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Phryganella acropodia Hertwig and Lesser, 1874, is one of the most common and abundant testate amoeba species. It represents the type species of the genus Phryganella Penard, 1902, which in turn is the type genus for the suborder Phryganellina (Arcellinida) Bovee, 1985, but despite its taxonomic importance it was not yet analyzed with molecular methods. We established two cultures of putative Phryganella acropodia, designed Phryganellina-specific primers, amplified SSU rDNA data and subjected these sequences to phylogenetic analyses. Morphological and genetic differences were found between both strains. With SSU rDNA phylogenetic analyses we confirm that Phryganella acropodia branches with Phryganella paradoxa Penard, 1902 and Cryptodifflugia Penard, 1890 in the Phryganellina. We thus give further evidence that pseudopodia morphology in the Arcellinida is a character of high taxonomic value, as suggested by Bovee and Jung when erecting the suborder Phryganellina. Moreover, we provide evidence for cryptic diversity and for the first time confirm the existence of a naked life stage in Arcellinida by molecular means., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Artodiscus saltans, first described by Penard (1890), has a unique morphology. Without genetic data it could not yet been reliably placed into a wider taxonomical context. We present morphological data for A. saltans from different aquatic habitats of four European countries. We subjected three cells of one strain from Germany to molecular analyses and, interestingly, obtained six different rDNA sequences. Phylogenetic analyses of these SSU rDNA sequences revealed that A. saltans branches close to the amoebozoan Multicilia marina (Variosea, Amoebozoa)., (Copyright © 2019 Elsevier GmbH. All rights reserved.)

The genus Kraken represents a distinct lineage of filose amoebae within the Cercozoa. Currently a single species, Kraken carinae, has been described. SSU rDNA phylogeny showed an affiliation to the Cercomonadida, branching with weak support at its base, close to Paracercomonas, Metabolomonas, and Brevimastigomonas. Light microscopical analyses showed several unique features of the genus Kraken, but ultrastructure data were lacking. In this study, K. carinae has been studied by electron microscopy, these data conjoined with a two-gene phylogeny were used to give more insight into the evolutionary relationship of the genus Kraken within Cercozoa. The data confirmed the absence of flagella, but also showed novel characteristics, such as the presence of extrusomes, osmiophilic bodies, and mitochondria with flat cristae. Surprising was the presence of single-tier scales which are carried by cell outgrowths, much of what is expected of the last common ancestor of the class Imbricatea. The phylogenetic analyses however confirmed previous results, indicating Kraken as a sister group to Paracercomonas in Sarcomonadea with an increased but still low support of 0.98 PP/63 BP. Based on the unique features of Kraken we establish the Krakenidae fam. nov. that we, due to contradictory results in morphology and phylogeny, assign incertae sedis, Monadofilosa., (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.)

We describe four new species of Flabellula, Leptomyxa and Rhizamoeba and publish new SSU rRNA gene and actin gene sequences of leptomyxids. Using these data we provide the most comprehensive SSU phylogeny of leptomyxids to date. Based on the analyses of morphological data and results of the SSU rRNA gene phylogeny we suggest changes in the systematics of the order Leptomyxida (Amoebozoa: Lobosa: Tubulinea). We propose to merge the genera Flabellula and Paraflabellula (the genus Flabellula remains valid by priority rule). The genus Rhizamoeba is evidently polyphyletic in all phylogenetic trees; we suggest retaining the generic name Rhizamoeba for the group unifying R. saxonica, R.matisi n. sp. and R. polyura, the latter remains the type species of the genus Rhizamoeba. Based on molecular and morphological evidence we move all remaining Rhizamoeba species to the genus Leptomyxa. New family Rhizamoebidae is established here in order to avoid paraphyly of the family Leptomyxidae. With the suggested changes both molecular and morphological systems of the order Leptomyxida are now fully congruent to each other., (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.)

