CHARACTER EVOLUTION IN POLYKRIKOID DINOFLAGELLATES1

A synthesis of available data on the morphological diversity of polykrikoid dinoflagellates allowed us to formulate a hypothesis of relationships that help explain character evolution within the group. Phylogenetic analyses of new SSU rDNA sequences from Pheopolykrikos beauchampii Chatton, Polykrikos kofoidii Chatton, and Polykrikos lebourae Herdman helped refine this hypothetical framework. Our results demonstrated that “pseudocolonies” in dinoflagellates evolved convergently at least three times independently from different Gymnodinium‐like ancestors: once in haplozoans; once in Ph. beauchampii; and at least once within a lineage containing Ph. hartmannii, P. kofoidii, and P. lebourae. The Gymnodiniales sensu stricto was strongly supported by the data, and the type species for the genus, namely Gymnodinium fuscum (Ehrenb.) F. Stein, formed the nearest sister lineage to a well‐supported Polykrikos clade. The best synapomorphy for the Polykrikos clade was the presence of two nuclei irrespective of zooid number. Two unidentified Gymnodinium species formed the nearest sister clade to Ph. beauchampii, which has four nuclei and four zooids per pseudocolony. The chain‐forming dinoflagellate G. catenatum L. W. Graham branched closely to the clade containing all members of Polykrikos and Pheopolykrikos, suggesting that an ancestral capacity toward chain formation existed before the evolution of pseudocolonies in this group. Our results also clarified the phylogenetic significance of nematocysts, ocelloids, and photosynthesis in reconstructing the evolution of polykrikoids and warnowiids. The molecular phylogenies exposed taxonomic problems associated with Polykrikos, Pheopolykrikos, and Gymnodinium, and suggested that a revision for some of these genera is warranted.     

Apical Groove Type and Molecular Phylogeny Suggests Reclassification of Cochlodinium geminatum as Polykrikos geminatum

Traditionally Cocholodinium and Gymnodinium sensu lato clade are distinguished based on the cingulum turn number, which has been increasingly recognized to be inadequate for Gymnodiniales genus classification. This has been improved by the combination of the apical groove characteristics and molecular phylogeny, which has led to the erection of several new genera (Takayama, Akashiwo, Karenia, and Karlodinium). Taking the apical groove characteristics and molecular phylogeny combined approach, we reexamined the historically taxonomically uncertain species Cochlodinium geminatum that formed massive blooms in Pearl River Estuary, China, in recent years. Samples were collected from a bloom in 2011 for morphological, characteristic pigment, and molecular analyses. We found that the cingulum in this species wraps around the cell body about 1.2 turns on average but can appear under the light microscopy to be >1.5 turns after the cells have been preserved. The shape of its apical groove, however, was stably an open-ended anticlockwise loop of kidney bean shape, similar to that of Polykrikos. Furthermore, the molecular phylogenetic analysis using 18S rRNA-ITS-28S rRNA gene cistron we obtained in this study also consistently placed this species closest to Polykrikos within the Gymnodinium sensu stricto clade and set it far separated from the clade of Cochlodinium. These results suggest that this species should be transferred to Polykrikos as Polykrikos geminatum. Our results reiterate the need to use the combination of apical groove morphology and molecular phylogeny for the classification of species within the genus of Cochlodinium and other Gymnodiniales lineages.     

Acquired phototrophy in Mesodinium and Dinophysis – A review of cellular organization,prey selectivity,nutrient uptake and bioenergetics

