Morphology and Molecular Phylogeny of a New Hypotrich Ciliate, Pseudourostyla Guizhouensis Sp. Nov. from Southern China, with No

View metadata,citationandsimilarpapersatcore.ac.uk This article isThis article by protected copyright. All rightsreserved. 10.1111/jeu.12428 differences this between and version the Version of Record. the through copyediting, typesetting, pagination and proofreading process, to whichmay lead This article acceptedhas been for publication andundergone fullpeer review buthasnotbeen described based Chinese ona (Guizhou)population. ciliate, hypotrich soil another 1972 Borror, that reveals data rDNA sequence SSU on based analysis phylogenetic Molecular rows. several into differentiates margin the that indicates regeneration pre 12 consisting zone adoral kineties; dorsal seven or six rows; marginal left four elongate fro nov.,collected morphology hypotrichThe ciliate, andmolecular phylogenyof asoil ABSTRACT Telephone/FAX number:53282898776;e +86 X. Chen, Telephone/FAX number:3125079364;e +86 F. Li, Correspondence Qingdao,Academy Sciences, 266071,China of e 210042,Nanjing China d c Department ofLife Natural Sciences, History Museum, SW7 London 5BD,UK Life Scienceand Technology, Xi’an University,Jiaotong Xi’an, 710049,Chi b University, Baoding, 071002,China a Yanbo Li (Foissner, 1987) Paiva al.,2012 et from nov. sp. Morphology andM al.Li et : Review Article type (Orcid LI : 0000 ID FENGCHAODR.

The Key Laboratory of Zoological Systemat Zoological of Laboratory Key The The Key Laboratory of Biomedical Information Engineering, Ministry of Education, School of of School Education, of Ministry Engineering, Information Biomedical of Laboratory Key The eatet f aie raim aooy n Pyoey Institu Phylogeny, and Taxonomy Organism Marine of Department – Nanjing Institute of Environmental Science, State Environmental Protection Administration, Administration, Protection Environmental State Science, Environmental of Institute Nanjing - 6 rna cri oe ucl irs 13 cirrus, buccal one cirri, frontal 16 transverse ventral cirri, and five to seven transverse cirri. Morphogenesis during physiological physiological during Morphogenesis cirri. transverse seven to five and cirri, ventral transverse

AcceptedCollege of HebeiUniversity, LifeSciences, Baoding 071002,China Article ---

a elliptical body elliptical Institute ofOcInstitute , ZhaoLyu

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m southern China, were investigated. investigated. were China, southern m - that the that 4866 en hn, with China, hern olecular b , Alan Warren

eanology, Chinese Academy of Qingdao Sciences, 266071,China

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T genus wo sp. nov. clusters with the type species type the nov.with sp. clusters P eiylotl franzi Hemicycliostyla hylogeny ofa - Hypotrichous Hypotrichous C 0002 Pseudourostyla

c (Ciliophora, Hypotricha) 180 , KexinZhou -

1986 al rows of each side originate from a common anlage that that anlage common a from originate side each of rows al – N 310 × 65 × 310 - ts n Chinese a on otes 1105) - – New H mail: - 20 midventral pairs, two frontoterminal cirri, two two cirri, frontoterminal two pairs, midventral 20 ics and Application, College of Life Sciences, Hebei Sciences, Life of College Application, and ics

mail: iliates fromiliates China

d is monophyletic. is – , Fengchao Li 85 µm in vivo; vivo; in µm 85 [email protected]

[email protected] ypotrich ypotrich Fise, 97 Pia t l, 2012 al., et Paiva 1987) (Foissner, Pseudourostyla

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C plto of opulation a iliate, iliate, & Xumiao Chen

invariably The morphological characters of characters morphological The .

Pseudourostyla cristata Pseudourostyla

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two right and three or three and right two e –

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guizhouensis guizhouensis sp. nov. has an an nov.has sp.

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CORE olwn te auatrrs ntutos Te S rN ws mlfe wt te eukaryotic the with isThis article by protected copyright. All rightsreser amplified was rDNA SSU The instructions. manufacturer’s the following Genomic DNAwas extracted potential each short, w procedure DNAgenomic The DNA PCR amplification extraction, Terminology mainly is accordingto Berger (2006). contour by depicted are structures ciliary (parental) old the processes, morphogenetic during occurring changes theTo illustratelucida. camera a of aid the with ciliature. ca and apparatus nuclear the reveal to used camera digital a using observed were cells Living Morphological investigations bacterialenrich the food to grains rice squeezed with water mineral containing dishes Petri in °C) 25 (about temperature room the employing by samples non soil the from emerge and excyst to stimulated About were mm. Ciliates laboratory. 1,300 is precipitation average annual the o and temperature °C 15 average annual The China. Province, Guizhou City, Tongren (27°34′ Zhonghuashan Soil andSampling cultivation MATERIALS AND METHODS data. ri subunit small on based investigated was species novel the of phylogeny guizhouensis Tamás, 1958 to P 2012; al. et Jung 2016; Foissner 2014; pelotensis levis (Jerka Hemicycliostyla (withi intrakinetal franzi of reorganizer ( group each of row rightmost the of right the to primordium common frommarginala arise ofmarginalgroups cirri the i.e., ofrows, mode formation Stok namely genera, ACCORDING C Keyw iliature; phylogeny; Pseudourostylidae;iliature; soil ciliate; rried out with an ocular micrometre. Drawings of stained specimens were performed at 1,250 × 1,250 at performed were specimens stained of Drawings micrometre. ocular an with out rried Hemicycliostyla - es, 1891 es, flooded Petri dish method described described method dish Petri flooded

samples were collected on 19 July 2014 from the floodplain of a small river near the village of village the near river small a of floodplain the from 2014 July 19 on collected were samples Th aaah, 1973, Takahashi, ords Accepted Article- Foissner, 1987), the the 1987), Foissner, Dziadosz, 1964) Borror,1964) 1972,Dziadosz, peet ae rprs h mrhlg o two of morphology the reports paper present e

contamination

Paiva and Silva and Paiva

el of cell as , . One of the most important diagnostic features diagnostic important most the of One . sp. nov. and H. marina

conducted as previous studies (Chen et al. 2017; Luo et al. 2017; Shao et al. 2014). al. et Shao 2017; al. et Luo 2017; al. et (Chen studies previous as conducted eiylotl franzi Hemicycliostyla o egr 20) te family the (2006), Berger to

. Hemicycliostyla o date, To n , namely , of - h nw species new the o) omto. Consequently formation. row)

Pseudourostyla ( 48 DP73 ,

. muscorum P. Kahl, 1932

and transferred to a 2 a to transferred and ″N, Hemicycliostyla Hemicycliostyla franzi

eight Neto, 2006, and 2006, Neto, , agnl os develop rows marginal H.

