Photosynthetic Dinoflagellates Research Articles

The host transcriptome remains unaltered during the establishment of coral–algal symbioses

Coral reefs are based on the symbiotic relationship between corals and photosynthetic dinoflagellates of the genus Symbiodinium. We followed gene expression of coral larvae of Acropora palmata and Montastraea faveolata after exposure to Symbiodinium strains that differed in their ability to establish symbioses. We show that the coral host transcriptome remains almost unchanged during infection by competent symbionts, but is massively altered by symbionts that fail to establish symbioses. Our data suggest that successful coral-algal symbioses depend mainly on the symbionts' ability to enter the host in a stealth manner rather than a more active response from the coral host.

ECTOCELLULAR GLUCOSIDASE AND PEPTIDASE ACTIVITY OF THE MIXOTROPHIC DINOFLAGELLATE PROROCENTRUM MINIMUM (DINOPHYCEAE)(1).

The activities of the enzymes α- and β-glucosidase, and leucine aminopeptidase were measured in cultures of the dinoflagellate Prorocentrum minimum (Pavill.) J. Schiller and in field samples collected during dinoflagellate blooms occurring in tributaries of the Chesapeake Bay, Maryland, USA. Activities were measured using fluorogenic artificial substrates and partitioned among the >5 μm size fraction, small microbes fraction (0.1-5 μm), and dissolved phase (<0.1 μm). P. minimum and most other photosynthetic dinoflagellates are >5 μm in size and thus can be separated from the small microbes fraction, which contains most bacteria. Little to no glucosidase activity was detected associated with the >5 μm size fraction in cultures or in field samples, with most of the activity (67% to 93% in cultures, 54% to 100% in field samples) in the small microbes size fraction for both α and β glucosidase. In contrast, 67% to 90% of the total leucine aminopeptidase (LAP) activity in cultures was measured in the >5 μm fraction. Within a culture, LAP activity in the size fraction containing P. minimum decreased in response to ammonium and urea additions, but not in response to nitrate. In field samples, LAP activity was positively correlated with dinoflagellate abundance and chl a, and negatively correlated with ammonium concentration. During blooms, up to 34% of LAP activity was associated with the >5 μm fraction, indicating that when abundant, dinoflagellates may make a substantial contribution to ectocellular LAP activity in the water column.

Azadinium spinosum gen. et sp. nov. (Dinophyceae) identified as a primary producer of azaspiracid toxins

Azaspiracids (AZAs) are a group of lipophilic marine biotoxins associated with human incidents of shellfish poisoning. During a research cruise to the North Sea, we analysed size-fractionated plankton for AZA by mass spectrometry and successfully isolated an AZA-producing dinoflagellate from the east coast of Scotland. As shown previously, an axenic culture of this dinoflagellate produces AZA 1, AZA 2 and an isomer of AZA 2. Here we give a taxonomic description of this new taxon Azadinium spinosum gen. et sp. nov., as a de novo producer of AZAs. Azadinium spinosum is a small (12–16 µm length and 7–11 µm width) peridinin-containing photosynthetic dinoflagellate with a superficial resemblance under light microscopy to gymnodinioids, but with a thin theca. The large nucleus is spherical and located posteriorly, whereas the single chloroplast is parietal, lobed, and typically extends into both the epi- and hyposome. The Kofoidian thecal tabulation is APC, 4′, 3a, 6″, 6C, 5?S, 6′″, 2″″. This plate pattern has an epithecal affinity to the Peridiniales and a hypothecal affinity to the Gonyaulacales, but is distinctly different from described dinoflagellate genera. The assignment of A. spinosum to the dinoflagellates is supported by molecular phylogenetic analysis of four genes, SSU rDNA, LSU rDNA (D1/D2 region), ITS and cytochrome oxidase (sub-unit 1) (COI). In agreement with the morphological description, phylogenetic analysis did not show any particularly close affiliation to the Peridiniales or Gonyaulacales, nor to any other dinoflagellate order represented in molecular databases. Consequently, we erected a new genus, Azadinium, for this taxon. However the ordinal affiliation of the genus is uncertain. This study represents the first description and confirmation of a new dinoflagellate species capable of producing AZA and is thus an important advance in surveillance programmes for toxigenic microalgae and toxins of human health significance.