Acquired phototrophy, i.e. the use of chloroplasts from ingested prey, can be found among some species of dinoflagellates and ciliates. The best studied examples of this phenomenon in these groups are within the ciliate genus Mesodinium and the dinoflagellate genus Dinophysis, both ecologically important genera with a worldwide distribution. Mesodinium species differ considerably in their carbon metabolism. Some species rely almost exclusively on food uptake, while other species rely mostly on photosynthesis. In Mesodinium with acquired phototrophy, a number of prey organelles in addition to chloroplasts may be retained, and the host ciliate has considerable control over the acquired chloroplasts; Mesodinium rubrum is capable of dividing its acquired chloroplasts and can also photoacclimate. In Dinophysis spp., the contents of ciliate prey are sucked out, but only the chloroplasts are retained from the ingested prey. Some chloroplast house-keeping genes have been found in the nucleus of Dinophysis and some preliminary evidence suggests that Dinophysis may be capable for photoacclimation. Both genera have been claimed to take up inorganic nutrients, including NO₃⁻, indicating that processes beyond photosynthesis have been acquired. M. rubrum seems to depend upon prey species within the Teleaulax/Plagioselmis/Geminigera clade of marine cryptophytes. Up until now, Dinophysis species have only been maintained cultured on M. rubrum as food, but other ciliates may also be ingested. Dinophysis spp. and M. rubrum are obligate mixotrophs, depending upon both prey and light for sustained growth. However, while M. rubrum only needs to ingest 1–2% of its carbon demand per day to attain maximum growth, Dinophysis spp. need to obtain about half of their carbon demand from ingestion for maximum growth. Both Mesodinium and Dinophysis spp. can survive for months in the light without food. The potential role for modeling in exploring the complex balance of phototrophy and phago-heterotrophy, and its ecological implications for the mixotroph and their prey, is discussed.     

Confirmation of the wide host range of Parvilucifera corolla (Alveolata,Perkinsozoa)

Marine parasites of the genus Parvilucifera have been described as endoparasitoids of dinoflagellates. Recently, the species Parvilucifera corolla was described, but its host range was not examined. Here, the host selectivity of P. corolla was screened, including 110 strains of dinoflagellates (24 genera) and other microalgal groups as potential hosts. Infections and the full life cycle of the parasitoid were observed in 73 strains (16 genera) of dinoflagellates. Parvilucifera corolla did not infect most chlorophytes, cryptophytes, chrysophytes, diatoms, haptophytes and raphidophytes but one strain of Pyramimonas (chlorophyte) was infected, although without viable sporangia. In Symbiodinium natans, a transition to the coccoid stage was induced above a certain parasite:host ratio. These results confirm P. corolla as a generalist parasitoid of dinoflagellates, with important differences in host range regarding other species of the genus.     

Ultrastructural and molecular phylogenetic delineation of a new order,the Rhizophydiales (Chytridiomycota)

In the order Chytridiales, Rhizophydium is a morphologically defined genus based upon the production of a monocentric, inoperculate, epibiotic sporangium, an endobiotic rhizoidal axis which branches, and an epibiotic resting spore. Despite its simple morphology, over 220 species of Rhizophydium have been described. Recent phylogenetic analyses using nuLSU rRNA (28 S rRNA) gene sequences of a geographically diverse sampling of Rhizophydium cultures revealed that the classical genus Rhizophydium is genetically more variable than previously understood and actually represents multiple genera. In the present study, we use zoospore ultrastructural characters and 28 S rRNA and 5.8 S ribosomal gene sequences of 96 isolates in culture to circumscribe the monophyletic Rhizophydium clade as a new order, Rhizophydiales. Correspondingly, zoospores of members of the Rhizophydiales exhibit a unique suite of ultrastructural character states that further define the order and distinguish it from the order Chytridiales. Molecular analyses reveal several strongly supported clades within the Rhizophydiales. Three of those clades encompass a broad range of isolates and are defined as new families Rhizophydiaceae, Terramycetaceae, and Kappamycetaceae. To resolve close relationships within Terramycetaceae, combined 28 S rRNA and ITS1–5.8 S–ITS2 sequences were analysed and details of zoospore ultrastructural character states determined, with two new genera, Terramyces and Boothiomyces, described. Two species formerly classified in Rhizophydium are transferred to the new genera. This work provides a framework for additional taxonomic revisions within the new order Rhizophydiales and compares genetic variation useful in defining genera, species, and populations within this lineage of chytrids. A broader sampling of representatives is needed before taxonomic decisions can be made for remaining clades within the Rhizophydiales.     

