assemblages changed temporally the shape dominance remained consistent suggesting shape as a robust morphological trait. Additionally, the study also revealed the existence of species-specific allelopathy on other microalgae by Amphidinium (potential harmful algae) indicating a type of adaptive strategy, in addition to eurytolerant capabilities, for its predominance in HT-RPs, which experiences varying environmental (light, temperature, and salinity) conditions. Research Highlights: Investigated factors influencing distribution of phytoplankton (species and shape-wise) in intertidal rock pools (RPs) of Anjuna rocky shores, Goa. Location of RPs determined nature of phytoplankton community, i.e., less diversity and high abundance in high-tide RPs than mid/low-tide RPs. Spherical-shaped dinoflagellates and elongated shape diatoms dominated phytoplankton community of high and mid/low tide RPs, respectively. Even though the species composition changed temporally the shape dominance remained consistent suggesting the latter as a robust morphological trait. Dominant dinoflagellate, Amphidinium caterae sensu stricto, also showed species-specific allelopathy on other phytoplankton. [ABSTRACT FROM AUTHOR]

The occurrence of benthic dinoflagellates, many of which are known to be toxic, is a critical concern for scientists, government officers, and people in the aquaculture, dining, and tourism industries. The interest in these dinoflagellates in countries with temperate climate is increasing because tropical or subtropical species introduced into temperate waters by currents are able to survive the winter season in the new environment owing to global warming. Recently, several species from the benthic dinoflagellate genera Amphidinium, Coolia, Ostreopsis, Gambierdiscus, and Prorocentrum have been reported in the waters of the South and East Sea of Korea. The advent of the benthic dinoflagellates in Korean waters is especially important because raw or slightly cooked seaweeds, which may harbor these benthic dinoflagellates, as well as raw fish, which can be potentially intoxicated by phytotoxins produced by some of these benthic dinoflagellates, are part of the daily Korean diet. The recent increase in temperature of Korean coastal waters has allowed for the expansion of benthic dinoflagellate species into these regions. In the present study, we reviewed the species, distribution, and toxicity of the benthic dinoflagellates that have been reported in Korean waters. We also provided an insight into the ecological and socio-economic importance of the occurrence of benthic dinoflagellates in Korean waters. [ABSTRACT FROM AUTHOR]

Background: Some dinoflagellates cause harmful algal blooms, releasing toxic secondary metabolites, to the detriment of marine ecosystems and human health. Our understanding of dinoflagellate toxin biosynthesis has been hampered by their unusually large genomes. To overcome this challenge, for the first time, we sequenced the genome, microRNAs, and mRNA isoforms of a basal dinoflagellate, Amphidinium gibbosum, and employed an integrated omics approach to understand its secondary metabolite biosynthesis. Results: We assembled the ~ 6.4-Gb A. gibbosum genome, and by probing decoded dinoflagellate genomes and transcriptomes, we identified the non-ribosomal peptide synthetase adenylation domain as essential for generation of specialized metabolites. Upon starving the cells of phosphate and nitrogen, we observed pronounced shifts in metabolite biosynthesis, suggestive of post-transcriptional regulation by microRNAs. Using Iso-Seq and RNA-seq data, we found that alternative splicing and polycistronic expression generate different transcripts for secondary metabolism. Conclusions: Our genomic findings suggest intricate integration of various metabolic enzymes that function iteratively to synthesize metabolites, providing mechanistic insights into how dinoflagellates synthesize secondary metabolites, depending upon nutrient availability. This study provides insights into toxin production associated with dinoflagellate blooms. The genome of this basal dinoflagellate provides important clues about dinoflagellate evolution and overcomes the large genome size, which has been a challenge previously. [ABSTRACT FROM AUTHOR]

