Biogeography and Character Evolution of the Ciliate Genus Euplotes (Spirotrichea, Euplotia), with Description of Euplotes curdsi sp. nov.
ABSTRACT: Ciliates comprise a diverse and ecologically important phylum of unicellular protists. One of the most specious and best-defined genera is Euplotes, which constitutes more than 70 morphospecies, many of which have never been molecularly tested. The increasing number of described Euplotes taxa emphasizes the importance for detailed characterizations of new ones, requiring standardized morphological observations, sequencing of molecular markers and careful comparison with previous literature. Here we describe Euplotes curdsi sp. nov., distinguishable by the combination of the following features: 45-65 ?m length, oval or elongated shape with both ends rounded, narrow peristome with 25-34 adoral membranelles, conspicuous paroral membrane, double-eurystomus dorsal argyrome type, 6-7 dorsolateral kineties and 10 frontoventral cirri. Three populations of the novel species have been found in brackish and marine samples in the Mediterranean and the White Sea. We provide the SSU rRNA gene sequences of these populations, and an updated phylogeny of the genus Euplotes. Using the molecular phylogenetic tree, we inferred aspects of the biogeographical history of the genus and the evolution of its most important taxonomic characters in order to provide a frame for future descriptions. Ultimately, these data reveal recurrent trends of freshwater invasion and highlight the dynamic, yet convergent, morphological evolution of Euplotes.
Project description:<h4>Background</h4>Ciliated protists, a huge assemblage of unicellular eukaryotes, are extremely diverse and play important ecological roles in most habitats where there is sufficient moisture for their survivals. Even though there is a growing recognition that these organisms are associated with many ecological or environmental processes, their biodiversity is poorly understood and many biotopes (e.g. soils in desert areas of Asia) remain largely unknown. Here we document an undescribed form found in sludge soil in a halt-desert inland of China. Investigations of its morphology, morphogenesis and molecular phylogeny indicate that it represents a new genus and new species, Parasincirra sinica n. g., n. sp.<h4>Results</h4>The new, monotypic genus Parasincirra n. g. is defined by having three frontal cirri, an amphisiellid median cirral row about the same length as the adoral zone, one short frontoventral cirral row, cirrus III/2 and transverse cirri present, buccal and caudal cirri absent, one right and one left marginal row and three dorsal kineties. The main morphogenetic features of the new taxon are: (1) frontoventral-transverse cirral anlagen II to VI are formed in a primary mode; (2) the amphisiellid median cirral row is formed by anlagen V and VI, while the frontoventral row is generated from anlage IV; (3) cirral streaks IV to VI generate one transverse cirrus each; (4) frontoventral-transverse cirral anlage II generates one or two cirri, although the posterior one (when formed) will be absorbed in late stages, that is, no buccal cirrus is formed; (5) the posterior part of the parental adoral zone of membranelles is renewed; (6) dorsal morphogenesis follows a typical Gonostomum-pattern; and (7) the macronuclear nodules fuse to form a single mass. The investigation of its molecular phylogeny inferred from Bayesian inference and Maximum likelihood analyses based on small subunit ribosomal DNA (SSU rDNA) sequence data, failed to reveal its exact systematic position, although species of related genera are generally assigned to the family Amphisiellidae Jankowski, 1979. Morphological and morphogenetic differences between the new taxon and Uroleptoides Wenzel, 1953, Parabistichella Jiang et al., 2013, and other amphisiellids clearly support the validity of Parasincirra as a new genus. The monophyly of the family Amphisiellidae is rejected by the AU test in this study.<h4>Conclusions</h4>The critical character of the family Amphisiellidae, i.e., the amphisiellid median cirral row, might result from convergent evolution in different taxa. Amphisiellidae are not monophyletic.