Parasitic dinoflagellates and ellobiopsids (Ellobiopsidae) of the coastal waters of the sea of Japan

Five species of parasitic dinoflagellates of the genera Protoodinium, Paulsenella, Duboscquella, Syndinium, and Dissodinium and two species of ellobiopsids of the genus Ellobiopsis are described. Figures and distribution data are provided. All species, except for Dissodinium pseudolunula Swift ex Elbrachter et Drebes, are first records for the Russia Far-Eastern seas.     

Molecular phylogeny of ocelloid-bearing dinoflagellates (Warnowiaceae) as inferred from SSU and LSU rDNA sequences

Background  

Dinoflagellates represent a major lineage of unicellular eukaryotes with unparalleled diversity and complexity in morphological features. The monophyly of dinoflagellates has been convincingly demonstrated, but the interrelationships among dinoflagellate lineages still remain largely unresolved. Warnowiid dinoflagellates are among the most remarkable eukaryotes known because of their possession of highly elaborate ultrastructural systems: pistons, nematocysts, and ocelloids. Complex organelles like these are evolutionary innovations found only in a few athecate dinoflagellates. Moreover, the taxonomy of warnowiids is extremely confusing and inferences about the evolutionary history of this lineage are mired by the absence of molecular phylogenetic data from any member of the group. In this study, we provide the first molecular phylogenetic data for warnowiids and couple them with a review of warnowiid morphological features in order to formulate a hypothetical framework for understanding character evolution within the group. These data also enabled us to evaluate the evolutionary relationship(s) between warnowiids and the other group of dinoflagellates with complex organelles: polykrikoids.     

Life cycle and molecular phylogeny of the dinoflagellates Chytriodinium and Dissodinium,ectoparasites of copepod eggs

The dinoflagellates Chytriodinium affine, C. roseum and Dissodinium pseudolunula are ectoparasites of crustacean eggs. Here, we present new observations regarding their life cycle based on coastal plankton samples and incubations and analyze their molecular phylogeny using the small subunit ribosomal RNA gene (SSU rDNA) as a marker. In contrast to the typical stages already documented for its life cycle, we observed that D. pseudolunula dinospores may exceptionally differentiate inside a globular cyst. Despite its parasitic life style, the cysts and dinospores of D. pseudolunula contain chlorophyll a. We obtained the first SSU rDNA sequences for the genera Chytriodinium (the type C. roseum and C. affine) and Dissodinium (D. pseudolunula). Classical taxonomical schemes have ascribed these genera to the order Blastodiniales. However, our SSU rDNA-based phylogenetic analysis shows that these ectoparasites form a clade in the Gymnodinium sensu stricto group, unarmored dinokaryotic dinoflagellates of the order Gymnodiniales. They branch in a subgroup composed of warnowiids, polykrikoids, the type of Gymnodinium, G. fuscum and G. aureolum. Although Chytriodinium and Dissodinium appear to be relatives based on SSU rDNA phylogeny, feeding and host specificity, their life cycles are substantially different. Based on these data we consider that the type of life cycle is a poor criterion for classification at the family level. We suggest that the morphology of the infective cell is probably the most reliable phenotypic characteristic to determine the systematic position of parasitic dinoflagellates.     

Are unusual plasmodesmata in the embryo-suspensor restricted to species from the genus Sedum among Crassulaceae?

