Exotic flagellates of coastal North Sea waters

Flagellate species have been shown to survive transocean passage by ballast water and the large dinoflagellateGymnodinium catenatum was introduced from Japanese to Tasmanian waters in this way.Gymnodinium mikimotoi—better known asGyrodinium aureolum—andFibrocapsa japonica as well asAlexandrium leeii are good candidates to have been introduced recently. Species which seem to have been introduced recently into the North Sea but apparently are transported from adjacent seas by currents into the region areGymnodinium chlorophorum andAlexandrium minutum. Species reported as introduced due to misidentifications areGymnodinium catenatum andLepidodinium viride. Under other names the speciesProrocentrum minimum, Prorocentrum redfieldii, andHeterosigma akashiwo have been known for a long time in the North Sea. The recent reports of threeChattonella species may be either due to introduction or they have been overlooked. The reasons why the introduction of flagellates into coastal North Sea waters is difficult to prove will be discussed.     

Morphology,phylogeny and toxin profiles of Gymnodinium inusitatum sp. nov., Gymnodinium catenatum and Gymnodinium microreticulatum (Dinophyceae) from the Yellow Sea,China

Four Gymnodinium species have previously been reported to produce microreticulate cysts. Worldwide, Gymnodinium catenatum strains are conservative in terms of larger subunit (LSU) rDNA and internal transcribed spacer region (ITS) sequences, but only limited information on the molecular sequences of other species is available. In the present study, we explored the diversity of Gymnodinium by incubating microreticulate cysts collected from the Yellow Sea off China. A total of 18 strains of Gymnodinium, from three species, were established. Two of these were identified as Gymnodinium catenatum and Gymnodinium microreticulatum, and the third was described as a new species, Gymnodinium inusitatum. Motile cells of G. inusitatum are similar to those of Gymnodinium trapeziforme, but they only share 82.52% similarity in LSU sequences. Cysts of G. inusitatum are polygonal in shape, with its microreticulate wall composed of approximately 14 concave sections. G. microreticulatum strains differ from each other at 69 positions (88.00% similarity) in terms of ITS sequences, whereas all G. catenatum strains share identical ITS sequences and belonged to the global populations. Phylogenetic analyses, based on LSU sequences, revealed that Gymnodinium species that produce microreticulate cysts are monophyletic. Nevertheless, the genus as a whole appears to be polyphyletic. Paralytic shellfish toxins (PSTs) were found in all G. catenatum strains tested (dominated by 11-hydroxysulfate benzoate analogs and N-sulfocarmaboyl analogs) but not in any of the G. microreticulatum and G. inusitatum strains. Our results support the premise that cyst morphology is taxonomically informative and is a potential feature for subdividing the genus Gymnodinium.     

Morphology and life cycle of Gymnodinium baicalense Ant. (Dinophyceae) from Lake Baikal

Living populations of Gymnodinium baicalense Ant. from Lake Baikal were studied. It has been shown that G. baicalense var. minor Ant. is a life stage of G. baicalense, whose species size is smaller than it is given in the diagnosis. Morphology of the living vegetative cells of G. baicalense is close to that of G. wigrense Wolosz. These species share ecological features. The vegetative cells form endogenous resting cysts. The sexual reproduction takes place in the period of the mass development of the species. The author believes that it is a relict species, which formed in the Ice age.     

Flagellar apparatus and nuclear chambers of the green dinoflagellate Gymnodinium chlorophorum

The green dinoflagellate Gymnodinium chlorophorum (BAH ME 100, the type culture) was reexamined with emphasis on the structure of the flagellar apparatus and nuclear envelope. Like other Gymnodinium species, G. chlorophorum possessed a nuclear fibrous connective linking the flagellar apparatus and the nucleus, albeit in a very reduced and unique form. Microtubules nucleated from the R3 flagellar root associated with the nuclear fibrous connective and terminated at the nucleus, a novel arrangement not known in any other dinoflagellate. Although overlooked by previous researchers, nuclear chambers were present in G. chlorophorum similar to those reported in Gymnodinium aureolum and Gymnodinium nolleri. In contrast to the type species of Gymnodinium, Gymnodinium fuscum, only one nuclear pore was present per chamber. The presence of a feeding tube (peduncle) suggests that G. chlorophorum is mixotrophic. Although the fine structure of G. chlorophorum revealed its affiliation to the Gymnodinium group the above discrepancies set it apart, indicating that it might belong in a different genus.     