CORRESPONDENCE BETWEEN COLD TOLERANCE AND TEMPERATE BIOGEOGRAPHY IN A WESTERN ATLANTIC SYMBIODINIUM (DINOPHYTA) LINEAGE(1).

Many corals form obligate symbioses with photosynthetic dinoflagellates of the genus Symbiodinium Freudenthal (1962). These symbionts vary genotypically, with their geographical distribution and abundance dependent upon host specificity and tolerance to temperature and light variation. Despite the importance of these mutualistic relationships, the physiology and ecology of Symbiodinium spp. remain poorly characterized. Here, we report that rDNA internal transcribed spacer region 2 (ITS2) defined Symbiodinium type B2 associates with the cnidarian hosts Astrangia poculata and Oculina arbuscula from northerly habitats of the western Atlantic. Using pulse-amplitude-modulated (PAM) fluorometry, we compared maximum photochemical efficiency of PSII of type B2 to that of common tropical Symbiodinium lineages (types A3, B1, and C2) under cold-stress conditions. Symbiont cultures were gradually cooled from 26°C to 10°C to simulate seasonal temperature declines. Cold stress decreased the maximum photochemical efficiency of PSII and likely the photosynthetic potential for all Symbiodinium clades tested. Cultures were then maintained at 10°C for a 2-week period and gradually returned to initial conditions. Subsequent to low temperature stress, only type B2 displayed rapid and full recovery of PSII photochemical efficiency, whereas other symbiont phylotypes remained nonfunctional. These findings indicate that the distribution and abundance of Symbiodinium spp., and by extension their cnidarian hosts, in temperate climates correspond significantly with the photosynthetic cold tolerance of these symbiotic algae.

Cellular mechanisms of Cnidarian bleaching: stress causes the collapse of symbiosis

Cnidarian bleaching is a breakdown in the mutualistic symbiosis between host Cnidarians, such as reef building corals, and their unicellular photosynthetic dinoflagellate symbionts. Bleaching is caused by a variety of environmental stressors, most notably elevated temperatures associated with global climate change in conjunction with high solar radiation, and it is a major contributor to coral death and reef degradation. This review examines the underlying cellular events that lead to symbiosis dysfunction and cause bleaching, emphasizing that, to date, we have only some pieces of a complex cellular jigsaw puzzle. Reactive oxygen species (ROS), generated by damage to both photosynthetic and mitochondrial membranes, is shown to play a central role in both injury to the partners and to inter-partner communication of a stress response. Evidence is presented that suggests that bleaching is a host innate immune response to a compromised symbiont, much like innate immune responses in other host-microbe interactions. Finally, the elimination or exit of the symbiont from host tissues is described through a variety of mechanisms including exocytosis, host cell detachment and host cell apoptosis.

“Windows of opportunity” for dinoflagellate blooms: Reduced microzooplankton net growth coupled to eutrophication

Links between eutrophication, plankton community structure, microzooplankton grazing and dinoflagellate abundance were investigated in two tributaries of the Chesapeake Bay, the Choptank and Patuxent Rivers (MD, USA). Sampling and experiments were conducted during the spring of consecutive dry (below average freshwater flow) and wet (above average freshwater flow) years. During the wet year (2003), dissolved inorganic nitrogen, phytoplankton, and copepod biomass, but not microzooplankton abundance, were greater than in the dry year. In 2003, but not 2002, small cell size photosynthetic dinoflagellates were abundant and blooms occurred in both rivers. Average potential microzooplankton grazing pressure on small dinoflagellates was spatially and temporally variable, but was not significantly different between years. Our data suggest that the variability in microzooplankton grazing pressure provided “windows of opportunity” for net growth of dinoflagellates in response to nutrient loading. The lack of net population growth of micrograzers in response to increases in dinoflagellate prey allowed dinoflagellate blooms to reach relatively high densities, however grazing also appeared to be important in limitation or demise of some blooms. We hypothesize that uncoupling of micrograzer–prey dynamics was partly due to strong top-down control by copepods of microzooplankton in the proportionately more eutrophic year, and perhaps also due to inhibition of microzooplankton grazing/growth once dinoflagellate densities are high.

