Multi-gene-based investigation on the molecular phylogeny of the hypotrichous family Strongylidiidae (Protista, Ciliophora), with notes on the ontogeny of a new genus and new species.

Ciliates in the subclass Hypotrichia have long been difficult to classify as they are one of the most polymorphic and highly differentiated groups, leading to their systematics remaining unresolved. Phylogenetic relationships within the hypotrich family Strongylidiidae have been ambiguous due to discordance between the morphological and genetic data. In this study, a new strongylidiid genus Heterouroleptus is established, mainly based on the novel mode of origin of the ventral cirral rows: left ventral cirral row (LVR) originates from frontal-ventral-transverse cirral anlagen (FVTA) III (anterior portion), IV (middle portion), and V (rear portion); right ventral cirral row comes from the entire FVTA VI. A new species, Heterouroleptus weishanensis gen. nov., sp. nov., is investigated along with the morphometric and molecular data from a population of Strongylidium wuhanense. Eight new sequences and nuclear gene markers (single-gene and multi-gene) are provided to analyze the phylogenetic relationships of strongylidiids, with the COI gene utilized to uncover further genetic information at species level and below. The results reveal that: (1) Strongylidiidae is monophyletic and has a close relationship with Dorsomarginalia; (2) Heterouroleptus gen. nov. forms a clade that is sister to all the other strongylidiids; (3) Hemiamphisiella Foissner, 1988 and Pseudouroleptus Hemberger, 1985 should not be synonyms, and both genera should be subdivided due to their variable morphological characteristics; (4) LVR originating from three anlagen is a plesiomorphy of Strongylidiidae. The discovery of the origin of the LVR not only contributes to the establishment of the genus Heterouroleptus, but also helps to improve the diagnosis of the family Strongylidiidae. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-024-00243-z.

Interleukin-33 improves the neurogenesis of neural stem cells in perinatal brain after hypoxia-ischemia.

Perinatal hypoxia-ischemia (HI) insult is an important cause of neonatal encephalopathy, and the effective therapeutic approaches are currently limited. Interleukin (IL)-33 acts as a member of the IL-1 superfamily and has been shown to be neuroprotective following experimental neonatal HI and adult stroke. Here, we explore the effect of IL-33 and its specific receptor ST2 axis on endogenous neurogenesis in neonatal brain after HI. ST2 was found on the surface of NSCs, and the expression of ST2 was further enhanced after HI challenge. Delivery of IL-33 obviously repopulated the size of NSC pool, whereas ST2 deficiency worsened the neurogenesis of NSCs in neonatal brain post HI insult. Further in vivo and in vitro studies showed IL-33 regulates the survival, proliferation and differentiation of NSCs through ST2 signaling pathways. Intriguingly, IL-33 facilitated translocation of Nrf2 from the cytoplasm to the nucleus, which is involved in neural differentiation of NSCs. These data demonstrate a critical role of IL-33/ST2 axis in regulation of endogenous neurogenesis of NSCs via activation of the Nrf2 signaling, which provide a new insight into the effect of IL-33 in neonatal brain following HI injury.

Field recommended concentrations of pyraclostrobin exposure disturb the development and immune response of worker bees (Apis mellifera L.) larvae and pupae.

The strobilurin fungicide pyraclostrobin is widely used to prevent and control the fungal diseases of various nectar and pollen plants. Honeybees also directly or indirectly contact this fungicide with a long-term exposure period. However, the effects of pyraclostrobin on the development and physiology of Apis mellifera larvae and pupae during continuous exposure have been rarely known. To investigate the effects of field-realistic concentrations of pyraclostrobin on honeybee survival and development, the 2-day-old larvae were continuously fed with different pyraclostrobin solutions (100 mg/L and 83.3 mg/L), and the expression of development-, nutrient-, and immune-related genes in larvae and pupae were examined. The results showed that two field-realistic concentrations of pyraclostrobin (100 and 83.3 mg/L) significantly decreased the survival and capped rate of larvae, the weight of pupae and newly emerged adults, and such decrease was a positive correlation to the treatment concentrations. qPCR results showed that pyraclostrobin could induce the expression of Usp, ILP2, Vg, Defensin1, and Hymenoptaecin, decrease the expression of Hex100, Apidaecin, and Abaecin in larvae, could increase the expression of Ecr, Usp, Hex70b, Vg, Apidaecin, and Hymenoptaecin, and decreased the expression of ILP1, Hex100 and Defensin1in pupae. These results reflect pyraclostrobin could decrease nutrient metabolism, immune competence and seriously affect the development of honeybees. It should be used cautiously in agricultural practices, especially in the process of bee pollination.