Yellow water in La Jolla Bay,California, July 1980. II. suppression of zooplankton grazing

Experiments were conducted to determine how much grazing pressure adult females of Calanus pacificus Brodsky, a dominant planktonic herbivore in local waters, exerted upon a bloom of the dino- flagellate, Gymnodinium flavum Kofoid & Swezy, in the waters off La Jolla in July, 1980. One set of females was presented with water collected from the chlorophyll maximum, and a second set was presented with Thalassiosira weissflogii Grunow, which is readily ingested by Calanus. Filtration rates upon the diatom were significantly higher (8.4 ml · copepod^?1 · h^?1) than upon the dinoflagellate (0.30 ml · copepod^?1 · h^?1). Calanus did not exert a significant grazing pressure upon the dinoflagellate bloom. Gut content analyses support this conclusion. The persistence of the bloom was probably due, in part, to the avoidance of Gymnodinium flavum by copepod grazers and to the consequent lack of grazing pressure.     

Woloszynskia stoschii andGymnodinium indicum,two new freshwater dinoflagellates from India: Morphology,reproduction and cytology

The distinctive morphological and karyological features of the two new species, collected from freshwater habitats in North India, are presented and compared with those of related taxa of the generaWoloszynskia andGymnodinium. Asexual reproduction takes place through zoospore formation and sexual reproduction through planozygotes inWoloszynskia stoschii.     

Gymnodinium smaydae n. sp., a New Planktonic Phototrophic Dinoflagellate from the Coastal Waters of Western Korea: Morphology and Molecular Characterization

The marine phototrophic dinoflagellate Gymnodinium smaydae n. sp. is described from cells prepared for light, scanning, and transmission electron microscopy. Also, sequences of the small (SSU) and large subunits (LSU) and the internal transcribed spacer region (ITS1–5.8S–ITS2) of ribosomal DNA were analyzed. This newly isolated dinoflagellate possessed nuclear chambers, nuclear fibrous connective, an apical groove running in a counterclockwise direction around the apex, and a major accessory pigment peridinin, which are four key features for the genus Gymnodinium. The epicone was conical with a round apex, while the hypocone was ellipsoid. Cells growing photosynthetically were 6.3–10.9 μm long and 5.1–10.0 μm wide, and therefore smaller than any other Gymnodinium species so far reported except Gymnodinium nanum. Cells were covered with polygonal amphiesmal vesicles arranged in 11 horizontal rows, and the vesicles were smaller than those of the other Gymnodinium species. This dinoflagellate had a sharp and elongated ventral ridge reaching half way down the hypocone, unlike other Gymnodinium species. Moreover, displacement of the cingulum was 0.4–0.6 × cell length while in other known Gymnodinium species it is less than 0.3 × cell length. In addition, the new species possessed a peduncle, permanent chloroplasts, pyrenoids, trichocysts, pusule systems, and small knobs along the apical furrow, but it lacked an eyespot, nematocysts, and body scales. The sequence of the SSU, ITS1–5.8S–ITS2, and LSU rDNA region differed by 1.5–3.8%, 6.0–17.4%, and 9.1–17.5%, respectively, from those of the most closely related species. The phylogenetic trees demonstrated that the new species belonged to the Gymnodinium clade at the base of a clade consisting of Gymnodinium acidotum, Gymnodinium dorsalisulcum, Gymnodinium eucyaneum, etc. Based on morphological and molecular data, we suggest that the taxon represents a new species, Gymnodinium smaydae n. sp.     

