Protoperidinium tricingulatum sp. nov. (Dinophyceae), a new motile form of a round,brown, and spiny dinoflagellate cyst

A small, broadly ovoidal and heterotrophic dinoflagellate containing round, brownish, and spiny cyst was found in the water column of Huibertsplaat in the Wadden Sea off the coast of the Netherlands. This dinoflagellate had these conspicuous morphological characters: a five‐sided first apical plate (1′), only three cingular plates, and an extremely small first antapical plate. Based on these morphological features, Protoperidinium tricingulatum Kawami, vanWezel, Koeman et Matsuoka is described as a new species. The flagellar pore of P. tricingulatum is covered with a small fin, which rises from the left side of the right sulcal plate to the large V‐shaped posterior sulcal plate. This feature suggests that P. tricingulatum is assigned to the Abé's Monovela Group. The cyst stage of P. tricingulatum was positively linked to the vegetative stage by comparison of the ribosomal 5.8S rDNA, internal transcribed spacers (ITS1 and ITS2). Living cysts of P. tricingulatum are round, brownish, and covered with many slender spines bearing capitate or cauliforate distal ends. The cyst also possesses a theropylic archeopyle formed by a slit corresponding to parasutures between three apical and two apical intercaraly plates. These morphological characters indicate that this species is morphologically related to two dinoflagellate cyst‐genera Islandinium and Echinidinium.     

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

Cellular 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.     

A further phylogenetic study of the heterotrophic dinoflagellate genus, Protoperidinium (Dinophyceae) based on small and large subunit ribosomal RNA gene sequences

The heterotrophic marine dinoflagellate genus Protoperidinium is the largest genus in the Dinophyceae. Previously, we reported on the intrageneric and intergeneric phylogenetic relationships of 10 species of Protoperidinium, from four sections, based on small subunit (SSU) rDNA sequences. The present paper reports on the impact of data from an additional 5 species and, therefore, an additional two sections, using the SSU rDNA data, but now also incorporating sequence data from the large subunit (LSU) rDNA. These sequences, in isolation and in combination, were used to reconstruct the evolutionary history of the genus. The LSU rDNA trees support a monophyletic genus, but the phylogenetic position within the Dinophyceae remains ambiguous. The SSU, LSU and SSU + LSU rDNA phylogenies support monophyly in the sections Avellana, Divergentia, Oceanica and Protoperidinium, but the section Conica is paraphyletic. Therefore, the concept of discrete taxonomic sections based on the shape of 1′ plate and 2a plate is upheld by molecular phylogeny. Furthermore, the section Oceanica is indicated as having an early divergence from other groups within the genus. The sections Avellana and Excentrica and a clade combining the sections Divergentia/Protoperidinium derived from Conica‐type dinoflagellates independently. Analysis of the LSU rDNA data resulted in the same phylogeny as that obtained using SSU rDNA data and, with increased taxon sampling, including members of new sections, a clearer idea of the evolution of morphological features within the genus Protoperidinium was obtained. Intraspecific variation was found in Protoperidinium conicum (Gran) Balech, Protoperidinium excentricum (Paulsen) Balech and Protoperidinium pellucidum Bergh based on SSU rDNA data and also in Protoperidinium claudicans (Paulsen) Balech, P. conicum and Protoperidinium denticulatum (Gran et Braarud) Balech based on LSU rDNA sequences. The common occurrence of base pair substitutions in P. conicum is indicative of the presence of cryptic species.     