Background: Acanthamoebidae is a "family" level amoebozoan group composed of the genera Acanthamoeba, Protacanthamoeba, and very recently Luapeleamoeba. This clade of amoebozoans has received considerable attention from the broader scientific community as Acanthamoeba spp. represent both model organisms and human pathogens. While the classical composition of the group (Acanthamoeba + Protacanthamoeba) has been well accepted due to the morphological and ultrastructural similarities of its members, the Acanthamoebidae has never been highly statistically supported in single gene phylogenetic reconstructions of Amoebozoa either by maximum likelihood (ML) or Bayesian analyses., Results: Here we show using a phylogenomic approach that the Acanthamoebidae is a fully supported monophyletic group within Amoebozoa with both ML and Bayesian analyses. We also expand the known range of morphological and life cycle diversity found in the Acanthamoebidae by demonstrating that the amoebozoans "Protostelium" arachisporum, Dracoamoeba jormungandri n. g. n. sp., and Vacuolamoeba acanthoformis n.g. n.sp., belong within the group. We also found that "Protostelium" pyriformis is clearly a species of Acanthamoeba making it the first reported sporocarpic member of the genus, that is, an amoeba that individually forms a walled, dormant propagule elevated by a non-cellular stalk. Our phylogenetic analyses recover a fully supported Acanthamoebidae composed of five genera. Two of these genera (Acanthamoeba and Luapeleameoba) have members that are sporocarpic., Conclusions: Our results provide high statistical support for an Acanthamoebidae that is composed of five distinct genera. This study increases the known morphological diversity of this group and shows that species of Acanthamoeba can include spore-bearing stages. This further illustrates the widespread nature of spore-bearing stages across the tree of Amoebozoa., Reviewers: This article was reviewed by Drs. Eugene Koonin, Purificacion Lopez-Garcia and Sandra Baldauf. Sandra Baldauf was nominated by Purificacion Lopez-Garcia, an Editorial Board member.

Myxomycetes or plasmodial slime molds are widespread and very common soil amoebae with the ability to form macroscopic fruiting bodies. Even if their phylogenetic position as a monophyletic group in Amoebozoa is well established, their internal relationships are still not entirely resolved. At the base of the most intensively studied dark-spored clade lies the order Echinosteliales, whose highly divergent small subunit ribosomal (18S) RNA genes represent a challenge for phylogenetic reconstructions. This is because they are characterized by unusually long variable helices of unknown secondary structure and a high inter- and infraspecific divergence. Current classification recognizes two families: the monogeneric Echinosteliaceae and the Clastodermataceae with the genera Barbeyella and Clastoderma. To better resolve the phylogeny of the Echinosteliales, we obtained three new small subunit ribosomal (18S) RNA gene sequences of Clastoderma and Echinostelium corynophorum. Our phylogenetic analyses suggested the polyphyly of the family Clastodermataceae, as Barbeyella was more closely related to Echinostelium arboreum than to Clastoderma, while Clastoderma debaryanum was the earliest branching clade in Echinosteliales. We also found that E. corynophorum was the closest relative of the enigmatic Semimorula liquescens, a stalkless-modified Echinosteliales. We discuss possible evolutionary pathways in dark-spored Myxomycetes and propose a taxonomic update., (© 2015 The Author(s) Journal of Eukaryotic Microbiology © 2015 International Society of Protistologists.)