Ontogenesis and systematic position of a new hypotrichous ciliate, Chaetospira sinica sp. nov., with an improved diagnosis of the poorly defined family Chaetospiridae Jankowski, 1985 (Protozoa, Ciliophora, Hypotrichia).

Ciliates are unique single-celled organisms that play important roles in ecological, environmental, evolutionary, and ontogenetic research. In the present study, phylogenetic analyses based on 18S rRNA gene sequence data reveal that Chaetospira sinica sp. nov. clusters with Stichotricha aculeata with strong to full support (97% ML, 1.00 BI), but is not closely related to members of Spirofilidae Gelei, 1929 to which Chaetospira and Stichotricha have previously been assigned. Phylogenetic analyses, together with morphological and morphogenetic data from Chaetospira sinica sp. nov., support the validity of family Chaetospiridae Jankowski, 1985. Chaetospira and Stichotricha are here assigned to the family Chaetospiridae, the improved diagnosis of which is as follows: non-dorsomarginalian Hypotrichia with flask-shaped body; oral region extending along narrow anterior neck region; lorica usually present; two ventral and two marginal cirral rows, all distinctly spiraled or obliquely curved; pretransverse and transverse cirri absent. The basic morphogenetic features in C. sinica sp. nov. can be summarized as: (1) the oral primordium for the opisthe develops de novo and the parental adoral zone is completely retained by the proter; (2) all ventral cirral anlagen and marginal anlagen developed intrakinetally; (3) three dorsal kineties anlagen formed intrakinetally in each daughter cell; and (4) macronuclear nodules fuse into a single mass. Exconjugant cells were also isolated and their morphologic and molecular data are provided.

Hypotrichidium tisiae (Gelei, 1929) Gelei, 1954: a unique hypotrichid ciliate having a highly specialized developmental pattern during binary division.

In sharp contrast to their pelagic relatives, the oligotrichs, the overwhelming majority of hypotrich ciliates inhabit the benthos. Only a few species, including those of the genus Hypotrichidium Ilowaisky, 1921, have adapted to a planktonic lifestyle. The ontogenetic mode of the highly differentiated ciliate, Hypotrichidium tisiae (Gelei, 1929) Gelei, 1954, is unknown. In this study, the interphase morphology and the ontogenetic process of this species are investigated. Accordingly, the previously unidentified ciliary pattern of Hypotrichidium is redefined. The main morphogenetic features are as follows: (1) The parental adoral zone of membranelles is inherited completely by the proter and the oral primordium of the opisthe arises in a deep pouch. (2) Five frontoventral cirral anlagen (FVA) are formed: FVA I contributes to the single frontal cirrus, FVA II-IV generate three frontoventral cirral rows, FVA V migrates and forms postoral ventral cirri. (3) All marginal cirral row anlagen develop de novo: each of the two left anlagen forms a single cirral row, while the single right anlage fragments into anterior and posterior parts. (4) Two dorsal kinety anlagen occur de novo, with the right one fragmenting to form kineties 2 and 3. (5) Two long caudal cirral rows are formed at the ends of dorsal kineties 1 and 3. On the basis of the morphogenetic features and phylogenetic analyses, the assignment of Hypotrichidium to the family Spirofilidae Gelei, 1929 within Postoralida is supported. The establishment of separate families for the slender "tubicolous" spirofilids and the highly helical spirofilids is also validated. Supplementary Information: The online version contains supplementary material available at 10.1007/s42995-022-00148-9.

Taxonomy and Molecular Phylogeny of Two New Urostylid Ciliates (Protozoa: Ciliophora) From Chinese Wetlands and Establishment of a New Genus.