COOLIA CANARIENSIS SP. NOV. (DINOPHYCEAE), A NEW NONTOXIC EPIPHYTIC BENTHIC DINOFLAGELLATE FROM THE CANARY ISLANDS(1).

A new photosynthetic dinoflagellate species, Coolia canariensis S. Fraga sp. nov., is described based on samples taken from tidal ponds on the rocky shore of the Canary Islands, northeast Atlantic Ocean. Its morphology was studied by LM and SEM. It is almost spherical and has a thick smooth theca with many scattered pores. Plate 1' is the biggest of the epithecal plates, and 7″ is twice as wide as it is long. Phylogeny inferred from the D1/D2 regions of the LSU nuclear rDNA of three strains of C.canariensis and several strains of other Coolia species, C.monotis, C.sp., showed that C.canariensis strains clustered in a well-supported clade distinct from the other species. No toxins were detected using mouse bioassay, liquid chromatography with Fluorescence detection (LC-FLD) or liquid chromatography-mass spectrometry (LC-MS). Its pigment composition is of the peridinin type of dinoflagellates. Together with this new species, many other strains of C.monotis from the Atlantic Ocean and Mediterranean Sea have been analyzed for toxin presence, and no evidence of toxin production related to yessotoxins (YTXs) was found, as was previously suggested for C.monotis from Australia.

Growth and grazing responses of the mixotrophic dinoflagellate Dinophysis acuminata as functions of light intensity and prey concentration

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 51:301-310 (2008) - DOI: https://doi.org/10.3354/ame01203 Growth and grazing responses of the mixotrophic dinoflagellate Dinophysis acuminata as functions of light intensity and prey concentration Sunju Kim1,5, Yi Gu Kang1, Hyung Seop Kim2, Wonho Yih1, D. Wayne Coats3, Myung Gil Park4,* 1Department of Oceanography, Kunsan National University, Gunsan, 573-701, Republic of Korea 2Gunsan Fisheries Office, NFRDI, Gunsan 573-882, Republic of Korea 3Smithsonian Environmental Research Center, PO Box 28, Edgewater, Maryland 21037, USA 4Laboratory of HAB Ecophysiology (LOHABE), Department of Oceanography, Chonnam National University, Gwangju, 500-757, Republic of Korea 5Present address: Smithsonian Environmental Research Center, PO Box 28, Edgewater, Maryland 21037, USA *Corresponding author. Email: mpark@chonnam.ac.kr ABSTRACT: Dinophysis acuminata, a photosynthetic marine dinoflagellate, possesses plastids of cryptophyte origin and causes diarrhetic shellfish poisoning (DSP). Recent work has shown D. acuminata to be a mixotroph that grows well when feeding on the photosynthetic ciliate Myrionecta rubra. Using established cultures, we examined the effects of light intensity and prey (M. rubra) concentration on growth and ingestion rates of D. acuminata. Growth rates increased with increasing prey concentration under continuous illumination of 60 µE m–2 s–1, with maximum mixotrophic growth (0.91 d–1) almost 5 times higher than growth in the absence of prey (0.19 d–1). The maximum ingestion rate of D. acuminata was 1296 pg C Dinophysis–1 d–1 (3.2 M. rubra cells Dinophysis–1 d–1) for data fitted to a Michaelis-Menten equation. Growth rate also increased with increasing light intensity, an effect even stronger when prey was supplied. Increased growth with increasing irradiance was accompanied by a corresponding increase in ingestion. While D. acuminata continued to grow in semi-continuous food-replete cultures at high (200 µE m–2 s–1) and low (10 µE m–2 s–1) light intensity, it failed to grow in darkness, despite the presence of prey. Our results suggest that D. acuminata is an obligate mixotroph that requires both light and prey for long-term survival. Results indicate that Dinophysis species are typically prey-limited in the field. KEY WORDS: Dinophysis acuminata · Myrionecta rubra · Growth · Grazing · Light · Mixotrophy · Phagotrophy Full text in pdf format PreviousCite this article as: Kim S, Kang YG, Kim HS, Yih W, Coats DW, Park MG (2008) Growth and grazing responses of the mixotrophic dinoflagellate Dinophysis acuminata as functions of light intensity and prey concentration. Aquat Microb Ecol 51:301-310. https://doi.org/10.3354/ame01203 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 51, No. 3. Online publication date: June 16, 2008 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2008 Inter-Research.