Heterolobosean amoebae are common and diverse members of soil protist communities. In this study, we isolated seven strains of amoebae from soil samples taken in Tibet (at high altitude), Sardinia and the Netherlands, all resembling to belong to a similar heterolobosean morphospecies. However, sequences of the small subunit (SSU) rDNA and internal transcribed spacers, including the 5.8S rDNA, revealed a high heterogeneity in the genus Allovahlkampfia to which six of the isolates belong. Some unnamed strains, of which the sequences had been published before, are also included within the genus Allovahlkampfia. One Allovahlkampfia isolated in the Netherlands harbors a twin-ribozyme, containing a His-Cys box, similar to the one found in strain BA of Allovahlkampfia. The other SSU rDNA sequence grouped in phylogenetic analyses with sequences obtained in environmental sequencing studies as sister to the genus Fumarolamoeba. This phylogenetic placement was supported by analyses of the 5.8S rDNA leading us to describe it as a new genus Parafumarolamoeba., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Amoebae able to form cytoplasmic networks or displaying a multiply branching morphology remain very poorly studied. We sequenced the small-subunit ribosomal RNA gene of 15 new amoeboid isolates, 14 of which are branching or network-forming amoebae (BNFA). Phylogenetic analyses showed that these isolates all group within the poorly-known and weakly-defined class Variosea (Amoebozoa). They are resolved into six lineages corresponding to distinct new morphotypes; we describe them as new genera Angulamoeba (type species Angulamoeba microcystivorans n. gen., n. sp.; and A. fungorum n. sp.), Arboramoeba (type species Arboramoeba reticulata n. gen., n. sp.), Darbyshirella (type species Darbyshirella terrestris n. gen., n. sp.), Dictyamoeba (type species Dictyamoeba vorax n. gen., n. sp.), Heliamoeba (type species Heliamoeba mirabilis n. gen., n. sp.), and Ischnamoeba (type species Ischnamoeba montana n. gen., n. sp.). We also isolated and sequenced four additional variosean strains, one belonging to Flamella, one related to Telaepolella tubasferens, and two members of the cavosteliid protosteloid lineage. We identified a further 104 putative variosean environmental clone sequences in GenBank, comprising up to 14 lineages that may prove to represent additional novel morphotypes. We show that BNFA are phylogenetically widespread in Variosea and morphologically very variable, both within and between lineages., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Two new species of the recently described genus Stenamoeba, named S. berchidia and S. sardiniensis were isolated from a single soil sample on Sardinia, Italy. Both share morphological features characteristic to Stenamoeba and form in phylogenetic analyses together with other Stenamoeba spp. a highly supported clade within the family Thecamoebidae. The ultrastructural investigation of Stenamoeba sardiniensis revealed the presence of cytoplasmic microtubule-organizing centers (MTOCs), located close to one of several dictyosomes found inside the cell. This is the first report of cytoplasmic MTOCs among Thecamoebidae. The presence of MTOCs is now shown in five of nine orders comprising the class Discosea and potentially could be a phylogenetic marker in this group. We re-isolated Stenamoeba limacina from German soils. This strain shows a similar morphology and an almost complete SSU rDNA sequence identity with the type strain of S. limacina originating from gills of fishes, collected in Czech Republic., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Myxomycetes (also called Myxogastria or colloquially, slime molds) are worldwide occurring soil amoeboflagellates. Among Amoebozoa, they have the notable characteristic to form, during their life cycle, macroscopic fruiting bodies, that will ultimately release spores. Some 1,000 species have been described, based on the macroscopic and microscopic characteristics of their fruiting bodies. We were interested in Physarum pusillum (Berk. & M.A. Curtis) G. Lister, a very common species described with two variants, each bearing such morphological differences that they could represent two distinct species. In order to test this, we observed key characters in a large selection of specimens attributed to P. pusillum, to its synonyms (in particular Physarum gravidum), and to related species. In addition, the small‐subunit ribosomal RNA gene was obtained from seven of these specimens. Based on these data, we provide a comprehensive phylogeny of the order Physarida (Eukaryota: Amoebozoa: Conosa: Macromycetozoa: Fuscisporidia). Morphology and phylogeny together support the reinstatement of P. gravidum Morgan 1896 with a neotype here designated, distinct from P. pusillum, here redefined. [ABSTRACT FROM AUTHOR]

About a thousand described species of Myxomycetes, colloquially named slime moulds, are present in most terrestrial habitats. Molecular environmental sampling has revealed that a significant proportion of sequences could not be assigned to known species, with some specific groups showing a high hidden diversity. One such group included Stemonitis and allied genera, with only a few closely related reference sequences from known species and many distantly related sequences from environmental sampling. To improve the phylogeny and help define genera in this lineage, we provided nine new small-subunit ribosomal RNA gene sequences of Stemonitis and allied genera, along with microscopy observations. We showed that the genus Stemonitis is polyphyletic, clearly separated into two groups, with the main distinguishing feature being spore ornamentation. The first group included the type species Stemonitis fusca, and was characterized by reticulated spores, while the second contained species with warted spores (e.g. S. flavogenita and S. axifera) and many undetermined environmental sequences. In addition, we show that the related genera Stemonitopsis, Stemonaria and Symphytocarpus were polyphyletic and that their circumscription was based on inconsistent characteristics. Instead, representatives of these genera clustered into the two clades according to their spore ornamentation. Our study provides a first insight into the genetic diversity of Stemonitis and allied genera, and proposes spore ornamentation as an evolutionary informative feature to distinguish genera in this group. [ABSTRACT FROM AUTHOR]

We describe a new species of the genus Lycodon Boie based on an adult male specimen from Khammouane Province, central Laos. Lycodon banksi sp. nov. is distinguished from its congeners by a combination of the following characters: (1) dorsal scales in 17-17-15 rows, dorsal scales on the anterior 2/3 of the body length smooth, the six central dorsal scale rows of the posterior 1/3 of the body length feebly keeled; (2) supralabials 8; (3) infralabials 10; (4) loreal in contact with the eye; (5) cloacal single; (6) ventral scales 241; (7) dorsal surface of body with 87 greyish yellow blotches; (8) ventral surface of body and tail uniformly grey cream. Based on the molecular comparison, Lycodon banksi sp. nov. is placed in a clade with other species previously considered to be members of the genus Dinodon. The new species is at least 9% genetically divergent from other species within this clade as shown by a fragment of the mitochondrial cytochrome b. This discovery increases the number of Lycodon species known from Laos to eleven. [ABSTRACT FROM AUTHOR]

