Aika Yamaguchi

A new heterotrophic dinoflagellate from the North-eastern Pacific, Protoperidinium fukuyoi: cyst–theca relationship, phylogeny, distribution and ecology

The cyst–theca relationship of Protoperidinium fukuyoi n. sp. (Dinoflagellata, Protoperidiniaceae) is established by incubating resting cysts from estuarine sediments off southern Vancouver Island, British Columbia, Canada, and San Pedro Harbor, California, USA. The cysts have a brown‐coloured wall, and are characterized by a saphopylic archeopyle comprising three apical plates, the apical pore plate and canal plate; and acuminate processes typically arranged in linear clusters. We elucidate the phylogenetic relationship of P. fukuyoi through large and small subunit (LSU and SSU) rDNA sequences, and also report the SSU of the cyst‐defined species Islandinium minutum (Harland & Reid) Head et al. 2001. Molecular phylogenetic analysis by SSU rDNA shows that both species are closely related to Protoperidinium americanum (Gran & Braarud 1935) Balech 1974. Large subunit rDNA phylogeny also supports a close relationship between P. fukuyoi and P. americanum. Three subgroups in total are further characterized within the Monovela group. The cyst of P. fukuyoi shows a wide geographical range along the coastal tropical to temperate areas of the North‐east Pacific, its distribution reflecting optimal summer sea‐surface temperatures of ~14–18 °C and salinities of 22–34 psu.

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.

Relationship between the dinoflagellate cyst Spiniferites pachydermus and Gonyaulax ellegaardiae sp. nov. from Izmir Bay, Turkey, and molecular characterization.

Here, we established the cyst‐motile stage relation‐ship for Spiniferites pachydermus through incubation of cysts with a characteristically microreticulate/perforate surface isolated from Izmir Bay in the eastern Aegean Sea of the eastern Mediterranean. The morphology of the motile stage was similar to Gonyaulax spinifera but had a different size, overhang, displacement and reticulations. Based on the distinct morphology of the cyst and morphological differences in motile cells, we assigned S. pachydermus from Izmir Bay to the new species Gonyaulax ellegaardiae. We elucidate the phylogenetic relationship of G. ellegaardiae through large and small subunit ribosomal DNA and show that it forms a clade with other species that belong to the G. spinifera complex.

Molecular Phylogeny of the Benthic Dinoflagellate Genus Amphidiniopsis and its Relationships with the Family Protoperidiniaceae

The genus Amphidiniopsis is a benthic (sand-dwelling) lineage of thecate dinoflagellates, containing 19 morphologically diverse species. Past work has shown that some Amphidiniopsis species form a clade with the sand-dwelling Herdmania litoralis as well as some planktonic species in the family Protoperidiniaceae (i.e. the Monovela group). Still, our contemporary knowledge regarding Amphidiniopsis is limited, compared to the Protoperidiniaceae. To this end, we obtained 18S rDNA data from seven Amphidiniopsis species and a part of the 28S rDNA from four Amphidiniopsis species, with the goal of improving our understanding of phylogenetic relationships among Amphidiniopsis and the Monovela group. Results from the molecular phylogenetic analyses showed that Amphidiniopsis spp., with the exception of A. cf. arenaria, H. litoralis, and members within the Monovela group formed a single clade. Within the clade, relationships among Amphidiniopsis spp. and the Monovela group were more complicated - some subclades contained both representatives of Amphidiniopsis and the Monovela group. Our study suggests that habitat (benthic or planktonic), as well as traditionally used, general morphological characteristics, do not reflect molecular phylogenetic relationships, and that the taxonomy of the sand-dwelling genus Amphidiniopsis, and the planktonic family Protoperidiniaceae, should be reconsidered simultaneously.

