MORPHOLOGY AND MOLECULAR PHYLOGENY OF A NEW MARINE SAND‐DWELLING PROROCENTRUM SPECIES,P. TSAWWASSENENSE (DINOPHYCEAE,PROROCENTRALES), FROM BRITISH COLUMBIA,CANADA1

A new marine benthic, sand‐dwelling Prorocentrum species from the temperate region of the Pacific coast of British Columbia, Canada, is described using LM and EM and molecular phylogenetic analyses. The cells have a broad oval shape, 40.0–55.0 μm long and 30.0–47.5 μm wide, and a wide U‐shaped periflagellar area on the right thecal plate. The left thecal plate consists of a straighter apical outline in the form of a raised ridge. Five to six delicate apical spines in the center of the periflagellar area are present. The nucleus is located in the posterior region of the cell, and a conspicuous pusule is located in the anterior region of the cell. The cells have golden‐brown chloroplasts with a compound, intrachloroplast pyrenoid that lacks a starch sheath. The thecal plates are smooth with round pores of two different sizes. The larger pores are arranged in a specific pattern of radial rows that are evenly spaced around the plate periphery and of irregular rows (or double rows) that form an incomplete “V” at the apical end of the plates. Large pores are absent in the center of the left and right thecal plates. The intercalary band is striated transversely and also has faint horizontal striations. Trichocysts and two types of mucocysts are present. The molecular phylogenetic position of Prorocentrum tsawwassenense sp. nov. was inferred using SSU rDNA sequences. This new species branched with high support in a Prorocentrum clade containing both benthic and planktonic species.     

<Emphasis Type="Italic">Prorocentrum vietnamensis</Emphasis> sp. nov. (Prorocentraceae Dinophyta): A new armored dinoflagellate from Vietnam coastal waters

TheProrocentrum species, which are primarily marine, are distributed worldwide and occur in ocean, neritic, and littoral environments.Prorocentrum is either a planktonic or benthic genus, with some species possibly being both. Until now, about 35 species had been reported including two that are fresh-water. Some species contain toxic substances. As one of many currently active studies, we collectedP. vietnamensis sp. nov. from Hon Mé, Gulf of Tongkin, Vietnam. Based on its cell shape, smooth valve surface, arrangement of valves and marginal pores, plus characteristics of the central pyrenoid, we propose thatP. vietnamensis is a new species. Its attributes include straight cell sides that are parallel to each other, as well as anterior and posterior areas with rounded ends that form a short, rod-shaped cell that is unique to this species.     

The dinoflagellate genus Prorocentrum along the coasts of the Mexican Pacific

We surveyed the dinoflagellate genus Prorocentrum Ehrenberg in Mexican Pacific waters, where it is rather common and sometimes causes red tides in coastal areas or shrimp farms. Material collected from Baja California and the Gulf of California was analyzed. Thirteen species were identified, all of them planktonic (although P. mexicanum is also epiphytic). All species are described by light microscopy, and most are also described by scanning electron microscopy; comments on morphology, taxonomy and distribution are made. Red tides were caused by P. dentatum, P. minimum and P. triestinum. Prorocentrum mexicanum and P. minimum were suspected of being toxic. Four species, previously reported in the Gulf of California, were not found. A total of 18 species, including the new records P. dactylum and P. lebourae have been to date reported from the Mexican Pacific.     

MORPHOLOGY AND MOLECULAR PHYLOGENY OF PROROCENTRUM CONSUTUM SP. NOV. (DINOPHYCEAE), A NEW BENTHIC DINOFLAGELLATE FROM SOUTH BRITTANY (NORTHWESTERN FRANCE)1