Project description:A hypotrichous ciliate, Paracladotricha salina n. g., n. sp., was discovered in hypersaline waters (salinity about 80‰) from Qingdao, China. Its morphology and some major ontogenetic stages were studied and the phylogenetic position was estimated using standard methods. Paracladotricha salina is characterized by a flexible, more or less slender body (size 50-120 × 20-35 ?m), a gonostomatid oral apparatus, one short and two long frontoventral rows, four macronuclear nodules, almost completely reduced dorsal kineties 1-3, and a loss of several parts of the ciliature, namely, the slightly shortened ciliary row of the adoral membranelles, the paroral, and the buccal, the postoral and pretransverse ventral, the transverse, and the caudal cirri. The ontogenesis is rather simple: anlage II of both filial products and anlage III of the opisthe originate de novo, while anlagen IV and V are formed within the parental rows. This combination of features requires the establishment of a new genus, Paracladotricha, which is, according to the morphological data, closely related to Schmidingerothrix and Cladotricha. The small-subunit rRNA gene was sequenced, indicating that P. salina is, as also demonstrated by the oral apparatus, a member of the gonostomatids. We provide a first, vague hypothesis about the phylogenetic relationships of the Gonostomatidae, Cladotrichidae, and Schmidingerotrichidae. However, since molecular data of the type species of these higher taxa are lacking, their validity and relationships remain obscure.
Project description:Cotterillia bromelicola nov. gen., nov. spec. was discovered in the tanks of the Mexican bromeliad Tillandsia heterophylla. Its morphology, ontogenesis, and 18S rDNA were studied with standard methods. Cotterillia has many cirral rows on both sides of the body. Uniquely, and thus used to diagnose the new genus Cotterillia, it has dorsal kineties originating de novo, producing neokinetal waves where the parental dorsal kineties reorganize to "combined rows", consisting of dorsal bristles anteriorly and of cirri posteriorly. Thus, up to four generations of bristles and cirri occur on the dorsal body surface. Cotterillia bromelicola has a gonostomatid body and adoral zone of membranelles, while the dense ciliature and the neokinetal waves resemble kahliellid hypotrichs. However, the de novo origin of anlage 1 and the molecular analyses show convincingly that Cotterillia belongs to the GonostomatidaeSmall and Lynn, 1985, for which an improved diagnosis is provided. Thus, neokinetal waves originated several times independently. The molecular differences between Trachelostyla, Gonostomum, and Cotterillia are small (? 5%) compared to their distinct morphologies and ontogeneses, suggesting that the 18S rDNA underestimates generic diversity. Our study emphasizes the need of combined morphological, ontogenetic, and molecular investigations to unravel the complex phylogeny and evolution of hypotrich ciliates.
Project description:An integrated approach considering both morphologic and molecular data is now required to improve biodiversity estimations and provide more robust systematics interpretations in hypotrichs, a highly differentiated group of ciliates. In present study, we document a new hypotrich species, Lamtostyla gui n. sp., collected from Chongming wetland, Shanghai, China, based on investigations using living observation, protargol staining, scanning and transmission electron microscopy, and gene sequencing. The new species is mainly recognized by having a short amphisiellid median cirral row composed of four cirri, three frontoventral cirri, three dorsal kinetids, four to eight macronuclear nodules, and small colorless cortical granules distributed as rosettes around dorsal bristles. Transmission electron microscope observation finds the associated microtubules of cirri and pharyngeal discs of L. gui are distinct from those in other hypotrichs. Morphogenesis of this species indicates that parental adoral membranelles retained intact or partial renewed is a potential feature to separate Lamtostyla granulifera-group and Lamtostyla lamottei-group. Phylogenetic analysis based on small subunit ribosomal RNA (rRNA) gene shows that this molecular marker is not useful to resolve phylogenetic relationships of the genus Lamtostyla, as well as many other hypotrichous taxa. We additionally characterize the internal transcribed spacers (ITS) region and the almost complete large subunit rRNA, which will be essential for future studies aimed at solving phylogenetic problems of Lamtostyla, or even the family Amphisiellidae. As a final remark, the critical screening of GenBank using ITS genes of our organism allows us to recognize a large amount of hypotrichous sequences have been misclassified as fungi. This observation suggests that hypotrichs could be frequently found in fungi-rich environment and overlooked by fungal specialists.