Hypotrich ciliates with evolutionary novelties are continually being discovered, challenging the current taxonomic system and attracting increased attention. In the present work, two new urostylid ciliates, Heterobakuella bergeri gen. nov., sp. nov. and Anteholosticha perezuzae sp. nov., isolated from Chinese wetland samples, were identified based on morphology and 18S rRNA gene sequences. Heterobakuella gen. nov. is defined by three frontal cirri, single buccal cirrus, one parabuccal cirrus, midventral complex composed of cirral pairs and one cirral row, one left and two right marginal cirral rows, transverse and pretransverse cirri present, caudal and frontoterminal cirri absent. Heterobakuella can be easily distinguished from the morphologically most similar genus, Apobakuella, mainly by the single buccal cirrus (vs. one buccal cirral row) and one parabuccal cirrus (vs. several parabuccal cirral rows originated from different anlagen). Phylogenetic analyses show that H. bergeri branches within the clade formed by Bergeriella ovata, Monocoronella carnea, Anteholosticha gracilis, and Neourostylopsis spp., rather than the clade represented by Apobakuella. The other species, A. perezuzae, is mainly characterized by a distinctly slender body shape with an average length:width ratio about 7, distinctively shaped biconcave and greenish cortical granules, as well as one or two pretransverse cirri. Phylogenetic analyses indicate the genus Anteholosticha is non-monophyletic.

New contributions to the taxonomy of urostylid ciliates (Ciliophora, Hypotrichia), with establishment of a new genus and new species.

Hypotrichia, one of the most complex and highly differentiated groups in Ciliophora, has been the object of extensive studies, especially in recent years. Nevertheless, methodological difficulties and insufficient faunistic studies have limited our understanding of their biodiversity and phylogeny. In this study, one novel urostylid ciliate, Pseudoholosticha zhaoi nov. gen., nov. spec. and two populations of Anteholosticha monilata (Kahl, 1928) Berger, 2003, type species of the latter genus, are studied using an integrative approach (live observation, protargol impregnation, scanning electron microscopy, and phylogenetic analysis) to provide further insights into the diversity, classification, and phylogeny of this group of ciliates. Pseudoholosticha nov. gen. can be separated from other morphologically similar genera mainly by the absence of buccal and caudal cirri. A key to 12 morphologically similar genera and illustrations of their cirral patterns are provided. The validation of the new genus and new species is supported by both morphological and phylogenetic analyses. The first 18S rRNA gene sequence of A. monilata, with detailed morphological data, provided a reliable clarification of A. monilata populations and corroborated the phylogenetic position of the type species of the polyphyletic genus Anteholosticha.

Reevaluation of the 'well-known' Paraurostyla weissei complex, with notes on the ontogenesis of a new Paraurostyla species (Ciliophora, Hypotrichia).

Two hypotrichous ciliates, Paraurostyla wuhanensis nov. spec. from Wuhan (China) and a new North American population of the Paraurostyla weissei complex, were studied based on live observations and protargol impregnation. Paraurostyla wuhanensis nov. spec. differs from the congeners by the combined features of having six or seven frontal cirri, 2-4 frontoventral cirri, 5-7 ventral rows, and yellow-greenish cortical granules. Ontogenesis proceeds as in the type species, except that fewer frontoventral cirri are formed in the new species. The morphology of the new population of the Paraurostyla weissei complex corresponds well with other American populations. In the phylogenetic trees based on the 18S rRNA gene, Paraurostyla sequences nest in a large clade together with Apoamphisiella and Notohymena. Monophyly of Paraurostyla is rejected by the results of the approximately unbiased test analyses. Morphological, morphogenetic, and phylogenetic analyses show a close relationship between the North American populations of the Paraurostyla weissei complex and Apoamphisiella, indicating that the taxonomic position of the former needs to be reassigned. A new combination, viz. Apoamphisiella polymicronucleata (Merriman, 1937) comb. nov., a reevaluation of the Paraurostyla weissei complex, and emended diagnoses of Paraurostyla and Apoamphisiella, are provided.

Transcriptome-wide study of the response of human trabecular meshwork cells to the substrate stiffness increase.