There is growing interest in finding microalgae species that efficiently convert dissolved nutrients contained in aquaculture effluents into highly valuable biomass. The different nitrogen forms that are present in aquaculture effluents are particularly concerning. This study demonstrated that the dinoflagellate Amphidinium carterae can acclimate to both combined and sole nitrogen sources such as nitrate, ammonium, and urea over a wide concentration range. As far as is known, it is the first time that a species of the genus Amphidinium has been successfully cultured with urea as the sole source of nitrogen. In the presence of 882 μM of nitrate, A. carterae tolerated urea concentrations up to 5000 μM. With respect to ammonium-N tolerance, it has been observed that it is lethal at concentrations higher than 441 μM. A robust laboratory experimental design was critical for accurately assessing this acclimation. Alternative N sources did not affect the production of high-value specific polyketide secondary metabolites from A. carterae, such as amphidinols, with an average concentration of 0.435 ± 0.038% biomass d.w. An analysis of the symbiotic microbial assemblages developed in a long-term A. carterae culture in an open raceway pond, and the fact that it is able to metabolize all three nitrogen sources simultaneously, supports the idea that this microalga has the potential to be successfully cultured with aquaculture effluents. [ABSTRACT FROM AUTHOR]

After a 1.5-year screening survey in the lagoons of Western Greece in order to isolate and culture sturdy species of microalgae for aquaculture or other value-added uses, as dictated primarily by satisfactory potential for their mass culture, five species emerged, and their growth was monitored in laboratory conditions. Amphidinium carterae, Nephroselmis sp., Tetraselmis sp. (var. red pappas), Asteromonas gracilis, and Dunaliella sp. were batch cultured using low (20 ppt), sea (40 ppt), and high salinity (50 or 60 or 100 ppt) and in combination with low (2000 lux) and high (8000 lux) intensity illumination. The results exhibited that all these species can be grown adequately in all salinities and with the best growth in terms of maximum cell density, specific growth rate (SGR), and biomass yield (g dry weight/L) at high illumination (8000 lux). The five species examined exhibited different responses in the salinities used, whereby Amphidinium clearly performs best in 20 ppt, far better than 40 ppt, and even more so than 50 ppt. Nephroselmis and Tetraselmis grow almost the same in 20 and 40 ppt and less well in 60 ppt. Asteromonas performs best in 100 ppt, although it can grow quite well in both 40 and 60 ppt. Dunaliella grows equally well in all salinities (20, 40, 60 ppt). Concerning the productivity, assessed as the maximum biomass yield at the end of the culture period, the first rank is occupied by Nephroselmis with ~3.0 g d.w./L, followed by Tetraselmis (2.0 g/L), Dunaliella (1.58 g/L), Amphidinium (1.19 g/L), and Asteromonas (0.7 g/L) with all values recorded at high light (8000 lux).

Coral reefs are some of the most important and ecologically diverse marine environments. At the base of the reef ecosystem are dinoflagellate algae, which live symbiotically within coral cells. Efforts to understand the relationship between alga and coral have been greatly hampered by the lack of an appropriate dinoflagellate genetic transformation technology. By making use of the plasmid-like fragmented chloroplast genome, we have introduced novel genetic material into the dinoflagellate chloroplast genome. We have shown that the introduced genes are expressed and confer the expected phenotypes. Genetically modified cultures have been grown for 1 year with subculturing, maintaining the introduced genes and phenotypes. This indicates that cells continue to divide after transformation and that the transformation is stable. This is the first report of stable chloroplast transformation in dinoflagellate algae.

Highlights • Multi-step extraction processes for carotenoids, fatty acids and APDs were assessed. • Each step was optimized for dinoflagellates using previously reported methodologies. • Total recoveries were 97% for carotenoids, 80% for fatty acids and 100% for APDs. Abstract Sustainable dinoflagellate microalgae-based bioprocess designed to produce secondary metabolites (SMs) with interesting bioactivities are attracting increasing attention. However, dinoflagellates also produce other valuable bioproducts (e.g polyunsaturated fatty acids, carotenoids, etc.) that could be recovered and should therefore be taken into account in the bioprocess. In this study, biomass of the marine dinoflagellate microalga Amphidinium carterae was used to assess and optimise three different methods in order to obtain three families of high-value biochemical compounds present in the biomass. The existing processes encompassed a multi-step extraction process for carotenoids, fatty acids and APDs individually and are optimized for the integral valorization of raw A. carterae biomass, with SMs being the primary target compounds. Total process recovery yields were 97% for carotenoids, 80% for total fatty acids and 100% for an extract rich in APDs (not purified). [ABSTRACT FROM AUTHOR]

Iriomoteolides-9a ( 1) and 11a ( 2), new 15- and 19-membered macrolides, respectively, have been isolated from the marine dinoflagellate Amphidinium species (strain KCA09052). Compounds 1 and 2 were obtained from the extracts of the algal cells inoculated in the PES and TKF seawater medium, respectively. The structures of 1 and 2 were assigned on the basis of detailed NMR analyses. Compounds 1 and 2 exhibited cytotoxic activity against human cervix adenocarcinoma HeLa cells. [ABSTRACT FROM AUTHOR]

