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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

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Marine Life science & Technology 2024年第3期6卷 442-461页

作者： Wenya song shijing Zhang Yuqing Li Honggang Ma Qiyu Li Xiaotian Luo khaled a.s.al-rasheid Hunter N.Hines Xiaoteng Lu Institute of Evolution&Marine Biodiversity Ocean University of ChinaQingdao 266003China Key Laboratory of Aquatic Biodiversity and Conservation of Chinese Academy of sciences Institute of HydrobiologyChinese Academy of SciencesWuhan 430072China Weishan special Aquaculture Base Jining 277600China Zoology Department King Saud University11451 RiyadhSaudi Arabia Harbor Branch Oceanographic Institute Florida Atlantic UniversityFort PierceFlorida 34946USA Department of Biology Shenzhen MSU-BIT UniversityShenzhen 518172China

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 *** relationships within the hypotrich family strongylidiidae have been ambiguous due to discordance between the morphological and genetic *** 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,Hetero-uroleptus weishanensis ***.,***.,is investigated along with the morphometric and molecular data from a population of strongylidium *** 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 *** results reveal that:(1)strongylidiidae is monophyletic and has a close relationship with Dorsomarginalia;(2)Heterouroleptus *** 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 *** 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.

关键词： Ciliates Cytochrome c oxidase subunit I gene Morphogenesis Phylogeny·Ribosomal RNA gene Taxonomy

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Evolutionary relationship and species separation of four morphologically similar stichotrichous ciliates (Protozoa,Ciliophora)

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Progress in Natural science:Materials International 2009年第5期19卷 581-586页

作者： John Clamp khaled a.s.al-rasheid Abdulaziz A.al-Khedhairy Department of Biology North Carolina Central University Zoology Department King Saud University

Phylogenetic and taxonomic studies on ciliate protists using molecular approaches have been demonstrated to be very reliable to form strong conclusions and results. In the present work, species separation of some morphologically similar stichotrichous ciliates, two species of Pseudokeronopsis and two species of Apokeronopsis, was reexamined using amplified ribosomal DNA restriction analysis (PCR-RFLP). Five of 10 restriction enzymes revealed species-specific polymorphic patterns, of which four similar stichotrichs could be significantly separated and identified. Among them, EcoR I offered almost no significantly different restriction fragment patterns, but the four species could be separated from one another and identified with Hae Ⅲ. Distinctly different restriction digestion haplotypes and simi-larity indices separated the species, and were used to construct a phylogeny. Phylogenies based on ITs2 nucleotide sequences and ITs2 secondary structures supported the separation of Pseudokeronopsis and Apokeronopsis using RFLP analysis, although three Pseudoker-onopsis carnea populations did not cluster together. In addition, phylogenetic analyses using multiple algorithms confirmed that these two genera formed two distinct groups within the urostylids.

关键词： Ciliophora Pseudokeronopsis Apokeronopsis PCR-RFLP ITs2 secondary structures

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Taxonomic description of a new marine ciliate,Euplotes qingdaoensis n. sp.(Ciliophora:Euplotida)

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Chinese Journal of Oceanology and Limnology 2014年第2期32卷 426-432页

作者：陈相瑞马洪钢 khaled a.s.al-rasheid school of Marine science Ningbo University Laboratory of Protozoology Institute of Evolution & Marine BiodiversityOcean University of China Zoology Department College of ScienceKing Saud University

The characteristics and infraciliature of a new euplotid ciliate, Euplotes qingdaoensis n. sp., discovered from the coastal waters of Jiaozhou Bay, northern China, were studied using live observation and silver impregnation. The new marine species is characterized by a slightly flattened body （90-110 um×70- 80 um, in vivo）, an adoral zone of 26-29 membranelles, 10 strong frontoventral and five transverse cirri, two caudal cirri, a single left marginal cirrus, seven dorsal kineties, a middle kinety with about 15 pairs of basal bodies, a silverline system of the double-eurystomus type, and a C-shaped macronucleus.