Elevated intraocular pressure, a major risk factor of glaucoma, is caused by the abnormal function of trabecular outflow pathways. Human trabecular meshwork (HTM) tissue plays an important role in the outflow pathways. However, the molecular mechanisms that how TM cells respond to the elevated IOP are largely unknown. We cultured primary HTM cells on polyacrylamide gels with tunable stiffness corresponding to Young's moduli ranging from 1.1 to 50 kPa. Then next-generation RNA sequencing (RNA-seq) was performed to obtain the transcriptomic profiles of HTM cells. Bioinformatics analysis revealed that genes related to glaucoma including DCN, SPARC, and CTGF, were significantly increased with elevated substrate stiffness, as well as the global alteration of HTM transcriptome. Extracellular matrix (ECM)-related genes were selectively activated in response to the elevated substrate stiffness, consistent with the known molecular alteration in glaucoma. Human normal and glaucomatous TM tissues were also obtained to perform RNA-seq experiments and supported the substrate stiffness-altered transcriptome profiles from the in vitro cell model. The current study profiled the transcriptomic changes in human TM cells upon increasing substrate stiffness. Global change of ECM-related genes indicates that the in vitro substrate stiffness could greatly affect the biological processes of HTM cells. The in vitro HTM cell model could efficiently capture the main pathogenetic process in glaucoma patients, and provide a powerful method to investigate the underlying molecular mechanisms.

Taxonomy, Ontogenesis and Evolutionary Relationships of the Algae-Bearing Ciliate Bourlandella viridis () comb. nov., With Establishment of a New Genus and New Family (Protista, Ciliophora, Hypotrichia).

Hypotrichs are the most complex and highly differentiated ciliate lineages and have great potential for evolutionary novelties. Problems in hypotrich systematics are mainly due to discordance between the morphological and genetic data (mainly small subunit rRNA gene). Species with morphologies that are characteristic of two or more higher rank taxa are probably a major contributing factor to these conflicts. The present study describes a Chinese population of a poorly known organism with numerous endosymbiotic zoochlorellae, the morphology of which corresponds well with the type population of Limnoholosticha viridis (Kahl, 1932) Li et al., 2017. Newly obtained information shows this species has a zigzag midventral cirral pattern that is diagnostic of urostylids, whereas the dorsal ciliature shares features (presence of dorsomarginal kinety and dorsal kinety 3 fragmentation) that are typical of oxytrichids. Molecular phylogenetic analyses reveal a close relationship with oxytrichids. An integrative approach combining morphological, morphogenetic, phylogenetic and ecological analyses indicates that L. viridis represents a new genus and new family which might be an intermediate form between uorstylids and dorsomarginalians. Thus, Bourlandellidae fam. nov. and Bourlandella gen. nov. are here established. Lastly, we speculate that phenotypic convergence and mixtrophy might confer on the new combination, Bourlandella viridis (Kahl, 1932) comb. nov., the ability to adapt to a wide range of environmental conditions.

Morphology, Morphogenesis, and Phylogeny of Urosoma caudata (Ehrenberg, 1833) Berger, 1999 (Ciliophora, Hypotrichia) based on a Chinese Population.

We report the morphology and morphogenesis of Urosoma caudata (Ehrenberg, 1833) Berger, 1999 based on in vivo observation and protargol impregnation and provide an improved diagnosis of U. caudata based on previous and current work. Urosoma caudata differs from its congeners mainly by the combination of the following features: tail-like posterior end, colorless cortical granules, and two macronuclear nodules. Urosoma caudata shares most of the ontogenetic features with its congeners: the oral primordium of the opisthe develops apokinetally, and the frontal-ventral-transverse cirral anlagen develop in five streaks. However, a unique morphogenetic characteristic is recognizable: the anlagen of three dorsal kineties occur de novo to the left of the parental structures differing from their intrakinetal origin in other Urosoma species. The first record of the 18S rRNA gene sequence for the species is also provided. Phylogenetic analyses based on 18S rRNA gene sequence data suggest that the genus Urosoma is a nonmonophyletic group.