Cyst‐theca relationship of a new dinoflagellate with a spiny round brown cyst,Protoperidinium lewisiae sp. nov., and its comparison to the cyst of Oblea acanthocysta

Round spiny brown cysts with apiculocavate processes were isolated from sediments of Lake Saroma, Japan, Changle Harbor, East China Sea, China, Jinzhou Harbor, Bohai Sea, China, and San Pedro Harbor, California, USA. Superficially similar round spiny brown cysts of the species, Oblea acanthocysta were, for comparison, restudied through light microscopy and scanning electron microscopy (SEM) and by sequencing of small subunit (SSU) and large subunit (LSU) rDNA obtained through a single cyst from Lake Saroma. These morphological measurements and SEM observations showed that the new cysts can be discriminated from O. acanthocysta by the archeopyle, number of processes, shape of process bases and its apiculocavate processes. Based on LSU sequences, the most closely related species was Protoperidinium monovelum, for which no cyst stage has been described so far. However, the thecal morphology of the specimens found in this study differed from P. monovelum in details of the sulcal plates and shape of apical pore and 2a plate. We therefore describe Protoperidinium lewisiae sp. nov., which can be found in estuarine subtropical to temperate waters of the Pacific Ocean.     

Heterocapsa psammophila sp. nov. (Peridiniales, Dinophyceae), a new sand-dwelling marine dinoflagellate

A new armored dinoflagellate species, Heterocapsa psammophila Tamura, Iwataki et Horiguchi sp. nov. is described from Kenmin‐no‐hama beach, Hiroshima, Japan using light and electron microscopy. This dinoflagellate possesses the typical thecal plate arrangement of the genus Heterocapsa, Po, cp, 5′, 3a, 7′′, 6c, 5s, 5′′′, 2′′′′; and the 3‐D body scales of Heterocapsa on the plasma membrane. The cell shape is ovoidal. The spherical nucleus and the pyrenoid are situated in the hypotheca and the epitheca, respectively. The ultrastructure of H. psammophila is typical of dinoflagellates and the pyrenoid is invaginated by cytoplasmic tubules. H. psammophila is distinguished from all other hitherto‐described Heterocapsa species by the cell shape, the relative position of the nucleus and pyrenoid and the structure of the body scale. The habitat and behavior of this new species in culture suggest that the organism is truly a sand‐dwelling species.     

Cyst-theca relationship and phylogenetic positions of Scrippsiella plana sp. nov. and S. spinifera (Peridiniales,Dinophyceae)

Species belonging to the dinophyte genus Scrippsiella are frequently reported in marine waters, but information on their distribution in brackish environments is limited. Here we describe a new species, S. plana, through incubation of non-calcified cysts from sediments collected in the South China Sea and Caspian Sea. The vegetative cells consist of a conical epitheca and a rounded hypotheca with the plate formula of Po, X, 4′, 3a, 7′′, 5C+t, 5S, 5′′′, 2′′′′. It differs from other Scrippsiella species by its flattened body in dorsoventral view and a small first anterior intercalary (1a) plate (half the size of plate 3a). Scrippsiella plana strains from the South China Sea and Caspian Sea share identical internal transcribed spacer (ITS) sequences, and show phenotypic plasticity and local adaptation in growth rate at various salinities, consistent with the environments in which they originated. In addition, two strains of S. spinifera were obtained by incubating ellipsoid cysts with calcareous spines from sediments collected along the Turkish and Hawaiian coast. They also share identical ITS sequences and differ from Duboscquodinium collinii (a parasite of tintinnids) only at two base pair positions (in the ITS2 region). Molecular phylogeny based on ITS and large subunit ribosomal DNA (LSU rDNA) sequences revealed that S. plana was nested within the Calciodinellum (CAL) clade and S. spinifera within the S. trochoidea (STR) clade. The phylogenetic position of ‘Peridinium’ wisconsinense is reported for the first time, which supports multiple transitions of the Peridiniales to freshwater.     

Plagiodinium ballux sp. nov. (Dinophyceae), a deep (36 m) sand dwelling dinoflagellate from subtropical Japan

A new species of marine sand‐dwelling dinoflagellate, Plagiodinium ballux N. Yamada, Dawut, R. Terada & T. Horiguchi is described from a deep (36 m) seafloor off Takeshima Island, Kagoshima Prefecture, Japan in the subtropical region of the northwest Pacific. The species is thecate and superficially resembles species of Prorocentrum, but possesses an extremely small epitheca. The cell varies from ovoid to a rounded square, and is small (15.0–22.5 μm in length) and laterally compressed. The thecal plates are smooth and the thecal plate arrangement (Po, 1′, 0a, 5″, 5C, 2S, 5?, 0p, 1″″) is similar to that of Plagiodinium belizeanum, the type species of the genus. Molecular phylogenetic analyses based on SSU rDNA and partial LSU rDNA reveal that the dinoflagellate is closely related to P. belizeanum, but it can be clearly distinguished by its size and cell shape. This suite of morphological and molecular differences leads to the conclusion that this deep benthic dinoflagellate represents a new species of the genus Plagiodinium.     