A new marine benthic dinoflagellate, Testudodinium magnum sp. nov., was described from a sand sample collected on the seafloor at a depth of 35 m off Mageshima Island, Kagoshima, Japan. The dinoflagellate possessed the characteristic features of the genus Testudodinium, but was distinguished by its extremely large size and pebbled dorsal surface of the hyposome, caused by the presence of numerous nodules. The cells of this dinoflagellate possessed a dominant sessile and a rare motile form that were morphologically distinct from each other: The sessile form was larger and the small rounded episome with longitudinal furrow was completely embedded in the hyposome; the motile form was smaller and the episome was only partly embedded in the hyposome. The ultrastructural investigations revealed the presence of a large, circular, starch-sheathed pyrenoid whose matrix was traversed by randomly arranged thylakoid lamellae. The chloroplasts formed a network radiating outward from the pyrenoid. Uniquely, this dinoflagellate possessed internal props - structures that spanned the thickness of the cell and were fibrous in nature. The props were numerous and passed through organelles and cell contents. Phylogenetic analyses based on the small-subunit ribosomal DNA gene sequences placed this dinoflagellate firmly within the Testudodinium clade with high support. On the basis of the morphological features and phylogenetic analyses, we concluded that this dinoflagellate was a new species in the genus Testudodinium. [ABSTRACT FROM AUTHOR]

Two new members of the amphidinol family, amphidinol A (1) and its 7-sulfate derivative amphidinol B (2), were isolated from a strain of Amphidinium carterae of Lake Fusaro, near Naples (Italy), and chemically identified by spectroscopic and spectrometric methods. Amphidinol A showed antifungal activity against Candida albicans (MIC = 19 µg/mL). Biosynthetic experiments with stable isotope-labelled acetate allowed defining the elongation process in 1. For the first time the use of glycolate as a starter unit in the polyketide biosynthesis of amphidinol metabolites was unambiguously demonstrated.

A novel linear polyketide, amphirionin-2 ( 1 ), with two unique hexahydrofuro[3,2- b ]furan moieties has been isolated from the cultivated algal cells of a benthic dinoflagellate Amphidinium sp. (strain KCA09051). The structure was elucidated on the basis of detailed analyses of 2D NMR data, and the absolute configuration of C-5 was determined by using modified Mosher’s method. Amphirionin-2 ( 1 ) exhibited potent cytotoxic activity against human colon carcinoma Caco-2 cells and human lung adenocarcinoma A549 cells. [ABSTRACT FROM AUTHOR]

Cells of five unarmoured kleptoplastidic dinoflagellates, Amphidinium latum , Amphidinium poecilochroum , Gymnodinium amphidinioides, Gymnodinium acidotum and Gymnodinium aeruginosum were observed under light and/or scanning electron microscopy and subjected to single-cell PCR. The SSU rDNA and the partial LSU rDNA of all the examined species were sequenced, and the SSU rDNA of G. myriopyrenoides was sequenced. Phylogenetic analyses revealed that the unarmoured kleptoplastidic species formed a monophyletic clade within the Gymnodinium -clade sensu Daugbjerg et al. (2000) . The sister taxa for this clade were Gymnodinium palustre and Spiniferodinium galeiforme , both of which possess brown-coloured chloroplasts. The results indicated that acquisition of kleptoplastidy in these unarmoured dinoflagellates was a single event and that these unarmoured kleptoplastidic dinoflagellates may have evolved from a form with permanent chloroplasts. Molecular trees suggested that the acquisition of kleptoplastidy took place in a marine habitat and later some species colonized the freshwater habitat. Because these unarmoured kleptoplastidic dinoflagellates are monophyletic and characterized by distinct morphological and cytological features (including the presence of the same type of apical groove, absence of nuclear chambers in the nuclear envelope, absence of genuine chloroplasts, and the possession of kleptochloroplasts), we propose the establishment of a new genus, Nusuttodinium , to accommodate all these dinoflagellates. [ABSTRACT FROM AUTHOR]