Morphology and phylogeny of four marine or brackish water spirotrich ciliates (Protozoa, Ciliophora) from China, with descriptions of two new species.

In this study, we investigated the morphology and molecular phylogeny of four marine or brackish spirotrichean ciliates found in China, namely: Caryotricha sinica sp. nov., Prodiscocephalus orientalis sp. nov., P. cf. borrori, and Certesia quadrinucleata. Caryotricha sinica is characterized by its small size, seven cirral rows extending posteriorly to about 65% of the cell length, and four transverse cirri. Prodiscocephalus orientalis differs from its congeners mainly by the number of cirri in the "head" region and on the ventral side. The SSU rDNA sequence of P. cf. borrori differs from that of other population of P. borrori by ca. 40 bp. Consequently, the nominal species P. borrori is considered to be a species-complex. New data are provided for Certesia quadrinucleata. The Chinese population of C. quadrinucleata, for example, has fewer left marginal cirri than the other populations for which such data are available. Phylogenetic analyses based on SSU rDNA sequence data show that the genus Caryotricha is monophyletic. All typical "discocephalids" with a discoid "head" form a strongly supported clade that is sister to the unstable uronychiids + pseudoamphisiellids clade within the Euplotia. The genus Certesia forms a sister group to the Euplotes clade, also within the Euplotia assemblage.

circRNAs Are Involved in the Rice-Magnaporthe oryzae Interaction.

Circular RNAs (circRNAs) play roles in various biological processes, but their functions in the rice (Oryza sativa) response to Magnaporthe oryzae remain elusive. Here, we demonstrate that circRNAs are involved in the rice-M. oryzae interaction using comparative circRNA-sequencing and transgenic approaches. We identified 2932 high-confidence circRNAs from young leaves of the blast-resistant accession International Rice Blast Line Pyricularia-Kanto51-m-Tsuyuake (IR25) and the blast-susceptible accession Lijiangxin Tuan Heigu (LTH) under M oryzae-infected or uninfected conditions; 636 were detected specifically upon M oryzae infection. The circRNAs in IR25 were significantly more diverse than those in LTH, especially under M. oryzae infection. Particularly, the number of circRNAs generated per parent gene was much higher in IR25 than in LTH and increased in IR25 but decreased in LTH upon M. oryzae infection. The higher diversity of circRNAs in IR25 was further associated with more frequent 3' and 5' alternative back-splicing and usage of complex splice sites. Moreover, a subset of circRNAs was differentially responsive to M oryzae in IR25 and LTH. We further confirmed that circR5g05160 promotes rice immunity against M oryzae Therefore, our data indicate that circRNA diversity is associated with different responses to M oryzae infection in rice and provide a starting point to investigate a new layer of regulation in the rice-M oryzae interaction.

Phylogeny of the ciliate family Psilotrichidae (Protista, Ciliophora), a curious and poorly-known taxon, with notes on two algae-bearing psilotrichids from Guam, USA.

BACKGROUND: The classification of the family Psilotrichidae, a curious group of ciliated protists with unique morphological and ontogenetic features, is ambiguous and poorly understood particularly due to the lack of molecular data. Hence, the systematic relationship between this group and other taxa in the subclass Hypotrichia remains unresolved. In this paper the morphology and phylogenetics of species from two genera of Psilotrichida are studied to shed new light on the phylogeny and species diversity of this group of ciliates. RESULTS: The 18S rRNA gene sequences of species from two psilotrichid genera were obtained. In the phylogenetic trees, the available psilotrichid sequences are placed in a highly supported clade, justifying the establishment of the family Psilotrichidae. The morphology of two little-known species, packed with green algae, including a new species, Hemiholosticha kahli nov. spec., and Psilotrichides hawaiiensis Heber et al., 2018, is studied based on live observation, protargol impregnation, and scanning electron microscopy. Both species are easily recognized by their green coloration due to the intracellular algae, and a comprehensive discussion as to the possible roles of the intracellular algae is provided. CONCLUSIONS: The 18S rRNA gene phylogeny supports the morphological argument that Hemiholosticha, Psilotrichides and Urospinula belong to the same family, Psilotrichidae. However, the single-gene analysis, not surprisingly, does not resolve the deeper relationships of Psilotrichidae within the subclass Hypotrichia. Two little-known psilotrichid genera with green algae were collected from the same puddle on the island of Guam, indicating a high species diversity and broader geographic distribution of this group of ciliates than previously supposed. Phylogenetic inferences from transcriptomic and/or genomic data will likely be necessary to better define the systematic position and evolution of the family Psilotrichidae. Further studies are also needed to clarify the role of the intracellular eyespot-bearing algae in these ciliates.

hnRNPDL extensively regulates transcription and alternative splicing.

