Dinoflagellate Cells Research Articles

Light scattering by Prorocentrum micans: a new method and results

Striking light-scattering behavior was observed from a marine dinoflagellate, Prorocentrum micans. Measurements of the angular dependence of the 16 Mueller matrix elements were performed on single cells with a polarization-modulation nephelometer by using a new method for cell immobilization. First the dinoflagellate cells were immobilized in a transparent silica gel containing alcohol, and then a second liquid was diffused into the gel to match the index of refraction of the gel network, thereby producing a transparent support medium that scatters less than one tenth the amount of light scattered by a single cell at 90 degrees . Measurements of scattering by a single cell revealed that all 16 matrix elements were significantly nonzero and different from each other. All matrix elements have an extremely rich, reproducible structure that is highly dependent on cell orientation. The matrix elements symmetrically across the diagonal were not equivalent. Striking features of the measurements are the large peak values of S(13), S(14), and other off-diagonal block elements. We believe that this is the first report of such scattering signals by single, suspended marine microorganisms.

Alongshore transport of a toxic phytoplankton bloom in a buoyancy current: Alexandrium tamarense in the Gulf of Maine

We examined the mechanisms controlling blooms of the toxic dinoflagellate Alexandrium tamarense Lebour and the concomintant patterns of shellfish toxicity in the southwestern Gulf of Maine, USA. During a series of cruises from 1987 to 1989, hydrographic parameters were measured to elucidate the physical factors affecting the distribution and abundance of dinoflagellates along this coast. In 1988 and 1989 when toxicity was detected in the southern part of this region, A. tamarense cells were apparently transported into the area between Portsmouth, New Hampshire, and Cape Ann, Massachusetts, in a coastally trapped buoyant plume. This plume appears to have been formed by the outflow from the Androscoggin and Kennebec Rivers. Flow rates of these rivers, hydrographic sections, and satellite images led us to conclude that the plume persisted for about a month, and extended alongshore for several hundred kilometers. The distribution of cells followed the position of the plume as it was influenced by wind and topography. When winds were downwelling-favorable (to the southwest), cells were moved alongshore to the south, and were held to the coast; when winds were upwelling-favorable (to the northeast),the plume sometimes separated from the coast, advecting the cells offshore. In 1987 when no plume was present, A. tamarense cells were scarce, and no toxicity was recorded at the southern stations. The alongshore advection of toxic cells within a coastally trapped buoyant plume can explain the details of the temporal and spatial patterns of shellfish toxicity along the coast. We hypothesize that (1) the source of the A. tamarense populations is in the north, possibly associated with the Androscoggin and Kennebec estuaries, that (2) toxicity patterns follow a predictable relationship with river flow volume and timing of flow peaks and that (3) wind stresses directly influence the distribution of low salinity water and the dinoflagellate cells. Local, in situ growth of dinoflagellates can be an important factor initiating toxic dinoflagellate blooms. However, these data demonstrate the significant role of alongshore transport of established populations of A. tamarense in controlling the location and timing of paralytic shellfish poisoning (PSP) outbreaks in May and June along the southwestern coast of the Gulf of Maine.

Comparison of the response of five larval fish species to the toxic dinoflagellate Alexandrium excavatum (Braarud) Balech

The response of fish larvae to the toxic dinoflagellate Alexandrium excavatum is reviewed based on published and new data from comparable experiments in which larvae were exposed to various concentrations of toxic A. excavatum (treatment) and nontoxic A. tamarense (control). In direct intoxication experiments (larvae fed toxic cells), severity of symptoms, response time and toxin-related mortality (7–96% · day −1) depended on species sensitivity, dinoflagellate strain toxicity and toxic cell concentration. In vectorial intoxication experiments (postlarvae fed tainted zooplankton), the mortality attributable to the toxins was lower and less variable. Elimination of the toxins by the zooplankton could limit the propagation of dinoflagellate toxins towards higher levels of the planktonic food web. It is concluded that the potential impact of A. excavatum blooms on fish survival in nature will be stronger during the phytophagous larval and the early postlarval stages when dinoflagellate cells are ingested directly. Along the northwest Atlantic seaboard, the threat to coastal fisheries from A. excavatum blooms is predicted to intensify from south to north, along a gradient of increasing toxicity of dinoflagellate strains.

