Table S1. Species for which ITS1-5.8S-ITS2 regions were newly sequenced in the present work.
Taxon / GB / ITS1-5.8S-ITS2 / ITS2Length (nt) / GC content (%) / Length (nt) / GC content (%)
Opisthonecta henneguyi / KF524430 / 427 / 34.66 / 167 / 31.14
Opisthonecta minima / KF524429 / 428 / 34.81 / 167 / 31.14
Vorticella aequilata Sa pop / KF524382 / 415 / 41.45 / 165 / 47.27
Vorticella aequilata Sc pop / KF524408 / 415 / 41.20 / 165 / 46.67
Vorticella aequilata Se pop1 / KF524392 / 415 / 41.45 / 165 / 47.27
Vorticella aequilata Se pop2 / KF524391 / 415 / 41.45 / 165 / 47.27
Vorticella aequilata Se pop3 / KF524388 / 415 / 41.45 / 165 / 47.27
Vorticella aequilata Se pop4 / KF524372 / 415 / 41.45 / 165 / 47.27
Vorticella aequilata Wh pop / KF524378 / 416 / 41.83 / 165 / 47.88
Vorticella aequilata-like Mh / KF524387 / 416 / 41.83 / 165 / 47.88
Vorticella aquadulcis? / KF524428 / 430 / 35.12 / 166 / 35.54
Vorticella astyliformis-like / KF524389 / 427 / 35.60 / 167 / 31.74
Vorticella campanula Ec pop / KF524425 / 425 / 35.76 / 168 / 33.33
Vorticella campanulaGe pop / KF524376 / 424 / 34.20 / 168 / 29.76
Vorticella campanula Ka pop / KF524426 / 425 / 35.76 / 168 / 33.33
Vorticella campanulaQd pop1 / KF524384 / 426 / 33.80 / 169 / 28.99
Vorticella campanula Qd pop2 / KF524385 / 425 / 33.83 / 169 / 28.99
Vorticella campanula Qd pop3 / KF524417 / 426 / 33.80 / 169 / 28.99
Vorticella campanula Qd pop4 / KF524418 / 426 / 33.80 / 169 / 28.99
Vorticella campanulaWh pop1 / KF524399 / 425 / 35.76 / 168 / 33.33
Vorticella campanulaWh pop2 / KF524400 / 425 / 35.76 / 168 / 33.33
Vorticella campanulaWh pop3 / KF524403 / 425 / 35.76 / 168 / 33.33
Vorticella campanula Wh pop4 / KF524419 / 426 / 33.80 / 169 / 28.99
Vorticella campanula Wh pop5 / KF524420 / 425 / 35.76 / 168 / 33.33
Vorticella campanula Wh pop6 / KF524421 / 426 / 33.80 / 169 / 28.99
Vorticella campanula-like Du pop / KF524413 / 426 / 33.80 / 169 / 29.59
Vorticella campanula-like Gz pop / KF524366 / 425 / 33.65 / 168 / 29.76
Vorticella chlorostigma / KF524422 / 420 / 36.67 / 169 / 38.46
Vorticella citrinaMh pop / KF524410 / 423 / 34.28 / 167 / 29.94
Vorticella citrinaWh pop / KF524402 / 423 / 34.75 / 167 / 29.94
Vorticella convallaria Ak pop / KF524406 / 423 / 35.22 / 167 / 32.34
Vorticella convallaria Fl pop / KF524407 / 423 / 35.22 / 167 / 32.34
Vorticella convallariaGz pop / KF524393 / 423 / 35.22 / 167 / 32.34
Vorticella convallariaNy pop / KF524404 / 423 / 35.22 / 167 / 32.34
Vorticella convallariavar compacta / KF524412 / 423 / 36.41 / 167 / 32.93
Vorticella elongata-like / KF524405 / 423 / 35.46 / 168 / 32.74
Vorticella fuscaGr pop1 / KF524365 / 421 / 37.53 / 167 / 35.93
Vorticella fusca Gr pop2 / KF524381 / 421 / 37.53 / 167 / 35.93