, using using

using using REDExtractthe O 109°15′

concha lympus pce hv be asge to assigned been have species

tks 1886, Stokes, , dif

and

was isolated, washed repeatedly repeatedly washed isolated, was

, P.cristatoides 42 guizhouensis guizhouensis

eeta itreec contrast interference ferential and genesequencing

Kh, 92 Bro, 1972, Borror, 1932) (Kahl, ) (Foissner, 1987) Paiva et al., 2012 (called (called 2012 al., et Paiva 1987) (Foissner, (Entz, 1884) Kahl, 1932 Kahl, 1884) (Entz, . The protargol silver staining method of Wilbert was of(1975) method staining silverprotargol The .

aiva and Silva and aiva ″ H. sphagni by , about E, P.subtropica Foissner ( Foissner Pseudourostylidae - ml microfuge tube wi tube microfuge ml Berger 2006 Berger ,

isolated fromisolated southern China.Themolecular soilin Pseudourostyla

Pia t l (02 taserd hs pce to species this transferred (2012) al. et Paiva , sp. nov.sp.

9 m bv sa ee) Ajahi Township, Aojiazhai level), sea above m 394 - ved. Jung et al., 2012, al., et Jung Stokes, 1886 taxonomy. n h uul anr o uotld, ht is that urostylids, for manner usual the in N esrmns f h sand pcmn were specimens stained the of Measurements 2016 - - Amp Tissue (Sigma, St. PCRKit MO) Louis, Neto 2006). F 2006). Neto

Chen et al., 2014 (Berger 2006; Chen et al. et Chen 2006; (Berger 2014 al., et Chen ) whereas new structures are shaded black shaded are structures new whereas was e was . Non .

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of the type type the of .

Kumar et al. (2010) reported that, in that, reported (2010) al. et Kumar

hypotrich , P

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(Berger 2006; H. franzi H. akwk, 99 opie three comprises 1979 Jankowski, clonal cultures were established at at established were cultures clonal seudourostyla orr 1972, Borror, kg 1 . nova P.

ive species ive microscopy and photographed photographed and microscopy th minimum volume of water. of volume minimum th P.dimo ed with

genus f ol a air was soil of

from single specimensfromsingle , bosomal DNA sequences sequences DNA bosomal

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Pseudourostyla

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have been assigned been have

and Foissner,2016 Pseudourostyla Pseudourostyla Trichototaxis - re i the in dried

Gellért and Gellért P . cristata cristata P.

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This article isThis article by protected copyright. All rightsreser typically2G). Locomotion byswimming mode crawling or (26 r (Fig length body 1A, margin (Fig. posteriorly body left near vacuole contractile one Usually 2B). (Fig. magnification low at dark cell rendering vacuoles food with filled Posterior inclusions. granular with packed and colorless Cytoplasm contractile. (Fig rounded broadly ends both 3 oftowidth ratio length Description. Pseudourostyla guizhouensis RESULTS be can Identifiers) LSID totheprefixhttps://zoobank.org/. Science (Life LSIDs ZooBank The r ICZN. the the Zoological ZooBank, for in registered of system been registration have Code contains online it International acts nomenclatural the amended and work the published The of requirements the and 2012), (ICZN Nomenclature to conforms article This Nomenclatural Acts were (Shimodaira andHasegawa 2001). taxa remaining the th using among relationships and MLunconstrained groups the to topologies compared was tree constrained the of reliability The unspecified. constrained the within relationships monophyletic is Pseudourostylidae 4.0 ( Gateway Science CIPRES the on Maximum trees sampled sampling a and generations 1,000,000 for chains four 2003) (AIC). Huelsenbeck 1, and (Ronquist Criterion 3.1.2 MrBayes of with Information performed matrix Akaike a with in model resulting manually, v.2. trimmed MrModeltest were ends the as selected 2004) 71 T Phylogenetic analyses al.201 et ( °C 72 at min 7 was extension final the s; 50 min 1 for °C 72 min, 1 for °C 56 s, 30 for °C 94 at Theampli amplificationerrors. Ex (Takarapolymerase TAC ACC TTC AGG TGC TTC (5' Forward primers universal ecognize the appending by browser web standard any through viewed information associated the and esolved he SSU rDNA sequence of sequence rDNA SSU he

other spirotrich ciliates from ciliates spirotrich other – (Tamura 2007) etal. towere used visualize tree topologies. n re t dtrie t determine to order In 40) ellipsoidal macronuclear nodules dispersed throughout cell, two to five micronuclei (Fig micronuclei five to two cell, throughout dispersed nodules macronuclear ellipsoidal 40)

Accepted Article. Apodiophrys ovalis Apodiophrys e approximately unbiased approximately e 6 , loosely and loosely , - ).

likelihood (ML) analysis was carried out online out carried was analysis (ML) likelihood the outgroup taxa. Regions that could not be aligned unambiguously were removed and and removed were unambiguously aligned be not could that Regions taxa. outgroup the

Body 180 Body

were discarded as burn as discarded were . 2

1B (Nyla

C); sometimes two contractile vacuoles positioned at about 35% and 70% of of 70% and 35% about at positioned vacuoles contractile two sometimes C); –

– , 2 D, irregularly Taq 310 × 65 × 310 – dr 04 sel 2004) nder 4:1, elongate , , Diophrys scutum Diophrys B

Takara – Pseudourostyla - sp. nov. AAC CTG GTT GATGTT CTG AAC ) Clres otcl rnls ca. granules, cortical Colorless D). fication cycles were as follows: 5 min at 94 °C, followed by 30 cycles cycles 30 by followed °C, 94 min at 5 follows: as were cycles fication . the the e ttsia poaiiy f h hptei that hypothesis the of probability statistical he

– hence the new name contained herein is available under that Code. Code. that under available is herein contained name new the hence

1A, 2A), dorsoventrally flattened about 2:1 about flattened dorsoventrally 2A), 1A,

ih w cletn cnl, n etnig neiry n one and anteriorly extending one canals, collecting two with distributed throughout the cortex cortex the throughout distributed 85 µm in vivo and 175 and vivo in µm 85 , Stamatakis et al. 2008). TreeView v1.6.6 (Page 1996) and MEGAand 1996) TreeView (Page 2008). v1.6.6 al. et Stamatakis (AU) test (Shimodaira 2002) implemented in the CONSEL v 0.1 v CONSEL the in implemented 2002) (Shimodaira test (AU) - ' ( 3') constrain GenBank database GenBank Biotechnology, Dalian Biotechnology, - in prior to constructing 50% majority rule consensus trees. consensus rule majority 50% constructing to prior in

elliptical inoutline This un e a. 2016 al. et Huang ce te T + (= I + GTR the ected

, work was published in a journal work waspublished ina withan ISSN ed

guizh Paradiophrys zhangi Paradiophrys

ML analyses were generated by PAUPMLby generated were analyses v.4.0 ouensis ved.

