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During a histopathological survey of Mytilus galloprovincialis in Galicia (NW Spain), microcells were observed infecting several organs of the symbiont copepod Mytilicola intestinalis. Positive results of PCR assay with specific primers for genus Mikrocytos and a clear signal of in situ hybridization with MACKINI-1 digoxigenin- labelled DNA probe (DIG-ISH) indicated a protozoan parasite of Mikrocytos genus. The ultrastructural study revealed intra and extracellular locations, polymorphic nuclei, intracellular round vesicles in the cytoplasm and absence of mitochondria. The present paper reports the characterization of the Mikrocytos sp. infecting M. intestinalis and proposes a novel species in the genus: Mikrocytos mytilicoli n. sp. A sequence of 18S-28S rDNA was obtained with 95.6% maximum identity (query cover 100%) with Mikrocytos mackini. Phylogenetic analysis showed that M. mytilicoli n. sp. and M. mackini share a common ancestor. However, comparison of the ITS1 rDNA region showed low similarity (75.8%) with M. mackini, which, combined with differences in ultrastructural details, host and geographic location, support the designation of a new species. This is the first description of a microcytid parasite of the genus Mikrocytos from a non-bivalve host., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Outbreaks of Marteilia cochillia have caused massive mortalities of common cockle, Cerastoderma edule, in some natural beds in Galicia (NW Spain) since 2012. The life cycle of Marteilia spp. is still unresolved and the most accepted hypothesis suggests that an additional host is involved. Researchers have assumed that sporangia are shed into the environment in the faeces, but details about this process have not been reported previously. Here, we report the massive liberation of Marteilia cochillia sporangia through the exhalant siphon into the environment, packaged as faeces. Using light microscopy observations on fresh samples, imprints and histology, we also describe a thick (ca. 5 µm) transparent envelope covering the sporangia that has not been reported previously. The massive release of encapsulated sporangia reported here ensures that millions of infective stages of M. cochillia cycle through the environment and become available for infection. The elucidation of the role played by the sporangia envelope would be of utmost importance for the understanding M. cochillia life cycle., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Rigid and persistent shells of microeukaryotes are widely used as bioindicators in ecological and paleontological studies. Drawing conclusions on ecological or evolutionary patterns depends strongly on the right taxonomic assignment of the observed species, however confusion is common. Especially in filose shelled amoebae it is often unclear whether species belong to the Imbricatea or Thecofilosea when only morphological data are collected. Molecular surveys shed light on their evolutionary relationship; based on these we propose a hypothesis how to differentiate doubtful species even light microscopically., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Thecofilosea is a class in Cercozoa (Rhizaria) comprising mainly freshwater-inhabiting algivores. Recently, numerous isolates of thecofilosean amoebae have been cultured and were characterized by an integrated morphological and molecular approach. As attempts to establish a culture of Lecythium mutabilis repeatedly failed, it was not yet investigated by molecular means. We isolated single cells of L. mutabilis directly from their habitat and successfully sequenced the V4 region of their SSU rDNA. Phylogenetic analyses showed that L. mutabilis is not directly related to the genus Lecythium and instead branches within the Fiscullidae (Tectofilosida, Thecofilosea). Accordingly, we transfer the species L. mutabilis to a novel genus Omnivora gen. nov., (© 2019 The Authors. Journal of Eukaryotic Microbiology published by Wiley Periodicals, Inc. on behalf of International Society of Protistologists.)

Sainouroidea is a molecularly diverse clade of cercozoan flagellates and amoebae in the eukaryotic supergroup Rhizaria. Previous 18S rDNA environmental sequencing of globally collected fecal and soil samples revealed great diversity and high sequence divergence in the Sainouroidea. However, a very limited amount of this diversity has been observed or described. The two described genera of amoebae in this clade are Guttulinopsis, which displays aggregative multicellularity, and Rosculus, which does not. Although the identity of Guttulinopsis is straightforward due to the multicellular fruiting bodies they form, the same is not true for Rosculus, and the actual identity of the original isolate is unclear. Here we isolated amoebae with morphologies like that of Guttulinopsis and Rosculus from many environments and analyzed them using 18S rDNA sequencing, light microscopy, and transmission electron microscopy. We define a molecular species concept for Sainouroidea that resulted in the description of 4 novel genera and 12 novel species of naked amoebae. Aggregative fruiting is restricted to the genus Guttulinopsis, but other than this there is little morphological variation amongst these taxa. Taken together, simple identification of these amoebae is problematic and potentially unresolvable without the 18S rDNA sequence., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

A phaeodarian morphotype, characterized by the feet surrounded with forked pedal spines with anchor-like structures, was collected in the subtropical North Pacific. Considering the morphological and phylogenetic uniqueness, this morphotype is described as Gazelletta kashiwaensis sp. nov. The distribution of this new species is possibly affected by the Kuroshio Current. The feeding behavior of living phaeodarians was first filmed: the present new species floated in the water column stretching "protoplasmic webs" and collected diatoms by repeating the expansion and retraction of "pseudopodium-like tentacles"., (© 2018 The Author(s) Journal of Eukaryotic Microbiology © 2018 International Society of Protistologists.)