Abstract: A novel linear polyketide, amphirionin-5 (1), has been isolated from the cultivated algal cells of the benthic dinoflagellate Amphidinium sp. (strain KCA09053), and the structure was elucidated on the basis of detailed analyses of 1D and 2D NMR data. Amphirionin-5 (1) resulted in a 282% increase in the proliferation of murine bone-marrow derived stromal ST-2 cells and 320% increase in the proliferation of murine osteoblastic MC3T3-E1 cells. [Copyright &y& Elsevier]

Abstract: Iriomoteolide-13a (1) has been isolated from a benthic dinoflagellate Amphidinium sp. (strain KCA09053) as a new 22-membered macrolide containing one hexahydrofuro[3,2-b]furan ring, one tetrahydropyran ring, two tetrahydrofuran rings, three one-carbon branches, and three hydroxyl groups including two hemiketals. The structure of 1 was assigned on the basis of a detailed 2D NMR analysis. Compound 1 exhibited cytotoxic activity against human cervix adenocarcinoma HeLa cells (IC50: 0.5 μg/mL). [Copyright &y& Elsevier]

Highlights: [•] Coolia malayensis is a common dinoflagellate species in New Zealand's sub-tropical northern waters. [•] New Zealand C. malayensis isolates reported as C. monotis are reclassified based on analysis of LSU rDNA sequence data. [•] The dinoflagellates Amphidinium thermaeum and Gambierdiscus cf. yasumotoi are new reports for Northland, New Zealand. [Copyright &y& Elsevier]

A new athecate dinoflagellate, Bispinodinium angelaceum N. Yamada et Horiguchi gen. et sp. nov., is described from a sand sample collected on the seafloor at a depth of 36 m off Mageshima Island, subtropical Japan. The dinoflagellate is dorsiventrally compressed and axi-symmetric along the sulcus. The morphology resembles that of the genus Amphidinium sensu lato by having a small epicone that is less than one third of the total cell length. However, it has a new type of apical groove, the path of which traces the outline of a magnifying glass. The circular component of this path forms a complete circle in the center of the epicone and the straight 'handle' runs from the sulcus to the circular component. Inside the cell, a pair of elongated fibrous structure termed here the 'spinoid apparatus' extends from just beneath the circular apical groove to a point near the nucleus. Each of two paired structures consists of at least 10 hyaline fibers and this is a novel structure found in dinoflagellates. Phylogenetic analyses based on the SSU and LSU RNA genes did not show any high bootstrap affinities with currently known athecate dinoflagellates. On the basis of its novel morphological features and molecular signal, we conclude that this dinoflagellate should be described as a new species belonging to a new genus. [ABSTRACT FROM AUTHOR]

A new athecate dinoflagellate, Bispinodinium angelaceum N. Yamada et Horiguchi gen. et sp. nov., is described from a sand sample collected on the seafloor at a depth of 36 m off Mageshima Island, subtropical Japan. The dinoflagellate is dorsiventrally compressed and axi-symmetric along the sulcus. The morphology resembles that of the genus Amphidinium sensu lato by having a small epicone that is less than one third of the total cell length. However, it has a new type of apical groove, the path of which traces the outline of a magnifying glass. The circular component of this path forms a complete circle in the center of the epicone and the straight 'handle' runs from the sulcus to the circular component. Inside the cell, a pair of elongated fibrous structure termed here the 'spinoid apparatus' extends from just beneath the circular apical groove to a point near the nucleus. Each of two paired structures consists of at least 10 hyaline fibers and this is a novel structure found in dinoflagellates. Phylogenetic analyses based on the SSU and LSU RNA genes did not show any high bootstrap affinities with currently known athecate dinoflagellates. On the basis of its novel morphological features and molecular signal, we conclude that this dinoflagellate should be described as a new species belonging to a new genus. [ABSTRACT FROM AUTHOR]