Diurnal vertical migration of Gymnodinium mikimotoi during a red tide in Hoketsu Bay, Japan

The depth, velocity and timing of the vertical migration ofGymnodinium mikimotoi were examined using fluorometric fieldobservations. Gymnodinium mikimotoi can migrate vertically {smalltilde}20 m daily at velocities of {small tilde}2.2 m h^–1     

Carotenoids of the dinophyceae

The carotenoids of the photosynthetic dinofiagellates Amphidinium carterae (two strains), Glenodinium sp.,Gymnodinium splendens, G. nelsoni and Gyrodinium dorsum have been investigated, quantitatively and qualitatively. Peridinin is the principal carotenoid in all species; also present are β-carotene, diadinoxanthin, dinoxanthin, pyrrhoxanthin, astaxanthin, peridininol, diatoxanthin and pyrrhoxanthinol. New structures have been assigned to dinoxanthin and pyrrhoxanthin while peridininol and pyrrhoxanthinol are new carotenoids not previously reported. A carotenoid glycoside, P-457, found in four species, is a hexoside. Dinoxanthin is the only, plausible biosynthetic precursor of peridinin that could be detected.     

Gymnodinium litoralis sp. nov. (Dinophyceae), a newly identified bloom-forming dinoflagellate from the NW Mediterranean Sea

Recurrent high-biomass blooms of a gymnodinioid species have been periodically recorded at different sites in the NW Mediterranean Sea (Catalan and Sardinian coast), causing intense discolorations of the water. In this study, several strains of the causative organism were isolated and subsequently studied with respect to the morphology of the vegetative cells and different life cycle stages, pigments profile, and molecular phylogeny. Based on phylogenetic analyses, the strains were placed within the Gymnodinium sensu stricto clade. The species possessed a horseshoe-shaped apical groove running anticlockwise around the apex and the major accessory pigment was identified as peridinin. These characteristics place the organism within the Gymnodinium genus, as defined today, although some other characteristics, such as vesicular chambers in the nuclear envelope and a nuclear fibrous connective were not observed. Morphologically, the isolates highly resemble Gyrodinium vorax (Biecheler) but major differences with the latter suggest that they comprise a new species, Gymnodinium litoralis sp. nov. The resting cyst of this species is described herein from field samples of the Catalan and Sardinian coast; pellicle cysts were observed in field samples and also in cultures. This species recurrently produces high biomass blooms (>10⁶ cell L⁻¹) in summer along several beaches and coastal lagoons in the NW Mediterranean Sea (L’Estartit, La Muga River mouth, and Corru S’Ittiri). Knowledge about its geographic distribution is limited, since the precise identification of G. litoralis from the field or fixed samples can be difficult. Therefore we expect that molecular studies will reveal a much wider distribution of the species.     

SYMBIOSIS IN THE PLANKTONIC FORAMINIFER,ORBULINA UNIVERSA,AND THE ISOLATION OF ITS SYMBIOTIC DINOFLAGELLATE,GYMNODINIUM BÉII SP. NOV.1

The symbiotic association of the spinose planktonic foraminifer, Orbulina universa, with the dinoflagellate, Gymnodinium béii sp. nov., was examined with light and electron microscopy, and the symbiont was isolated into unialgal culture. The intact association is characterized by a diurnal movement of the symbionts from the distal regions of the spines during the day, to perialgal vacuoles within the host cytoplasm at night. This diurnal migration involves a daily endo- exocytotic cycle. Gymnodinium béii is non-motile and spindle-shaped within the host, whereas it is motile and gymnodinoid in shape when in culture. Ultrastructural examination revealed two or more stalked pyrenoids penetrated by lamellae, a typical dinokaryon nucleus and no trichocysts. A distinct ‘flange’projects over the sulcus from the hypocone. The swimming behavior of this dinoflagellate was characterized by intermittent darting events. Swimming speeds during a dart reached velocities of 770 μm. s^?1 as compared to a mean, non-darting swimming velocity of 126 μm. s^?1. Gymnodinium béii is eurythermal and division rates ranged between 0.16 and 0.65 divisions day^?1 for culture temperatures between 6.5 and 25° C respectively.     