Ultrastructure of the marine benthic dinoflagellate Plagiodinium belizeanum (Dinophyceae) from the southeast Pacific island of Okinawa, Japan

Abstract: We isolated Plagiodinium belizeanum into clonal culture from the Pacific island of Okinawa (Japan) and characterized it using a combination of light microscopy, scanning electron microscopy, transmission electron microscopy and 18S/28S ribosomal (r) gene sequences. Although molecular phylogenetic analyses of 18S rDNA and 28S rDNA sequences were unable to resolve the phylogenetic position of P. belizeanum within dinoflagellates, the ultrastructural data provided some new traits for the species. For instance, double-membrane-bound vesicles, distinct from the mitochondria, were interpreted as autolysosomes containing electron-dense virus particles. The thecal plate pattern was Po 1′ 0a 5′′ 5(6)C 4S 5′′′ 0p 1′′′′, which is slightly different from the original description in having an additional epithecal plate and four sulcal plates. The laterally flattened cells were 22–34 μm long, 11–13 μm deep, and 15–18 μm wide and contained a peridinin-type plastid with lobes radiating from a central pyrenoid that lacked starch sheaths and was traversed by stacks of thylakoids. This isolate represents the first record of the species in Japan, and the new ultrastructural and DNA sequence data were used to emend the species description.

Pentaplacodinium saltonense gen. et sp. nov. (Dinophyceae) and its relationship to the cyst-defined genus Operculodinium and yessotoxin-producing Protoceratium reticulatum

Strains of a dinoflagellate from the Salton Sea, previously identified as Protoceratium reticulatum and yessotoxin producing, have been reexamined morphologically and genetically and Pentaplacodinium saltonense n. gen. et sp. is erected to accommodate this species. Pentaplacodinium saltonense differs from Protoceratium reticulatum (Claparède et Lachmann 1859) Bütschli 1885 in the number of precingular plates (five vs. six), cingular displacement (two widths vs. one), and distinct cyst morphology. Incubation experiments (excystment and encystment) show that the resting cyst of Pentaplacodinium saltonense is morphologically most similar to the cyst-defined species Operculodinium israelianum (Rossignol, 1962) Wall (1967) and O. psilatum Wall (1967). Collections of comparative material from around the globe (including Protoceratium reticulatum and the genus Ceratocorys) and single cell PCR were used to clarify molecular phylogenies. Variable regions in the LSU (three new sequences), SSU (12 new sequences) and intergenic ITS 1-2 (14 new sequences) were obtained. These show that Pentaplacodinium saltonense and Protoceratium reticulatum form two distinct clades. Pentaplacodinium saltonense forms a monophyletic clade with several unidentified strains from Malaysia. LSU and SSU rDNA sequences of three species of Ceratocorys (C. armata, C. gourreti, C. horrida) from the Mediterranean and several other unidentified strains from Malaysia form a well-supported sister clade. The unique phylogenetic position of an unidentified strain from Hawaii is also documented and requires further examination. In addition, based on the V9 SSU topology (bootstrap values >80%), specimens from Elands Bay (South Africa), originally described as Gonyaulax grindleyi by Reinecke (1967), cluster with Protoceratium reticulatum. The known range of Pentaplacodinium saltonense is tropical to subtropical, and its cyst is recorded as a fossil in upper Cenozoic sediments. Protoceratium reticulatum and Pentaplacodinium saltonense seem to inhabit different niches: motile stages of these dinoflagellates have not been found in the same plankton sample.

Molecular Phylogeny and Morphology of Haplozoon ezoense n. sp. (Dinophyceae): A Parasitic Dinoflagellate with Ultrastructural Evidence of Remnant Non-photosynthetic Plastids

This study describes a novel species of Haplozoon, H. ezoense n. sp., a dinoflagellate parasite isolated from the intestines of Praxillella pacifica (Polychaeta). Trophonts (feeding stages) of H. ezoense n. sp. were isolated and studied with scanning and transmission electron microscopy, and molecular phylogenetic analyses was performed using 18S rDNA and 28S rDNA. Trophonts had an average length of 120μm, and were linear, forming a single longitudinal row comprising a trophocyte with a stylet, an average of 14 gonocytes (width=10μm), and bulbous cells that we concluded were likely sporocytes. The surface of H. ezoense n. sp. was covered with projections of the amphiesma. Sections viewed under TEM revealed multiple triple membrane-bound organelles reminiscent of relic non-photosynthetic plastids within the gonocytes. Haplozoon ezoense n. sp., H. praxillellae, and H. axiothellae formed a well-supported clade in the 18S rDNA datasets. The sequences of H. ezoense n. sp. differed from H. praxillellae, a species of Haplozoon isolated from the same host species in the Northeast Pacific, at 88/1,748 bases; and 155/1,752 bases from H. axiothellae. Concatenated 18S rDNA and 28S rDNA datasets were unable to resolve the deeper relationships of Haplozoon in the context of dinoflagellates.