RNA binding proteins (RBPs) are key players of genome regulation. Here we report the transcriptome study of HnRNP D-Like protein, which belongs to the hnRNP family. We used RNA-seq to analyze the global transcript level and alternative splicing on hnRNPDL shRNA-treated cells and control. Sh-hnRNPDL extensively increased in the expression of genes involved in female pregnancy, cell apoptosis, cell proliferation and cell migration. HnRNPDL regulated alternative splicing of hundreds of genes enriched in transcription regulation and signaling pathways including NOD-like receptor signaling, Notch signaling, and TNF signaling. This study provides the first transcriptome-wide analysis of hnRNPDL regulation of gene expression, which adds to the understanding of critical hnRNPDL functions.

The Little-known Freshwater Metopid Ciliate, Idiometopus turbo (Dragesco and Dragesco-Kernéis, 1986) nov. gen., nov. comb., Originally Discovered in Africa, Found on the Micronesian Island of Guam.

Class Armophorea Lynn, 2004 includes two orders of mainly free-living anaerobic ciliates, Armophorida Jankowski, 1980 and Metopida Jankowski, 1980 and, a third, the exclusively endosymbiotic Clevelandellida. Kahl described the majority of free-living metopid species early in the 20th century. Excepting Jankowski in the 1960's, little further interest was shown in this group. Metopus turbo Dragesco and Dragesco-Kernéis, 1986, from a West African pond, was more recently described. Although not explicitly described as endemic, Dragesco believed in some degree of endemism of sub-Saharan ciliates. Our discovery of M. turbo on Guam, Micronesia was unexpected. Metopus turbo was identified by live observation, protargol impregnation, and scanning electron microscopy, providing reasonable evidence of conspecificity when compared with the original description. Outstanding morphologic features include the stout shape and transversely situated bandform macronucleus. The morphology of Metopus turbo differs markedly from that of the Metopus/Brachonella clade and is distant from Metopus es in phylogenetic analyses. The 18S rDNA sequence of the Guam M. turbo differs from that of M. es, type species of Metopus, by >8% and shows distinct morphologic differences from the genus Atopospira to which it is sister. These facts suggest that transfer to a new genus, Idiometopus gen. nov. is indicated.

New contribution to the species-rich genus Euplotes: Morphology, ontogeny and systematic position of two species (Ciliophora; Euplotia).

The morphology, ontogeny and phylogeny of two Euplotes species, E. estuarinus sp. nov. and a population of E. platystoma Dragesco and Dragesco-Kernéis, 1986, both collected from tropical brackish waters in south China, were investigated based on living morphology, ciliary pattern and molecular data. Euplotes estuarinus sp. nov. is small (about 60 × 40 µm in vivo), has a dargyrome of the double-eurystomus type, and the transverse cirri are arranged in two groups, with two left and three right ones. The original description of the poorly known species, E. platystoma, is brief, and the species was never investigated using live observation and molecular methods Hence, we provided a detailed redescription. Some stages of their morphogenesis were observed which proceed in the same pattern as in their congeners. The new species E. estuarinus sp. nov. clusters with E. curdsi, differing only by 1 bp in their SSU rRNA gene sequences, which is likely due to the recent speciation event and the limited resolution of the SSU rRNA gene at species level in this group as the two species are clearly morphologically distinct.

Morphological and Molecular Redefinition of Euplotes platystoma Dragesco & Dragesco-Kernéis, 1986 and Aspidisca lynceus (Müller,1773) Ehrenberg, 1859, with Reconsideration of a "Well-known" Euplotes Ciliate, Euplotes harpa Stein, 1859 (Ciliophora, Euplotida).

