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The dinoflagellate genus Bysmatrum encompasses five epibenthic or tide-pool species and has been characterized by separated anterior intercalary plates. In the present study, we obtained six strains of Bysmatrum from the South China Sea and French Atlantic coast by isolating single cells/cysts from plankton and sediment samples. All strains were examined with light microscopy and scanning electron microscopy. Based on morphological observations, three strains were identified as Bysmatrum subsalsum, characterized by the elongated and rectangular first and a hexagonal second anterior intercalary plate. They differ from each other in the number of sulcal lists and the configuration of the first anterior intercalary plate. One strain was identified as Bysmatrum gregarium and the other two as Bysmatrum granulosum. The cyst-theca relationship of B. subsalsum from the French Atlantic was established by incubation of the cyst, and the geochemical composition of the cyst wall was measured through micro-Fourier transform infrared spectroscopy. Bysmatrum subsalsum from Malaysia shows a bright red stigma in the sulcal area under light microscopy, which was confirmed with transmission electron microscopy: it was identified as a type B eyespot. Small subunit ribosomal DNA (SSU rDNA), partial large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer (ITS) sequences were obtained from all six strains. The maximum likelihood and Bayesian inference analysis based on concatenated SSU, ITS and LSU sequences revealed that Bysmatrum is monophyletic and nested within Peridiniales. Our strains of B. subsalsum form a new ribotype in the molecular phylogeny (designated as ribotype B). The genetic distance based on ITS sequences among Bysmatrum species ranged from 0.34 to 0.47 and those genetic distances at the intraspecific level of B. subsalsum could Please note that this is an author-produced PDF of an article accepted for publication following peer review. The definitive publisher-authenticated version is available on the publisher Web site. reach 0.41, supporting the possibility of hidden crypticity within B. subsalsum.

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.

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The first chapter of this special issue introduces the proceedings of two workshops concerning the cystbased genus Spiniferites Mantell 1850. The historical background of the cyst-based genus Spiniferites, its closely related genera and the theca-based genus Gonyaulax Diesing 1866 is presented here.

In the current circumscription, the Thoracosphaeraceae comprise all dinophytes exhibiting calcified coccoid cells produced during their life-history. Species hitherto assigned to Ensiculifera and Pentapharsodinium are mostly based on the monadoid stage of life-history, while the link to the coccoid stage (occasionally treated taxonomically distinct) is not always resolved. We investigated the different life-history stages and DNA sequence data of Ensiculifera mexicana and other species occurring in samples collected from all over the world. Based on concatenated ribosomal RNA gene sequences Ensiculiferaceae represented a distinct peridinalean branch, which showed a distant relationship to other calcareous dinophytes. Both molecular and morphological data (particularly of the coccoid stage) Please note that this is an author-produced PDF of an article accepted for publication following peer review. The definitive publisher-authenticated version is available on the publisher Web site.revealed the presence of three distinct clades within Ensiculiferaceae, which may include other dinophytes exhibiting a parasitic life-history stage. At a higher taxonomic level, Ensiculiferaceae showed relationships to parasites and endosymbionts (i.e., Blastodinium and Zooxanthella) as well as to dinophytes harbouring diatoms instead of chloroplasts. These unexpected phylogenetic relationships are corroborated by the presence of five cingular plates in all such taxa, which differs from the six cingular plates of most other Thoracosphaeraceae. We herein describe Ensiculiferaceae, emend the descriptions of Ensiculifera and Pentapharsodinium, erect Matsuokaea and provide several new combinations at the species level.

It is well known that modern resting cysts with morphologies matching those of species of the fossil genus Spiniferites germinate into motile cells of the genus Gonyaulax. Different Spiniferites species have been connected to a single Gonyaulax species, raising the question of whether they are over-classified. Through germination experiments of cysts with the morphological features of four species of Spiniferites, viz. S. bentorii, S. hyperacanthus, S. ramosus and S. scabratus, we established cyst-theca relationships. Cysts with the morphology of S. bentorii gave rise to vegetative, motile cells of Gonyaulax nezaniae sp. nov., which is characterized by two stout antapical spines. Cysts with S. hyperacanthus and S. ramosus morphologies germinated into Gonyaulax whaseongensis and G. spinifera, respectively. Cysts with S. scabratus morphology lacked a ventral pore and were attributed to Gonyaulax cf. spinifera. Gene sequences for SSU, LSU and/or ITS-5.8S rRNA were obtained from these four species, and from cysts with the morphology of Spiniferites belerius, S. mirabilis, S. lazus, Spiniferites cf. bentorii and Please note that this is an author-produced PDF of an article accepted for publication following peer review. The definitive publisher-authenticated version is available on the publisher Web site.Tectatodinium pellitum. The maximum likelihood and Bayesian inference analyses based on LSU and SSU rRNA gene sequences revealed that cysts assignable to Spiniferites formed a polyphyletic group, intermingled with Tectatodinium, Bitectatodinium, Ataxiodinium and Impagidinium, whereas Gonyaulax species appeared as monophyletic. From our results we inferred the phylogenetic positions of S. bentorii, S. mirabilis, S. lazus, S. scabratus, Tectatodinium pellitum and Gonyaulax digitale for the first time, supporting the idea that Spiniferites species are not over-classified and each of them may correspond to different Gonyaulax species.