The genus Penardeugenia DEFLANDRE 1958 accommodates a single species which was described as Pamphagus bathybioticus PENARD 1904. Although this species seems to be well characterized in its description, it has never been recorded again. Despite its possession of silica scales, it was assigned to the scale-lacking Chlamydophryidae (Thecofilosea, Cercozoa). We here present morphological data of three isolates. One of them was successfully cultured and enabled the amplification of its SSU rDNA sequence. Molecular analyses revealed the evolutionary origin of Penardeugenia in the scale-bearing flagellate class Thaumatomonadida, which is backed up by morphological similarities of their scales. We consider Penardeugenia to represent a closely related genus to Thaumatomastix, which apparently lost its ability to form flagella. We describe two new species from Germany, P. huelsmannii and P. hausmannii. [ABSTRACT FROM AUTHOR]

Summary -- Three new species of the genus Sectonema collected from natural habitats in Vietnam are studied, described and illustrated, including line drawings, LM and/or SEM pictures. Sectonema birrucephalum sp. n. is characterised by its 2.73-4.35 mm long body, lip region 18-20 µm broad and offset by deep constriction, odontostyle 10.0-11.5 µm long on its ventral side, 659-989 µm long neck, pharyngeal expansion occupying 63-68% of total neck length, uterus a simple tube, 221-277 µm long, pars refringens vaginae present, V = 54-56, tail short (31-43 µm, c = 85-111, c' = 0.6-0.8) and rounded, spicules 72-75 µm long, and four or five irregularly spaced ventromedian supplements beyond the range of the spicules. Sectonema buccociliatum sp. n. is distinguished by its 2.00-2.46 mm long body, lip region offset by constriction, 19-20 µm broad and bearing perioral cilia-like structures, odontostyle 13-14 µm long at its ventral side, 530-625 µm long neck, pharyngeal expansion occupying 62-69% of total neck length, uterus a simple tube, 116-152 µm long, pars refringens vaginae present, V = 56-62, tail short (23-31 µm, c = 72-104, c = 0.6-0.8) and rounded, spicules 56-68 µm long, and 3-5 spaced and weakly developed ventromedian supplements beyond the range of the spicules. Sectonema ciliatum sp. n. is characterised by its 2.79-3.13 mm long body, lip region offset by constriction, 21-22 µm broad and bearing perioral cilia-like structures, odontostyle 14-15 µm long at its ventral side, 699-722 µm long neck, pharyngeal expansion occupying 60% of total neck length, uterus a simple tube, 201-244 µm long, pars refringens vaginae present, V = 52-53, tail short (33-35 µm, c = 82-92, c' = 0.6-0.7) and rounded, spicules 70-72 µm long, and three or four spaced and weakly developed ventromedian supplements beyond the range of the spicules. Molecular data obtained for S. ciliatum sp. n. and the derived evolutionary tree show a close phylogenetic relationship with other species of the genus. [ABSTRACT FROM AUTHOR]

Testate amoebae have been frequently studied by protistologists, but still little information is available on some groups like the Chlamydophryidae. These amoebae are difficult to culture and therefore quantitative information on their morphology, phylogeny and ecology is scarce. We isolated and cultured a small testate amoeba from an agricultural field at Müncheberg near Berlin, Germany. Morphological analyses revealed it to be a new species of the genus Lecythium . We describe Lecythium terrestris sp. nov. and present its morphology, mycophagous and algivorous feeding habits and its ability to form cell aggregates by fusion. Using small-subunit ribosomal RNA gene phylogeny, we could confirm the phylogenetic position of the genus Lecythium among the Cercozoa where it groups closely to Pseudodifflugiidae (Tectofilosida). [ABSTRACT FROM AUTHOR]

Amoebae of the genus Cochliopodium are characterized by a tectum that is a layer of scales covering the dorsal surface of the cell. A combination of scale structure, morphological features and, nowadays, molecular information allows species discrimination. Here we describe a soil species Cochliopodium plurinucleolum n. sp. that besides strong genetic divergence from all currently described species of Cochliopodium differs morphologically by the presence of several peripheral nucleoli in the nucleus. Further, we unambiguously show that the Golgi attachment associated with a dictyosome in Cochliopodium is a cytoplasmic microtubule organizing center (MTOC). Last, we provide detailed morphological and molecular information on the sister clade of C. plurinucleolum, containing C. minus, C. minutoidum, C. pentatrifurcatum and C. megatetrastylus. These species share nearly identical sequences of both, small subunit ribosomal RNA and partial Cox1 genes, and nearly identical structure of the scales. Scales of C. pentatrifurcatum differ, however, strongly from scales of the others while sequences of C. pentatrifurcatum and C. minus are nearly identical. These discrepancies urge for future sampling efforts to disentangle species characteristics within Cochliopdium and to investigate morphological and molecular patterns that allow reliable species differentiation. [ABSTRACT FROM AUTHOR]