Research Note: Molecular phylogenetic affiliations of Dissodinium pseudolunula, Pheopolykrikos hartmannii, Polykrikos cf. schwartzii and Polykrikos kofoidii to Gymnodinium sensu stricto species (Dinophyceae)

Partial large subunit ribosomal DNA (LSU rDNA) sequences of Dissodinium pseudolunula Swift ex Elbrächter et Drebes, Pheopolykrikos hartmannii (Zimmermann) Matsuoka et Fukuyo, Polykrikos cf. schwartzii Bütschli and Polykrikos kofoidii Chatton were analyzed to reveal their phylogenetic status. The four athecate dinoflagellate species fell within the clade of Gymnodinium sensu G. Hansen et Moestrup supported by high likelihood values without apparent phylogenetic relationships to neither Gymnodinium species nor each other. Their genetic affiliations to typical Gymnodinium species were morphologically supported by the loop-shaped apical grooves in Pheopolykrikos and Polykrikos species and the Gymnodinium -like motile stage at least once during pleomorphic life cycles in D. pseudolunula and Ph. hartmannii .     

Ankistrodinium armigerum sp. nov. (Dinophyceae), a new species of heterotrophic marine sand‐dwelling dinoflagellate from Japan and Australia

A new heterotrophic sand‐dwelling dinoflagellate, Ankistrodinium armigerum K. Watanabe, Miyoshi, Kubo, Murray et Horiguchi sp. nov., is described from Ishikari Beach, Hokkaido, Japan and Port Botany, NSW, Australia. The dinoflagellate is laterally compressed, possessing a short triangular epicone and a large sac‐like hypocone. It possesses a right‐handed cingulum and a deeply‐incised sulcus. The sulcus descends towards the posterior of the cell where it becomes much deeper and wider, resulting in a bilobed ventral side to the hypocone, with a greater excavation of the left lobe than the right. In addition, the right lobe of the hypocone is shorter than the left lobe, which allows a partial view of the left sulcal wall when the cell is viewed from its right side. The sulcus ascends in the epicone to form an apical groove. The apical groove is linear but terminates in an ellipsoid fashion and its extremity approaches, but does not form a closed loop with the apical end of the linear portion. The dinoflagellate possesses two distinct size classes of trichocysts. The large trichocysts are located in the posterior part of the cell, while small trichocysts are distributed throughout the cell. The dinoflagellate shares morphological characteristics with the heterotrophic sand‐dwelling dinoflagellate, Ankistrodinium semilunatum, the type species of the genus. These include a laterally compressed cell, a right‐handed cingulum, a deeply‐incised sulcus and the same basic structure to the apical groove. Molecular phylogenetic analyses based on small and large subunits of rDNA showed that in both trees, A. semilunatum and A. armigerum formed a robust clade, suggesting that these two species are closely related. Because no organism with the characteristics of this species exists and because this species is closely related to A. semilunatum, we concluded that this species should be described as a second species of the genus Ankistrodinium.     

Alexandrium camurascutulum sp. nov. (Dinophyceae): a new dinoflagellate species from New Zealand

A new species, Alexandrium camurascutulum sp. nov. MacKenzie et Todd, is described from specimens collected from Tasman Bay and the Marlborough Sounds New Zealand. These small (26–28 μm long × 21–24 μm wide) cells can be discriminated from other species in the Alexandrium minutum group by three distinctive morphological features. The sixth pre-cingular plate (6′′) is up to 1.6 times wider than high and the left side of the plate is concave resulting in a markedly ‘hooked’ appearance. In all specimens observed, the first apical plate (1′) does not directly connect with the apical pore plate (Po) and the posterior sulcal plate (S.p.) is markedly different from the usual A. minutum form and may contain a posterior attachment pore (pap) connected to the right side plate margin. The cells may or may not have an anterior attachment pore (aap) in the apical pore plate (Po). The cells display a prominent list along the left sulcal margin and the thecal surface is perforated with numerous areolated pores. A. camurascutulum sp. nov. has been observed occasionally over a number of years in coastal waters of the northern South Island of New Zealand. There is circumstantial evidence that suggests it is not toxic.     