Microtubular spindle and centrosome structures during the cell cycle in a dinoflagellate Crypthecodinium cohnii B.: an immunocytochemical study

In order to determine if a mitotic spindle organizing center is present in dinoflagellate cells, we used a library of 12 monoclonal antibodies obtained by immunizing mice with isolated human centrosomes. When tested by immunofluorescence on cryosections of the dinoflagellate Crypthecodinium cohnii B., a positive labeling was obtained with three of these antibodies. In interphase cells, the anti-centrosome antibodies labeled structures located either in the cell periphery, corresponding probably to both basal bodies (i.e. kinetosomes) and in the perinuclear area. In the latter case, two punctate structures were observed near the nuclear envelope. They have never been described, either in light, or in electron microscopic studies of dinoflagellates. We have designated them as centrosome-like structures. A microtubular desmose reacting positively with anti-tubulin Ab was also visible, linking kinetosomes and centrosome-like structures. During mitosis, the double punctate structures were observed at the poles of the nucleus. Double immunolabeling with tubulin and anti-centrosome Ab was also carried out and strongly suggested that in mitotic cells, centrosome-like structures, located at the poles of the mitotic spindle, were associated with microtubular bundles and probably organize and polarize them. These data indicate the existence of centrosome-like structures in C. cohnii cells and the strong conservation of some centrosomal epitopes from dinoflagellates to human. One of the antibodies (CTR 210) recognized by immunoblotting, a single protein band at 72 kDa from a total protein extract. The direct demonstration that this protein is located at the centrosome-like structures and at the kinetosomes deserves further study.

Influence of dinoflagellate cell toxicity on uptake and loss of paralytic shellfish toxins in the northern quahog Mercenaria mercenaria

Influence of dinoflagellate cell toxicity on uptake and loss of paralytic shellfish toxins in the northern quahog Mercenaria mercenaria

Influence of dinoflagellate cell toxicity on uptake and loss of paralytic shellfish toxins in the northern quahog Mercenaria mercenaria

Influence of dinoflagellate cell toxicity on uptake and loss of paralytic shellfish toxins in the northern quahog Mercenaria mercenaria

Nucleolar localization of rRNA coding sequences in Prorocentrum micans Ehr. (dinomastigote, kingdom Protoctist) by in situ hybridization

To define the molecular mechanisms of ribosome biogenesis and to find out in which nucleolar compartment transcription of rDNA occurs, we have performed in situ hybridization (ISH) of RNase-treated cryosections using biotinylated rRNA coding sequences as a probe and the eukaryotic dinoflagellate nucleolar system as a model. Recent data from ISH of eukaryotic ribosomal genes by electron microscopy (EM) has so far failed to establish a consensus which clearly defines the function of the three compartments of the nucleolus. Dinomastigote protoctists are the only known eukaryotes whose chromatin is totally devoid of nucleosomes. Their chromosomes remain permanently condensed during the entire cell cycle and active nucleoli arise from an unwound part of some of the otherwise compact chromosomes. In this work, DNA-DNA hybrids were detected either by fluorescent avidin or by indirect immunogold staining procedures in EM; this is the first use of cryosections to detect hybrids in EM not only in the nucleolus sensu lato but also in a dinomastigote cell. Coding sequences of ribosomal genes were detected both in the periphery of the nucleolar organizer region (NOR), which corresponds to the unwound part of the nucleolar chromosome, and in the proximal part of the fibrillo-granular (FG) region. These results suggest that the rRNA gene transcription predominantly occurs at the periphery of the NOR where the coding sequences are located. A predictive model summarizes and allows discussions and comparisons with other eukaryotes in which nucleolar mechanisms were previously studied. This leads to the conclusion that dinoflagellate cells constitute an excellent model for the study of the functional structure of the eukaryotic nucleolus. In addition, the high level of labeling obtained on cryosections suggests that this procedure should be preferred for future ISH work.

LEPIDODINIUM VIRIDE GEN. ET SP. NOV. (GYMNODINAIALES, DINOPHYTA), A GREEN DINOFLAGELLATE WITH A CHLOROPHYLL A‐ AND B‐CONTAINING ENDOSYMBIONT1,2

ABSTRACTA further description of the green dinoflagellate, strain Y‐100, with a chlorophyll a‐ and b‐containing endosymbiont is given with special emphasis on the morphology of the host cell. The host dinoflagellate cell is unarmored and has a gymnodinoid overall appearance. The theca or amphiesma basically consists of the outer membrane and flattened thecal vesicles in which no thecal plates are developed. Unusual hand basket‐shaped scales cover the entire cell surface together with a layer of mucilaginous material. These findings suggest that the organism is a new member of the Gymnodiniaceae (Gymnodiniales). We propose the name Lepidodinium viride gen. et sp. nov. The ultrasturcture of the host cell is typical of the dinoflagellates; however, the organism has 1) an unusual cytoplasmic projection that may be a homologue of the peduncle, 2) a single membrane‐bounded body containing membranous sheets, closely situated next to the endosymbiont, and 3) an electron opaque network‐forming appendage surrounding the transverse flagellum. Name of these features have been found in other dinoflagellates.