Toxigenic algae and associated phycotoxins in two coastal embayments in the Ebro Delta (NW Mediterranean)

Harmful Algal Bloom (HAB) surveillance is complicated by high diversity of species and associated phycotoxins. Such species-level information on taxonomic affiliations and on cell abundance and toxin content is, however, crucial for effective monitoring, especially of aquaculture and fisheries areas. The aim addressed in this study was to determine putative HAB taxa and related phycotoxins in plankton from aquaculture sites in the Ebro Delta, NW Mediterranean. The comparative geographical distribution of potentially harmful plankton taxa was established by weekly field sampling throughout the water column during late spring–early summer over two years at key stations in Alfacs and Fangar embayments within the Ebro Delta. Core results included not only confirmed identification of HAB taxa that are common for the time period and geographical area, but also provided evidence of potentially new taxa. At least 25 HAB taxa were identified to species level, and an additional six genera were confirmed, by morphological criteria under light microscopy and/or by molecular genetics approaches involving qPCR and next generation DNA pyrosequencing. In particular, new insights were gained by the inclusion of molecular techniques, which focused attention on the HAB genera Alexandrium, Karlodinium, and Pseudo-nitzschia. Noteworthy is the discovery of Azadinium sp., a potentially new HAB species for this area, and Gymnodinium catenatum or Gymnodinium impudicum by means of light microscopy. In addition, significant amounts of the neurotoxin domoic acid (DA) were found for the first time in phytoplankton samples in the Ebro Delta. While the presence of the known DA-producing diatom genus Pseudo-nitzschia was confirmed in corresponding samples, the maximal toxin concentration did not coincide with highest cell abundances of the genus and the responsible species could not be identified. Combined findings of microscopic and molecular detection approaches underline the need for a synoptic strategy for HAB monitoring, which integrates the respective advantages and compensates for limitations of individual methods.     

Comparative study of Gymnodinium mikimotoi and Gymnodinium aureolum, comb. nov. (=Gyrodinium aureolum) based on morphology, pigment composition, and molecular data

Light and electron microscopy, nuclear-encoded LSU rDNA sequences, and pigment analyses were performed on five geographically separate isolates of Gymnodinium mikimotoi. The morphological variation between the isolates equals that found within the isolates. The nuclear-encoded LSU rDNA sequences were nearly identical in all isolates, and molecular analyses using maximum likelihood, parsimony, and neighbor joining showed the geographical isolates as an unresolved clade. Based on the available data it is concluded that the European isolates, formerly identified as Gyrodinium aureolum , Gyrodinium cf. aureolum , or Gymnodinium cf. nagasakiense , are conspecific with the Japanese Gymnodinium mikimotoi. An isolate from the Pettaquamscutt River, USA, is suggested to represent what Hulburt (1957) described as Gyrodinium aureolum. The LSU rDNA sequence data and ultrastructural characters in this isolate closely resemble those of Gymnodinium fuscum , the type species of Gymnodinium , and Gyrodinium aureolum Hulburt is therefore renamed Gymnodinium aureolum (Hulburt) G. Hansen, comb. nov.     

Phylogenetic study of Gymnodinium dorsalisulcum comb. nov. from tropical Australian coastal waters (Dinophyceae)

A new species of the dinoflagellate genus Gymnodinium Stein, previously considered a member of Katodinium Fott, is characterized from two marine benthic habitats in tropical northern Australia. Gymnodinium dorsalisulcum comb. nov. was found to be very abundant at times, and in culture produced large quantities of mucus. We analyzed two regions of ribosomal DNA from this species (partial large subunit and complete small subunit sequences), using Bayesian analysis and phylogenetic models appropriate to alignments of ribosomal RNA genes. We compared it to eight species of the ‘true’Gymnodinium clade and to other dinoflagellates. The results show that it is a member of the Gymnodinium clade, and is closely related to Gymnodinium impudicum and G. chlorophorum. Katodinium was originally defined as having cells with an epitheca that is much larger than the hypotheca. However, this character is clearly inadequate, and the genus requires a re‐investigation to determine the apomorphies of the type species.     