Project description:Oligotrich ciliates are common marine microplankters, but their biodiversity and evolutionary relationships have not been well-documented. Morphological descriptions and small subunit rRNA gene sequences of two new species representing two new strombidiid genera, Sinistrostrombidium cupiformum gen. nov., sp. nov. and Antestrombidium agathae gen. nov., sp. nov. are presented, and their taxonomy and molecular phylogeny are analyzed. Sinistrostrombidium gen. nov. is characterized by a sinistrally spiraled girdle kinety and a longitudinal ventral kinety. Antestrombidium gen. nov. is distinguished by tripartite somatic kineties (circular and ventral kineties plus dextrally spiraled girdle kinety). Sinistrostrombidium and Antestrombidium branched separately from one another in phylogenetic trees, clustering with different clades of strombidiids. The new genera added to the diversities of ciliary patterns and small subunit rRNA gene sequences in strombidiids leads to presentation of a new hypothesis about evolution of the 12 known strombidiid genera, based on ciliary pattern and partly supported by molecular evidence. In addition, our new morphological and molecular analyses support establishment of a new order Lynnellida ord. nov., characterized by an open adoral zone of membranelles without differentiation of anterior and ventral membranelles, for Lynnella, but we remain unable to assign the genus to a subclass with confidence.
Project description:The conjugation of Halteria grandinella was studied in protargol preparations. The isogamontic conjugants fuse partially with their ventral sides to a homopolar pair. The first maturation division generates dramatic transformations: (i) the partners obtain an interlocking arrangement; (ii) the number of bristle kineties decreases from seven to four in each partner; and (iii) the right conjugant loses its buccal membranelles, the left the whole adoral zone. The remaining collar membranelles arrange around the pair's anterior end and are shared by both partners; finally, the couple resembles a vegetative specimen in size and outline. The vegetative macronucleus fragments before pycnosis. The micronucleus performs three maturation divisions, but only one derivative each performs the second and third division. The synkaryon divides twice, producing a micronucleus, a macronucleus anlage, and two disintegrating derivatives. Scattered somatic kinetids occur during conjugation, but disappear without reorganization. An incomplete oral primordium originates in both partners. The conjugation of Halteria grandinella resembles in several respects that of hypotrich spirotrichs; however, the majority of morphological, ontogenetical, and ultrastructural features still indicates an affiliation with the oligotrich and choreotrich spirotrichs. Accordingly, the cladistic analysis still contradicts the genealogy based on the sequences of the small subunit rRNA gene.
Project description:A new species of Protocruzia, isolated from the deep-sea Pacific Ocean (>3,000-m depth) in the vicinity of the Mariana Trench, is described based on morphological and molecular data. The systematic status of the ciliate genus Protocruzia has long been highly ambiguous, and Protocruzia species have been assigned to an independent class until recently. In the present study, we described Protocruzia marianaensis sp. n. as a small (25–32 × 14–17 μm in vivo) drop-shaped ciliate, with longitudinal furrows along the ciliary rows on the right side, six adoral membranelles, eight somatic kineties, and one macronucleus comprising 7–11 nuclear globules. Phylogenetic analyses inferred from small subunit rRNA gene sequences revealed that seven Protocruzia species in the phylogenetic tree formed a fully supported clade representing an independent class. Protocruzia marianaensis sp. n. was established to be most closely related to Protocruzia adhaerens, with a sequence similarity of 96.64%, and was found to be able to survive at both atmospheric pressure and hydrostatic pressure of 320 bar, thereby indicating effective barotolerance.
