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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 relationships of the common armored dinoflagellate Protoperidinium oblongum were re-investigated by incubation experiments and molecular phylogenetic analysis. Living cysts provided for incubation experiments were collected from several places in Japan, including Omura Bay, West Japan and Lake Saroma in Hokkaido, North Japan. One hundred and four cysts were incubated and 18 motile cells were germinated from these cysts. To clarify their morphological characteristics both cysts and germinated thecate cells were observed, especially the archeopyle type, the number and shape of anterior intercalary plates, and the shape and development of apical and antapical horns. In order to provide molecular phylogenetic analysis, germinated cells from incubated cysts were examined for their LSU rDNA sequences. Results reveal that three morphologically different cysts produced three morphologically different thecate cells, which were previously known as Protoperidinium oblongum var. latidorsale, Protoperidinium oblongum var. inaequale, and Protoperidinium oblongum var. symmetricum. The molecular phylogenetic analysis demonstrated that the Oceania group in the genus Protoperidinium includes these three varieties as well as Protoperidinium divergens, Protoperidinium claudicans, and Protoperidinium steigingerae. This group is separated from Protoperidinium sensu stricto, and the three plankton forms are phylogenetically separate and independent species. Based on these facts, two new species, Protoperidinium quadrioblongum Sarai, Kawami et Matsuoka, the new name for Protoperidinium oblongum var. symmetricum and Protoperidinium paraoblongum Sarai, Kawami et Matsuoka for Protoperidinium oblongum var. inaequale are described.

In this paper we describe a new species, Archaeperidinium saanichi sp. nov. within the Archaeperidinium minutum Jörgensen 1912 species complex. We examined the morphological variation of the cyst and motile stage by incubation experiments from sediment samples collected in coastal British Columbia (Canada), and compared it to closely related species. The theca of A. saanichi is differentiated from related species by overall size, the asymmetry of the intercalary plates and the right-sulcal plate (S.d.) not touching the cingulum. We provide a key to differentiate all closely related species. A. saanichi can be readily distinguished from A. minutum by a distinctively large cyst with a broad 2a type archeopyle and regularly spaced processes with relatively broad bases and aculeate process tips. Molecular phylogenetic analyses of large and small subunit (LSU and SSU) rDNA sequences demonstrated a close affinity of this species to A. minutum; however, the relatively high level of sequence conservation in dinoflagellate rDNA sequences made these particular markers inadequate for distinguishing one species from the other. Sediment-trap data suggest that A. saanichi has a preference for cooler temperatures and lowered salinities.

SUMMARY A small, broadly ovoidal and heterotrophic dinoflagel- late 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, Protope- ridinium 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 Abe'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. tricingula- tum 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.

SUMMARY Cellular and body scale structure of a new armored dinoflagellate Heterocapsa huensis, collected from Hue, Vietnam were investigated. Morphology of motile cell was observed by light, fluorescent and scanning electron microscopy, and body scale structure was examined by whole mounts of transmission electron microscopy. Cells of H. huensis were ellipsoid with a spherical nucleus located in the posterior and multiple pyrenoids located above the nucleus; this arrangement was similar to that of Heterocapsa pygmaea. Transmission electron microscopy revealed ultrastructure of the body scales consisted of a rounded triangular basal plate and three-dimensional ornaments. Structure of the basal plate resembles that of Heterocapsa illdefina; however, the number of the peripheral spine is different from that of H. illdefina and this structure has never been reported from Heterocapsa species. A new Heterocapsa species, H. huensis Iwataki et Matsuoka sp. nov., is described based on positions of organelles and body scale ultrastructure.

We have re-examined the cyst–motile relationships of two Polykrikos species; P. schwartzii and P. kofoidii (Dinophyceae), based on the literature, incubation experiments, and molecular phylogenetic analysis. The longitudinal furrows on the hypocone of P. kofoidii differentiate it from P. schwartzii. Differences in surface ornamentations on the cysts of P. schwartzii and P. kofoidii were considered as important morphological features to differentiate these two species. Many researchers accepted that the cyst of P. schwartzii was characterized by reticulate ornaments and P. kofoidii by separate, rod-like processes. However, encystment/excystment experiments carried out in previous studies clarified that the P. kofoidii cyst has coarse reticulate ornaments, not rod-like processes. Further subsequent observations on these relationships have indicated that rod-like processes develop not on cysts of P. kofoidii but on cysts of P. schwartzii. In combination with morphological observations, phylogenetic analyses of the small subunit and large subunit rDNA sequences, directly collected from vegetative cells and living cysts of P. kofoidii and P. schwartzii confirmed these findings on the cyst motile relationships and thus the criteria for the identification of cysts required revision. Furthermore morphologically intermediate forms, which sometimes occur, are identical to the cysts of P. schwartzii based on the molecular data of a single cell PCR technique for living cysts.