Molecular phylogeny of the benthic dinoflagellate Cabra matta (Dinophyceae) from Okinawa, Japan

Abstract Cabra is a genus of marine heterotrophic thecate dinoflagellates with a peculiar cell shape and thecal plate tabulation. Previous studies suggested it was related to the benthic genus Roscoffia based on morphological features. A previous molecular phylogenetic study using 18S rDNA showed that Roscoffia capitata and members of the family Podolampadaceae formed a well-supported monophyletic group; however, the classification of Cabra remains uncertain. In this study, the first molecular phylogenetic analysis based on 18S rDNA sequence for the genus was performed using specimens of Cabra matta collected in Okinawa, Japan. Molecular phylogeny showed that C. matta formed a clade with R. capitata and the planktonic oceanic members of Podolampadaceae including Podolampas spp. and Blepharocysta sp. This result supported previous studies that inferred the close affinity between Cabra, podolampadaceans and R. capitata based on a very narrow and long 1′ plate, and the diminutive 2′ and 3′ plates, the larger...

The affiliation of Hexasterias problematica and Halodinium verrucatum sp. nov. to ciliate cysts based on molecular phylogeny and cyst wall composition

Species in the genera Hexasterias and Halodinium have been recorded over the last decades as acritarchs in palynological and/or plankton studies. In paleoenvironmental studies, these resting stages are often interpreted as indicators of freshwater input. The biological affinity of these genera has never been definitely established. Here, a new species, Halodinium verrucatum sp. nov., is described and molecular evidence (single specimen SSU and LSU rDNA sequencing) reveals that both this new species and Hexasterias problematica, collected from sediment samples in the Skagerrak and Baltic Sea, are resting stages of prorodontid ciliates. Additionally, infrared spectroscopic analysis (micro-FTIR) of Hexasterias problematica and Halodinium spp. specimens indicates a carbohydrate-based composition of the cyst wall with evidence for nitrogen-containing functional groups. A similar composition is recorded for tintinnid loricae, which further supports the placement of Hexasterias and Halodinium as ciliate cysts, and the composition is consistent with the heterotrophic nature of ciliates. The morphologically similar species Radiosperma corbiferum has a comparable composition, suggesting a similar ciliate affinity and indicating the utility of micro-FTIR in understanding acritarch affinity. Hexasterias problematica typically occurs in coastal waters from temperate to arctic regions. Halodinium verrucatum sp. nov. is observed in temperate estuarine sediments in the northern hemisphere.

Molecular Phylogeny of the Marine Planktonic Dinoflagellate Oxytoxum and Corythodinium (Peridiniales, Dinophyceae)

The dinoflagellate genera Oxytoxum and Corythodinium that account for more than fifty species are widespread in warm oceans. These genera have been considered synonyms and thecal plate designations varied among authors. Several planktonic and sand-dwelling genera have been placed within the Oxytoxaceae. We obtained the first molecular data based on small subunit (SSU) rRNA gene sequences of Oxytoxum and Corythodinium, including the type species (O. scolopax and C. tessellatum) and C. frenguellii and C. cristatum. The three species of Corythodinium branched together a strong support [bootstrap (BP) of 98%]. This formed a sister clade with moderate support (BP 75%) with O. scolopax that supported the generic split. Oxytoxaceae should exclusively remain for Oxytoxum and Corythodinium, as an independent group, unrelated to any other known dinoflagellate. Oxytoxum was characterized by spindle-shaped cells with an anterior narrow epitheca, an apical spine and little cingular displacement. Corythodinium exhibits relatively broad cell shapes, with wider epitheca and greater cingular displacement, and an obovate or pentangular anterior sulcal plate that noticeably indented the epitheca. This suggested the need of new combinations for species that were described as Oxytoxum and possessed the characteristics of Corythodinium.