The Crassulaceae family comprises mainly herbaceous leaf succulents, some of which have an ornamental value. During embryogenesis, they produce a suspensor with a giant polyploid basal cell. It has recently been shown that in Sedum acre and S. hispanicum this cell has compound plasmodesmata with an unusual dome of electron-dense material associated on the cell's side. These compound plasmodesmata differ from the typical ones occurring in other angiosperms. In this study, the hypothesis was tested that the unusual plasmodesmata in the embryo-suspensor are a feature not only restricted to species from the genus Sedum, but are also found in other Crassulaceae genera. Suspensors of example species from the genera Sempervivum and Jovibarba, which have vegetative morphologies quite different from Sedum and which are placed in the traditional classification into another subfamily, were first examined using an electron microscope. It was found that the unusual compound plasmodesmata in the suspensor are not only restricted to species from the genus Sedum but are also found in species from other Crassulaceae genera (Sempervivum arachnoideum and Jovibarba sobolifera). It should be noted that some ultrastructural features of compound plasmodesmata in the analyzed genera (e.g. the character of the wall with plasmodesmata, plasmodesmata diameter or occurrence of the electron-dense material) are different from the suspensor plasmodesmata recorded in species from the Sedum genus. We found that in Sempervivum arachnoideum the size of the plasmodesmata diameter varies according to the micropylar-chalazal axis of the embryo. This is the first report of variation in the diameter of the plasmodesmata within the embryo of angiosperms. Further study will be needed to show whether compound plasmodesmata occur in other Crassulaceae clades, whether they are a stable feature at the genus level in this family, and also whether they have evolved several times or only once in Crassulaceae.     

Effects of the bacterial algicide IRI-160AA on cellular morphology of harmful dinoflagellates

The algicide, IRI-160AA, induces mortality in dinoflagellates but not other species of algae, suggesting that a shared characteristic or feature renders this class of phytoplankton vulnerable to the algicide. In contrast to other eukaryotic species, the genome of dinoflagellates is stabilized by high concentrations of divalent cations and transition metals and contains large amounts of DNA with unusual base modifications. These distinctions set dinoflagellates apart from other phytoplankton and suggest that the nucleus may be a dinoflagellate-specific target for IRI-160AA. In this study, morphological and ultrastructural changes in three dinoflagellate species, Prorocentrum minimum, Karlodinium veneficum and Gyrodinium instriatum, were evaluated after short-term exposure to IRI-160AA using super resolution structured illumination microscopy (SR-SIM) and transmission electron microscopy (TEM). Exposure to the algicide resulted in cytoplasmic membrane blebbing, differing chloroplast morphologies, nuclear expansion, and chromosome expulsion and/or destabilization. TEM analysis showed that chromosomes of algicide-treated K. veneficum appeared electron dense with fibrous protrusions. In algicide-treated P. minimum and G. instriatum, chromosome decompaction occurred, while for P. minimum, nuclear expulsion was also observed for several cells. Results of this investigation demonstrate that exposure to the algicide destabilizes dinoflagellate chromosomes, although it was not clear if the nucleus was the primary target of the algicide or if the observed effects on chromosomal structure were due to downstream impacts. In all cases, changes in cellular morphology and ultrastructure were observed within two hours, suggesting that the algicide may be an effective and rapid approach to mitigate dinoflagellate blooms.     

Phylogenetic status of Xylaria subgenus Pseudoxylaria among taxa of the subfamily Xylarioideae (Xylariaceae) and phylogeny of the taxa involved in the subfamily

To infer the phylogenetic relationships of Xylaria species associated with termite nests within the genus Xylaria and among genera of the subfamily Xylarioideae, β-tubulin, RPB2, and α-actin sequences of 131 cultures of 114 species from Xylaria and 11 other genera of the subfamily were analyzed. These 11 genera included Astrocystis, Amphirosellinia, Discoxylaria, Entoleuca, Euepixylon, Kretzschmaria, Nemania, Podosordaria, Poronia, Rosellinia, and Stilbohypoxylon. We showed that Xylaria species were distributed among three major clades, TE, HY, and PO, with clade TE—an equivalent of the subgenus Pseudoxylaria—encompassing exclusively those species associated with termite nests and the other two clades containing those associated with substrates other than termite nests. Xylaria appears to be a paraphyletic genus, with most of the 11 genera submerged within it. Podosordaria and Poronia, which formed a distinct clade, apparently diverged from Xylaria and the other genera early. Species of Entoleuca, Euepixylon, Nemania, and Rosellinia constituted clade NR, a major clade sister to clade PO, while those of Kretzschmaria were inserted within clade HY and those of Astrocystis, Amphirosellinia, Discoxylaria, and Stilbohypoxylon were within clade PO.     