Organic-walled resting cysts of Protoperidinium parthenopes Zingone & Montresor 1988 were collected from a sediment trap in Omura Bay, western Japan. The cysts are spherical and pale brown in color. The cyst wall has two layers: a thick endophragm with granulate surface, and a thin periphragm. Three complete and incomplete parasutures appear on the surface of the endocyst. The archeopyle formed on the endocyst is basically saphopylic and compound with some combination of complete and incomplete parasutures. The cyst of Protoperidinium parthenopes closely resembles the cyst of Protoperidinium americanum (Gran & Braarud 1935) Balech 1974, but differs in the shape of the periphragm, the cyst diameter, and in the archeopyle.

In Gamak Bay, Paralytic Shellfish Poisoning (PSP) was first detected from seafoods in 2003, however the toxin source is unknown yet. In this study, we report potential PSP producers of toxic dinoflagellates, describing morphology and abundance of cysts isolated from surface sediment of Gamak Bay. The most abundant type in these cysts was characterized with ellipsoidal and transparent wall identical to Alexandrium catenella and/or A. tamarense. Germination experiment of the cysts revealed that all motile cells germinated were morphologically identified as A. tamarense. This result suggests that A. tamarense may relate to PSP contaminations in Gamak Bay. Moreover, bottom water temperature in Gamak Bay is favorable for germination of A. tamarense cysts. Further studies are required to carry out the PSP monitoring for preventing the risk of PSP events that may outbreak in future at Gamak Bay.

Phylogenetic relationships among chain-forming Cochlodinium species, including the harmful red tide forming dinoflagellate Cochlodinium polykrikoides, were investigated using specimens collected from coastal waters of Canada, Hong Kong, Japan, Korea, Malaysia, MA©xico, Philippines, Puerto Rico, and USA. The phylogenetic tree inferred from partial (D1-D6 regions) large subunit ribosomal RNA gene (LSU rDNA) sequences clearly differentiated between C. polykrikoides and a recently described species, Cochlodinium fulvescens. Two samples collected from the Pacific coasts of North America (British Columbia, Canada and California, USA) having typical morphological characters of C. fulvescens such as the sulcus located in the intermediate region of the cingulum, were closely related to C. fulvescens from western Japan in the phylogenetic tree. Cochlodinium polykrikoides formed a monophyletic group positioned as a sister group of the C. fulvescens clade with three well-supported sub-clades. These three clades were composed of (1) East Asian, including specimens collected from Hong Kong, western Japan, and southern Korea, (2) Philippines, from Manila Bay, Philippines and Omura Bay, Japan, and (3) American/Malaysian, from the Atlantic coasts of USA, the Pacific coast of MA©xico, Puerto Rico, and Borneo Island, Malaysia. Each of these clades is considered to be a so-called "ribotype" representing the population inhabiting each region, which is distinguished based on ribosomal RNA gene sequences in the species despite similarities in their morphological characters. © 2007 Elsevier B.V. All rights reserved.

The morphology of an unarmored chain-forming harmful dinoflagellate Cochlodinium polykrikoides and its similar species such as Cochlodinium catenatum, Cochlodinium fulvescens, and Cochlodinium convolutum was carefully observed, emphasizing the single cell stage for clarifying taxonomically important morphological features. To differentiate C. polykrikoides from C. convolutum, the shape and the position of the nucleus are useful characters. C. polykrikoides also differs from C. fulvescens in being smaller in size, possessing many rod-shaped chloroplasts and having the sulcus running just below the cingulum on the dorsal surface. Careful observation of the ichnotype of C. catenatum suggests that C. catenatum sensu Kofoid and Swezy collected from off La Jolla, CA, USA, is not identical to C. catenatum sensu Okamura and is probably a different species, in having no chloroplasts and a nucleus positioned at the center of the cell. In addition, C. polykrikoides has many morphological features in common with C. catenatum sensu Okamura except for slightly elongate cells and is probably a junior synonym of this species., Harmful Algae, 7(3), pp.261-270; 2008

SUMMARY 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.