Discovery of a novel type of body scale in the marine dinoflagellate,Amphidinium cupulatisquama sp. nov. (Dinophyceae)

A new species of Amphidinium, A. cupulatisquama Tamura et Horiguchi, from sand samples from Ikei Island, Okinawa Prefecture in subtropical Japan, is described based on light, scanning and transmission electron microscopy and the partial sequencing of the large subunit rDNA gene. The species has a typical morphology for the genus, but is distinguished from previously described species by having a combination of the following characteristics: (i) a relatively large cell (over 30 µm in length); (ii) possessing an eyespot on the dorsal side of the cingulum; (iii) the longitudinal flagellum emerging from a point close to the cingulum; (iv) cell division taking place in the motile phase; and (v) possessing body scales. This is the third species of this genus to possess body scales. The body scales of A. cupulatisquama are uniform and cup‐shaped in side view and elliptical in face view. Their dimensions are 136.4 nm by 91.0 nm by 81.8 nm high. In side view, the scale is seen to have a thick lower half and a thin upper half. This scale type is very different from those of previously reported Amphidinium species (HG114 and HG115). The molecular tree indicated that A. cupulatisquama and the two other strains of body scale‐bearing Amphidinium are distantly related within the Amphidinium clade.     

Laciniporus arabicus gen. et sp. nov. (Dinophyceae,Peridiniales), a new thecate,marine, sand‐dwelling dinoflagellate from the northern Indian Ocean (Arabian Sea)1

A new thecate, photosynthetic, sand‐dwelling marine dinoflagellate, Laciniporus arabicus gen. et sp. nov., is described from the subtidal sediments of the Omani coast in the Arabian Sea, northern Indian Ocean, based on detailed morphological and molecular data. Cells of L. arabicus are small (16.2–30.1 μm long and 13.1–23.2 μm wide), dorsoventrally compressed, with a small apical flap‐shaped projection pointing to the left. The thecal plate pattern is distinguished by minute first precingular plate and sulcus, which extends into the epitheca, with large anterior and right sulcal plates. The Kofoidian thecal tabulation is Po, X, 4′, 2a, 7′′, 6c, 6s, 5′′′, 2′′′′. Morphologically, the revealed plate pattern has an affinity to the Peridiniales, and LSU rDNA based phylogenetic analyses placed L. arabicus within the Thoracosphaeraceae, close to calcareous‐cyst producing scrippsielloids, predatory pfiesteriaceans, and photosynthetic freshwater peridinioids Chimonodinium lomnickii and Apocalathium spp. However, the thecal plate arrangement of L. arabicus differs noticeably from any currently described dinoflagellates, and the species stands out from closely related taxa by extensive differences in physiology and ecology.     

Morphology,ultrastructure and molecular phylogeny of cyst-producing Caladoa arcachonensis gen. et sp. nov. (Peridiniales,Dinophyceae) from France and Indonesia