关键词： Euplotes infraciliature morphology new species taxonomy

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Phylogenetic position of three Condylostoma species (Protozoa, Ciliophora, Heterotrichea) inferred from the small subunit rRNA gene sequence

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Progress in Natural science:Materials International 2008年第9期18卷 1089-1093页

作者： khaled a.s.al-rasheid saleh A.al-Farraj saleh A.al-Qurishy Zoology Department King Saud University Riyadh 11451 Saudi Arabia

The systematically poorly known ciliate genus Condylostoma was erected by Vincent in *** 10 morphotypes have been reported, but any molecular investigations concerning this group so far are *** this work, the small subunit ribosomal RNA (ss rRNA) gene of three marine Condylostoma species was sequenced, by which the phylogenetic trees were constructed by distance-matrix, maximum parsimony and Bayesian inference *** results show that (1) all the trees have similar topologies with high supports;(2) Condylostoma is mostly related to the genus Condylostentor;and (3) three Condylostoma species as well as Condylostentor auriculatus cluster together and form a sister group with other *** is moderately consistent with the assessment of phylo-genetic relationships of Condylostoma-related heterotrichs from the morphological *** phylogenetic relationship of some other related heterotrichs, Peritromus, Folliculina, stentor and Blepharisma, has been also discussed.

关键词： Marine ciliates Condylostoma ss rRNA Phylogeny Heterotrichs

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Phylogeny of subclass scuticociliatia(Protozoa,Ciliophora) using combined data inferred from genetic,morphological,and morphogenetic evidence

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Chinese Journal of Oceanology and Limnology 2010年第4期28卷 778-784页

作者：伊珍珍王艳刚林晓凤 khaled a.s.al-rasheid 宋微波 Laboratory of Protozoology Key Laboratory of Ecology and Environmental Science in Guangdong Higher EducationSouth China Normal University Laboratory of Protozoology Ocean University of China Zoology Department King Saud University

Gene sequence-based genealogies of scuticociliates are different from those produced by morphological *** this reason,11 representative scuticociliates and two ambiguously related genera were chosen to test the ability of combined phylogenetic analyses using both gene sequences and morphological/morphogenetic *** of both the ssrRNA gene sequences and the combined datasets revealed a consistent branching *** the terminal branches and the order level relationships were generally well resolved,the family level relationships remain ***,two other trees based on ITs1-5.8s-ITs2 region sequences and morphological/morphogenetic characters showed limited information,due to a lack of informative sites in these two *** data suggest,however,that the combined analysis of morphological/morphogenetic characters and gene sequences did produce some changes to the phylogenetic estimates of this group.

关键词： Ciliophora scuticociliatia phylogenetic analyses combined data

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Insights into the phylogeny of sporadotrichid ciliates (Protozoa, Ciliophora: Hypotricha) based on genealogical analyses of multiple molecular markers

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Chinese Journal of Oceanology and Limnology 2011年第1期29卷 96-102页

作者：胡晓燕胡晓钟 khaled a.s.al-rasheid saleh A.al-FARRAJ 宋微波 Laboratory of Protozoology Institute of Evolution and Marine BiodiversityOcean University of China Department of Zoology Natural History Museum Zoology Department King Saud University

The sporadotrichid ciliates are an especially diverse group. A number of investigators have studied the morphological, morphogenetic, and molecular relationships among members of this group. Despite this, a consistent classification is still lacking and several important questions about the phylogenetic relationships within this group remain unsolved. To improve our understanding of these relationships, we constructed phylogenetic trees using the nucleotide sequences of the small-subunit rRNA (ssrRNA) gene and amino acid sequences of actin I and α-tubulin. Analyses of ssrRNA gene sequences indicated that: 1) the sporadotrichida sensu Lynn (2008) and the Oxytrichidae are polyphyletic; 2) the Uroleptus species, which are classified to urostylids, formed a sister group with the oxytrichids; 3) Halteria grandinella, which is grouped morphologically with oligotrich species, clustered within the oxytrichids. These results are congruent with previous studies based on ssrRNA gene sequences. However, the amino acid sequences of actin I and α-tubulin yielded different topologies. The main results are: 1) in all phylogenetic trees, the genus Oxytricha was paraphyletic; 2) Uroleptus was sister to a subset of Urostyla and Holosticha, albeit with low supporting values; 3) Halteria grandinella was separated distantly from the Oxytrichidae in trees inferred from actin I amino acid sequences but clustered with oligotrichids in the α-tubulin analysis. The inconsistency among the trees inferred from these different molecular markers may be caused by rapidly accumulated genetic characterizations of ciliates. Further studies with additional molecular markers and sampling of more taxa are expected to better address the relationships among sporadotrichids.