Characterization of azaspiracids in plankton size-fractions and isolation of an azaspiracid-producing dinoflagellate from the North Sea

Azaspiracids (AZAs) are a group of lipophilic polyether toxins implicated in incidents of shellfish poisoning in humans, particularly in northern Europe. In an attempt to establish the biogeographical distribution of AZA toxins, their association with plankton size-fractions, and to confirm the identity of the causative species responsible for human poisoning, a month-long oceanographic study was undertaken in coastal North Sea waters. The occurrence and abundance of AZA analogues was measured by on board triple quadrupole mass spectrometry coupled to liquid chromatography (LC-MS/MS). In size-fractionated plankton samples collected by net tows (20 μm mesh-size), by pumping from discrete depths and from Niskin entrapment bottle casts to fixed depths, AZA-1 was consistently the major azaspiracid component. In eastern Scottish coastal waters, the highest amounts of AZA-1 in net tow samples were in the 50–200 μm fractions, with lesser amounts detected in the >200 μm and 50–20 μm fractions. At these stations, the 50–200 μm fractions were rich in the ciliate Favella ehrenbergii. Cells of F. ehrenbergii isolated by microcapillary indeed contained AZA-1, but isolated cells grown and fed the non-toxic dinoflagellate Scrippsiella trochoidea for one week failed to contain any detectable AZA-1—evidence that F. ehrenbergii is merely a vector for AZA. Detailed analysis of plankton from Niskin bottle samples from around the North Sea typically showed highest amounts of AZA in the 3–20 μm fraction. From this fraction, a large number of crude cultures were established and subsequently screened for the presence of AZAs. A small photosynthetic thecate dinoflagellate, provisionally designated as strain 3D9, was isolated by microcapillary and brought into pure culture. This dinoflagellate strain was found to produce AZA-1, AZA-2 and an isomer of AZA-2. Sequence comparisons by molecular genetic techniques also indicated that this genotype was present in field samples rich in AZA. This discovery of a novel causative dinoflagellate for AZA toxicity essentially explains the lack of correlation of AZA with the abundance and distribution of the previously postulated culprit species Protoperidinium crassipes. We instead propose that such large phagotrophic dinoflagellates can act as an AZA vector following grazing upon a proximal source, such as the dinoflagellate 3D9 strain.

Calcification and associated physiological parameters during a stress event in the scleractinian coral Stylophora pistillata

High calcification rates observed in reef coral organisms are due to the symbiotic relationship established between scleractinian corals and their photosynthetic dinoflagellates, commonly called zooxanthellae. Zooxanthellae are known to enhance calcification in the light, a process referred as “light-enhanced calcification”. The disruption of the relationship between corals and their zooxanthellae leads to bleaching. Bleaching is one of the major causes of the present decline of coral reefs related to climate change and anthropogenic activities. In our aquaria, corals experienced a chemical pollution leading to bleaching and ending with the death of corals. During the time course of this bleaching event, we measured multiple parameters and could evidence four major consecutive steps: 1) at month 1 (January 2005), the stress affected primarily the photosystem II machinery of zooxanthellae resulting in an immediate decrease of photosystem II efficiency, 2) at month 2, the stress affected the photosynthetic production of O 2 by zooxanthellae and the rate of light calcification, 3) at month 3, there was a decrease in both light and dark calcification rates, the appearance of the first oxidative damage in the zooxanthellae, the disruption of symbiosis, 4) and finally the death of corals at month 6.