CCT GCC AGT GCC CCT frequency of 10 of frequency The ,

using the online program Muscle 3.7 ( 3.7 Muscle program online the using

– , ,

rately fastrately on substrate. ; ) was used to minimize the possibility of possibility the minimize to used was )

300 × 50 × 300 with parallel, margins lateral less more or sp. nov. sp.

using RAxML using aein neec (BI) inference Bayesian eln t l 18) Hg fidelity High 1988). al. et Medlin

.68 + (= G + 0.5618)

, and ,

0.5 which was aligned to the sequences of sequences the to aligned was – (Fig. 632 0 generations. The first 25% of 25% first The generations. 0 - 88 µm 88 3') and Reverse (5' Reverse and 3')

m n diameter, in µm

Uronychia multicirrus Uronychia 1E, - hrces Te program The characters. was run with two with run was HPC2 on XSEDE (8.0.24) (8.0.24) XSEDE on HPC2 after after 2E, F). On average 33 33 average On F). 2E, , very flexible but not but flexible very ,

.57 a t as 0.4527) cell protargol staining, staining, protargol

portion usually portion

nlss was analysis the - ifcl to difficult TGA TCC TGATCC .

. Internal

he best best he sets of sets family Edgar Edgar

were

Gao . The Taq

1 G , ,

This article isThis article by protected copyright. All rightsreser onto and cirrus, parabuccal one usually cirri, buccal zig a in pairs midventral 12 About bicorona. impregnation, protargol after optically and curved slightly endoral and paroral mark question of number The 1). swimming. Table when axis body main 3C; about rotating and debris (Fig. on fast moderately crawling staining by observed usually Locomotion protargol were micronuclei after (three size uncertain in is micronuclei μm 5 × 10 approximately indiameter, aboutcolorless, 0.6μm irregularly. scattering 166 amoeba testate prey, the captured possibly of full specimens 3A, (Fig. protargolelongate after staining, population. (Guizhou) Chinese the of Description franzi Hemicycliostyla by AU ( tests genealogies the in (DQ019318). 4). (Fig. branches (ML on clade supported indicated algorithms both from values ha 45.2%. of content analyses. guizhouensis phylogenetic and sequences rDNA SSU the replace (Fig to posteriorly and anteriorly extend then intrakinetally, develop anlagen three cirral marginal one right and left contribute streaks 1 two (Fig. cirri last frontoterminal two the forms streak last each, the and cirrus each, cirrus pretransverse transverse one generate streaks seven to five last corona the (Fig. 1) Phy dorsal and cilia aboutlength 5µm long. S rows. marginal right two invariably and left pre two the to almost extending 13 and cirri, frontoterminal 12 two cirrus, cilia buccal Single with long. µm cirri, of pairs eight to six comprising bicorona a forming cirri Frontal posteriorly. intersect optically and curved 1 cilia with 1) (Table membranelles ve t

he proximal he anterior regeneration. siological . Adoral zone of membranelles occupying about 27% of cell length cell of 27% about occupying membranelles of zone Adoral 57 of composed specimens, fixed in length body of 38% about membranelles of zone Adoral

i o svn ih ma right seven or six

1 1 or four or osl ufc and surface dorsal iia topologies similar

Accepted ArticleI

H and , 2J). ,

2 F). One contractile vacuole left of buccal vertex (Fig. 3F). (Fig. vertex buccal of left vacuole contractile One F).

I); 2) the the 2) I);

the buccal cirrus, buccal the corona

P left and invariably two right new marginal rows (Fig. 2I); marginalnew(Fig. rows right two invariably and left

p nv ( nov. sp. The genera The -

shaped = 0.012)

portion of the parental adoral zone is resorbed and replaced by new membranelles new by replaced and resorbed is zone adoral parental the of portion , and t and , The phylogenetic trees inferre trees phylogenetic The

and the and frontal

, with 55 with , 100%, BI 1.00), and 1.00), BI 100%,

(Foissner, 1987) etal.,2012 Paiva . eBn acsin ubr X340 i 177 p og n hs G+C a has and long bp 1,727 is KX139470) number accession GenBank ;

therefore Hemicycliostyla emntn wt dra bite (i. 3E) (Fig. bristles dorsal with terminating gnl os ih neir oto o otrot agnl os extending rows marginal outermost of portion anterior with rows rginal hypothesis that hypothesis

- undulating membranes undulating Two reorganizers were Tworeorganizers midventral

rows develop from develop rows streak extending posteriorly extending – ing -

66 membranelles 66 transverse ventral cirri. Five to seven transverse cirri. Three or four four or Three cirri. transverse seven to Five cirri. ventral transverse elliptical in ou in elliptical Pharyn . ,

s III to III s snl tplg fo te L re s rsne wt support with presented is tree ML the from topology single a 2 – 1 -

5 transverse

, µm long. Undulating membranes relatively short, slightly slightly short, relatively membranes Undulating long. µm

Pseudourostyla these geal fibers distinct after protargol staining, extending extending staining, protargol after distinct fibers geal and – P.guizhouensis - 1 forms other cirri in bicorona and bicorona in cirri other forms 1 zag row extending to mid to extending row zag ix or s or ix

tline, slightly narrowed at both ends and very flexible very and ends both at narrowed slightly tline, two or three frontoterminal cirri. Seven or eight left eight or Seven cirri. frontoterminal three or two

d from the SSU rDNA sequences using ML and BI, and ML using rDNA sequences SSU the from d a three intersecting

. Body about 260 × 80 μm in vivo and 250 × 9 × 250 and vivo in μm 80 × 260 about Body

common

U , (FVT) (

found streak streak ved. Fig. 3 Fig. ndulating membranes in membranes ndulating even longitudinal dorsal kineties of almost cell cell almost of kineties dorsal longitudinal even h SU DA eune of sequence rDNA SSU The

genera

cirr , I)

(Fig. II and

. Eleven to 15 to Eleven . unit of unit Difflugia type the to sister is nov. sp.

(Fig. 3D, (Fig. – forms the second cirrus in the in cirrus second the forms al al n ny n ot f ih specimens). eight of out one only in form a monophy a form – 195 ellipsoidal

Trichototaxis 20 midventral pairs in a zig a in pairs midventral 20 1 streak I forms I streak H Pseudourostyla

– anlage on each side, which form form which side, each on anlage J , ;

(Fig. 3A, (Fig. P

2 - and K agnl os cuy large occupy rows marginal body (Fig. 3D). Two three 3D). or (Fig. body osterior I

, J , ). Pharyngeal fibers distinct fibers Pharyngeal ). (in protargol preparations) protargol (in 4) the new dorsal kineties kineties dorsal new the 4) frontal cirri arranged in a in arranged cirri frontal ); 2 );

are Oxytricha macronuclear nodules,

) letic group is rejected is group letic

the leftmost the . In these specimens: specimens: these In . F). Cortical gran Cortical F). midventral pairs, the the pairs, midventral cell

distinctly separate distinctly p. om fully a form spp. H portion portion , Pseudourostyla