Chlorotic streak is a global disease of commercial sugarcane (Saccharum spp. hybrids). The disease is transmitted by wet soil, water, as well as in diseased planting material. Although first recognized almost 90 years ago and despite significant research effort, the identity of the causal agent has been elusive. Metagenomic high throughput sequencing (HTS) facilitated the discovery of novel protistan ribosomal and nuclear genes in chlorotic streak-infected sugarcane. These sequences suggest a possible causal agent belonging to the order Cercomonadida (Rhizaria, phylum Cercozoa). An organism with morphological features similar to cercomonads (=Cercomonadida) was isolated into pure axenic culture from internal stalk tissues of infected sugarcane. The isolated organism contained DNA sequences identical to those identified in infected plants by HTS. The DNA sequences and the morphology of the organism did not match any known species. Here we present a new genus and species, Phytocercomonas venanatans, which is associated with chlorotic streak of sugarcane. Amplicon sequencing also supports that P. venanatans is associated with this disease. This is the first reported member from Cercomonadida showing a probable pathogenic association with higher plants.

Precise and sufficiently detailed morphological taxonomy is vital in biology, for example in the accurate interpretation of ecological and palaeoecological datasets, especially in polar regions, where biodiversity is poor. Testate amoebae on the Antarctic Peninsula (AP) are well-documented and variations in their population size have recently been interpreted as a proxy for microbial productivity changes in response to recent regional climate change. AP testate amoeba assemblages are dominated by a small number of globally ubiquitous taxa. We examine morphological variation in Corythion spp. across the AP, finding clear evidence supporting the presence of two morphospecies. Corythion constricta (Certes 1889) was identified on the AP for the first time and has potentially been previously misidentified. Furthermore, a southerly trend of decreasing average test size in Corythion dubium (Taránek 1881) along the AP suggests adaptive polymorphism, although the precise drivers of this remain unclear, with analysis hindered by limited environmental data. Further work into morphological variation in Corythion is needed elsewhere, alongside molecular analyses, to evaluate the potential for (pseudo)cryptic diversity within the genus. We advocate a parsimonious taxonomical approach that recognises genetic diversity but also examines and develops accurate morphological divisions and descriptions suitable for light microscopy-based ecological and palaeoecological studies., (Copyright © 2017 The Author(s). Published by Elsevier GmbH.. All rights reserved.)