Feeding mechanism, prey specificity and growth in light and dark of the plastidic dinoflagellate Karlodinium armiger

The food uptake mechanism and prey specificity of the most recently described member of the ichthyotoxic photosynthetic dinoflagellate genus Karlodinium (K. armiger) was studied. K. armiger extracts the contents of prey through an inconspicuous feeding tube (peduncle), but may also ingest whole prey cells. This species is omnivorous, ingesting prey from all major groups of marine protists. K. armiger displays a searching pre-capture behavior with attraction to prey cells and formation of feeding aggregates. In batch cultures, growth rates in the light without food were low (0.01 to 0.10 d -1 ), but when the culture medium was enriched with soil extract, initial growth rate increased (0.19 d -1 ); it further increased (0.60 d -1 ) when fed the cryptophyte Rhodomonas marina in the light (170 µmol photons m 2 s -1 ). R. marina was also ingested in the dark, but did not support posi- tive growth rates and survival. Thus, K. armiger is an omnivorous obligate phototrophic mixotroph which seems to obtain a growth-essential substance or growth factor through phagotrophy.

Heat stress causes inhibition of the de novo synthesis of antenna proteins and photobleaching in cultured Symbiodinium

Coral bleaching, caused by heat stress, is accompanied by the light-induced loss of photosynthetic pigments in in situ symbiotic dinoflagellate algae (Symbiodinium spp.). However, the molecular mechanisms responsible for pigment loss are poorly understood. Here, we show that moderate heat stress causes photobleaching through inhibition of the de novo synthesis of intrinsic light-harvesting antennae [chlorophyll a-chlorophyll c(2)-peridinin-protein complexes (acpPC)] in cultured Symbiodinium algae and that two Clade A Symbiodinium species showing different thermal sensitivities of photobleaching also show differential sensitivity of this key protein synthesis process. Photoinhibition of photosystem II (PSII) and subsequent photobleaching were observed at temperatures of >31 degrees C in cultured Symbiodinium CS-73 cells grown at 25-34 degrees C, but not in cultures of the more thermally tolerant control Symbiodinium species OTcH-1. We found that bleaching in CS-73 is associated with loss of acpPC, which is a major antennae protein in Symbiodinium. In addition, the thermally induced loss of this protein is light-dependent, but does not coincide directly with PSII photoinhibition and is not caused by stimulated degradation of acpPC. In cells treated at 34 degrees C over 24 h, the steady-state acpPC mRNA pool was modestly reduced, by approximately 30%, whereas the corresponding synthesis rate of acpPC was diminished by >80%. Our results suggest that photobleaching in Symbiodinium is consequentially linked to the relative susceptibility of PSII to photoinhibition during thermal stress and occurs, at least partially, because of the loss of acpPC via undefined mechanism(s) that hamper the de novo synthesis of acpPC primarily at the translational processing step.

Molecular Characterization of Iron-Containing Superoxide Dismutases in the Heterotrophic Dinoflagellate Crypthecodinium cohnii