J l structures old ); 3) the new the 3) ); - pattern, i.e. pattern, P.cristata -

cirr zag i anterior anterior n some some n 4 us in in us

ules ules

row – – μm 70 15 15 d , ,

This article isThis article by protected copyright. All rightsreser Indian specimens Guizhou Indian matches which length, (Guizhou) Chinese the of zone adoral habitats. terrestrial in all Furthermore, cirri. caudal of absence the and surface, cell the the characterist Indian and 2006) (Guizhou) (Guizhou) Chinese of Comparison paper.specimens present in more A Hunan T franzi Hemicycliostyla ne generate in 2012 al. et Jung righ the within appears which anlage single a from formed are in that to similar is process This parental rightmost the of dedifferentiation the marginalt rows; from originated probably which of each side, each the of splitting 1 (Fig. reorganizer longitudinal the From available. not comparison. Morphogenetic 2006; Kahl1932) 1988). Wiackowskimuscroum 2014; al. et (Chen Pseudourostyla 9 22 and nov. sp. as similarly rows, marginal remaining two The Lak the and separated guizhouensis congeners its and taxa. related with comparison Morphological Pseudourostyla guizhouensis DISCUSSION 3D, (Fig. cirri transverse eight extending almost (Fig. entire cell length 3E, to Five surface. cell of portion – he

25 – P. cristata cristata P. outermost marginal rows composed of dorsal bristles, marginal rows occupying a large of a portion marginaloccupying rows bristles, dorsal ofmarginal composedoutermost rows 3 Chinese ). 6 ), left marginal rows ( rows marginal left ), Accepted Article specimens specimens province

Furthermore, detailed description and illustration and description detailed specimens

from those species with four or more or four with species those from

by (Berger 2006; Foissner 1987; Kumar et al. 2010); 3) both the Chinese (Guizhou) and the the and (Guizhou) Chinese the both 3) 2010); al. et Kumar 1987; Foissner 2006; (Berger population w dorsal kineties w dorsal ics, i.e., the numbers of marginal rows and macronuclear nodules, macronuclear and rows marginal of numbers the i.e., ics, e Biwa populations), Biwa e

P.nova

p nv i ta i ivral has invariably it that is nov. sp. ouain of population and the numbers of numbers the

whereas , ).

hese anlagen then split and form three left and two right new marginal rows (Fig. 2I). 2I). (Fig. marginalrows new right two and left three form and split then anlagen hese (Shen et al et (Shen

Further

possess possess (39%); 2) the numbers of buccal cirri, transverse cirri, and marginal rows in the the in rows marginal and cirri, transverse cirri, buccal of numbers the 2) (39%); H P.

r peetd n al 2. Table in presented are are similar to those in the African the in those to similar are population ,

Pseudourostyla cristatoides are the numbers of frontal cirri ( cirri frontal of numbers the are J

There are, however, minor differences among these three populations: three these among differences minor however, are, There ), we speculate that the new marginal cirral rows develop from one anlage on anlage one from develop rows cirral marginal new the that speculate we ), . guizhoue P. of

its (Foissner, 1987) etal.,2012 Paiva

similarities of the cortical development in corticaldevelopmentsimilarities the of

eiylotl franzi Hemicycliostyla h fotl ir in cirri frontal the 0.6 length in the African the in length .

P.guizhouensis

1992) three or four vs. four or three eiylotl franzi Hemicycliostyla extend Very early reorganizers of reorganizers early Very

sp. nov. buccal cirri buccal µm s (Kumar et al. 2010) in the ciliary pattern and other unique unique other and pattern ciliary the in 2010) al. et (Kumar s

P.cristata P.cristatoides

- r that are , but only a only but , sized, spherical cortical granules, whereas the African the whereas granules, cortical spherical sized, nsis ing

population pce, namely species,

anteriorly and posteriorly anteriorly(Chen andposteriorly al.2010; et specimens p nv hs bcrn ta i tpcl n h genus the in typical is that bicorona a has nov. sp.

(one . guizhouensis P. h dra knt algn rgnt i prna rw and rows parental in originate anlagen kinety dorsal the and

s L sp. nov.. sp.

). of Chinese population are population Chinese of . nova P. invariably brief characterization brief The major morphological features features morphological major The

right marginal rows right

vs. P. h ke The two

with African and Indian Indian and African with

specimens ace wl te fia (osnr 97 Berger 1987; (Foissner African the well matches cristatoides

d of nine

imorpha

a firstly was ved.

ih mria rw ad hs t a be can it thus and rows marginal right

The main differenc main The Hemicycliostyla franzi Hemicycliostyla r arne i a ige o o monocorona or row single a in arranged are specimens 12 . muscorum P. ) and transverse cirri ( cirri transverse and ) dsigihn faue of feature distinguishing y

marginal cirral anlagen of a middle stage stage middle a of anlagen cirral marginal

two Pseudourostyla –

16 vs. 4 vs. 16 p nv cn e itnuse from distinguished be can nov. sp. (30%), whereas it is much longer in the in longer much is it whereas (30%), , ),

P.levis ,

and macronuclear nodules nodules macronuclear and this is, the marginal rows of each side each of marginalrows the is, this tmost parental row (Chen et al. 2010; al. et (Chen row parental tmost found

, bu , these three populations were found were populations three these

J , i.e. ,

– P . he o fu dorsal four or Three ). the present species present the , 6 t .

P.pelotensis ),

guizhouensis differ from tho from differ and n subtropical in ,

midventral pairs (13 pairs midventral P. provided based on based provided guizhouensis es cristata population . nova P. between

occupies 27% of body of 27% occupies

5 an – 7 vs. 8 vs. 7

of the new species new the of anterior portion of portion anterior ,

, have two right right two have , (both the neotype the (both Jung 2012 etal. and sp.nov.

se in the Indian Indian the in se P.guizhouensis Pseudourostyla .

soils

sp. nov. sp. was The Chinese Chinese The P.subtropica – 15 ( specimens

) ( ) rm the from

provided. with that Guizhou kineties – – Berger 20 vs. 20 40 vs. 40 1) easily

were were the P. ).

This article isThis article by protected copyright. All rightsreser China (27°34′ locality. Type and kineties.dorsal 26 left four or Three cirri. transverse seven to 13 cirri, frontoterminal two cirrus, buccal one irregularly distributed Cell margin. body left near vacuole Diagnosis. Pseudourostyla guizhouensis Genus Family Pseudourostylidae Jankowski, 1979 Jankowski, Urostylida Order 1979 SpirotricheaClass Bütschli, 1889 TAXONOMIC SUMMARY in pattern congeners morphogenetic the Furthermore, cirri. caudal largei.e., data, morphological the with well corresponds which support, high to moderate with species revise of species Trichototaxis marina m during in anlage common a from but origin rows cirral marginal 1) include: These genera. di more a suggest also processes morphogenetic the Furthermore 2014). al. et (Lu findings previous with consistent is which clades, separate three into of placement familial (Kumarreported 2012). 2010;et etal. Paiva the support not sphagni did data molecular Trichototaxis and morphogenetic row, marginal a possess i.e. pattern, cirral of species new tha Two at tentative. lacking were genera two latter the of pattern to assigned ( midventral row cirral a marginalConsequently left bicorona, one than a more and only, having pairs by cirral of characterized composed complex are Pseudourostylidae family the of Members Phy thewhereas African specimens have al. 4)theis Paiva et pharyngealwall 2012); 1.0 possesses AU test AU logeny here, presented tree rDNA SSU the In

study, present the In body,

Accepted familythe Pseudourostylidae. Article Pseudourostyla

Stokes, 1886, has since been redescribed, and an Indian population of population Indian an and redescribed, been since has 1886, Stokes,