The marine killers: Dinoflagellates in estaurine and coastal waters

Records of massive fish kills and paralytic shellfish poisoning (PSP) in Europe and North America go back to the 17th century. But, it was not until the 1940s when the relationship between PSP, red tide and toxic dinoflagellateGonyaulax was established. Recent records show that PSP and related poisons caused by toxic dinoflagellates in coastal waters and estuaries, are a world-wide problem. Diarrhetic shellfish poisoning (DSP) and neurotoxic poisoning (NSP), believed earlier as bacterial or viral infections are now shown to be caused by other toxic dinoflagellates such asDinophysis. The shellfish most often involved in the poisoning are mussels and clams. Other dinoflagellates,Gyrodinium, occasionally cause massive fish kills in vast coastal areas, resulting in fishery and economic losses. Factors promoting toxic dinoflagellate bloom development and PSP/DSP outbreaks are not fully understood. In previous studies, temperature was considered as the principal factor influencing dinoflagellate blooming. Recent studies showed that other factors such as salinity, sunlight, freshwater runoff and water stability are also important. Pollution from land drainage and sewage discharge in inshore waters were also implicated. Current knowledge indicates that although chemical and biotic factors are important forin-situ growth of dinoflagellate cells, convergence by thermal and tidal fronts is essential for cell accumulation and bloom development. Advances in physical oceanographic research, modelling and remote sensing enabled the detection of fronts and bordering eddies with high precision. There is a potential for an increased use of these technological advances in predicting and monitoring the bloom development. The present paper overviews the history and distribution of toxic dinoflagellates, and the physical factors influencing bloom development and PSP/DSP outbreaks. Future research needs to improve the predictability and control of this world-wide hazard are also discussed.

Assessment of Ciguatera Dinoflagellate Populations: Sample Variability and Algal Substrate Selection

Preliminary assessment is made of two key components in ciguatera ecology. First, we examined the numerical variability of Gambierdiscus toxicus as an epiphyte on the macroalgae, Dictyota and Galaxaura. Variability was examined by a statistical bootstrap technique to determine the minimum number of samples required to adequately estimate the abundance of G. toxicus at one station and to test for statistically significant differences between two stations. A minimum of 10 replicates were needed at the relatively low G. toxicus abundance found at our study site. Second, we demonstrated the feasibility of conducting controlled laboratory experiments to assess the short-term colonization behavior of G. toxicus on selected macroalgae offered in varying mass and surface area ratios. To assess G. toxicus abundance and distribution, the number of cells per unit of host alga must be standardized. We show that contradictory conclusions can be reached depending on whether the number of dinoflagellate cells is normali...

SPECIALIZATION OF ENDOPLASMIC RETICULUM ARCHITECTURE IN RESPONSE TO A BACTERIAL SYMBIOSIS IN PERIDINIVM BALTICUM (PYRRHOPHYTA)1

ABSTRACT Transmission elrctron microscopy (TEM) and rapid freeze‐fracture analyses were used to provide data for a more accurate description of the relationship of Peridinium balticum (Levander) Lemmermann to endosymbiotic bacteria present in the dinoflagellate's cytoplasm. Bacteria were found in all of the cells observed and were present in both gamete and zygote life cycle stages. Analysis of micrographs showed that bacteria were tightly clustered in the posterior portion of the dinoflagellate cell in numbers of 50‐200/ section. Rapid freeze‐fracture replicas re‐vealed a specialization of the endoplasmic reticulum (ER). ER surrounded each bacterium and consisted of a series of infoldings which contacted the bacterial plasma membrane in several places. Extensions of ER were common between adjacent bacteria. We have interpreted the development and specialization of endoplasmic reticulum of the dinoflagellate host as an example of a cellular response of one organism to another, supportive of the serial endosymbiotic theory. Intramembrane particles (IMP) were seen on both protoplasmic (P) and exoplasmic (E) faces of the bacterial plasma membrane. Aquisition of the bacterial symbiont and the effects of long term culturing are discussed.