A new marine benthic Prorocentrum species from sandy habitats of South Brittany (northwestern France), P. consutum sp. nov., is described using LM and SEM and molecular phylogenetic analyses. Cells have a subcircular to broadly ovoid shape and are plainly flattened. They are 57–61 μm long and 52–55 μm wide. A central pyrenoid is present, and the kidney‐shaped nucleus is positioned in the posterior region. In right valve view, the periflagellar area is deeply excavated, and the left valve forms a prominent apical ridge. The periflagellar area consists of nine platelets, and a small narrow collar is present around the flagellar pore. The ornamentation of this new species is very peculiar and is characterized by a ring of round areolae located at the periphery of the valves, each areola containing three or four pores. Apart from this ring of areolae, the cell surface is smooth and with scattered pores. Pores are not present in the center of the right or left valve. The intercalary band is generally narrow and faintly striated horizontally. The molecular phylogenetic position of P. consutum sp. nov. was inferred using SSU and LSU rDNA. In both analyses, this species branched with high support in the clade comprising species with a symmetric shape and appeared to be a sister group to that formed by P. lima and other tropical benthic species, such as P. arenarium, P. belizeanum, P. hoffmannianum, and P. maculosum.     

Molecular Phylogeny and Ultrastructure of Caliculium glossobalani n. gen. et sp. (Apicomplexa) from a Pacific Glossobalanus minutus (Hemichordata) Confounds the Relationships Between Marine and Terrestrial Gregarines

Gregarines are a diverse group of apicomplexan parasites with a conspicuous extracellular feeding stage, called a “trophozoite”, that infects the intestines and other body cavities of invertebrate hosts. Although the morphology of trophozoites is very diverse in gregarines as a whole, high degrees of intraspecific variation combined with relatively low degrees of interspecific variation make the delimitation of different species based on trophozoite morphology observed with light microscopy difficult. The coupling of molecular phylogenetic data with comparative morphology has shed considerable light onto the boundaries and interrelationships of different gregarine species. In this study, we isolated a novel marine gregarine from the hepatic region of a Pacific representative of the hemichordate Glossobalanus minutus, and report the first ultrastructural and molecular data from any gregarine infecting this distinctive group of hosts. Molecular phylogenetic analyses of an SSU rDNA sequence derived from two single‐cell isolates of this marine gregarine demonstrated a strong and unexpected affiliation with a clade of terrestrial gregarines (e.g. Gregarina). This molecular phylogenetic data combined with a comparison of the morphological features in previous reports of gregarines collected from Atlantic representatives of G. minutus justified the establishment of a new binomial for the new isolate, namely Caliculium glossobalani n. gen. et sp. The molecular phylogenetic analyses demonstrated a clade of terrestrial gregarines associated with a sequence acquired from a marine species, which suggest that different groups of terrestrial/freshwater gregarines evolved independently from marine ancestors.     

北部湾海洋原甲藻的形态特征及其系统进化分析

为探讨广西北部湾海洋原甲藻(Prorocentrum micans)的形态特征及其系统进化意义,利用光学显微镜、分子生物学方法,对海洋原甲藻BBW-01 藻株的形态特征进行了描述,并分析了其分子系统进化关系.结果表明,各地理株系的海洋原甲藻的形态特征相似,仅在细胞大小上存在差异.海洋原甲藻BBW-01 与采自于广东大亚湾的海洋原甲藻形态特征最为接近,其壳板后端的7 个呈“V”字形对称排列的大孔可作为海洋原甲藻鉴定的重要指标.18S rDNA 序列同源检索和系统进化分析表明,海洋原甲藻BBW-01 与源自中国南海的海洋原甲藻的亲缘关系最近,并与其他2 株海洋原甲藻聚成一支,属于浮游、兼性浮游类原甲藻.因此,对赤潮原因种的准确识别有助于预防和减轻赤潮对海洋环境和人类带来的危害.     

PHYLOGENETIC ANALYSIS OF NINE SPECIES OF PROROCENTRUM (DINOPHYCEAE) INFERRED FROM 18S RIBOSOMAL DNA SEQUENCES, MORPHOLOGICAL COMPARISONS, AND DESCRIPTION OF PROROCENTRUM PANAMENSIS, SP. NOV.