Superoxide dismutases (SODs) are a family of antioxidant enzymes that catalyse the degradation of toxic superoxide radicals in obligate and facultative aerobic organisms. Here, we report the presence of a multi-copy gene family encoding SODs in the heterotrophic dinoflagellate Crypthecodinium cohnii. All the genes identified (sod1 to sod17) have been cloned and sequenced, and shown to encode potentially functional dimeric iron-containing SOD isozymes. Our data revealed a considerable molecular heterogeneity of this enzyme in C. cohnii at both genomic and transcriptional levels. The C. cohnii SOD1, overexpressed in Escherichia coli, was active and its structure obtained by homology modeling using X-ray crystal structures of homologues exhibited the typical fold of dimeric FeSODs. Phylogenetic studies including 110 other dimeric FeSODs and closely related cambialistic dimeric SOD sequences showed that the C. cohnii SODs form a monophyletic group and have all been acquired by the same event of horizontal gene transfer. It also revealed a dichotomy within the C. cohnii SOD sequences that could be explained by an ancestral sod gene duplication followed by subsequent gene duplications within each of the two groups. Enzyme assays of SOD activity indicated the presence of two FeSOD activities in C. cohnii cell lysate whereas MnSOD and Cu/ZnSOD were not detected. These activities contrasted with the SOD repertoire previously characterized in photosynthetic dinoflagellates. To explain these differences, a hypothetical evolutionary scenario is proposed that suggests gains and losses of sod genes in dinoflagellates.

Spatiotemporal Patterns of Subtidal Benthic Microalgal Biomass and Community Composition in Galveston Bay, Texas, USA

Many Gulf of Mexico estuaries have low ratios of water volume to bottom surface area, and benthic processes in these systems likely have a major influence on system structure and function. The purpose of this study was to determine the spatiotemporal distribution of biomass and community composition of subtidal benthic microalgal (BMA) communities in Galveston Bay, TX, USA, compare BMA community composition and biomass to phytoplankton in overlying waters, and estimate the potential contribution of BMA to the trophodynamics in this shallow, turbid, subtropical estuary. The estimates of BMA biomass (mean = 4.21 mg Chl a m−2) for Galveston Bay were within the range of the reported values for similar Gulf of Mexico estuaries. BMA biomass in the central part of the bay was essentially homogeneous, whereas biomass at the seaward and upper bay ends of the transect were significantly lower. Peridinin, fucoxanthin, and alloxanthin were the three carotenoids with the highest concentrations, with fucoxanthin having the highest mean concentration (1.82 mg m−2). The seaward and landward ends of the transect differed from the central region of the bay with respect to the relative abundances of chlorophytes, cyanobacteria, and photosynthetic bacteria. Benthic microalgal community composition also showed a gradual shift over time due to changes in the relative abundances of photosynthetic bacteria, cryptophytes, dinoflagellates, and cyanobacteria. Major changes in community composition occurred in the spring months (March to April). On an areal basis, BMA biomass in Galveston Bay occurred at minor concentrations (16.5%) relative to phytoplankton. Furthermore, the concentrations of carotenoid pigments for phytoplankton and BMA (fucoxanthin, alloxanthin, and zeaxanthin) were correlated (r = 0.48 to 0.61), suggesting a close linkage between microalgae in the water column and sediments. The contribution of BMA to the primary productivity of the deeper waters (>2 m) of Galveston Bay is probably very small in comparison to shallower waters along the bay margins. The significant similarities in the community composition of phytoplankton and BMA illustrate the potential importance of deposition and resuspension processes in this turbid, shallow estuary.

Cochlodinium fulvescens sp. nov. (Gymnodiniales, Dinophyceae), a new chain‐forming unarmored dinoflagellate from Asian coasts

SUMMARYCellular morphology and the phylogenetic position of a new unarmored photosynthetic dinoflagellate Cochlodinium fulvescens Iwataki, Kawami et Matsuoka sp. nov. were examined by light microscopy and molecular phylogenetic analyses based on partial large subunit ribosomal DNA (LSU rDNA) and small subunit ribosomal DNA (SSU rDNA) sequences. The cells of C. fulvescens closely resemble C. polykrikoides, one of the most harmful red tide forming dinoflagellates, due to it possessing a cingulum encircling the cell approximately twice, a spherical nucleus positioned in the anterior part of the cell and an eyespot‐like orange pigmented body located in the dorsal side of the epicone, as well as formation of cell‐chains. However, this species is clearly distinguished from C. polykrikoides based on several morphological characteristics, namely, cell size, shape of chloroplasts and the position of narrow sulcus situated in the cell surface. The sulcus of C. fulvescens is located at the intermediate position of the cingulum in the dorsal side, whereas that of C. polykrikoides is situated immediately beneath the cingulum. LSU rDNA phylogenies indicated that C. fulvescens is clearly distinct from, but closely related to C. polykrikoides among dinoflagellates.