. T rhgnss in orphogenesis iooa a ivnrl ope cmoe o cra pis only pairs cirral of composed complex midventral a bicorona,

Trichototaxis

of the familyof the Pseudourostylidae and a the the reject he monophyly of ody – bicorona, midventral pairs, two or more marginal rows on each side, and side, each on marginalrows more or two pairs, midventral bicorona, , (Chen et al. 2007; Lu et al. 2014). In addition, the type species o species type the addition, In 2014). al. et Lu 2007; al. et (Chen 48

2.0 × 1.5 µm 1.5 × 2.0 he gnr, namely genera, three ilg o Zogusa, oiza Twsi, oge Ct, uzo Province, Guizhou City, Tongren Township, Aojiazhai Zhonghuashan, of Village Pseudourostylidae by Berger (2006) who noted, however, that details of the cirral cirral the of details that however, noted, who (2006) Berger by Pseudourostylidae ″N, , 180 s

–

T. marina T. 40 macronuclear 40 macronuclear an nodules the the

109°15′

– (Lu et al. 2014). al. et (Lu

(loosely spaced) (loosely 310 × 65 × 310 Borror, 1972 possibility that the family Pseudourostylidae is monophyletic. is Pseudourostylidae family the that possibility , T Pseudourostyla guizhouensis Pseudourostyla s . - 42 sized, ellipsoida sized, Pseudourostyla

sp. nov.

Lu et al., 2014 al., et Lu tagnatilis Pseudourostyla ″ Pseudourostyla E – Trichototaxis 85 µm 85 ), soil sample. ), soil

many

with colorless, spherical (ca. (ca. spherical colorless, with However, typethe for datamolecular morphogeneticand since . 57 .

Hemicycliostyla in v in tks 19, r nt vial, t wo it available, not are 1891, Stokes,

– about 1 μm (Berger longrods about 2006;

70 without peculiarities , ivo Pseudourostyla l cortical granules which can be extruded (Berger 2006; (Berger extruded be can which granules cortical l

and –

is ae ic be dsrbd icuig eal o their of details including described, been since have hra te fs it mr ta oe as in mass one than more into fuse they whereas mass single a into fuse nodules macronuclear the 2) ; adoral membranelles; 12 membranelles; adoral

20 d two tofive micronuclei. , elongate elliptical in outline, in elliptical elongate ,

thus well

stant relationship among the three pseudourostylid pseudourostylid three the among relationship stant

T. songi T. invariably Pseudourostyla guizhouensis midventral pairs, two pre two pairs, midventral ved. ate time t

sp. nov. clustered with nov.clustered sp.

from individual anlagen in anlagen individual from

Pseudourostyla

supported Chen ; ,

. guizhouensis P.

w rgt agnl rows. marginal right two so Hemicycliostyla ,

in the their placement in this family was was family this in placement their et al., et 0.5

by our analyses.

Chinese (Guizhou) µm across) cortical granules cortical across) µm 2007. Although both species both Although 2007. – 16 and ,

cirri forming a bic a forming cirri

transverse cirri transverse and more than one left left one than more and

, p nv rsmls its resembles nov. sp.

usually one contractile one usually

and Foissner 1987)

f other

l b peaue to premature be uld sp. nov. Trichototaxis Hemicycliostyla, H. franzi franzi H.

Trichototaxis Hemicycliostyla

Pseudourostyla

the absence of absence the Differences in Differences Berger 2006). Berger i or Six

specimens , has been has

and five and , .

orona,

seven

were

fall

H. H. a , , , ,

This article isThis article by protected copyright. All rightsreser G.2012. Min, S. & M. K. Park, H., J. Jung, CommissionICZN =International on Zoological Nomenclatur Bo X., Luo J., Huang, W.,Song, Xu, F.,J., Gao, Li, colpo und Venezuelafrom and hypotriche Ciliophora) (Protozoa, ciliates semiterrestrial TerrestrialW.and Foissner,2016. bekannte wenig und Neue 1987. W. Foissner, throughput. high and accuracy high with alignment sequence multiple Muscle: 2004. C. R. Edgar, Chen a of sequence gene and morphogenesis Morphology,Y. 2010. Kusuoka, & X. Hu, Z., Li, X., Chen, new two of phylogeny and Morphology 2017. C. C. John, & M. Miao, C., Shao, X., Zhao, L., Chen, Hypotricha). (Ciliophora, hy freshwater new A 2007. X. Urostyloidea Hu, & X. Shi, H., Chen, the of Monograph 2006. H. Berger, LITERATURECITED Partnering alsogratefulare to China ma BBRSC the preparing in a help their for and Huang, Jie Dr. (14967611D)and Shao Chen Dr. University Award. We thank Hebei at Application and Systematics of Provinc Hebei of Foundation Science Natural 2015T80753), the and 2014M550378 (no. Foundation Science Postdoctoral China the 31401954), and by supported was study This ACKNOWLEDG ZooBank LSID. taxon Etymology. number NHMUK2017.1.6. with London Museum, History Natural the in deposited is (LFC2014080101C) slide paratype Asecond China. China, of University Protozoology,Ocean of Laboratory the of collection slid paratype one and F) 1E, (Fig. (LFC2014080101A) T p slides. ype Hebei Provincial Education Department (ZD2015038), the Key Laboratory of Zoological Zoological of Laboratory Key the (ZD2015038), Department Education Provincial Hebei Phylogenet. Phylogenet. Evol. Hypotrichia). Ciliophora: (Protozoa: Trachelostylidae and Amphisiellidae families ciliate and cyrtostrombidiids groups: dominant two on strombidiids (Protozoa, Ciliophora). emphasis with ciliates oligotrich of analysis Nucleic Acids Res., methodspublication. of 10, 21and78ofthe Galápagos. Galápagos. Ciliophora) aus BödenundMoosen. Microbiol. phylogeny Japan. ciliate, water fresh fromHypotrichia) north ciliates, ,

had been from China(Ciliophora Stichotrichina). Hangzhou, 85:750

AcceptedMiao X., Article Eur. J. Protistol.,

The Sterkiellasinica –

810.

, ,

discovered. poagl ld wt te ooye specimen holotype the with slide protargol A

61:594 Denisia, Denisia, M., Ma M.,

f w baks uotld iits (Protist, ciliates urostylid brackish two of specific epithet specific urn: lsid:zoobank.org:urn: 4EB3962C pub: MENTS

the the ,

anonymous for reviewers 101:101

rad W. urland, – 32:1792 , Pseudour

610. International CodeInternational Nomenclature ofZoological H., Al H., 35:1 1.

Bull. Zool. Bull.