关键词： sporadotrichid ciliates phylogeny ssrRNA actin I alpha-tubulin amino acid sequences

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Molecular Phylogenetic Lineage of Plagiopogon and Askenasia(Protozoa, Ciliophora) Revealed by Their Gene sequences

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Journal of Ocean University of China 2015年第4期14卷 724-730页

作者： LIU An YI Zhenzhen LIN Xiaofeng HU Xiaozhong saleh A.al-FARRAJ khaled a.s.al-rasheid Laboratory of Protozoology Institute of Evolution & Marine Biodiversity Ocean University of China Laboratory of Protozoology School of Life Science South China Normal University Zoology Department College of Science King Saud University

Prostomates and haptorians are two basal groups of ciliates with limited morphological characteristics available for tax- onomy. Morphologically, the structures used to identify prostomates and haptorians are similar or even identical, which generate heavy taxonomic and phylogenetic confusion. In present work, phylogenetic positions lineage of two rare genera, Plagiopogon and Askenasia, were investigated. Three genes including small subunit ribosomal RNA gene （hereafter ssU rDNA）, internal transcribed spacer region （ITs region）, and large subunit ribosomal RNA gene （LsU rDNA） were analyzed, 10 new sequences five species each. Our findings included 1） class Prostomatea and order Haptorida are multiphyletic; 2） it may not be appropriate to place order Cyclotriehiida in subclass Haptoria, and the systematic lineage of order Cyclotrichiida needs to be verified further; 3） genus Plagio- pogon branches consistently within a clade covering most prostomes and is basal of clade Colepidae, implying its close lineage to Prostomatea; and 4） Askenasia is phylogenetically distant from the subclass Haptoria but close to classes Prostomatea, Plagiopylea and Oligohymenophorea. We supposed that the toxicyst of Askenasia may be close to taxa of prostomes instead of haptorians, and the dorsal brush is a more typical morphological characteristics of haptorians than toxicysts.

关键词： Plagiopogon Askenasia multi-gene phylogeny ssU rDNA ITs region LsU rDNA

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Reconsideration of the phylogenetic positions of three stichotrichous genera Holosticha,Anteholosticha and Pseudokeronopsis (spirotrichea:Ciliophora) inferred from complete ssU rRNA gene sequences

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Progress in Natural science:Materials International 2009年第6期19卷 769-773页

作者： Mann Kyoon shin alan Warren khaled a.s.al-rasheid Abdulaziz A.al-Khedhairy saud al-Arifi Department of Biology University of Ulsan Department of Zoology The Natural History Museum 3. Zoology DepartmentKing Saud University

The small subunit rRNA (ssU rRNA) gene for six marine stichotrichs, Pseudokeronopsis carnea, P. flava, Holosticha heterofoissneri, H. diademata, H. bradburyae and Anteholosticha manca, was sequenced and characterized. Using this molecular information, the phylogenetic positions of three related genera, Pseudokeronopsis, Holosticha and Anteholosticha were determined. The results indicate that the family Urostylidae is clearly separated from the family Pseudokeronopsidae. The present phylogenetic analyses unambiguously placed Pseudokeronopsis close to Holosticha, and support the conclusion that these genera should be regarded as the members of the order Urostylidae within the stichotrich clade. Furthermore, the results of this study also support the monophyly of the genus Pseud-okeronopsis, the redefinition of Holosticha s. str. and the placement of Anteholosticha in a clade separate from Holosticha.