Cercozoa are a highly diverse protist phylum in soils and in the phyllosphere of plants. Many families are still poorly described and the vast majority of species are still unknown. Although testate amoebae are among the better-studied protists, only little quantitative information exists on the morphology, phylogeny and ecology of cercozoan Rhogostomidae. We cultured four different strains of Rhogostoma spp. isolated from Arabidopsis leaves, agricultural soil and rhizosphere soil of Ocimum basilicum and Nicotiana sp. We describe Rhogostoma epiphylla sp. nov. and R. cylindrica sp. nov. and present their morphology, studied their food spectra in food range experiments and obtained two SSU rDNA gene sequences resulting in an updated thecofilosean phylogeny. Short generation times, desiccation resistance and the ability to prey on a wide range of algae and yeasts from the phyllosphere were seen as crucial traits for the phyllosphere colonization by Rhogostoma. In contrast, the soil-dwelling R. cylindrica did not feed on eukaryotes in our experiment., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Although testate amoebae have attracted great interest of protistologists for more than a century, some groups, especially those with a hyaline, organic test (=theca) are still poorly known. One of those is the genus Lecythium Hertwig et Lesser, 1874. Only recently Lecythium spp. were characterized by morphological and molecular means, but data on the type species Lecythium hyalinum Hertwig et Lesser, 1874, was still lacking. In this study, we screened for L. hyalinum in freshwater samples of Germany and the Netherlands. Four different isolates of L. hyalinum and one novel species were cultured and characterized by light microscopy. Phylogenetic analyses based on the small ribosomal subunit (SSU) RNA gene show that the genus Lecythium forms two robust clades, one forming a sister group to the Rhizaspididae/Pseudodifflugiidae clade (Tectofilosida), the other branching within 'Novel Clade 4' (Tectofilosida). We untangle this polyphyly by establishing Fisculla gen. nov. and the Fiscullidae fam. nov. for the former of these two clades., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Background: Genome degradation of host-restricted mutualistic endosymbionts has been attributed to inactivating mutations and genetic drift while genes coding for host-relevant functions are conserved by purifying selection. Unlike their free-living relatives, the metabolism of mutualistic endosymbionts and endosymbiont-originated organelles is specialized in the production of metabolites which are released to the host. This specialization suggests that natural selection crafted these metabolic adaptations. In this work, we analyzed the evolution of the metabolism of the chromatophore of Paulinella chromatophora by in silico modeling. We asked whether genome reduction is driven by metabolic engineering strategies resulted from the interaction with the host. As its widely known, the loss of enzyme coding genes leads to metabolic network restructuring sometimes improving the production rates. In this case, the production rate of reduced-carbon in the metabolism of the chromatophore., Results: We reconstructed the metabolic networks of the chromatophore of P. chromatophora CCAC 0185 and a close free-living relative, the cyanobacterium Synechococcus sp. WH 5701. We found that the evolution of free-living to host-restricted lifestyle rendered a fragile metabolic network where >80% of genes in the chromatophore are essential for metabolic functionality. Despite the lack of experimental information, the metabolic reconstruction of the chromatophore suggests that the host provides several metabolites to the endosymbiont. By using these metabolites as intracellular conditions, in silico simulations of genome evolution by gene lose recover with 77% accuracy the actual metabolic gene content of the chromatophore. Also, the metabolic model of the chromatophore allowed us to predict by flux balance analysis a maximum rate of reduced-carbon released by the endosymbiont to the host. By inspecting the central metabolism of the chromatophore and the free-living cyanobacteria we found that by improvements in the gluconeogenic pathway the metabolism of the endosymbiont uses more efficiently the carbon source for reduced-carbon production. In addition, our in silico simulations of the evolutionary process leading to the reduced metabolic network of the chromatophore showed that the predicted rate of released reduced-carbon is obtained in less than 5% of the times under a process guided by random gene deletion and genetic drift. We interpret previous findings as evidence that natural selection at holobiont level shaped the rate at which reduced-carbon is exported to the host. Finally, our model also predicts that the ABC phosphate transporter (pstSACB) which is conserved in the genome of the chromatophore of P. chromatophora strain CCAC 0185 is a necessary component to release reduced-carbon molecules to the host., Conclusion: Our evolutionary analysis suggests that in the case of Paulinella chromatophora natural selection at the holobiont level played a prominent role in shaping the metabolic specialization of the chromatophore. We propose that natural selection acted as a "metabolic engineer" by favoring metabolic restructurings that led to an increased release of reduced-carbon to the host.

The thecate filose amoeba Paulinella chromatophora is a good model organism for understanding plastid organellogenesis because its chromatophore was newly derived from an alpha-cyanobacterium. Paulinella chromatophora was the only known photosynthetic Paulinella species until recent studies that suggested a species level of diversity. Here, we described a new photosynthetic species P. micropora sp. nov. based on morphological and molecular evidence from a newly established strain KR01. The chromatophore genome of P. micropora KR01 was fully determined; the genome was 976,991bp in length, the GC content was 39.9%, and 908 genes were annotated. A pairwise comparison of chromatophore genome sequences between strains KR01 and FK01, representing two different natural populations of P. micropora, showed a 99.85% similarity. Differences between the two strains included single nucleotide polymorphisms (SNPs) in CDSs, which resulted in 357 synonymous and 280 nonsynonymous changes, along with 245 SNPs in non-coding regions. Indels (37) and microinversions (14) were also detected. Species diversity for photosynthetic Paulinella was surveyed using samples collected from around the world. We compared our new species to two photosynthetic species, P. chromatophora and P. longichromatophora. Phylogenetic analyses using four gene markers revealed three distinct lineages of photosynthetic Paulinella species including P. micropora sp. nov., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Several distantly related, phagotrophic microeukaryotes share the remarkable ability to perforate foreign cell walls in a well-defined pattern to acquire protoplast material as food. The underlying cellular processes, especially the local application of cell wall degrading agents, are still unexplored. We examined the distribution of F-actin and alpha-tubulin in the algivorous, viridiraptorid amoeboflagellates Orciraptor agilis and Viridiraptor invadens over their life histories using phalloidin conjugates and immunolocalization. During attack, both species form distinctive, F-actin-rich structures at the contact zone to the algal prey cell, which exactly match the species-specific cell wall perforations and resemble invadopodia and podosome rosettes of mammalian cells to a certain extent. Furthermore, F-actin is involved in the extraction of plastid material by Orciraptor and in prey cell invasion by Viridiraptor (here, F-actin localizes to a characteristic hyaline channel, which surrounds the streaming cytoplasm). The digestive-reproductive stages of viridiraptorids display a highly ordered microtubular cytoskeleton, whereas distinct phalloidin-positive actin structures could not be detected. We discuss the presumed function of F-actin during perforation and penetration of the algal cell wall by viridiraptorids, and compare the cytoskeleton architecture of these protoplast feeders to amoeboflagellates from different eukaryotic supergroups., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