46:43

west China. sp. nov sp. D., Warren, A., Yi,Warren,Y.S. A., D., Gao, & the Natural Science Foundation of China (project numbers: 31172063 numbers: (project China of Foundation Science Natural the –

– 912. - – 110. Rasheid

guizhouensis 1797. ostyla cristata ostyla

. Go F & a, S. Gao, & F. Gao, A., –

60.

. and.

Nomencl., , Mol. Phylogenet.Evol. Mol.

K.A.S., Xu K.A.S., Zool. Beitr. (N. F.), Syst. BiodiversSyst. Rubrioxytricha tsinlingensisRubrioxytricha Morphology, morphogenesis, a Morphology,morphogenesis, refers

(Ciliophora, Urostylida) from the ancient Lake Biwa, Biwa, Lake ancient fromthe Urostylida) (Ciliophora, the the

e 69:161

( critical comments. ved. C2017201200 , to

the .

potrichous ciliate, potrichous -

., e (LFC2014080101B) are deposited in the the in deposited are (LFC2014080101B) e & Lin & – DD5F Acta Protozool 169. 15:131 2016. Guizhou Province Guizhou

31:187 Ciliophora e. 2012. Amendment of 8, 9, Articles

, 2016. ,

X. 480B 105:141 Multigene – ), 142. 2014. – the Natural Science Foundation Science Natural the

282. Multi -

891D sp.nov. (Protozoa, Ciliophora,

akd by marked

, ., to expand andrefine –

Morphology,and ontogeny

150. Hypotricha). 46:131 dide Ciliaten (Protozoa: (Protozoa: Ciliaten dide Trichototaxis songi Trichototaxis - - gene - nd molecular phylogeny molecular nd based phylogeny of the the of phylogeny based 67302C6724C7. , China, where the new the where China, ,

- – based phylogenetic phylogenetic based 138. Monogr. Biol., Biol., Monogr. the the n n circle ink in

nuscript. We nuscript. J. Eukaryot. Eukaryot. J. registration

sp. n. n. sp. Mol.

This article isThis article by protected copyright. All rightsreser 1975. N. Wilbert, Wiackowski,1988. K. 2007. S. Kumar, & M. Nei, J., Dudley, K., Tamura, Ara 2008. J. P.Hoover,Rougemont, A., & Stamatakis, selection. tree phylogenetic of test unbiased approximately An 2002. H. Shimodaira, Conse 2001. M. Hasegawa, & H. Shimodaira, Y.,Shen, B. Song, J., Liu, a of ontogeny and phylogeny Molecular 2014. H. Berger, W.& Song, Q., Zhang, L., Li, C., Shao, P.F.J. Ronquist, Huelsenbeck, & 2003. Silva N., B. Borges, S., T. Paiva, Silva & T.S. Paiva, M. 1996.TreePage, R.D. View: phylogenetic onpersonal Anapplication to display computers. trees Untersuchunge Taxonomische 1996. E. Aescht, & R. Oberschmidleitner, Nylander, 2. version MrModeltest, 2004. J.A. S. Stickel, J., H. Elwood, L., Medlin, Al Y., Pan, Z., Yi, F., Gao, X., Luo, Hu, India: C., Shao, Park, F., National Gao, Valley X., Silent Lu, the of Ciliates 2010. R. G. Sapra, & K. Kamra, S., Kumar, Spi 3. (Infusoria) Ciliata oder Wimpertiere I: Protozoa oder Urtiere 1932. A. Kahl, analysis (MEGA)version4.0. software servers. Comput. Appl Ober österreichs Belebtschl aus Protozoa) (Ciliophora, Uppsala. amplified 16S eukaryotic (Hypotrichida, Ciliophora). genus the of phylogeny Urostylida): ciliatea newpsammophilic from Brazil. southern the genus new a of Ciliophora). (Protozoa, establishment the with ciliates, hypotrichous brackish new two of phylogeny Protistol., ciliate, marine new a of phylogeny urostyloid hypotrichsof regionanote the on with habitat. the Dtl., ofEastcoast Sea,SouthKorea. ciliate, water brackish new a of selection. selection. Press,of p.150 China,Science Beijing. Microbiol genus, ciliate new Accepted64:171 51:492 models. Article 25:399 Syst. Biol.,

– – 179. 508. Bioinformatics, Bioinformatics,

Hemicycliostyla. Int. J.Syst.Evol.Microbiol., 50:524 ., Bioinformatics, –

. Biosci., 61:371 650. Hemicycliostyla sphagni Hemicycliostyla

ie ebset Tcnk e Poaglmrgain ü Ciliaten. für Protargolimprägnation der Technik verbesserte Eine

- ., Neto, I. D. 2006. D. I. Neto,

–

4:3 Morphology and morphogenesis of a new species in the genus the in species new a ofmorphogenesis and Morphology

537. 57:758

– Paracladotricha salina Paracladotricha – 380.

& 30. 12:357

Zool. J.Linn. Soc.,

- Gu, M. 1992. Protozoa. Protozoa. 1992. M. Gu, like rRNA

– 19:1572 771.

17:1246 J. Nat. (London),Hist. X. – - Neto, I. Neto, 358.

& Zootaxa, Pseudourostylacristatoides

- – & Pseudourostyla pelotensis Pseudourostyla arj SA & arn A 21. Taxo 2017. A. Warren, & S.A. Farraj, ln W. Alan,

1574. - M – Trichototaxismarina

coding regions.coding 1247. Sogin, M. L. 1988. L. M. Sogin,

rBAYES 3: Bayesian Phylogenetic inference under mixed under inference Phylogenetic Bayesian 3: rBAYES ä

Mol. Biol.Mol. Evol., D. & Harada, M. L. 2012. Morphology and 18 and Morphology 2012. L. M. Harada, & D. (Ciliophora, Hypotricha) from Brazil with redefinition of redefinition with fromBrazil Hypotricha) (Ciliophora, mmen ober mmen

– 3334:42 151 (in 151 (in Chinese).

: o assig the assessing for l: 179:475

n. g., n. sp. (Ciliophora, (Ciliophora, sp. n. g., n. 04 Mrhlg, opoeei ad molecular and morphogenesis Morphology, 2014. Evolutionary biology Uppsala biology University,Evolutionary centre, ved. In: –

54. pid bootstrap algorithm for the RAxMLWeb the for algorithm bootstrap pid

22:1085 –

Yin,animals soil W. Subtropical (ed.), al. et EA: oeua eouinr genetics evolutionary Molecular MEGA4: Gene 491. sterreichischer K sterreichischer

24:1596 62:229

, 71:491 The characterization of enzy of characterization The n. sp. (Ciliophora, Urostylida). (Ciliophora, sp. n. –

1094.

sp. n., from Songjiho lagoon on the the on lagoon Songjiho from n., sp. sp. nov. (Ciliophora, Stichotrichia, nov.(Ciliophora, sp. Acta Protozool.,

– ofdne f hlgntc tree phylogenetic of confidence – 241. 1599. – Zootaxa, 499.