Chlorophyllb-containing oxygenic photosynthetic prokaryotes: Oxychlorobacteria (Prochlorophytes)

The major ultrastructural and pigment characteristics of three chlorophyllb-containing photosynthetic prokaryote genera,Prochloron, Prochlorothrix, andProchlorococcus, are summarized. Aspects of their ecology are reviewed, as are also the major findings of comparisons among the three genera, and between them and the cyanobacteria, based on analyses of nucleotide sequences. The information summarized is discussed to assess how closely the three genera are related and whether they form a natural taxonomic grouping within the cyanobacteria. It is concluded that, although much of the evidence points to substantial differences among the three genera, the convenience of keeping them together as a group, namely, Oxychlorobacteria (formerly Prochlorophyta), outweighs certain inconsistencies revealed by analysis of a range of objective criteria. Current views on the possible significance of the group to considerations of the evolution of green chloroplasts are also presented.     

Molecular characterization and morphology of Cochlodinium strangulatum,the type species of Cochlodinium,and Margalefidinium gen. nov. for C. polykrikoides and allied species (Gymnodiniales,Dinophyceae)

Photosynthetic species of the dinoflagellate genus Cochlodinium such as C. polykrikoides, one of the most harmful bloom-forming dinoflagellates, have been extensively investigated. Little is known about the heterotrophic forms of Cochlodinium, such as its type species, Cochlodinium strangulatum. This is an uncommon, large (∼200 μm long), solitary, and phagotrophic species, with numerous refractile bodies, a central nucleus enclosed in a distinct perinuclear capsule, and a cell surface with fine longitudinal striae and a circular apical groove. The morphology of C. polykrikoides and allied species is different from the generic type. It is a bloom-forming species with single, two or four-celled chains, small cell size (25–40 μm long) with elongated chloroplasts arranged longitudinally and in parallel, anterior nucleus, eye-spot in the anterior dorsal side, and a cell surface smooth with U-shaped apical groove. Phylogenetic analysis based on LSU rDNA sequences revealed that C. strangulatum and C. polykrikoides/C. fulvescens formed two distally related, independent lineages. Based on morphological and phylogenetic analyses, the diagnosis of Cochlodinium is emended and C. miniatum is proposed as synonym of C. strangulatum. The new genus Margalefidinium gen. nov., and new combinations for C. catenatum, C. citron, C. flavum, C. fulvescens and C. polykrikoides are proposed.     

Phlebotomine sand flies (Diptera: Psychodidae) of the Maghreb region: A systematic review of distribution,morphology, and role in the transmission of the pathogens

BackgroundPhlebotomine sand flies (Diptera: Psychodidae) are important vectors of various human and animal pathogens such as Bartonella bacilliformis, Phlebovirus, and parasitic protozoa of the genus Leishmania, causative agent of leishmaniases that account among most significant vector-borne diseases. The Maghreb countries Mauritania, Morocco, Algeria, Tunisia, and Libya occupy a vast area of North Africa and belong to most affected regions by these diseases. Locally varying climatic and ecological conditions support diverse sand fly fauna that includes many proven or suspected vectors. The aim of this review is to summarize often fragmented information and to provide an updated list of sand fly species of the Maghreb region with illustration of species-specific morphological features and maps of their reported distribution.Materials and methodsThe literature search focused on scholar databases to review information on the sand fly species distribution and their role in the disease transmissions in Mauritania, Morocco, Algeria, Tunisia, and Libya, surveying sources from the period between 1900 and 2020. Reported distribution of each species was collated using Google Earth, and distribution maps were drawn using ArcGIS software. Morphological illustrations were compiled from various published sources.Results and conclusionsIn total, 32 species of the genera Phlebotomus (Ph.) and Sergentomyia (Se.) were reported in the Maghreb region (15 from Libya, 18 from Tunisia, 23 from Morocco, 24 from Algeria, and 9 from Mauritania). Phlebotomus mariae and Se. africana subsp. asiatica were recorded only in Morocco, Ph. mascitti, Se. hirtus, and Se. tiberiadis only in Algeria, whereas Ph. duboscqi, Se. dubia, Se. africana africana, Se. lesleyae, Se. magna, and Se. freetownensis were reported only from Mauritania. Our review has updated and summarized the geographic distribution of 26 species reported so far in Morocco, Algeria, Tunisia, and Libya, excluding Mauritania from a detailed analysis due to the unavailability of accurate distribution data. In addition, morphological differences important for species identification are summarized with particular attention to closely related species such as Ph. papatasi and Ph. bergeroti, Ph. chabaudi, and Ph. riouxi, and Se. christophersi and Se. clydei.     