We documented the morphology, infraciliature, silverline system, and molecular data of two euplotid species isolated from China, including two populations of the poorly known Euplotes platystoma Dragesco & Dragesco-Kernéis, and the previously well described Aspidisca lynceus (Müller, ) Ehrenberg, 1830. Based on the information available, an improved diagnosis of Euplotes platystoma is given, including: a narrow adoral zone with 44-68 membranelles, 10 frontoventral, 5 transverse, 2 left marginal and 2 caudal cirri, 11-13 dorsal kineties with 17-25 dikinetids in the mid-dorsal row, and dorsal silverline system of the double-eurystomus type. The Chinese population of Aspidisca lynceus closely resembles previously described populations. Phylogenetic analyses inferred from SSU rDNA sequences show that E. platystoma is closely related with E. neapolitanus, and the internal position of A. lynceus within this genus is still not robust. A reconsideration of the "well-known" Euplotes harpa and a comparison of all SSU rDNA sequences of E. harpa in GenBank are provided. We speculate that the sequences available from GenBank under the name of E. harpa are very likely from misidentified materials, that is, the identity of the species currently associated with the SSU rDNA of this "well-known" form in molecular databases requires further confirmation.

A New Subspecies of Oxytricha granulifera (Hypotrichia: Oxytrichidae) from Mexico, with Notes on its Morphogenesis and Phylogenetic Position.

The genus Oxytricha Bory de Saint-Vincent in Lamouroux, Bory de Saint-Vincent and Deslongchamps, 1824 comprises about 38 species distributed worldwide and has been considered to be a nonmonophyletic group. Based on living observations, protargol preparations, and a small subunit ribosomal RNA (SSU rRNA) gene sequence, we describe a new subspecies Oxytricha granulifera chiapasensis n. subsp. This new taxon is morphologically characterized by undulating membranes basically in a Stylonychia-pattern, six dorsal kineties, size in vivo ca. 60-120 × 20-40 µm, 21-30 right and 21-31 left marginal cirri, 22-29 adoral membranelles, and spherical cortical granules arranged in longitudinal rows on the dorsal side. In terms of the SSU rRNA gene sequence, the new subspecies differs from populations of O. granulifera from GENBANK by 7-35 nucleotides. Phylogenetic analyses showed that Oxytricha granulifera gene sequences were nested into three groups, with the new subspecies included in one of them. Oxytricha granulifera chiapasensis n. subsp. is different from Oxytricha granulifera granulifera Foissner and Adam, 1983 and Oxytricha granulifera quadricirrata Blatterer and Foissner, 1988 based on: (i) undulating membranes in Stylonychia-pattern, (ii) formation of a sixth dorsal kinety during morphogenesis, (iii) the adoral membranelles number, and (iv) inhabiting freshwater habitats.

The diverse morphogenetic patterns in spirotrichs and philasterids: Researches based on five-year-projects supported by IRCN-BC and NSFC.

In the five years 2012-2016, the ciliate research group at Ocean University of China and their collaborators have performed several investigations on the morphogenesis of ciliated protists during binary division. Multiple samples were collected from 17 cities and cortical development studied in 42 species belonging to 32 genera and 13 families (Amphisiellidae, Euplotidae, Kahliellidae, Oxytrichidae, Philasteridae, Pseudokeronopsidae, Pseudourostylidae, Schmidingerotrichidae, Spirofilidae, Strobilidiidae, Uroleptidae, Uronychiidae and Urostylidae). Among these, 12 genera were investigated morphogenetically for the first time, revealing some unusual pattern formations and allowing four new genera to be established: Heterokeronopsis, Apobakuella, Parabistichella and Apoholosticha. The objective of this review is to: 1) summarize the morphogenetic studies supported by the IRCN-BC and NSFC projects during these five years; 2) summarize the patterns of development and document deviations from normal morphogenetic events within a group; 3) discuss how studies on morphogenesis have helped to advance understanding in the three dimensions of biodiversity, i.e. taxonomy, genetics and function; and 4) suggest potential future directions for the morphogenetic study of ciliated protists.