Dinoflagellates are a diverse protist group possessing many unique traits. These include (but are not limited to) expansive genomes packaged into permanently condensed chromosomes, photosynthetic or cryptic plastids acquired vertically or horizontally in serial endosymbioses, and a ruffle-like transverse flagellum attached along its length to the cell. When reconstructing character evolution, early branching lineages with unusual features that distinguish them from the rest of the group have proven useful for inferring ancestral states. The Noctilucales are one such lineage, possessing relaxed chromosomes in some life stages and a trailing, thread-like transverse flagellum. However, most of the cellular and molecular data for the entire group come from a single cultured species, Noctiluca scintillans , and because its phylogenetic position is unresolved it remains unclear if these traits are ancestral or derived. Here, we use single cell transcriptomics to characterize three diverse Noctilucales genera: Spatulodinium, Kofoidinium , and a new lineage, Fabadinium gen. nov. We also provide transcriptomes for undescribed species in Amphidinium and Abediniales, critical taxa for clarifying the phylogenetic position of Noctilucales. Phylogenomic analyses suggests that the Noctilucales are sister to Amphidinium rather than an independent branch outside the core dinoflagellates. This topology is consistent with observations of shared characteristics between some members of Noctilucales and Amphidinium and provides the most compelling evidence to date that the unusual traits within this group are derived rather than ancestral. We also confirm that Spatulodinium plastids are photosynthetic and of ancestral origin, and show that all non-photosynthetic Noctilucales retain plastid genes indicating a cryptic organelle., (© The Author(s) 2022. Published by Oxford University Press on behalf of National Academy of Sciences.)

SUMMARY A new genus of sand-dwelling photosynthetic dinoflagellate, Testudodinium Horiguchi, Tamura, Katsumata et A. Yamaguchi is proposed based on Testudodinium testudo (Herdman) Horiguchi, Tamura, Katsumata, et A. Yamaguchi comb. nov. (Basionym: Amphidinium testudo Herdman) and a new species in this new genus, Testudodinium maedaense Katsumata et Horiguchi sp. nov. is described. Amphidinium corrugatum is also transferred to this genus, making a new combination T. corrugatum (Larsen et Patterson) Horiguchi, Tamura et A. Yamaguchi. These three species are similar to the members of the genus Amphidinium in having an extremely small episome and a dorsoventrally flattened cell body. They are, however, distinguished from the genus Amphidinium seusu stricto by the possession of a distinct longitudinal furrow in the middle of ventral side of the episome. Phylogenetic trees based on small subunit (SSU) rDNA revealed that all three of these Testudodinium species formed a robust clade and, although statistical support is not high, the tree suggests Testudodinium clade is not closely related to Amphidinium seusu stricto clade. The morphological differences together with molecular data support the establishment of a new genus for A. testudo and its related species. [ABSTRACT FROM AUTHOR]

The occurrence of harmful epiphytic dinoflagellates is of concern to scientists, the aquaculture industry, and government due to their toxicity not only to marine organisms but also to humans. There have been no studies on the abundance of the epiphytic dinoflagellates in Korean waters. We explored the presence of epiphytic dinoflagellates in the coastal waters off Jeju Island, southwestern Korea. Furthermore, we measured the abundance of epiphytic dinoflagellates on the thalli of 24 different macroalgae, collected from five different locations in October 2009. Five epiphytic dinoflagellate genera Amphidinium, Coolia, Gambierdiscus, Ostreopsis, and Prorocentrum were found. These five genera were observed on the thalli of the macroalgae Chordaria flagelliformis, Martensia sp., Padina arborescens, and Sargassum sp., while none were observed exceptionally on Codium fragile. The abundance of Ostreopsis spp. was highest on Derbesia sp. (8,660 cells/g wet weight), while that of Gambierdiscus spp. was highest on Martensia sp. (4,870 cells/g-ww). The maximum abundances of Amphidinium spp., Coolia spp., and Prorocentrum spp. were 410, 710, and 300 cells/g-ww, respectively. The maximum abundance of Coolia spp., Gambierdiscus spp., and Ostreopsis spp. obtained in the present study was lower than for other locations reported in literature. The results of the present study suggest that the presence and abundance of epiphytic dinoflagellates may be related to the macroalgal species of the coastal waters of Jeju Island. [ABSTRACT FROM AUTHOR]