Sequences of 18S rRNA genes were obtained from eight species of Prorocentrum Ehrenberg: P. minimum (Pavillard) Schiller, P. mexicanum Osorio Tafall, P. emarginatum Fukuyo, P. lima (Ehrenberg) Dodge, P. arenarium Faust, P. maculosum Faust, P. concavum Fukuyo, and P. panamensis, sp. nov. Prorocentrum panamensis is a new species of tropical dinoflagellate isolated from a benthic coral reef on the Pacific coast of Panama and described here using scanning electron microscopy. Cells are heart shaped, 46–52 μm long and 43–46 μm wide. The valve surfaces are areolate except in the central area. Pores of 0.15 μm in diameter are scattered in areolae, mainly around the periphery of the cell. The right valve has a specific ovoid depression with numerous appressed pores; we named this structure the sieve-like depression. The periflagellar area is nearly ovoid, located in a shallow depression, and almost equally set into both valves. It is unornamented (no apical expansion) but has numerous depressions in platelets. The flagellar and auxiliary pores are different in size and shape. The intercalary band is transversally striated. Phylogenetic relationships of gonyaulacoid, peridinioid, gymnodinioid, and prorocentroid dinoflagellates were inferred from complete 18S rDNA sequences. Two distinct phylogenetic analyses are presented for armored and unarmored Dinophyceae in an attempt to make the phylogenetic relationships between these different kinds of organisms clearer. The Prorocentrales appear to have a common origin, although two groups of Prorocentrum spp. are apparent. The first group includes benthic, symmetrical species (P. lima, P. arenarium, P. maculosum, and P. concavum). The second group contains planktonic and bentho-planktonic species (P. micans Ehrenberg, P. minimum, P. mexicanum, and P. panamensis sp. nov.). Genetic distances between species within these two groups were high; however, the divergence between the two groups seems to have occurred late in dinoflagellate evolution. In addition, the bentho-planktonic P. emarginatum appeared distantly related to both groups; however,its 18S rDNA sequence shares specific nucleotide substitutions with the two groups, suggesting an older origin of this species compared to the others. A morphological interpretation of this phylogenetic analysis is made on the basis of the specific structure of the periflagellar area. Finally, genetic data and morphological observations support the hypothesis that the genus Prorocentrum is rather heterogeneous; several species could be considered to constitute distinct genera.     

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.     

Ultrastructure and Systematics of Two New Species of Dinoflagellate,Paragymnodinium Asymmetricum sp. nov. and Paragymnodinium Inerme sp. nov. (Gymnodiniales,Dinophyceae)1

The genus Paragymnodinium currently includes two species, P. shiwhaense and P. stigmaticum, that are characterized by mixotrophic nutrition and the possession of nematocysts. In this study, two new dinoflagellates belonging to this genus were described based on observations using LM, SEM, and TEM together with a molecular analysis. Cells of P. asymmetricum sp. nov., isolated from Nha Trang Beach, Vietnam, were 7.9–12.6 μm long and 4.7–9.0 μm wide. The species showed no evidence of feeding behavior and was able to sustain itself phototrophically. Paragymnodinium asymmetricum shared many features with P. shiwhaense, including presence of nematocysts, absence of an eyespot, and a planktonic lifestyle, but was clearly distinguished by the asymmetric shape of the hyposome, possession of a single chloroplast, and its nutritional mode. Cells of P. inerme sp. nov., isolated from Jogashima, Kanagawa Pref, Japan, were 15.3–23.7 μm long and 10.9–19.6 μm wide. This species also showed no evidence of feeding behavior. Paragymnodinium inerme was similar to cells of P. shiwhaense in shape and planktonic lifestyle, but its nutritional mode was different. The presence of incomplete nematocysts was also a unique feature. A phylogenetic analysis inferred from concatenated SSU and LSU rDNA sequences recovered the two dinoflagellates in a robust clade with Paragymnodinium spp., within the clade of Gymnodinium sensu stricto. This evidence, together with their morphological similarities, made it reasonable to conclude that these two dinoflagellates are new species of Paragymnodinium.     