Vorticella fuscaWh / KF524367 / 421 / 39.43 / 167 / 40.72
Vorticella gracilisSc pop / KF524414 / 423 / 35.46 / 168 / 32.74
Vorticella gracilisDu pop / KF524409 / 423 / 35.46 / 168 / 32.74
Vorticella gracilis Gz pop1 / KF524423 / 423 / 35.70 / 168 / 33.33
Vorticella gracilis Gz pop2 / KF524370 / 423 / 35.46 / 168 / 33.33
Vorticella gracilisWh pop1 / KF524401 / 423 / 35.70 / 168 / 33.33
Vorticella gracilisWh pop2 / KF524373 / 423 / 35.70 / 168 / 33.33
Vorticella gracilis-like Gr pop1 / KF524394 / 423 / 35.70 / 168 / 33.33
Vorticella gracilis-like Gr pop2 / KF524395 / 423 / 35.70 / 168 / 33.33
Vorticella infusionum / KF524368 / 427 / 35.36 / 167 / 31.74
Vorticella infusionum-like / KF524380 / 430 / 37.44 / 166 / 37.35
Vorticella microstoma-like / KF524379 / 431 / 34.57 / 167 / 31.14
Vorticella microstoma ATCC / KF524396 / 428 / 35.28 / 167 / 31.14
Vorticella natans Gz / KF524369 / 416 / 41.59 / 165 / 47.27
Vorticella natansCa / KF524383 / 416 / 41.59 / 165 / 47.27
Vorticella similisCh / KF524398 / 422 / 36.97 / 167 / 35.33
Vorticella similisDu / KF524397 / 422 / 36.97 / 167 / 35.33
Vorticella similisMh / KF524411 / 423 / 36.41 / 167 / 32.93
Vorticella similis Qd pop1 / KF524416 / 423 / 34.75 / 167 / 30.54
Vorticella similis Qd pop2 / KF524386 / 423 / 34.75 / 167 / 30.54
Vorticella sp2 / KF524415 / 423 / 35.22 / 167 / 31.14
Vorticella sp3 / KF524390 / 421 / 37.53 / 167 / 35.93
Vorticella sp11 / KF524371 / 421 / 37.29 / 168 / 37.50
Vorticella sp12 / KF524377 / 423 / 34.99 / 167 / 31.74
Vorticella sp18 / KF524424 / 415 / 41.69 / 165 / 47.27
Vorticellasp 23 / KF524374 / 423 / 34.75 / 167 / 32.34
Vorticellasp. / KF524375 / 432 / 35.19 / 167 / 32.93
Vorticella striata? / KF524427 / 427 / 35.60 / 167 / 31.74
Table S2. Genetic distance of ITS1-5.8S-ITS2 sequences between samples of speciescollected more than once.
Species / No. of populations and Collection Localities / Genetic distance (%)Vorticellasimilis / 5; China, USA / 4.23
Vorticellacitrina / 2; China; USA / 3.06
Vorticellafusca / 3; Austria, China / 1.60
Vorticella campanula / 13; Austria, China, Japan / 2.89
Vorticella campanula-like / 2; China, USA / 2.24
Vorticella aequilata / 7; Austria, China, USA / 0.48
Vorticella gracilis / 6; China, USA / 0.32
Vorticella gracilis-like / 2; Austria / 0.00
Vorticella natans / 2; China, USA / 0.16
Vorticellaconvallaria / 4; China, USA / 0.00
Table S3. Morphometric data of samples in clade II based on measurements of 10–30 cells from each one. Silverlines are transversely oriented pellicular ridges evident as surface features of the cell after staining with silver nitrate; P–TB, number of silverlines from peristome to trochal band; TB–S, number of silverlines from trochal band to scopula.