The species composition of dinoflagellates in epiphytic assemblages at 0.5-3 m depth in Sobol Bight (Peter the Great Bay in the Sea of Japan) was examined in August-October 2006. A total of 29 dinoflagellate species were found. Planktonic species contributed 54% and benthic and benthic-planktonic species together contributed 36% to the total number of dinoflagellate species. The latter group was represented by 13 species of eight genera, including five potentially toxic species. The occurrences of Cabra matta Murray et Patterson, Ostreopsis cf. siamensis Schmidt, O. cf. ovata Fukuyo, and Prorocentrum fukuyoi Murray et Nagahama are recorded for the seas of Russia for the first time. Amphidinium carterae Hulburt is the first record for the Far Eastern seas and Pseudothecadinium campbellii Hoppenrath et Selina for the Sea of Japan. Morphological descriptions of dinoflagellates that are new records for the study region or potentially toxic species are given. One to ten species of benthic and benthic-planktonic dinoflagellates per macrophyte were found. In all, 13 species of red algae, 9 brown algae, 3 green algae, and 1 species of seagrass were examined. O. cf. ovata, O. cf. siamensis, and P. fukuyoi were the most frequent and abundant. [ABSTRACT FROM AUTHOR]

Abstract: Dinoflagellate species isolated from the green calcareous seaweed, Halimeda sp. J.V. Lamouroux, growing in Rarotongan lagoons, included Gambierdiscus australes Faust & Chinain, Coolia monotis Meunier, Amphidinium carterae Hulburth, Prorocentrum lima (Ehrenberg) Dodge, P. cf. maculosum Faust and species in the genus Ostreopsis Schmidt. Isolates were identified to species level by scanning electron microscopy and/or DNA sequence analysis. Culture extracts of G. australes isolate CAWD149 gave a response of 0.04pg P-CTX-1 equiv. per cell by an N2A cytotoxicity assay (equivalent to ca 0.4pg CTX-3C cell−1). However, ciguatoxins were not detected by LC-MS/MS. Partitioned fractions of the cell extracts potentially containing maitotoxin were found to be very toxic to mice after intraperitoneal (i.p.) injection. A. carterae was also of interest as extracts of mass cultures caused respiratory paralysis in mice at high doses, both by i.p. injection and by oral administration. The Rarotongan isolate fell into a different clade to New Zealand A. carterae isolates, based on DNA sequence analysis, and also had a different toxin profile. As A. carterae co-occurred with G. australes, it may contribute to human poisonings attributed to CTX and warrants further investigation. A crude extract of C. monotis was of low toxicity to mice by i.p. injection, and an extract of Ostreopsis sp. was negative in the palytoxin haemolysis neutralisation assay. [ABSTRACT FROM AUTHOR]

Identification of novel dinoflagellate taxa through molecular analysis is hindered by lack of well-defined basal lineages. To address this issue, we attempted to reassess the phylogenetic status of Oxyrrhis marina Dujard. as well as other potentially basal taxa. The analysis was based on two newly established premises: (1) editing density of mitochondrial cob and cox1 mRNA increases from basal to later diverging lineages; (2) nuclear-encoded mRNA in dinoflagellates is trans-spliced to receive a 22 bp spliced leader (SL) at the 5′-end. We analyzed these two genetic traits in O. marina, Noctiluca scintillans (Macartney) Kof. et Swezy , Heterocapsa triquetra (Ehrenb.) F. Stein, H. rotundata (Lohmann) Ge. Hansen, Amphidinium carterae Hulburt, and A. operculatum Clap. et J. Lachm. Surprisingly, no editing was detected in cob and cox1 mRNAs in these lineages, except for a small number of editing events in Amphidinium. However, nuclear-encoded mRNAs in these species contained the SL sequence at the 5′-end, indicative of SL RNA trans-splicing. These findings, together with the recent cob- cox1-18S rRNA three-gene phylogeny, suggest the following: (1) O. marina is a basal dinoflagellate; (2) Heterocapsa, Amphidinium, and Noctiluca likely are also early diverging lineages of dinoflagellates, and the position of Heterocapsa is inconsistent with literature and needs further investigation; and (3) the presence of the 22 bp SL and mitochondrial (mt) mRNA editing can be considered a landmark of dinoflagellate splits. [ABSTRACT FROM AUTHOR]

Abstract: A new member of amphidinols (AM7) possessing a polyene-polyhydroxyl structure with the shortest carbon backbone and sulfate ester was isolated from the cultured dinoflagellate, Amphidinium klebsii. AM7 showed hemolytic and antifungal activities. The structure was elucidated on the basis of 2D NMR data in combination with CID MS/MS experiments. [Copyright &y& Elsevier]