Towards a new classification of the Arthoniales (Ascomycota) based on a three-gene phylogeny focussing on the genus Opegrapha

A multi-locus phylogenetic study of the order Arthoniales is presented here using the nuclear ribosomal large subunit (nuLSU), the second largest subunit of RNA polymerase II (RPB2) and the mitochondrial ribosomal small subunit (mtSSU). These genes were sequenced from 43 specimens or culture isolates representing 33 species from this order, 16 of which were from the second largest genus, Opegrapha. With the inclusion of sequences from GenBank, ten genera and 35 species are included in this study, representing about 18 % of the genera and ca 3 % of the species of this order. Our study revealed the homoplastic nature of morphological characters traditionally used to circumscribe genera within the Arthoniales, such as exciple carbonization and ascomatal structure. The genus Opegrapha appears polyphyletic, species of that genus being nested in all the major clades identified within Arthoniales. The transfer of O. atra and O. calcarea to the genus Arthonia will allow this genus and family Arthoniaceae to be recognized as monophyletic. The genus Enterographa was also found to be polyphyletic. Therefore, the following new combinations are needed: Arthonia calcarea (basionym: O. calcarea), and O. anguinella (basionym: Stigmatidium anguinellum); and the use of the names A. atra and Enterographa zonata are proposed here. The simultaneous use of a mitochondrial gene and two nuclear genes led to the detection of what seems to be a case of introgression of a mitochondrion from one species to another (mitochondrion capture; cytoplasmic gene flow) resulting from hybridization.     

A taxonomic note on the genus Prevotella: Description of four novel genera and emended description of the genera Hallella and Xylanibacter

The genus Prevotella comprises 55 species with validly published, and correct, names (at June 2021) that are phenotypically, ecologically and functionally diverse. This study used a range of comparative genome approaches (marker gene-based genome phylogeny, core genome phylogeny, average amino acid identity, percentage of conserved proteins and clade-specific marker genes) to identify large differences between the 53 species for which genomes are available, as well as two effectively published yet not validly named species and four novel species. These differences were consistent between the various analysis methods and justify the separation of Prevotella into multiple genera. While the distribution across 19 ecosystem types was unique for each species and inconsistent within clades, the functional repertoire based on the presence/absence of both PFAMs and CAZy families revealed distinct clustering based on the proposed genera. Based on the integration of all results, we propose the reclassification of species previously assigned to the genus Prevotella into seven genera, including four novel genera for which the names Segatella, Hoylesella, Leyella and Palleniella are proposed. In addition to the reclassification of Prevotella, this work describes four novel species, Hallella faecis, Xylanibacter rodentium, Xylanibacter muris, and Palleniella intestinalis.     

On New Findings of a Rare Sea Snake <Emphasis Type="Italic">Leioselasma czeblukovi</Emphasis> Kharin, 1984, with Remarks on Species Composition and Taxonomical Position of the Genus <Emphasis Type="Italic">Leioselasma</Emphasis> Lacepede, 1804, (Serpentes: Hydrophiidae)

Information is presented about a finding of two specimens of a rare sea snake Leioselasma czeblukovi, which was known until now only from the holotype and a paratype, and also about the taxonomical position of the genus Leioselasma within the group of genera Hydrophis sensu lato and the species composition of the latter. An identification key for species is presented.