Variation in photosynthetic rates among individual cells of a marine dinoflagellate

Variation in photosynthetic rates among individual cells of a marine dinoflagellate

Isolation and characterization of low molecular weight carbohydrates dissolved in seawater

Chemical characterization and quantitative determination of dissolved low molecular weight carbohydrates in Mikawa Bay near Nagoya, Japan were conducted. The water samples were collected during the period of algal bloom of the dinoflagellate Prorocentrum minimum on 31 May 1980. Low molecular weight carbohydrates in seawater samples from depths of 1 and 6 m were first retained on a charcoal column and then eluted with aqueous ethanol. The carbohydrates obtained were permethylated and then isolated into each of the components by thin layer chromatography. The sugars isolated were characterized by gas chromatography (GC), combined gas chromatography and mass spectrometry (GC-MS), proton nuclear magnetic resonance spectroscopy ( 1H-NMR) and some chemical analyses. Laminaribiose, laminaritriose, sucrose, raffinose, 2-O-α- d- glucopyranosylglycerol , 1-O-β- d- galactopyranosylglycerol 6-O-α- d- galactopyranosyl-1-O-β- d- galactopyranosylglycerol were fully characterized and quantified with ranges from 2.3 to 27.7 μg l −1 and from 0.5 to 17.8 μg l −1. These low molecular weight carbohydrates were also identified, with some difference in their relative abundance, in particulate matter consisting mainly of dinoflagellate cells collected on the same occasion from this bay. These results indicate that low molecular weight carbohydrates dissolved in seawater are directly derived from those of phytoplankton through extracellular release or cell lysis.

Scintillation and autoradiographic studies on 63 nickel uptake in Pseudomonas tabaci.

Scintillation studies on the uptake of 63Ni2+ by Pseudomonas tabaci demonstrate an incorporation of approximately 2.5 nmol per 10(10) bacterial cells, in medium containing 12 nmol (8.3 microCi) per ml. Over 80% of the incorporated Ni2+ is lost from the cells during washing, fixation and dehydration with ethanol. The remaining insoluble (bound) 63Ni2+ has the highest level in cells fixed in acetic acid/ethanol (0.4 nmol/10(10) cells), with smarter amounts in paraformaldehyde- and glutaraldehyde-fixed cells. The radioactive level in aldehyde-fixed cells represents a total Ni2+ uptake of about 10(-18) g or 10(4) atoms per cell. Light- and electron-microscope autoradiography corroborated the scintillation studies in demonstrating a higher retention of label by cells fixed in acetic acid/ethanol, possibly reflecting a higher retention of medium Mr proteins with this type of fixation. High-resolution electron-microscope autoradiography involving gold latensification with physical development demonstrated a clear localization of silver grains to the central nucleoid region (seen most clearly over the discrete nucleoid of aldehyde-fixed cells) and within this to the chromatin (seen most clearly over the condensed chromatin of acetic acid/ethanol-fixed cells). It is suggested that the incorporated 63Ni2+ labels mainly central, genetically inactive DNA, while peripheral, actively transcribing DNA has little associated radioactivity. The pattern of cation association seen in this bacterium shows a number of close similarities to the situation seen in dinoflagellate cells.

Patterns of DNA Synthesis and Cell Division in Marine Dinoflagellates1

ABSTRACTChanges in mean cell size, DNA and cell density were monitored at 6‐h intervals for 72 h in populations of six species (eight clones) of marine dinoflagellates to determine the temporal relationships between the cell cycle events of DNA replication and cytokinesis. Batch cultures were maintained at 15 or 20°C on a 12‐h light: 12‐h dark photoperiod. Cell densities and size frequency distributions were determined conductimetrically and the amount of DNA within populations was measured fluorometrically. A variety of intra‐ and interspecific relationships were observed, ranging from parallel phasing of cell cycle processes to variations which involved the temporal uncoupling of DNA synthesis from the phased pattern of cell division which is characteristic of dinoflagellate cell cycles. Daily growth rates of individual populations varied from 0.05 (Gymnodinium nelsoni) to 2.08 (Amphidinium carteri) cell divisions day‐1 and DNA doubling rates ranged from 0 to 1.14 day‐1. Mean doubling rates for DNA were usually 30–40% lower than those for cells. The degree of difference in these rates and the amount of variability evident in cell cycle sequences may be major factors in determining the rate and extent of development of dinoflagellate populations in nature.

Photosynthetically-Competent Phytoplankton from the Aphotic Zone of the Deep Ocean

Comparison of 2 water samples, one collected from 10 m, the other from the aphotic zone (1000 m) on the Costa Rica Dome in the eastern tropical Pacific Ocean, revealed the presence in both samples of pigmented cells of several diatoms, dinoflagellates and coccoid organisms. Measurements of carbon assimilation rates in temperature-controlled incubators across a light gradlent demonstrated that the assimilation number (mg C [mg Chl a]-' h-') of the 1000 m sample was about 0.8, slmilar to that of the 10 m sample. The ratios of RuBP carboxylase to other carboxylating enzymes were also similar between 10 m and the aphotic zone. However, the initial slope a and the inhlbltion parameter P were considerably higher for the deep sample than for the 10 m sample. Possible mechanisms by which these viable algae reached the aphotic zone are discussed.