The recent emendation ofAmphidinium(Dinophyceae), which now only consists of species with minute left-deflected epicone, has left more than 100 species without a clear generic affiliation. In the present study, a strain identified as one of the species with a divergent epicone type,Amphidinium britannicum(Herdman) Lebour, and six strains resemblingA. britannicumbut smaller in size were examined by light, scanning and transmission electron microscopy and by sequence analyses of nuclear-encoded partial large subunit ribosomal DNA to establish their phylogeny.Amphidinium britannicumwas not closely related to other genera included in the molecular phylogenetic analyses, but formed a highly supported clade in Bayesian analysis together with the six small-sized strains. The six strains also formed a highly supported clade, consisting of two closely related, albeit distinct, clades. Light and scanning electron microscopy did not reveal significant differences between the vegetative motile cells; however, cells about to undergo mitosis developed longitudinal grooves on the hypocone in one of the clades but not in the other. Both clades differed substantially fromA. britannicumin partial large subunit ribosomal DNA as well as in size and shape. Based on morphological similarity and partial large subunit ribosomal DNA evidence, we erect the new genus,Togulagen. nov. with the emended type speciesTogula britannica(Herdman) comb. nov. Based on differences in division pattern and partial large subunit ribosomal DNA gene divergence we further describe the speciesTogula compacta(Herdman) comb. nov. andTogula jollasp. nov. [ABSTRACT FROM AUTHOR]

Members of Amphidinium Claparède and Lachmann constitute a major part of sand-dwelling benthic dinoflagellates worldwide. The genus is traditionally defined by its small epicone size, not exceeding one third of the total cell length. It has long been suspected that this functional definition does not reflect phylogeny, yet the problem of identifying the type species A. operculatum and closely related species has until now hindered attempts to redefine the genus. In this study 12 Amphidinium species were examined using phylogenetic analyses based on nuclear-encoded, partial, large subunit (LSU) rDNA, with a further six Amphidinium species being included in a morphological cladistic analysis. The species selected represented taxa with a range of morphological dissimilar epicone forms. Both cladistic analysis and analyses based on partial LSU rDNA revealed that Amphidinium species with minute left-deflected epicones formed a monophyletic clade that included the type species. Amphidinium species with other epicone types were found to be unrelated to this clade. The type species A. operculatum was identified based on general cell shape and size, position of a dark organelle previously defined as a stigma, and origin of the sulcus. The description of A. elegans by Grell and Wohlfarth-Bottermann was found to be identical to it. A species fitting the original description of A. operculatum was cultured and included in the analyses. Based on cladistic and molecular analyses, it grouped together with all other species with minute left-deflected epicones, and this group constitutes the true genus Amphidinium sensu stricto. An emendation of the genus definition is presented. [ABSTRACT FROM AUTHOR]

We present an easy and sensitive method for measuring thiamine and its phosphate esters in small biological samples of microalgae (Amphidinium carterae Hulburt and Nitzschia microcephala Grun). The method consists of extraction of thiamine and its derivatives in acid solution, followed by liquid chromatography with fluorescence detection. The detection limit is as low as 15 fmol of thiamine. For comparison to microalgae, the method has been applied to evaluate thiamine levels in the crustacean Artemia salina Leach and is suitable for nutritional studies of the food web of the Baltic salmon, which suffers from thiamine deficiency. This method of HPLC analysis can be readily utilized to follow uptake and interconversion of thiamine and its phosphate esters in many micro- and macroalgae. [Copyright &y& Elsevier]

After a 1.5-year screening survey in the lagoons of Western Greece in order to isolate and culture sturdy species of microalgae for aquaculture or other value-added uses, as dictated primarily by satisfactory potential for their mass culture, five species emerged, and their growth was monitored in laboratory conditions. Amphidinium carterae, Nephroselmis sp., Tetraselmis sp. (var. red pappas), Asteromonas gracilis, and Dunaliella sp. were batch cultured using low (20 ppt), sea (40 ppt), and high salinity (50 or 60 or 100 ppt) and in combination with low (2000 lux) and high (8000 lux) intensity illumination. The results exhibited that all these species can be grown adequately in all salinities and with the best growth in terms of maximum cell density, specific growth rate (SGR), and biomass yield (g dry weight/L) at high illumination (8000 lux). The five species examined exhibited different responses in the salinities used, whereby Amphidinium clearly performs best in 20 ppt, far better than 40 ppt, and even more so than 50 ppt. Nephroselmis and Tetraselmis grow almost the same in 20 and 40 ppt and less well in 60 ppt. Asteromonas performs best in 100 ppt, although it can grow quite well in both 40 and 60 ppt. Dunaliella grows equally well in all salinities (20, 40, 60 ppt). Concerning the productivity, assessed as the maximum biomass yield at the end of the culture period, the first rank is occupied by Nephroselmis with ~3.0 g d.w./L, followed by Tetraselmis (2.0 g/L), Dunaliella (1.58 g/L), Amphidinium (1.19 g/L), and Asteromonas (0.7 g/L) with all values recorded at high light (8000 lux). [ABSTRACT FROM AUTHOR]