Project description:BACKGROUND: There are thousands of very diverse ciliate species from which only a handful mitochondrial genomes have been studied so far. These genomes are rather similar because the ciliates analysed (Tetrahymena spp. and Paramecium aurelia) are closely related. Here we study the mitochondrial genomes of the hypotrichous ciliates Euplotes minuta and Euplotes crassus. These ciliates are only distantly related to Tetrahymena spp. and Paramecium aurelia, but more closely related to Nyctotherus ovalis, which possesses a hydrogenosomal (mitochondrial) genome. RESULTS: The linear mitochondrial genomes of the hypotrichous ciliates Euplotes minuta and Euplotes crassus were sequenced and compared with the mitochondrial genomes of several Tetrahymena species, Paramecium aurelia and the partially sequenced mitochondrial genome of the anaerobic ciliate Nyctotherus ovalis. This study reports new features such as long 5'gene extensions of several mitochondrial genes, extremely long cox1 and cox2 open reading frames and a large repeat in the middle of the linear mitochondrial genome. The repeat separates the open reading frames into two blocks, each having a single direction of transcription, from the repeat towards the ends of the chromosome. Although the Euplotes mitochondrial gene content is almost identical to that of Paramecium and Tetrahymena, the order of the genes is completely different. In contrast, the 33273 bp (excluding the repeat region) piece of the mitochondrial genome that has been sequenced in both Euplotes species exhibits no difference in gene order. Unexpectedly, many of the mitochondrial genes of E. minuta encoding ribosomal proteins possess N-terminal extensions that are similar to mitochondrial targeting signals. CONCLUSION: The mitochondrial genomes of the hypotrichous ciliates Euplotes minuta and Euplotes crassus are rather different from the previously studied genomes. Many genes are extended in size compared to mitochondrial genes from other sources.
Project description:Cilia are highly conserved organelles present in almost all types of eukaryotic cells, and defects in cilia structure and/or function are related to many human genetic disorders. Single-celled ciliated protists, which possess diverse types of cilia, are remarkable model organisms for studying cilia structures and functions. Euplotes vannus is a representative ciliate with many intriguing features; for example, it possesses extensively fragmented somatic genomes and a high frequency of + 1 programmed ribosomal frameshifting. However, the molecular mechanisms underlying these remarkable traits remain largely unknown, mainly due to the lack of efficient genetic manipulation tools. Here, we describe the first application of a morpholino-based strategy to knockdown gene expression in E. vannus. Through interfering with the function of two ciliary genes, ZMYND10 and C21ORF59, we showed that these two genes are essential for the ciliary motility and proliferation of E. vannus cells. Strikingly, both ZMYND10- and C21ORF59-knockdown cells developed shorter cilia in the ventral cirri, a special type of ciliary tuft, suggesting a novel role for these genes in the regulation of cilia length. Our data provide a new method to explore gene function in E. vannus, which may help us to understand the functions of evolutionarily conserved cilia-related genes as well as other biological processes in this intriguing model.
Project description:Using standard methods, we studied the morphology and ontogenesis of a German Leptopharynx costatus costatus. This population makes two morphs: microstomes with a size of 40 × 25 ?m, about 190 basal bodies, and 5 ?m wide oral basket; and macrostomes with a size of 55 × 40 ?m, about 264 basal bodies, and 15 ?m wide oral basket. Because the identity is threatened, this population is designated as the neotype of L. costatus costatus. Ontogenesis is complex due to the preoral kineties and the postoral complex produced by kineties 9 and 10. Stomatogenesis is mixokinetal: the opisthe membranelles 1 and 2 are formed by the oral primordium, whereas membranelle 3 is produced by the posterior portion of somatic kinety 1. The nasse kinetosomes are generated by the anterior portion of the oral primordium. Preoral kineties 1 and 3 develop de novo, while kinety 2 originates by intrakinetal proliferation of kinety 8; preoral kinety 4 is produced by the postoral complex, thus being a somatic kinety. Kinety 6 has two anterior kinetids in line with kinety 7. These observations require changes in the descriptive morphology, support the classification of Leptopharynx into the Microthoracidae, and sustain the nonmonophyly of the Nassophorea.