Phytoplankton ecology of a barrier island estuary: Great South Bay, New York

The phytoplankton ecology of Great South Bay, New York, was studied over a 1-year period. The study area, a large barrier island estuary (coastal lagoon with estuarine circulation), was characterized by high levels of inorganic nutrients, high turbidity and a shallow euphotic zone (<2 m). Net annual primary production by phytoplankton was high—450 g C m −2 year −1—and accounted for approximately 85% of the total ecosystem primary production. Chlorophyll a-specific productivity was dependent on mean photic zone light intensity in areas of the bay <1 m in depth from September 1979 through June 1980; 65–95% of the total light extinction in those areas was attibutable to suspended solids. Nitrogenous nutrient concentration did not limit phytoplankton productivity. Diatom and dinoflagellate cell densities varied greatly over time, while cryptomonad and chlorophyte species were abundant throughtout the year. Chlorophytes of 2–4 μm (‘small forms’) were numerically dominant, and contributed approximately half of the total phytoplankton biomass. Dilution of bay water by intruding ocean water appeared to control the spatial distribution of chlorophyll a on the south side of the bay; in other areas, growth appeared to exceed the rate of dilution by flushing. Waters entrained in eelgrass beds were significantly higher in salinity and mean photic zone light intensity, and had lower phytoplankton standing stock and depth-integrated primary production than control areas; waters in the sediment plume of active clamdigging boats were statistically similar to control areas with respect to water quality and phytoplankton community characteristics.

Identification of three different anti-4S RNA sera associated with autoimmune disease

Analysis of antibodies obtained from 10 patients with myositis, 20 with systemic lupus erythematosus and 24 with scleroderma revealed that 2 patients with myositis and one with systemic lupus erythematosus produced antibodies precipitating 4S RNAs. Each of the 3 anti-4S RNA sera precipitated a different subset of the total 4S RNA population. Two sera precipitated deproteinized or gel extracted 4S RNA from Novikoff hepatoma or Dinoflagellate cells. The other serum did not precipitate deproteinized RNA and may precipitate a 4S RNA in a small RNP particle. The reactive 4S RNAs are extensively modified, which suggests that they may be tRNAs, tRNA precursors or previosly unknown RNA species.

Interaction of the Circadian Cycle with the Cell Cycle in Pyrocystis fusiformis

Dividing pairs or single cells of the large dinoflagellate, Pyrocystis fusiformis Murray, were isolated in capillary tubes and their morphology was observed over a number of days, either in a light-dark cycle or in constant darkness. Morphological stages were correlated with the first growth stage, G(1), DNA synthesis, S, the second growth stage, G(2), mitosis, M, and cytokinesis, C, segments of the cell division cycle. The S phase was identified by measuring the nuclear DNA content of cells of different morphologies by the fluorescence of 4', 6-diamidino-2-phenylindole dichloride.Cells changed from one morphological stage to the next only during the night phase of the circadian cycle, both under light-dark conditions and in continuous darkness. Cells in all segments of the cell division cycle displayed a circadian rhythm in bioluminescence. These findings are incompatible with a mechanism for circadian oscillations that invokes cycling in G(q), an hypothesized side loop from G(1). All morphological stages, not only division, appear to be phased by the circadian clock.

X-ray microanalysis of unfixed chromatin in dinoflagellate cells prepared by a monolayer cryotechnique

The fine structural appearance and elemental composition of unfixed chromatin is described for cells of the dinoflagellate Prorocentru micans prepared by a freeze-drying technique. This involves rapid freezing, cryo-dehydration and resin infiltration of a monolayer of cells dispersed over a resin base. The fine structure of the nucleus appears quite different from chemically fixed and dehydrated cells. In stained sections, the chromosomes are seen as pale areas containing diffuse chromatin, and are surrounded by electron-dense nucleoplasm which has a reticulate substructure. X-ray microanalysis reveals the presence of high levels of chromatin-associated Ca and transition metals Fe, Ni, Cu and Zn, in accordance with previous observations on chemically processed cells. Calculation of elemental mass fractions by on-line computer demonstrates an overall ratio of one divalent cation per two phosphorus groups (or per two nucleotides). This ratio is similar to that obtained previously for chemically fixed chromatin, and shows that the precise overall association of metals is not primarily determined by the process of fixation.