Gymnodinium acidotum Nygaard is a freshwater dinoflagellate that is known to harbor a cryptomonad endosymbiont whose chloroplasts give the organism an overall blue-green color. The ultrastructure of G. acidotum was examined with particular attention being given to the three dimensional nature of the flagellar apparatus. The flagellar apparatus is composed of two functional basal bodies that are slightly offset and lie at an angle of approximately 90° to one another. As in other dinoflagellates the transverse basal body is associated with a striated, fibrous root that extends from the proximal end of the basal body to the transverse flagellar opening. At least one microtubular root extends from the proximal end of the transverse basal body, and a multi-membered longitudinal microtubular root is associated with the longitudinal basal body. The most striking feature of the flagellar apparatus of G. acidotum is the large fibrous connective that extends from the region of the proximal ends of the basal bodies to the cingulum on the dorsal side of the cell. A similar structure has been reported from only one other dinoflagellate, Amphidinium cryophilum Wedernayer. Wilcox, and Graham. The presence of this structure as well as similarities in external morphology suggest that these two species may be more closely related to each other than either is to other gymnodinioid taxa. The taxonomic importance of dinoflagellate flagellar apparatus components is discussed. [ABSTRACT FROM AUTHOR]

The sub-thecal microtubular cytoskeleton of Amphidinum rhynchocephalum Anissimowa was investigated using indirect immunofluorescence microscopy and transmission electron microscopy. The majority of sub-checal microtubules are longitudinally oriented and radiate from one of two sub-thecal transverse microtubular bands that lie adjacent to the anterior and posterior edge of the cingulum. Both transverse bands consist of 3-5 microtubules and are loop shaped with one end adjacent to the cell's right edge of the sulcus and the other end adjacent to the fibrous ventral ridge. The posterior transverse microtubular band (PTB) defines the posterior edge of the cingulum and gives rise to numerous posteriorly directed longitudinal microtubular bundles that consist of 1-3 microtubules per bundle. These bundles end at the posterior end of the cell. The PTB also gives rise to the cingular longitudinal microtubules that underlie the cingular groove and terminate at the anterior transverse microtubular band (ATB). The ATB defines the anterior edge of the cingulum and loops around the base of the epicone. This band gives rise to anteriorly directed longitudinal microtubular bundles that terminate in the small epicone of the cell. The longitudinal microtubular root of the flagellar apparatus is directed posteriorly and lies immediately beneath the theca but is distinct from the subthecal microtubule system. A narrow fibrous ridge is ventrally located to the cell's left between the exit apertures of the transverse and longitudinal flagella. In this position, the ventral ridge lies between and also associated with three microtubules that are distinct from other cytoskeletal microtubules. Our results demonstrate that the majority of sub-thecal microtubules originate from one of two microtubular bands associated with the cingulum. The possible role of the fibrous ventral ridge and its associated microtubules is also discussed. [ABSTRACT FROM AUTHOR]

The dinoflagellate Amphidinium cryophilum sp. nov is one of the few gymnodinians to be studied at the ultra structural level. It resembles other dinoflagellates in the structure of the nucleus, thrichocyts, storage materials, flagella, mitochondrta, and microbodies. Other features of A. cryphilum less commonly observed in related organisms include a network of small interconnected vesicles, a system of large, peripheral vacuoles, chloroplasts bound by two rather than three membranes, an accumulation body, thylakoid-associated plastoglabidi, a vesiculated nuclear envelope, a complex tubular pusule, striated flagellar collars, collared pits, and a peduncle. The occurrence of a peduncle, a structure implicated in photography, in this autographic organism is noteworthy. The ultrastructure of the peduncle of A. cryophilum differs significantly from that reported in another dinoflagellate. [ABSTRACT FROM AUTHOR]