For over a century testate amoebae have been a favoured group of interest for protistologists, however there is still an endless amount of unanswered questions. The genus Plagiophrys, Claparède and Lachmann 1859, is still one of the unresolved mysteries as it comprises species with high morphological diversity of which no molecular data are available. To shed light on the phylogeny and taxonomy of Plagiophrys we (a) cultured four isolates of three Plagiophrys morphospecies and provided morphological observations (b) obtained three new SSU RNA gene sequences and conducted phylogenetic analyses of the Thecofilosea and (c) did intensive literature research, showing that Plagiophrys is polyphyletic. We partially untangle this polyphyly by combining several of its species with the genus Rhizaspis, Skuja 1948. Furthermore, we establish Sacciforma gen. nov. to accommodate P. sacciformis as it groups within the formerly known Rhizaspididae, which do not comprise our isolates of Rhizaspis (and therefore were renamed Rhogostomidae) since Rhizaspis it groups with maximum support as a sister-group to the Pseudodifflugiidae., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

The morphology of tests ("shells") representing populations of the small testate amoeba Cyphoderia laevisPenard, 1902 (Cercozoa, Euglyphida) from Canada and Hungary was quantified. Despite often considerable variation the cross-sectional shape of tests, multivariate statistical analysis confirmed the general uniformity of test morphology within populations and between populations at both local (regional, inter- and intra-watershed) scales and over inter-continental scales (Canada and Hungary). Corrections are suggested to recently published misidentifications of C. laevis as Corythionella georgiana Nicholls. The differences between these two taxa are subtle, but significant, and are emphasized here to help preclude similar errors in the future. Schaudinnula nanaBadewitz, 2003 is shown to be a superfluous synonym for Cyphoderia laevis. Cyphoderia truncataSchulze, 1875 has some similarities to C. laevis, but its status remains unresolved. Some preliminary reservations are expressed about the classification of Cyphoderia laevis within the genus Cyphoderia owing mainly to its Corythionella-like scale structure and other characteristics of its test that appear to be outside the "usual" range found in other Cyphoderia species., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Two heterotrophic flagellate strains were isolated from marine sediment samples off eastern Canada and Korea. These new isolates are indistinguishable by light microscopy from the unclassified protist Cyranomonas australis. The organisms are ovoid-shaped cells, 3.5-6μm long, laterally compressed, and somewhat flexible. They have two unequal flagella, about 1.1-2.5 times body length. Typically, the cells show a gliding motility and do not exhibit any amoeboid form or pseudopodia. 18S rDNA phylogenies clearly indicate that the isolates can be assigned to the taxon Filosa, within Cercozoa. The isolates are closest to an environmental sequence (CYSGM-16; 99% identity). Cyranomonas, CYSGM-16, and uncultured eukaryote RM1-SGM46 form a clade with strong statistical supports, here called novel clade CU (Cyranomonas plus Uncultured eukaryotes). This clade may be sister to the order Marimonadida. The novel clade CU and the Marimonadida have been detected only in marine habitats. Our findings suggest that C. australis may not belong to any previously described family within Filosa and Cercozoa., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

The term 'filose amoebae' describes a highly polyphyletic assemblage of protists whose phylogenetic placement can be unpredictable based on gross morphology alone. We isolated six filose amoebae from soils of two European countries and describe a new genus and species of naked filose amoebae, Kraken carinae gen. nov. sp. nov. We provide a morphological description based on light microscopy and small subunit rRNA gene sequences (SSU rDNA). In culture, Kraken carinae strains were very slow-moving and preyed on bacteria using a network of filopodia. Phylogenetic analyses of SSU sequences reveal that Kraken are core (filosan) Cercozoa, branching weakly at the base of the cercomonad radiation, most closely related to Paracercomonas, Metabolomonas, and Brevimastigomonas. Some Kraken sequences are >99% similar to an environmental sequence obtained from a freshwater lake in Antarctica, indicating that Kraken is not exclusively soil dwelling, but also inhabits freshwater habitats., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

A new phaeodarian species, characterized by the presence of long developed side branches recurved proximally and distally on the surface of its radial tube, was described as Auloscena pleuroclada. This new species was only collected from the layers below the 250 m depth in the Sea of Japan. They have never been found in the shallower layers (above 250 m) of this sea or in other investigated areas. The distribution of the present new species is presumably restricted within the deep water of this area, and this species could be a specific phaeodarian adapted to the deep-sea environment., (© 2015 The Author(s) Journal of Eukaryotic Microbiology © 2015 International Society of Protistologists.)