Euchlorocystis gen. nov. and Densicystis gen. nov., Two New Genera of Oocystaceae Algae from High‐altitude Semi‐saline Habitat (Trebouxiophyceae,Chlorophyta)

The Oocystaceae family is generally considered to contain common freshwater eukaryotic microalgae, and few are reported living in semi‐saline habitats. Our latest ecological survey in Qinghai Lake and Angzicuo Lake, both large, closed, high‐altitude, semi‐saline lakes located on the Qinghai‐Tibet plateau in China, revealed Oocystaceae species as a dominant group among plankton. Since limited knowledge exists about semi‐saline species in the Oocystaceae family, a taxonomical study was carried out using morphological and phylogenetic methods. Using this approach, four new strains of Oocystaceae were identified and successfully cultured in the lab. Molecular results correlated with morphological characters and resolved these species into at least three genera. A new genus, Euchlorocystis, with type species Euchlorocystis subsalina, is described here as having the distinctive morphology of multiple pyrenoids per chloroplast among Oocystaceae, and an independent phylogenetic position at the base of the Oocystaceae. Similarly, the genus Densicystis, with type species Densicystis glomerata, is newly proposed here as having a unique colony morphology of dozens or hundreds of little cells tightly embedded in ellipsoid to round mucilage masses. Oocystis marina, originally described from the Baltic Sea, was also identified in Qinghai Lake and Angzicuo Lake and phylogenetically positioned in the semi‐saline clade of the Oocystaceae. The result that a marine species was detected in the closed inland lakes implies a further need to reevaluate the origins of these species.     

Phylogenetic study of benthic, spine-bearing prorocentroids, including Prorocentrum fukuyoi sp. nov.

Species of prorocentroid dinoflagellates are common in marine benthic sediment and epibenthic habitats, as well as in planktonic habitats. Marine planktonic prorocentroids typically possess a small spine in the apical region. In this study, we describe a new, potentially widely distributed benthic species of Prorocentrum, P. fukuyoi sp. nov., from tidal sand habitats in several sites in Australia and from central Japan. This species was found to possess an apical spine or flange and was sister species to P. emarginatum. We analyzed the phylogeny of the group including this new species, based on large subunit (LSU) rDNA sequences. The genus contained a high level of divergence in LSU rDNA, in some cases among sister taxa. P. fukuyoi and P. emarginatum were found to be most closely related to a clade of generally planktonic taxa. Several morphological features may constitute more informative synapomorphies than habitat in distinguishing clades of prorocentroid species.     

Morphology and phylogenetic position of two novel marine gregarines (Apicomplexa,Eugregarinorida) from the intestines of North‐eastern Pacific ascidians

Eugregarine apicomplexans parasitize marine, freshwater and terrestrial invertebrates, and have lifecycles involving trophozoites (feeding stages) with complex morphologies and behaviour. The genus Lankesteria refers to marine aseptate eugregarines that parasitize ascidians. We described the surface ultrastructure of two new gregarine species, L. chelyosomae sp. n. and L. cystodytae sp. n. that inhabit the intestines of Chelyosoma columbianum and Cystodytes lobatus, respectively, collected from the North‐eastern Pacific Ocean. Apart from inhabiting different hosts and major differences in the cell size of L. chelyosomae sp. n. (mean length 182 µm) and L. cystodytae sp. n. (mean length 70 µm), the morphology of both gregarine species was quite similar. The trophozoites ranged from elliptoid to obdeltoid in shape and were brownish in colour. The nucleus was situated at the anterior end of the cell just behind a pointed mucron. A dense array of epicytic knobs was present over the entire surface of trophozoites in both species, and longitudinal epicytical folds were only weakly developed. We also sequenced the small subunit rDNA from the gregarines collected from both hosts, which supported the establishment of two new Lankesteria species. Phylogenetic analyses of the new DNA sequences and those derived from other alveolates, demonstrated that both new species clustered in a strongly supported clade consisting of other Lankesteria species, Lecudina species, and some environmental sequences. These morphological and molecular phylogenetic data suggested that improved knowledge of gregarine diversity could lead to the recognition of more than one distinct clade (genus) of gregarines within ascidian hosts.     

Marinamoeba thermophila, a new marine heterolobosean amoeba growing at 50 °C

Two amoeba strains were isolated from marine sediment taken at the same place with 18 months interval from a region of the sea floor heated by extended submarine hot springs and fumaroles. These thermophilic amoebae grow at temperatures up to 50 °C. Sequences of the internal transcribed spacer demonstrated that the two strains belong to the same species and are different from any genus for which sequences are known. Phylogeny using small subunit ribosomal RNA places the amoeba in the Heterolobosea. Their closest relatives are the hypersaline flagellate Pleurostomum flabellatum and the hypersaline amoeba Tulamoeba peronaphora. The freshwater amoeboflagellate genera Naegleria and Willaertia belong to the same phylogenetic clade in the Vahlkampfiidae. The new marine species does not transform into flagellates. It forms cysts, which are round to ellipsoidal with few pores. Because of their unique place in the molecular phylogenetic tree, and because there is no morphologically identical species found in the literature, these isolates are considered to be a new species and a new genus, Marinamoeba thermophila.     