关键词： Urostylidae Clade Evolution Taxonomy

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On four epibiotic peritrichous ciliates(Protozoa,Ciliophora)found in Lake Weishan Wetland:morphological and molecular data support the establishment of a new genus,Parapiosoma ***.,and two new species

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Marine Life science & Technology 2023年第3期5卷 337-358页

作者： Tong Wu Ting Cheng Xiao Cao Yaohan Jiang khaled a.s.al-rasheid alan Warren Zhe Wang Borong Lu Marine College Shandong UniversityWeihai 264209China Institute of Evolution and Marine Biodiversity Ocean University of ChinaQingdao 266003China Zoology Department College of ScienceKing Saud UniversityRiyadh 11451Saudi Arabia Department of Life sciences Natural History MuseumLondon SW75BDUK Weishan Fishery Development service Center Jining 277600China

During a study on the diversity of ciliated protists in Lake Weishan Wetland,the largest wetland in northern China,four epibiotic sessilid peritrichs were isolated from aquatic host *** of them,i.e.,Epistylis cambari Kellicott,1885 and Epistylis lwof Fauré-Fremiet,1943,were known species whereas the other two,i.e.,Parapiosoma typicum ***.,*** Orborhabdostyla gracilis ***.,are new to *** new genus Parapiosoma *** characterized by its branched non-contractile stalk,everted peristomial lip,obconical macronucleus and transverse *** species are assigned to the new genus,namely Parapiosoma typicum *** Parapiosoma gasterostei(Fauré-Fremiet,1905)***,*** *** recognized by its goblet-shaped zooids,single-layered peristomial lip,dichotomously branched stalk,and infundibular polykinety 3(P3)containing three equal-length *** gracilis *** characterized by its slender zooid,curved macronucleus,and three equal-length rows in infundibular *** diagnoses and redescriptions of *** and *** are provided including,for the frst time,data on the ciliature of *** analyses based on ssU rDNA,ITs1-5.8s rDNA-ITs2,and LsU rDNA sequence data strongly support the assertion that the family Epistylididae comprises morphospecies with diferent evolutionary lineages and indicate that Parapiosoma *** represent a new taxon at family level.

关键词： Ectoparasite Parapiosoma gen.nov. sessilid peritrichs systematics

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New considerations on the phylogeny of sessilida(Protista:Ciliophora:Peritrichia)based on multiple-gene information,with emphasis on colonial taxa

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Marine Life science & Technology 2022年第4期4卷 569-583页

作者： Zhe Wang Xiaoting Feng Lifang Li khaled a.s.al-rasheid alan Warren Chundi Wang Zemao Gu Marine College Shandong UniversityWeihai264209China College of Fisheries/shuangshui shuanglüInstitute Huazhong Agricultural UniversityWuhan430070China Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention Wuhan430070China Department of Zoology College of ScienceKing Saud UniversityRiyadh11451Saudi Arabia Department of Life sciences Natural History MuseumLondonSW75BDUK

The subclass Peritrichia,containing two orders sessilida and Mobilida,is a major group of ciliates with worldwide distribution and high species *** studies have investigated the phylogeny of peritrichs;however,the evolutionary relationships and classification of some families and genera within the sessilida remain *** the present study,we isolated and identified 22 peritrich populations representing four families and six genera and obtained 64 rDNA sequences to perform phylogenetic analyses and assess their systematic *** character reconstruction was also carried out to infer evolutionary routes within the *** results indicate:(1)family Vaginicolidae is monophyletic and acquisition of the typical peritrich lorica represents a single evolutionary divergence;(2)core epistylidids evolved from a Zoothamnium-like ancestor and experienced spasmoneme loss during evolution;(3)Campanella clusters with species in the basal clade and shows stable morphological differences with other epistylidids,supporting its assignment to a separate family;(4)the structure of the peristomial lip may be a genus-level character rather than a diagnostic character for discriminating Epistylididae and Operculariidae,thus a redefinition of Operculariidae should be carried out when more species have been investigated;(5)some characters,such as lifestyle(solitary or colonial),spasmoneme and living habit(sessile or free-swimming),evolved repeatedly among sessilids indicating that species with non-contractile stalks or that are free-swimming have multiple evolutionary routes and might derive from any sessilid lineage without a *** close phylogenetic relationships of some morphologically distinct sessilids imply that the diagnoses of some genera and families should be improved.

关键词： Ancestral state reconstruction Ciliates Morphology Peritrichs Phylogenetic analyses

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