The heterotrophic armored dinoflagellate Protoperidinium thulesense has an unusual combination of morphological characters, i.e. the thecal plate arrangement of the motile cell resembles a typical Protoperidinium, whereas the shape and archeopyle of the cyst are like the diplopsalids. We have re-examined the cyst-motile relationship of P. thulesense by cyst incubation and thecal plate analysis together with a molecular phylogenetic study based on small subunit (SSU) rDNA sequences. Five isolates of P. thulesense, including motile cells and cysts, and three Protoperidinium and three diplopsalid species were examined by using the single cell PCR method. The thecal plate arrangement of the motile cells isolated from field samples and those germinated from cysts were identical. The plate formula was: Po, X, 3′, 3a, 7″, 3c+t, 4s, 5′″, 2″″. The cysts of P. thulesense were round and brown with a theropylic archeopyle, and rather similar to the diplopsalid species, Diplopsalis lenticula, D. lebourae, Got...

Oblea acanthocysta of the diplopsalid group is described as a new species from Omura Bay, West Japan. The motile cells are subspherical, have a large sulcal list at the left margin, and are characterized by the plate formula, Po, X, 3′, 1a, 6″, 3c+t, 6s?, 5?, 2″″. The species resembles Oblea torta in shape and plate distribution, but is smaller and differs in the shape of plate 1′ and position of plate 1a. Resting cysts of O. acanthocysta are spherical and pale brown in color, possess many hollow acuminate spines, and have a theropylic archeopyle. In the SSU rRNA gene sequences of three cells of O. acanthocysta and two cells of O. torta, no intraspecific base substitutions were detected within either species. Based on phylogenetic analyses, the O. acanthocysta clade is included in the diplopsalids clade together with other diplopsalid species such as O. torta, Diplopsalis lebourae, Diplopsalopsis bomba and Gotoius excentricus. The sequences of O. acanthocysta are different from those of O. torta in 154 base pair substitutions, but O. acanthocysta and O. torta have a very close phylogenetic relationship., Plankton and Benthos Research, 1(4), pp.183-190; 2006

HcRNAV is a single-stranded RNA (ssRNA) virus that specifically infects the bivalve-killing dinoflagellate, Heterocapsa circularisquama. HcRNAV strains are grouped into 2 types (UA and CY), based on intra-species host specificity and the amino acid sequence of the major capsid protein (MCP). In the present study, we report the isolation of novel HcRNAV clones (n=51) lytic to the H. circularisquama strains, HU9433-P, HCLG-1, 05HC05 and 05HC06. HcRNAV34, HcRNAV109, HcRNAV641, and HcRNAV659, which displayed lytic activity against the strains, HU9433-P, HCLG-1, 05HC05, and 05HC06, respectively, were selected as typical virus clones and were intensively examined. The infection intensity of each host-virus combination was analyzed by examining the algicidal activity, detecting the intracellular replication of the viral RNA as well as the appearance of host cells with a morphologically abnormal nucleus post-infection. Interestingly, the strains, 05HC05 and 05HC06, were markedly sensitive to HcRNAV641 and HcRNAV659, respectively. Tertiary structural modeling predicted 4 unique amino acid (aa) substitutions in HcRNAV659-MCP to be exposed to an ambient water environment, which contributed towards determining its infection specificity. Neighbor-joining analysis of MCP aa sequences from HcRNAV clones revealed 3 clades, namely, the CY type and the UA1 and UA2 subtypes. The HcRNAV clones lytic to HCLG-1 (ex. HcRNAV109), HU9433-P and 05HC05 (ex. HcRNAV34), and 05HC06 (ex. HcRNAV659) were categorized into CY type, UA1 and UA2 subtypes, respectively. The present study highlights the complexity of the H. circularisquama-HcRNAV host-virus system, i.e., clonal variation, microbial control, and ecology in a natural algal population.

To investigate the distribution of Heterocapsa including a harmful species H. circularisquama, cells were surveyed in three Vietnamese coasts, Ha Long Bay, Hue and Phu Quoc Island in 2006. Cells of Heterocapsa were detected from Lang Co Lagoon and adjacent coast in Hue. Cell of the species possesses thecal plates corresponding to the typical of Heterocapsa. Under TEM, body scales approximately 500 nm in diameter are observed. The scale structure is similar to that of H. illdefina, however, number of the marginal spine is different. We concluded it is an undescribed Heterocapsa species possessing a new body scale ultrastructure.