The freshwater testate filose amoeba Paulinella chromatophora is the sole species in the genus to have plastids, usually termed "chromatophores", of a Synechococcus/Prochlorococcus-like cyanobacterial origin. Here, we report a new marine phototrophic species, Paulinella longichromatophora sp. nov., using light and electron microscopy and molecular data. This new species contains two blue-green U-shaped chromatophores reaching up to 40 μm in total length. Further, the new Paulinella species is characterized by having five oral scales surrounding the pseudostomal aperture. All trees generated using three nuclear rDNA datasets (18S rDNA, 28S rDNA, and the concatenated 18S + 28S rDNA) demonstrated that three photosynthetic Paulinella species (two freshwater species, P. chromatophora and Paulinella strain FK01, and one marine species, P. longichromatophora) congruently formed a monophyletic group with strong support (≥ 90% of ML and ≥ 0.90 of PP), but their relationship to each other within the clade remained unresolved in all trees. P. longichromatophora, nevertheless, clustered consistently together with Paulinella strain FK01 with very low support, but the clade received strong support in plastid phylogenies. Phylogenetic analyses inferred from plastid-encoded 16S rDNA and a concatenated dataset of plastid 16S+23S rDNA demonstrated that chromatophores of all photosynthetic Paulinella species were monophyletic. The monophyletic group fell within a cyanobacteria clade having a close relationship to an α-cyanobacterial clade containing Prochlorococcus and Synechococcus species with very robust support (100% of ML and 1.0 of PP). Additionally, phylogenetic analyses of nuclear 18S rDNA and plastid 16S rDNA suggested divergent evolution within the photosynthetic Paulinella population after a single acquisition of the chromatophore. After the single acquisition of the chromatophore, ancestral photosynthetic Paulinella appears to have diverged into at least two distinct clades, one containing the marine P. longichromatophora and freshwater Paulinella strain FK01, the other P. chromatophora CCAC 0185., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Background: Division of double-membraned plastids (primary plastids) is performed by constriction of a ring-like division complex consisting of multiple plastid division proteins. Consistent with the endosymbiotic origin of primary plastids, some of the plastid division proteins are descended from cyanobacterial cell division machinery, and the others are of host origin. In several algal lineages, complex plastids, the "secondary plastids", have been acquired by the endosymbiotic uptake of primary plastid-bearing algae, and are surrounded by three or four membranes. Although homologous genes for primary plastid division proteins have been found in genome sequences of secondary plastid-bearing organisms, little is known about the function of these proteins or the mechanism of secondary plastid division., Results: To gain insight into the mechanism of secondary plastid division, we characterized two plastid division proteins, FtsZD-1 and FtsZD-2, in chlorarachniophyte algae. FtsZ homologs were encoded by the nuclear genomes and carried an N-terminal plastid targeting signal. Immunoelectron microscopy revealed that both FtsZD-1 and FtsZD-2 formed a ring-like structure at the midpoint of bilobate plastids with a projecting pyrenoid in Bigelowiella natans. The ring was always associated with a shallow plate-like invagination of the two innermost plastid membranes. Furthermore, gene expression analysis confirmed that transcripts of ftsZD genes were periodically increased soon after cell division during the B. natans cell cycle, which is not consistent with the timing of plastid division., Conclusions: Our findings suggest that chlorarachniophyte FtsZD proteins are involved in partial constriction of the inner pair of plastid membranes, but not in the whole process of plastid division. It is uncertain how the outer pair of plastid membranes is constricted, and as-yet-unknown mechanism is required for the secondary plastid division in chlorarachniophytes.

Phaeodarians are a group of widely distributed marine cercozoans. These plankton organisms can exhibit a large biomass in the environment and are supposed to play an important role in marine ecosystems and in material cycles in the ocean. Accurate knowledge of phaeodarian classification is thus necessary to better understand marine biology, however, phylogenetic information on Phaeodaria is limited. The present study analyzed 18S rDNA sequences encompassing all existing phaeodarian orders, to clarify their phylogenetic relationships and improve their taxonomic classification. The monophyly of Phaeodaria was confirmed and strongly supported by phylogenetic analysis with a larger data set than in previous studies. The phaeodarian clade contained 11 subclades which generally did not correspond to the families and orders of the current classification system. Two families (Challengeriidae and Aulosphaeridae) and two orders (Phaeogromida and Phaeocalpida) are possibly polyphyletic or paraphyletic, and consequently the classification needs to be revised at both the family and order levels by integrative taxonomy approaches. Two morphological criteria, 1) the scleracoma type and 2) its surface structure, could be useful markers at the family level., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