Species / Body Length (µm) / Width of Body (µm) / Silverlines P–TB / Silverlines TB–S / Shape; orientation of MaVorticella striata? / 35-40 / 30-35 / — / — / C; transverse
Vorticella infusionum / 35-40 / 30-40 / 35-37 / 7-8 / C; transverse
Vorticella astyliformis-like / 25-30 / 20-25 / 33-35 / 4-6 / C; transverse
Vorticella infusionum-like / 40-50 / 25-35 / 33-34 / 10-11 / C; transverse
Vorticella microstoma-like / 35-40 / 25-35 / 26-33 / 4-6 / C; longitudinal
Vorticella aquadulcis? / 25-30 / 20-25 / — / — / C ; transverse
Opisthonecta henneguyi / 90-110 / 45-55 / 60-65 / 42-45 / C; transverse
Opisthonecta minima / 60-70 / 35-50 / 35-39 / 15-17 / C; transverse
Species / 1 / 2 / 3 / 4 / 5 / 6 / 7 / 8 / 9 / 10
1 / Opisthonecta henneguyi / —
—
—
2 / Opisthonecta minima / 0.195
1.337
1.818 / —
—
—
3 / Vorticella infusionum / 0.522
3.429
2.436 / 0.521
2.027
1.829 / —
—
—
4 / Vorticella astyliformis-like / 0.522
4.144
2.436 / 0.456
2.752
1.829 / 0.065
0.671
0.000 / —
—
—
5 / Vorticella striata? / 0.522
3.429
2.436 / 0.521
2.027
0.602 / 0.260
0.000
1.212 / 0.325
0.671
1.212 / —
—
—
6 / Vorticella microstoma ATCC / 0.522
3.406
3.061 / 0.456
2.722
1.212 / 0.325
0.000
1.829 / 0.260
0.667
1.829 / 0.065
0.000
0.602 / —
—
—
7 / Vorticella infusionum-like / 1.047
8.393
11.259 / 1.113
9.140
9.184 / 1.313
9.232
8.468 / 1.312
8.452
8.468 / 1.313
9.232
8.468 / 1.312
9.941
9.165 / —
—
—
8 / Vorticella microstoma-like / 1.244
8.584
5.593 / 1.310
8.618
3.682 / 1.444
9.361
4.315 / 1.378
8.582
4.315 / 1.444
9.361
3.057 / 1.378
9.349
2.439 / 1.642
3.311
9.132 / —
—
—
9 / Vorticellasp. / 1.442
6.910
5.593 / 1.376
7.660
4.952 / 1.509
7.705
5.593 / 1.443
7.716
5.593 / 1.509
7.705
4.319 / 1.443
7.676
4.952 / 1.642
6.152
9.047 / 1.375
6.155
4.952 / —
—
—
10 / Vorticella aquadulcis? / 2.109
7.745
10.404 / 2.108
8.472
8.355 / 2.242
8.544
9.047 / 2.175
8.556
9.047 / 2.242
8.544
9.047 / 2.175
8.489
9.748 / 2.649
6.906
8.381 / 2.043
7.000
9.739 / 2.243
1.976
11.808 / —
—
—
Table S4. Genetic distance between sequences of SSU rDNA (upper) gene, ITS1 (middle) and ITS2 (lower) regions of species within clade II,determined by Kimura two-parameter distance method (Kimura, 1980) and expressed as percentages.
Table S5. Taxa sampled in the present study.