The dinoflagellate order Peridiniales encompasses several well circumscribed families. However, the family level of some genera, such as Bysmatrum and Vulcanodinium, has remained elusive for many years. Four Peridinium-like strains were established from the Atlantic coast of France and North Sulawesi, Indonesia through cyst germination or isolation of single cells. The cyst-theca relationship was established on specimens from the French Atlantic. Their morphologies were examined using light, scanning and transmission electron microscopy. The cells were characterized by a much larger epitheca relative to the hypotheca, a large anterior sulcal (Sa) plate deeply intruding the epitheca and a small first anterior intercalary plate. The plate formula was identified as Po, cp, X, 4′, 3a, 7′′, 6C, 5S, 5′′′, 2′′′′, shared by Apocalathium, Chimonodinium, Fusiperidinium and Scrippsiella of the family Thoracosphaeraceae but the configuration of Sa plate and anterior intercalary plates is different. Transmission electron microscopy showed that the eyespot was located within a chloroplast comprising two rows of lipid globules and thus belongs to type A. All four strains were classified within a new genus Caladoa as C. arcachonensis gen. et sp. nov. Small subunit ribosomal DNA (SSU rDNA), partial large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer ribosomal DNA (ITS rDNA) sequences were obtained from all strains. Genetic distance based on ITS rDNA sequences between French and Indonesian strains reached 0.17, suggesting cryptic speciation in C. arcachonensis. The maximum likelihood and Bayesian inference analysis based on concatenated data from SSU and LSU rDNA sequences revealed that Caladoa is monophyletic and closest to Bysmatrum. Our results supported that Caladoa and Bysmatrum are members of the order Peridiniales but their family level remains to be determined. Our results also support that Vulcanodinium is closest to the family Peridiniaceae.     

Ultrastructure and molecular phylogenetic position of a new marine sand-dwelling dinoflagellate from British Columbia,Canada: Pseudadenoides polypyrenoides sp. nov. (Dinophyceae)

Two monospecific genera of marine benthic dinoflagellates, Adenoides and Pseudadenoides, have unusual thecal tabulation patterns (lack of cingular plates in the former; and no precingular plates and a complete posterior intercalary plate series in the latter) and are thus difficult to place within a phylogenetic framework. Although both genera share morphological similarities, they have not formed sister taxa in previous molecular phylogenetic analyses. We discovered and characterized a new species of Pseudadenoides, P. polypyrenoides sp. nov., at both the ultrastructural and molecular phylogenetic levels. Molecular phylogenetic analyses of SSU and LSU rDNA sequences demonstrated a close relationship between P. polypyrenoides sp. nov. and Pseudadenoides kofoidii, and Adenoides and Pseudadenoides formed sister taxa in phylogenetic trees inferred from LSU rDNA sequences. Comparisons of morphological traits, such as the apical pore complex (APC), demonstrated similarities between Adenoides, Pseudadenoides and several planktonic genera (e.g. Heterocapsa, Azadinium and Amphidoma). Molecular phylogenetic analyses of SSU and LSU rDNA sequences also demonstrated an undescribed species within Adenoides.     

Taxonomy and phylogeny of a new kleptoplastidal dinoflagellate, Gymnodinium myriopyrenoides sp. nov. (Gymnodiniales, Dinophyceae), and its cryptophyte symbiont

A new kleptoplastidal dinoflagellate, Gymnodinium myriopyrenoides sp. nov., was described using light microscopy, electron microscopy and phylogengetic analysis based on partial LSU rDNA sequences. Cells were dorsiventrally flattened, elongate-elliptical in ventral view. There was no displacement of the cingulum encircling the anterior part of the cell. The cingulum was curved posteriorly at the terminal junction with the sulcus. The sulcus was generally narrow but expanded in the posterior end. The epicone possessed an apical groove made of one and one-half counterclockwise revolutions. Phylogenetic analysis based on LSU rDNA showed that the sequence of G. myriopyrenoides was included in the Gymnodiniales sensu stricto clade and had special affinities with the species Amphidinium poecilochroum and Gymnodinium acidotum, which also harbor kleptochloroplasts. Phylogenetic analysis based on plastid-encoded SSU rDNA and ultrastructural observations suggested that the symbionts of G. myriopyrenoides were cryptophytes of the genus Chroomonas or Hemiselmis. Organelles including the nucleus, the nucleomorph, mitochondria, Golgi bodies and large chloroplasts remained in the cytoplasm of the symbionts, but not the periplast, ejectosomes or flagellar apparatus. The symbiotic level of G. myriopyrenoides was estimated to be a relatively early stage in the unarmored kleptoplastidal dinoflagellates.