We describe 11 new species of Thaumatomonadida using light and electron microscopy and rDNA gene sequences (18S, ITS1, 5.8S, ITS2). We found clear distinctions between major clades in molecular and morphological traits that support now splitting Thaumatomastix into three genera: new marine genera Ovaloplaca (oval plate-scales) and Thaumatospina (triangular plate-scales), both with distinctive radially-symmetric bobbin-based spine-scales, restricting Thaumatomastix to freshwater species with putatively non-homologous eccentric-spine scales and thicker triangular plate-scales. New genus Scutellomonas lacks spine-scales, having oval plate-scales with deeply-dished upper tier as in Ovaloplaca, with which it forms a clade having short/absent anterior cilium. Cowlomonas gen. n. is possibly naked. We describe two new Allas species, two new Thaumatomonas, and one new Reckertia species, and transfer R. hindoni to Thaumatomonas. Triangular-scaled Reckertia has varied plate-scales and ciliary scales. Thaumatomonas rDNA trees reveal two clades: zhukovi/seravini (predominantly triangular scales); coloniensis/oxoniensis/lauterborni/constricta/solis (scales mostly oval). We hypothesise that the ancestor of Thaumatomonadidae had radially-symmetric bobbin-based spine-scales and triangular plate-scales, bobbin-based spine-scales being lost in one lineage and eccentric-spine scales evolving in Thaumatomastix. Bobbin-based spine-scales arguably evolved from triangular plate-scales and single-tier ciliary scales (Ovaloplaca and Reckertia only) from plate-scale rudiments. We present a unified scheme for scale evolution and development in Imbricatea., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Most euglyphids, a group of testate amoebae, have a shell that is constructed from numerous siliceous scales. The euglyphid Paulinella chromatophora has photosynthetic organelles (termed cyanelles or chromatophores), allowing it to be cultivated more easily than other euglyphids. Like other euglyphids, P. chromatophora has a siliceous shell made of brick-like scales. These scales are varied in size and shape. How a P. chromatophora cell makes this shell is still a mystery. We examined shell construction process in P. chromatophora in detail using time-lapse video microscopy. The new shell was constructed by a specialized pseudopodium that laid out each scale into correct position, one scale at a time. The present study inferred that the sequence of scale production and secretion was well controlled., (© 2014 The Author(s) Journal of Eukaryotic Microbiology © 2014 International Society of Protistologists.)

Euglyphid testate amoebae are a highly conspicuous group of Cercozoa whose systematics is based mainly on the shape and ultrastructure of the shell. However, only a couple of species have been studied with molecular methods. As a consequence, there are still some genera whose classification remains uncertain. Amongst those are Sphenoderia and Trachelocorythion, two genera with diverging ecological requirements that share a collar composed of small scales around the aperture. We demonstrate here with a molecular and morphological approach that they are closely related, and propose a new family, Sphenoderiidae fam. nov. to group these species. Some species share almost similar morphology in spite of being genetically distantly related (Sphenoderia minuta and S. pseudominuta sp. nov.), underlining the importance of combining ultrastructural and morphological data when describing new species of protists. In addition, we describe here Sphenoderia valdiviana sp. nov., a new species isolated from Southern Chile temperate rainforests., (Copyright © 2013 Elsevier GmbH. All rights reserved.)

Cryptophyte and chlorarachniophyte algae are transitional forms in the widespread secondary endosymbiotic acquisition of photosynthesis by engulfment of eukaryotic algae. Unlike most secondary plastid-bearing algae, miniaturized versions of the endosymbiont nuclei (nucleomorphs) persist in cryptophytes and chlorarachniophytes. To determine why, and to address other fundamental questions about eukaryote-eukaryote endosymbiosis, we sequenced the nuclear genomes of the cryptophyte Guillardia theta and the chlorarachniophyte Bigelowiella natans. Both genomes have >21,000 protein genes and are intron rich, and B. natans exhibits unprecedented alternative splicing for a single-celled organism. Phylogenomic analyses and subcellular targeting predictions reveal extensive genetic and biochemical mosaicism, with both host- and endosymbiont-derived genes servicing the mitochondrion, the host cell cytosol, the plastid and the remnant endosymbiont cytosol of both algae. Mitochondrion-to-nucleus gene transfer still occurs in both organisms but plastid-to-nucleus and nucleomorph-to-nucleus transfers do not, which explains why a small residue of essential genes remains locked in each nucleomorph.

We report a new naked cercozoan flagellate, Esquamula lacrimiformis n. g., n. sp., collected from a sandy beach in Japan. Its cells were 4.5-11.3 μm in length and 3.9-8.8 μm in width and possess two unequal flagella. Cells move in a smooth gliding motion and have a trailing long posterior flagellum. Phylogenetic analyses with small and large subunit ribosomal RNA genes revealed that E. lacrimiformis forms a novel lineage within the Thaumatomonadida, the members of which are flagellates with siliceous scales. However, our light and electron microscopic observations indicated that E. lacrimiformis cells do not possess any siliceous structures. Furthermore, other morphological characteristics, such as the shape of the extrusomes and the structural arrangement of the microbody, were clearly different from those of previously described thaumatomonads. On the basis of a combination of these morphological observations and our phylogenetic analyses, we conclude that E. lacrimiformis should be treated as a new species of a new genus and placed into a new family, Esquamulidae n. fam., under Thaumatomonadida., (© 2012 The Author(s) Journal of Eukaryotic Microbiology © 2012 International Society of Protistologists.)