Taxon / Localities / DateOpisthonecta henneguyi / Austria, Salzburg, urban pond / 07/19/2009
Opisthonecta minima / Austria, Salzburg, urban pond / 07/19/2009
Vorticella aequilata Sa pop / Austria, S Bruck an der Großglocknerstraße, eutrophic pool / 07/17/2009
Vorticella aequilata Sc pop / United States, NC, Sampson Co., 6.45 km SSE Newton Grove, small creek at US701 / 02/23/2008
Vorticella aequilata Se pop1 / Austria, Seewalchen, swimming area on S shore of Wallersee / 07/12/2009
Vorticella aequilata Se pop2 / Austria, Seewalchen, puddle in road near S shore Wallersee / 07/20/2009
Vorticella aequilata Se pop3 / Austria, Seewalchen, swimming area on S Shore Wallersee / 07/12/2009
Vorticella aequilata Se pop4 / Austria, Seekirchen, Small stream through center of town / 07/12/2009
Vorticella aequilata Wh pop / China, Wuhan, SW shore DonghuLake, enclosed research area at Institute of Hydrobiology floating laboratory / 05/25/2009
Vorticella aequilata-like Mh pop / United States, NC, Morehead city, Craven Co., 17 km WSW Havelock, swampy area alongside Great Lake Road / 08/12/2008
Vorticella aquadulcis? / Japan, LakeBiwa, KarasumaPeninsula / 07/21/2010
Vorticella astyliformis-like / Austria, Mattsee marshy area along margin of lake / 07/12/2009
Vorticella campanula Ec pop / Japan, LakeBiwaMuseum, Ecopond / 07/21/2010
Vorticella campanula Ge pop / Austria, Gegend, bog at edge of path in woods / 07/25/2009
Vorticella campanula Ka pop / Japan, LakeBiwa, KarasumaPeninsula / 07/21/2010
Vorticella campanulaQd pop1 / China, W Qingdao, 1.2 km E Lijiazhuang, pond receiving outflow from Jihongtan Reservoir / 05/18/2009
Vorticella campanula Qd pop2 / China, Qingdao, drainage ditch near Jihongtan Reservoir / 05/18/2009
Vorticella campanula Qd pop3 / China, Qingdao, LuxunPark, lake / 05/10/2008
Vorticella campanula Qd pop4 / China, Qingdao, ZhongshanPark, lake / 05/11/2008
Vorticella campanulaWh pop1 / China, Wuhan, WuhanBotanical Garden, Lotus Pond / 05/26/2009
Vorticella campanulaWh pop2 / China, Wuhan, Sand Lake / 05/31/2009
Vorticella campanulaWh pop3 / China, Wuhan, fish culturing pond / 05/30/2009
Vorticella campanula Wh pop4 / China, Wuhan, MulanMountain / 06/11/2008
Vorticella campanula Wh pop5 / China, Wuhan, Donghu Lake near main gate of WuhanUniversity / 05/23/2008
Vorticella campanula Wh pop6 / China, Wuhan, Mulan Moutain / 06/11/2008
Vorticella campanula-like Du pop / United States, NC, Durham, Ellerbe Creek / 01/27/2008
Vorticella campanula-like Gz pop / China, S Guangzhou, 1.1 km SSW Shibi, canal at DazhouBridge / 06/13/2009
Vorticella chlorostigma / United States, NC, Morehead city, pond / 08/11/2008
Vorticella citrina Mhpop / United States, NC, Morehead city, Craven Co., 14.3 km WSW Havelock, CroatanNational Forest, W shore GreatLake / 08/12/2008
Vorticella citrina Wh pop / China, Wuhan, SW shore DonghuLake, boating facility / 05/25/2009
Vorticella convallaria Akpop / United States, Alaska, ToolikLake / 07/28/2006
Vorticella convallariaFl pop / United States, Florida, Gilchrist Co., 12.3 km SW Bell, Hart Springs / 08/02/2002
Vorticella convallariaGz pop / China, Guangzhou, SouthChinaBotanical Garden, Bamboo Grove pond / 06/10/2008
Vorticella convallariaNy pop / United States, NY, OtiscoLake, planktonic cyanobacterial clumps / 07/02/2001
Vorticella convallaria var compacta / United States, NC, Cartaret Co., Morehead city,urban pond / 08/11/2008
Vorticella elongata-like / United States, Tennessee, Great Smoky Mountain N.P. / 09/06/2009