Fukuyoa paulensis gen. et sp. nov., a New Genus for the Globular Species of the Dinoflagellate Gambierdiscus (Dinophyceae)

The marine epiphytic dinoflagellate Gambierdiscus is a toxicologically important genus responsible for ciguatera fish poisoning, the principal cause of non-bacterial illness associated with fish consumption. The genus currently contains species exhibiting either globular or anterior-posteriorly compressed morphologies with marked differences in cell shape and plate arrangement. Here we report a third globular, epiphytic and tychoplanktonic species from the coasts of Ubatuba, Brazil. The new species can be distinguished from G. yasumotoi and G. ruetzleri by its broader first apical plate that occupies a larger portion of the epitheca. Accordingly, phylogenetic trees from small subunit (SSU) and large subunit (LSU) ribosomal DNA sequences also showed strongly supported separation of the new species from the G. yasumotoi / G. ruetzleri group albeit with short distance. The molecular phylogenies, which included new sequences of the planktonic species Goniodoma polyedricum, further indicated that the globular species of Gambierdiscus formed a tight clade, clearly separated (with strong bootstrap support) from the clade of lenticular species including the type for Gambierdiscus. The morphological and molecular data in concert support the split of Gambierdiscus sensu lato into two genera. Gambierdiscus sensu stricto should be reserved for the species with lenticular shapes, highly compressed anterioposteriorly, with short-shank fishhook apical pore plate, large 2'' plate, low and ascending cingular displacement, and pouch-like sulcal morphology. The new genus name Fukuyoa gen. nov. should be applied to the globular species, slightly laterally compressed, with long-shank fishhook apical pore plate, large 1'' plate, greater and descending cingular displacement, and not pouch-like vertically-oriented sulcal morphology. Fukuyoa contains the new species Fukuyoa paulensis gen. et sp. nov., and F. yasumotoi comb. nov. and F. ruetzleri comb. nov.     

Fine structure and 18S rDNA phylogeny of a marine araphid pennate diatom Plagiostriata goreensis gen. et sp. nov. (Bacillariophyta)

A marine araphid pennate diatom Plagiostriata goreensis is described from the sand grains of Goree Island, Dakar, Republic of Senegal, based on observations of fine structure of its frustule. The most striking feature of the species is its striation, which is angled at approximately 60° across the robust sternum. The other defining features of the species are its one highly reduced rimoportula and apical pores located at both ends of the valve margin. In the 18S rDNA phylogeny, the species appears as a member of a ‘small‐celled clade’ of araphid pennate diatoms that consist of Nanofrustulum, Opephora and Staurosira. The results of the phylogenetic analyses suggest that the distinct characters of the diatom; namely, oblique striae and apical pores, may have been acquired independently. However, it remains unclear whether the rimoportula of P. goreensis is a reduced state or P. goreensis acquired its morphologically curious rimoportula independently after the loss of an ancient rimoportula at the root of the small‐celled clade.     

Renewed sampling of inland aquatic habitats in southern Africa yields two novel freshwater crab species (Decapoda: Potamonautidae: Potamonautes)

A recent survey of the freshwater streams of the Mecula and Yao mountains in the Niassa province of Mozambique resulted in the discovery of a new freshwater crab species. This species is genetically and morphologically distinct from described species from Mozambique or its neighbouring countries, and is described as P otamonautes bellarussus sp. nov. In addition, a new semi‐terrestrial burrowing freshwater crab P otamonautes flavusjo sp. nov. from the Highveld of the Mpumalanga province in South Africa is described based on unique genetic and morphological characters. The phylogenetic affinities of the two new species in relation to the described eastern and southern African Potamonautes species is determined and the biogeographic implications are discussed. © 2014 The Linnean Society of London