Stable oxygen isotopes of Thoracosphaera heimii (Dinophyceae) in relationship to temperature; a culture experiment

To establish a relationship between temperature and the stable oxygen isotopic composition ( δ 18O) of vegetative cysts of the photosynthetic calcareous dinoflagellate cyst Thoracosphaera heimii, two unicellular cultures of T. heimii have been cultured under different temperatures by using a temperature gradient box. There is a clear relationship between temperature variance and the isotopic composition of T. heimii cysts according to the relationship: T (°C) = − 6.827 ( δ 18O c– δ 18O w)–3.906 ( R = 0.921), with c = calcite and w = water. Within this paper we are the first to discuss the possible vital effects that might cause an offset between the temperature–isotope relationship found for T. heimii calcite and that of equilibrium inorganic calcite precipitation. No indication for strong kinetic effects as result of fast calcite precipitation can be found. We observed a positive relationship between δ 18O c– δ 18O w and ambient mediumwater pH. We speculate that this might be the result of the presence of external carbonate anhydrase, which is common in photosynthetic dinoflagellates. The efficiency of this enzyme increases rapidly between pH 7.5 to 9, which could result in an increase in CO 2 uptake relative to HCO 3 − with increasing pH. We furthermore discuss the possibility of T. heimii using respirative carbon at least as part of its carbon source for calcite precipitation, which can be based on the light values of δ 18O c– δ 18O w and δ 13C c– δ 13C DIC (DIC = dissolved inorganic carbon) found in this and previous studies on the isotopic composition of calcareous dinoflagellates. The results of this study as well as the broad geographic distribution of T. heimii, its stable position within the water column, its presence in the geological record since the Late Cretaceous and its resistance against dissolution compared to other plankton groups underlines the potential for a wide usability of the oxygen isotope composition of T. heimii as palaeotemperature proxy for the deeper parts of the photic zone.

Morphology and Phylogeny of the Pseudocolonial Dinoflagellates Polykrikos lebourae and Polykrikos herdmanae n. sp.

Both the photosynthetic and heterotrophic forms of the only known marine benthic (sand-dwelling) species of Polykrikos, namely P. lebourae, were investigated using light and electron microscopy and molecular phylogenetic analyses. The pseudocolonies usually contained eight integrated zooids and two nuclei. Pseudocolonies consisting of four or five zooids and one nucleus were observed for the first time for this species; some of these reduced pseudocolonies contained plastids, while others were heterotrophic and contained taeniocyst-nematocyst complexes. The ultrastructure of the plastids in P. lebourae did not conform to the organization of thylakoids and enveloping membranes present in the peridinin-containing plastids of other photosynthetic dinoflagellates (i.e. stacks of 3 thylakoids and 3 outer membranes). Instead, the plastids in P. lebourae had thylakoids arranged in pairs and appeared to be enveloped by only two membranes. Molecular phylogenetic data using small subunit rDNA demonstrated that the photosynthetic and heterotrophic forms of P. lebourae represent two distinct clades. The more inclusive clade containing both forms of P. lebourae was most closely related to heterotrophic polykrikoids, namely P. kofoidii. These results led us to conclude that the photosynthetic and heterotrophic forms of P. lebourae are in fact two distinct lineages, and the heterotrophic form is described here as Polykrikos herdmanae n. sp.