Hypotrichia). lä

ranlagen. Beitr. ranlagen. ü n oy n molecular and nomy

1247:43 e eng Ciliaten einige ber

49:339 rotricha. Pseudourostyla Mikrokosmos,

– J. Eukaryot. J. yt Biol., Syst. 58. – 364. matically

S rDNA S Tierwelt

Naturk. Eur.J.

This article isThis article by protected copyright. All rightsreser bar scale The phylogenies. BI and for eachspecies. ML the in differ that tw to corresponds topologies indicates circles. (*) solid asterisk with marked The are branches (100%/1.00) supported Fully (BI). inference Bayesian for non of relationships 4 Figure μm. P nodules; macronuclear 1 kineties the indicates arrowhead cirr cirri, transverse parabuccal the indicates (arrow pattern ciliary the show to specimen fiber pharyngeal impregnated ( arrow cirri, buccal the indicates (arrowhead (arrow ( vacuole. contractile the protargol 3 Figure 1 rows; marginal bars=Scale 60μm. right of anlage RMA, nodules; macronuclear Ma, rows; LMA cirri; frontoterminal FTC, vacuoles; contractile CV, membranelles; anlage the cirrus, micronucleiandthe indicate andarrowhead Arrows pattern. ciliary the showing specimen, (E D). (C, vacuoles arrang irregularly contractile the and (B) inclusions ( staining protargol 2 Figure 1 =60μm.bars membranes; undulating of anlage UMA, cirri; transverse TC, 2; 1, rows marginal right 2, 1, RMR rows; marginal right of anlage RMA, PT,cirri; paroral; pretransverse of anlage LMA, cirri; frontoterminal FTC, 1 LMR marginal rows; cirri; frontal FC, endoral; E, vacuole; contractile reorganizer, indicates ne and arrow double mid a of ( (arrowheads). micronuclei and nodules, macronuclear pattern, ciliary c of ( staining 1 Figure FIGUR Difflugia ria granules ortical

h cnrcie vacuoles. contractile the -

parametric bootstrap values for maximum likel maximum for values bootstrap parametric

Acceptedhead Article - respectively. (I, J) Ventral and dorsal views of a mid a of views dorsal Ventraland J) (I, respectively. arrowhead indicate E LEGENDS E - ) and the contractile vacuole. ( vacuole. contractile the and ) – M staining ( staining opooy of Morphology stage stage Morphology

htmcorps of Photomicrographs 4; E, endoral; FC, frontal cirri; FTC, frontoterminal cirri; LMR, left marginal rows; Ma, rows; marginal left LMR, cirri; frontoterminal FTC, cirri; frontal FC, endoral; E, 4; – ) ( s). aximum of I ( . ). (A) Ventral view of a typical individual. (B individual. typical a Ventralof (A) view ).

H) s. Z, drl oe f ebaels CV membranelles; of zone adoral AZM, us). nuaig ebae, ro idcts h bca cru) AM aoa zn of zone adoral AZM, cirrus). buccal the indicates arrow membranes, undulating , E) D,

eraie, roha indicates arrowhead reorganizer, . s ed c ed o substitutions per 100 nucleotide positions. GenBank accession numbers are given are numbers accession GenBank positions. nucleotide 100 per substitutions o

suorsya guizhouensis Pseudourostyla ( al sae f omto of formation of stage Early (arrows). B , G F,

G - likelihood (ML) tree based on SSU rDNA sequences showing the phylogenetic phylogenetic the showing rDNAsequences SSU on based tree (ML) likelihood –

ortical granules (arrow granules ortical –

E, G E, eta ad osl iw o te ae pcmn o hw h ciliar the show to specimen same the of views dorsal and Ventral – J f hns pplto of population Chinese of ) 3, left marginal rows 1 marginalrows left 3, . ( ). ,

C) eta ad osl iw f h hltp seie, hwn te general the showing specimen, holotype the of view dorsal and Ventral paroral; RMR paroral; s suorsya guizhouensis Pseudourostyla –

A) the frontalthe K E Ln darm f dor of diagram Line (E) osl iw showing view Dorsal ( J ). ( ).

) eta ve o a yia seie. (B specimen. typical a of view Ventral

rnvre ir. ( cirri. Transverse A) Pseudouros wly cirrus. formed zoneofmembranelles;AZM, adoral buccal CV,

V entral view of a typical individual. ( individual. typical a of Ventralview - midventral , G)

right marginal rows; TC, transverse c transverse TC, rows; marginal right Showing the cortical granules (arrows) which are darkly are which (arrows) granules cortical the Showing mark head tyla guizhouensis guizhouensis tyla – 3; Ma, macronuclear nodule; MP, nodule; macronuclear Ma, 3; pairs midventral

s the parabuccal cirrus). ( cirrus). parabuccal the s s). ( s). p nov. sp. -

h oa piodu (ros. ( (arrows). primordium oral the transverse cirralstreak transverse eiylotl franzi Hemicycliostyla the the ihood (ML) and the posterior probability values probability posterior the and (ML) ihood the ved. K G,H) , sal surface showing the irregularly arranged arranged irregularly the showing surface sal

frontal

arncer nodules macronuclear L

p nv i vv ( vivo in nov. sp. F , )

– Ventral and dorsal view of an interphase interphase an of view dorsal Ventraland e ta ad osl iw o te same the of views dorsal and Ventral (bold). Numbers near nodes are the the are nodes near Numbers (bold). D) Showing arrangement and variability and arrangement Showing D) ) - stage reorganizer (arrowhead indicates indicates (arrowhead reorganizer stage , - Showing the dorsal surface and the the and surface dorsal the Showing midventral otatl vcoe D 1 DK vacuole; contractile p nv i vivo in nov. sp. – H, I H, 7, dorsal kineties dorsal 7, –

- n. ( D) Showing the granular granular the Showing D) rnvre irl streak cirral transverse F) A ) n io ( vivo in alg o lf marginal left of anlage , B)

– – V entral and dorsal view dorsal Ventraland

J S 1 kineties dorsal 7,

D ) , D

howing ingested food ingested howing

Ventral of view a late ) irri. Scale bars = 60 = bars Scale irri. and orsal view orsal

n atr protargol after and

( I) A A the

– , hwn the Showing F F micronuclei ) and after after and )

ad after and ) 1 – the , –

pattern y 4, dorsal dorsal 4, showing 7. Scale Scale 7. buccal left – ; P,; 7. 7. I ,

This article isThis article by protected copyright. All rightsreser cirral marginal Accepted left in Cirri row 1,number cirral marginal left in Cirri rows, number cirral marginal Left numberMidventral pairs, number cirri, ventral Pretransverse Transverse cirri,number Frontoterminal cirri,number Frontal cirri, number cirri,Buccal number Article membranelles,Adoral number length by occupied AZM (%) body of Proportion zoneLength ofadoral Body, width Body, length Character Hemicycliostyla franzi 1. Table

a opoerc aa of data Morphometric

(Hf)

fromChin

Species Pg Pg Hf Pg Hf Pg Pg Hf Pg Hf Pg Hf Pg Hf Pg Hf Pg Hf Pg Hf Pg Hf Pg Hf Pg Pseudourostyla guizhouensis guizhouensis Pseudourostyla

55 57 23 29 36 50 65 50 160 175 Min 20 22 7 3 8 13 2 5 5 2 2 11 12 2 1

66 70 32 42 86 110 115 88 290 300 Max 32 36 8 4 14 20 2 8 7 3 2 15 16 3 1

ved.