Phylogenetic relations of the dinoflagellate Gymnodinium baicalense from Lake Baikal

Freshwater dinoflagellates still remain poorly studied by modern biological methods. This lack of knowledge prevents us from understanding the evolution and colonization patterns of these ecologically important protists. Gymnodinium baicalense is the most abundant, and possibly endemic, planktonic dinoflagellate from the ancient Lake Baikal. This dinoflagellate species blooms in the spring under the ice. This study analyzed the origin of this Baikalian dinoflagellate using three markers (two ribosomal and one mitochondrial DNA). It was found that this species is a true member of the order Gymnodiniales and has close relatives in the glacial melt waters of the Arctic Ocean. It seems that G. baicalense has diversified relatively recently from the arctic marine gymnodinioids. These results shed light on dinoflagellate biogeography and their colonizations in Lake Baikala biodiversity hotspot.     

INVESTIGATION OF 4-METHYL STEROLS FROM CULTURED DINOFLAGELLATE ALGAL STRAINS

The marine dinoflagellates Prorocentrum micans, Gonyaulax polyedra, Gymnodinium sp., and Alexandrium tamarense, collected from the Adriatic Sea during red-tide blooms, were cultured to investigate the 4-methyl sterol constituents. To ascertain a possible influence of cell age on the 4-methyl sterol content, for one strain (Gymnodinium sp.)we investigated the composition of these constituents at exponential and stationary growing phases. The lipid material extracted with acetone from the lyophilized algal samples was fractionated by thin-layer chromatography. The 4-methyl sterols recovered from the layer were converted into the corresponding OTMS derivatives. Nine of 11 constituents were identified by gas chromatography and gas chromatography-mass spectrometry; only two minor constituents were characterized by their gas chromatographic parameters. All free methyl sterols identified in the algal samples had been detected previously in various dinoflagellates. The 4-methyl sterol fractions generally contained very few constituents. Except for the Gymnodinium sp. sample, collected at the exponential growing phase (GyD2 exp), which contains 4,24-dimethylcholestan-3-ol as a unique constituent, dinosterol was the major component. Moreover, 4,24-ethylcholestan-3-ol was also an important constituent of both Prorocentrum and Gonyaulax strains, whereas considerable amounts of dinostanol characterized all the Gymnodinium sp. strains. In addition, the latter contained several minor constituents such as 4-methylcholestan-3-ol, 4,24-dimethylcholesta-22-en-3-ol, and 4-methyl-24-ethylcholestan-3-ol. 4-Methyl-24-methylene-cholestan-3-ol was a constituent of the Gymnodinium sp. sample, collected at the stationary growing phase (GyD2 stat)only, whereas 4-methylgorgostanol was identified only in the Alexandrium tamarense Gt4 strain. Except for 4-methyl-24-ethylcholesta-8(14)-en-3-ol, all the methyl sterol constituents from our algae show a saturated polynuclear system. The pathways by which side-chain modifications occur in dinoflagellate 4-methyl sterols are considered, and a map of the fragmentation pattern of the trimethylsilyl-4-methyl sterols under electronic impact is also reported.     

Histone-like protein and chromatin structure in the wall-less dinoflagellate Gymnodinium nelsoni

Basic nuclear proteins from the wall-less dinoflagellate Gymnodinium nelsoni were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS). One major histone-like protein with a molecular weight of about 10 000 was present in acid extracts of whole nuclei and chromatin isolated from growing cultures. In addition, two minor components of 17 000 and 13 000 daltons were also noted. Chromatin fibers spread by the microcentrifugation technique showed no indication of a subunit structure, but instead appeared as smooth threads with a diameter of about 6.5 nm.     

Gymnodinium helveticum Penard F. Achroum Skuja a case of phagotrophy

Gymnodinium helveticum f. achroum in Blelham Tarn was occasionally noted to exhibit phagotrophy. Various algae were observed inside Gymnodinium cells. The seeding of an experimental enclosure with dead Lycopodium spores resulted in some of the Gymnodinium cells containing a spore.

From the results, it is suggested that this dinoflagellate exhibits non-selective and non-predatory phagotrophy.