This study aimed to investigate the relationship between the size structure of testate amoebae in distinct habitats, i.e. plankton, aquatic macrophytes and aquatic sediment. The samples were taken from a floodplain lake of the upper Paraná River. The assumptions we strived to scrutinize were that (i) larger mean sizes of testate amoebae would be recorded in the sediment of the lake; and (ii) temporally, smaller individuals would be registered during the high water period in all habitats. The sampling was done monthly, from April 2007 to March 2008, in triplicates for each habitat. Testate amoebae were represented by individuals sized between 20 and 400 μm. The smaller individuals predominated in plankton samples, while in the aquatic sediment the larger ones were chiefly represented. These differences were probably associated with metabolic activities, i.e. the energy needs of these unicellular organisms, in each habitat. Two-way ANOVA yielded significant differences between hydrological periods. During the high water period, the increase in rainfall and consequently in water flow, decreased the stability of the system and increased turbulence and water column circulation. Therefore, environmental stability seems to be one of the main factors driving the temporal variation in the size structure of these specific organisms., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

A culture of Thaumatomastix was isolated from a sediment sample collected in Oslofjorden and established as a monospecific strain (UIO286). Based on this culture, light and transmission electron microscopy and phylogenetic analyses were carried out. Thaumatomastix species are confined within the order Thaumatomonadida of the class Imbricatea and phylum Cercozoa. They are heterotrophic and their cell bodies are covered with silica scales. Observations of thin sections as well as whole mounts indicate that the morphology and ultrastructure of UIO286 is identical to T. salina, which was initially described from salt pools in Denmark. Detailed examination revealed some new features such as the presence of pseudopodia and silica deposition vesicles producing spine scales. The phylogeny presented here includes ribosomal DNA sequences from both imbricatean cultures and environmental samples. The 18S rDNA phylogenetic tree suggests that (i) Thaumatomastix is paraphyletic within the Thaumatomonadida clade, (ii) there is no close affinity between T. salina and other cultured and sequenced strains, but it is closely related to a sequence obtained from environmental DNA; we propose the present strain to serve as a reference culture of Thaumatomastix species and T. salina. Further, we discuss the distribution, habitats, and evolution of scale formation among euglyphids and thaumatomonads., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Cercomonads are important components of microbial food webs in soils and aquatic sediments. Here, we investigated the general morphology, behaviour, life cycle and 18S rDNA phylogeny of cercomonad cultures from a German grassland soil habitat. We describe ten new species including two new genera from 23 strains. Three Cercomonas, two Eocercomonas and three Paracercomonas species are described. Based on large phylogenetic distance and distinct morphology, we erect two novel clade B genera near the root of the cercomonad tree. Nucleocercomonas nov. gen. bears a number of characters unusual for cercomonads: Its anterior flagellum is extremely long, it mostly does not glide, and in its most frequent life stage the cell body does not attach to the substratum, but produces unattached pseudopodia. Furthermore, it has a unique nucleus with a peripheral nucleolus that attaches to the nuclear envelope opposite the basal body connection. Metabolomonas nov. gen. is extremely metabolic. It is characterized by a very high beating frequency of the anterior flagellum, fast gliding, rapid changes in shape and strong cytoplasmic streams. A new genus Brevimastigomonas is erected for the previously described species Paracercomonas anaerobica. The general morphology of cercomonad species often does not correspond with their phylogenetic position: closely related species may have a very different morphology., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

The evolution of plastids from cyanobacteria is believed to represent a singularity in the history of life. The enigmatic amoeba Paulinella and its 'recently' acquired photosynthetic inclusions provide a fascinating system through which to gain fresh insight into how endosymbionts become organelles.The plastids, or chloroplasts, of algae and plants evolved from cyanobacteria by endosymbiosis. This landmark event conferred on eukaryotes the benefits of photosynthesis--the conversion of solar energy into chemical energy--and in so doing had a huge impact on the course of evolution and the climate of Earth 1. From the present state of plastids, however, it is difficult to trace the evolutionary steps involved in this momentous development, because all modern-day plastids have fully integrated into their hosts. Paulinella chromatophora is a unicellular eukaryote that bears photosynthetic entities called chromatophores that are derived from cyanobacteria and has thus received much attention as a possible example of an organism in the early stages of organellogenesis. Recent studies have unlocked the genomic secrets of its chromatophore 23 and provided concrete evidence that the Paulinella chromatophore is a bona fide photosynthetic organelle 4. The question is how Paulinella can help us to understand the process by which an endosymbiont is converted into an organelle.