61.3 63.1 27.0 38.0 68.0 91.4 93.8 71.8 249.4 243.3 Mean 26.4 25.6 7.8 3.3 11.7 17.1 2.0 6.6 6.1 2.8 2.0 13.6 13.9 2.7 1.0

62 63 28 38 70 90 103 75 268 247 Med 26 25 8 3 12 17 2 7 6 3 2 14 14 3 1

p nov. sp.

3.8 4.5 0.0 4.8 9 17.2 19.2 9.9 42.5 39.0 SD 3.9 4.3 — — 1.8 2.0 0.0 1.0 0.8 0.4 0.0 1.4 1.2 0.5 0.0 14.

(Pg) (Pg)

6.2 7.1 10.3 12.5 22.1 18.9 20.5 13.8 17.0 16.0 CV 14.6 16.7 — — 15.0 11.8 0.0 14.9 12.4 14.4 0.0 10.3 8.7 19.3 0.0

and

6 18 8 18 8 18 8 18 8 18 n 15 15 8 18 8 18 18 7 18 6 18 8 18 6 18 pplto of population a

This article isThis article by protected copyright. All rightsreser b a deviation.standard minimum; Min, mean; arithmetic Mean, maximum; All measurements in Micronuclei, width Micronuclei, length Micronuclei, number Macronucleus, width Macronucleus, length nodules, number Macronuclear kineties,Dorsal number rows 2,number cirral marginal right in Cirri rows 1,number in Cirri number rows, cirral marginal Right row 4,number cirral marginal left in Cirri row 3,number cirral marginal left in Cirri row 2,number

S Data based on Acceptedtatistically senseless. Article

ih mria cirral marginal right

randomly selected

µm

. AZM, adoral zone of membranelles; zoneof CV,. AZM,adoral coefficient Max, ofvariation in%; Pg Pg Pg Pg Hf Pg Hf Pg Hf Pg Hf Pg Hf Pg Pg Pg Hf Pg ,

protargol 26 3 6 22 20 6 2 2 16 3 5 3 2 3 4 5 8 166

stained specimens.

40 4 7 37 35 7 2 12 30 5 9 3 5 7 7 16 16 195

ved. 32.8 3.5 6. 9 30.4 28.0 6.6 2.0 5.0 21.1 3.7 6.2 — 3.7 4.6 5.5 10.2 12.4 177.3 n

, number number ,

32 4 7 30 28 7 2 4 21 4 6 — 4 4 5 10 13 175

f pcmn ivsiae; SD, investigated; specimens of 3.9 — — 4.3 4.7 — 0.0 4.1 4.7 0.7 1.3 — 0.8 0.7 1.1 3.3 2.5 9.7

11.8 — — 14.0 16.7 — 0.0 82.8 22.3 18.8 20.4 — 21.0 15.7 20.3 32.5 20.2 5.5

18 8 18 15 15 8 18 5 15 18 18 1 18 8 18 8 18 7

by AZM (%) occupied length body of Proportion size Body size Body Character Data source number Micronuclei, number nodules, Macronuclear number kineties, Dorsal Transverse cirri, number marginal Right number rows, marginal Left rows, number number pairs, Midventral cirri, number Frontal cirri, number Buccal number membranelles, Adoral This article isThis article by protected copyright. All rightsreser b a All measurements in T

after after protargol staining vivo in

Data combining thebicoronaData combining a C able 2.

Accepted Article alculated alculated from the

Morphometric comparison of

µm. Present Present (24 (26 33 (67 (56 2 (33 (13 17 (12 14 1 (57 63 38 50 175 65 180 n. sp. guizhouensis P. two morphs.two

– – 88 – 85 – – – or or or 0 × 300 × 310 5) 7)

– – – – AZM, 7) 4)

20) 16) 70) 40) paper

nd midventralro adoral zoneofmembranellesadoral

8 5 4 (on 4 average) 21.5 (18 22 1 88 40 40 181 59 221 cristata (neotype) P. (2006) (1996 ner Oberschmidleit (35 (27 41 – – or or or

10 6 – –

111 – 147 – – 5 2 and 303 324 7)

);

– –

Pseudourostyla guizhouensis

55) 26)

Aescht Aescht Berger

× ×

Chen et al. al. (2010) et Chen (25 (15 28 8 7 5 5 (17 22 (20 23 1 (80 89 40 100 270 90 280 Biwa) cristata P. ws. – – or or

10 10 – 130 – – – – 6 220 380 × 400 9)

– – – –

ved.

36) 25) 24) 94)

(Lake (Lake

× ; /, data not; /, data available. 25 150 (neotype) P. (2006) Berger ca. 3 43 7 7 4 5 19 / 1 67 38 192

or or

– 100 100 –

× 8

0 × 300

levis levis

sp. n 246 （ P. levis al. (2010) et Kumar (46 (58 74 7 7 5 5 34 / 1 (79 87 40 58 189 76

– ov Indian

13 – – – 42 83 – – 100) . × × 266 9)

b – with

92) ）

other 65 250 muscroum P. (2006) Berger (1932 Kahl / / / 8 2 2 / / ca. 9 / 31 /

–

– 15 95 –

350

)

;

Pseudourostyla

× (2014) al. et Chen (24 (9 16 (78 (78 2 2 (22 29 (44 1 (52 58 35 60 200 45 140 P. nova

– – 125 – 55 – – – – – 375 × 200 8) – 9) 12) 6)

25 – –

36) 64)

） ×

(2006) and Silva Paiva / (62 88 7 5 5 4 (10 13 9 1 (38 47 36 51 145 × 80 190 P.

species. – – – – – pelotensis

11 9 8 6 13 –

70 –

195 -

Neto Neto – – –

114) 15) 54)

× (2012) J / (30 78 11 (10 (69 (44 (56 (17 21 (20 24 1 (84 97 33 65 210 85 220 P. cristatoides

n e al. et ung

– – 125 – 125 – – – or or 290 × 265 12) 7) – – – – –

13)

106) 25) 30) 115)

× Chen et al. al. (2014) et Chen (24 114 (68 11 (8 7 5 7 (14 19 (26 32 1 (104 131 25 160 300 120 300 P. subtropica – – – 12 9 13 –

33 – – – – – 360 600 × 200 450 – 6)

14) – –

– 28) 44)

219) –

173)

× (2016) Foissner 3 67 2 4 4 5 (68 (69 2 31 22 21 106 25 120 P. dimorpha – – – or or or 9 – 9 6 – – – –

160 47 38 57 – 65 – – – 3 6 9 × 198 × 230 11) 12)

This article isThis article by protected copyright. All rightsreser Accepted Article ved.