Did the peridinin plastid evolve through tertiary endosymbiosis? A hypothesis

Most photosynthetic dinoflagellates harbour the peridinin plastid. This plastid is surrounded by three membranes and its characteristic pigments are chlorophyll c and the carotenoid peridinin. The evolutionary origin of this peculiar plastid remains controversial and is hotly debated. On the recently published tree of concatenated plastid-encoded proteins, dinoflagellates emerge from within the Chromista (clade containing cryptophytes, heterokonts, and haptophytes) and cluster specifically with Heterokonta. These data inspired a new version of the ‘chromalveolate’ model, according to which the peridinin plastid evolved by ‘descent with modification’ from a heterokont-like plastid that had been acquired from a rhodophyte by an ancestral chromalveolate. However, this model of plastid evolution encounters serious obstacles. Firstly, the heterokont plastid is surrounded by four membranes, which means that the ancestral peridinin plastid must have lost one of these primary membranes. However, such a loss could be traumatic, because it could potentially disturb protein import into and/or within the plastid. Secondly, on the phylogenetic tree of Dinoflagellata and Heterokonta, the first to diverge are not plastid, but heterotrophic, aplastidal taxa. Thus, when accepting the single origin of the heterokont and peridinin plastids, we would have to postulate multiple plastid losses, but such a scenario is highly doubtful when the numerous non-photosynthetic functions of plastids and their existence in heterotrophic protists, including parasitic lineages, are considered. Taking these obstacles into account, we suggest an alternative interpretation of the concatenated tree of plastid-encoded proteins. According to our hypothesis, the peridinin plastid evolved from a heterokont alga through tertiary endosymbiosis.

Kleptoplasty in an Antarctic dinoflagellate: caught in evolutionary transition?

Photosynthetic dinoflagellates contain a diverse collection of plastid types, a situation believed to have arisen from multiple endosymbiotic events. In addition, a number of heterotrophic (phagotrophic) dinoflagellates possess the ability to acquire chloroplasts temporarily by engulfing algae and retaining their chloroplasts in a functional state. These latter relationships typically last from a few days to weeks, at which point the chloroplasts lose function, are digested and replaced with newly acquired plastids. A novel and abundant dinoflagellate related to the icthyotoxic genera Karenia and Karlodinium was recently discovered by us in the Ross Sea, Antarctica. Sequencing of its plastid small subunit ribosomal gene indicated that it did not share evolutionary history with the plastids of Karenia or Karlodinium, but was closely related to the free-living haptophyte Phaeocystis antarctica, a species that often dominates phytoplankton blooms in the Ross Sea. Chloroplast uptake was observed to occur rapidly (within 2 days), with retention in cultures being long-lived (several months) but not permanent. The dinoflagellate was also incapable of growing indefinitely in continuous darkness with algae as prey. Our findings may indicate an emerging endosymbiotic event yielding a dinoflagellate that is presently neither purely phototrophic nor purely heterotrophic, but occupies a niche juxtaposed between these contrasting nutritional modes.

Sterols of the Syndinian Dinoflagellate Amoebophrya sp., a Parasite of the Dinoflagellate Alexandrium tamarense (Dinophyceae)

Several harmful photosynthetic dinoflagellates have been examined over past decades for unique chemical biomarker sterols. Little emphasis has been placed on important heterotrophic genera, such as Amoebophrya, an obligate, intracellular parasite of other, often harmful, dinoflagellates with the ability to control host populations naturally. Therefore, the sterol composition of Amoebophrya was examined throughout the course of an infective cycle within its host dinoflagellate, Alexandrium tamarense, with the primary intent of identifying potential sterol biomarkers. Amoebophrya possessed two primary C(27) sterols, cholesterol and cholesta-5,22Z-dien-3beta-ol (cis-22-dehydrocholesterol), which are not unique to this genus, but were found in high relative percentages that are uncommon to other genera of dinoflagellates. Because the host also possesses cholesterol as one of its major sterols, carbon-stable isotope ratio characterization of cholesterol was performed in order to determine whether it was produced by Amoebophrya or derived intact from the host. Results indicated that cholesterol was not derived intact from the host. A comparison of the sterol profile of Amoebophrya to published sterol profiles of phylogenetic relatives revealed that its sterol profile most closely resembles that of the (proto)dinoflagellate Oxyrrhis marina rather than other extant genera.