Chlorarachniophytes are enigmatic marine unicellular algae that acquired photosynthesis by secondary endosymbiosis. Chlorarachniophytes are unusual in that the nucleus of the engulfed algal cell (a green alga) persists in a miniaturized form, termed a nucleomorph. The nucleomorph genome of the model chlorarachniophyte, Bigelowiella natans CCMP621, is 373 kilobase pairs (kbp) in size, the smallest nuclear genome characterized to date. The B. natans nucleomorph genome is composed of three chromosomes, each with canonical eukaryotic telomeres and sub-telomeric ribosomal DNA (rDNA) operons transcribed away from the chromosome end. Here we present the complete rDNA operon and telomeric region from the nucleomorph genome of Lotharella oceanica CCMP622, a newly characterized chlorarachniophyte strain with a genome ∼610 kbp in size, significantly larger than all other known chlorarachniophytes. We show that the L. oceanica rDNA operon is in the opposite chromosomal orientation to that of B. natans. Furthermore, we determined the rDNA operon orientation of five additional chlorarachniophyte strains, the majority of which possess the same arrangement as L. oceanica, with the exception of Chlorarachnion reptans and those very closely related to B. natans. It is thus possible that the ancestral rDNA operon orientation of the chlorarachniophyte nucleomorph genome might have been the same as in the independently evolved, red algal-derived, nucleomorph genomes of cryptophytes. A U2 small nuclear RNA gene was found adjacent to the telomere in Gymnochlora stellata CCMP2057 and Chlorarachnion sp. CCMP2014. This feature may represent a useful evolutionary character for inferring the relationships among extant lineages., (© 2010 The Author(s). Journal of Eukaryotic Microbiology © 2010 International Society of Protistologists.)

This study aimed to investigate the testate amoebae (Arcellinida and Eugliphida) species diversity in plankton, macrophytes and aquatic sediment samples from a shallow lake of the Upper Paraná River floodplain. Samples were carried out from April 2007 to March 2008. We recorded 89 taxa, belonging to 10 families. Eighty-two taxa were found in the aquatic sediment, 71 in the macrophytes and 53 in the plankton. Highest values of alpha diversity were observed in the aquatic sediment. Although the plankton had the highest number of accidental species, accessory and constant species were also observed in this habitat. Most of the species classified as constant for the plankton belonged to the genus Arcella. Most of the constant species in the macrophytes and aquatic sediment belonged to the genus Difflugia. This study supports the idea that the presence of these protists in the plankton should not be attributed only to stochastic processes because (i) the species diversity recorded in this habitat was remarkably high in relation to the total biodiversity of the lake, and (ii) we also recorded frequent and constant species in the plankton., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

Clautriavia is a genus of uncertain taxonomic affinity that was initially described as gliding cells with one prominent trailing flagellum and a mid-ventral groove. The genus has been classified either with euglenids on the basis of similar paramylon-like granules or with cercozoans, specifically Protaspis spp., on the basis of general similarities in cell morphology and behavior. We isolated and cultivated a novel species of Clautriavia, namely C. biflagellata n. sp., from marine sand samples collected from the west coast of Vancouver Island, Canada and characterized this isolate with high resolution microscopy (LM, SEM, and TEM) and small subunit (SSU) rDNA sequence. The gliding cells of C. biflagellata n. sp. were round to oval in outline (12-20 microm wide and 15-20 microm long), dorsoventrally flattened, and capable of engulfing other eukaryotic cells (e.g., diatoms). The cells possessed two recurrent flagella of unequal length that emerged from a subapical pit within a ventral depression: the longer prominent flagellum was about 2X the cell length; the shorter flagellum was inconspicuous and was confined to the ventral depression. Molecular phylogenetic analyses demonstrated that C. biflagellata n. sp. branched strongly within the Cercozoa, but was only distantly related to Protaspis spp. Instead, C. biflagellata n. sp. branched closely with the recently established Auranticordida clade, consisting of Auranticordis quadriverberis and Pseudopirsonia mucosa. This position was concordant with our ultrastructural data, which demonstrated several features shared by A. quadriverberis and C. biflagellata n. sp. that are not present in Protaspis spp.: (1) a dense distribution of pores on the cell surface; (2) a distinct layer of muciferous bodies immediately beneath the cell surface; (3) a robust microtubular root attached to the anterior end of the nucleus; (4) the absence of a thick cell covering; and (5) the absence of conspicuously condensed chromosomes., (Copyright 2009 Elsevier GmbH. All rights reserved.)