Vorticella fusca Gr pop1 / Austria, 0.6km NE Fraham, S margin of Grabensee / 07/12/2009
Vorticella fusca Gr pop2 / Austria, Salzburg, S margin of Grabensee / 07/12/2009
Vorticella fusca Wh pop / China, Wuhan, farming plot on NW margin of SandLake / 05/31/2009
Vorticella gracilis Scpop / United States, NC, Sampson Co., 6.45 km SSE Newton Grove, small creek at US701 / 02/23/2008
Vorticella gracilis Du pop / United States, NC, Beaufort Co., Durham Creek at Tunstall Swamp Rd. / 08/13/2008
Vorticella gracilis Gz pop1 / China, Guangzhou, South China Normal University, pond / 06/16/2009
Vorticella gracilis Gz pop2 / China, Guangzhou, SouthChinaBotanical Garden, Bamboo grove / 06/10/2008
Vorticella gracilisWh pop1 / China, Boating faculty on Donghu road, Wuhan / 05/25/2009
Vorticella gracilis Wh pop2 / China, Wuhan, SW shore DonghuLake near WuhanUniversity / 05/25/2009
Vorticella gracilis-like Gr pop1 / Austria, S Bruck an der Großglocknerstraße, Wallackhaus area, stream, moss debris / 07/10/2009
Vorticella gracilis-like Gr pop2 / Austria, S Bruck an der Großglocknerstraße, Wallackhaus area, stream, moss debris / 07/10/2009
Vorticella infusionum / Austria, Bayerham, puddles near manure and hay compost pile / 07/12/2009
Vorticella infusionum-like / Austria, Bayerham, puddles near manure and hay compost pile / 07/12/2009
Vorticella microstoma-like / China, Wuhan, Wuhan Bontanical garden, ephemeral pool / 05/26/2009
Vorticella microstoma ATCC / ATCC / —
Vorticella natansCapop / America, Carrboro, a farm pond / 08/12/2008
Vorticella natans Gz pop / China, S Guangzhou, 1.1 km SSW Shibi, canal at DazhouBridge / 06/13/2009
Vorticella similisCh pop / United States, NC, Chapel hill, Orange Co., Chapel Hill, flooded area on margin of Bolin Creek / 01/13/2009
Vorticella similisDu pop / United States, NC, Durham, flooded area along Al Buehler Cross Country trail, Duke University / 02/17/2009
Vorticella similis Mh pop / United States, NC, Morehead city, Cartaret Co., drainage ditch alongside US70 / 08/14/2008
Vorticella similis Qd pop1 / China, Qingdao, Jihongtan Reservior, ditch / 05/18/2009
Vorticella similis Qd pop2 / China, W Qingdao, 1.2 km E Lijiazhuang, pond receiving outflow from Jihongtan Reservoir / 05/18/2009
Vorticella sp2 / United States, NC, Durham, flooded area along Al Buehler Cross Country trail, Duke University / 12/08/2008
Vorticella sp3 / Austria, 0.6 km NE Fraham, S. shore of Grabensee / 07/12/2009
Vorticella sp11 / China, S Guangzhou, 1.1km SSW Shibi, canal at Dazhou bridge / 06/13/2009
Vorticella sp12 / China, 15.7 km SSE Guangzhou, 1.5km NW Xiecun, irrigation pond / 06/13/2009
Vorticella sp18 / Japan, Hokkaido, Lake Toufutsu / 08/12/2009
Vorticella sp 23 / United States, NC, Bladen Co., Colly Swamp at NC53 / 02/23/2008
Vorticella sp. / China, Wuhan, SW shore DonghuLake, enclosed research area at Institute of Hydrobiology floating laboratory / 05/25/2009
Vorticella striata? / Japan, Nagano Pref., Ogouchi / 03/23/2009
Figure S1. The section of the alignment of ITS2 sequences from all samples corresponds to helix III, which is the most conserved of the three helices in the secondary structure of the region. Structures on the right show the predicted secondary structure of Helix III for each clade and subclade.
Figure S2. Photomicrographs of species in clade II. Gross morphology of V. striata? (a), V. infusionum (b), V. astyliformis-like (c), Opisthonecta minima (d), O. henneguyi (e), V. infusionum-like (f), V. microstoma-like (g) and V. aquadulcis? (h) from life and after silver staining.