<p>Winter-summer succession of unicellular eukaryotes in a meso-eutrophic coastal system.</p><p>Urania Christaki, Konstantinos Ar. Kormas, Savvas Genitsaris, Clément Georges, Télesphore Sime-Ngando, Eric</p><p>Viscogliosi, Sébastien Monchy</p><p>Supplementary Material</p><p>1 Table S1: OTUs representing >0. 1 % in all samples No. of Closest relative, (% similarity) Closest cultured relative (% OTU similar tags Putative affiliation Sample [GenBank accession No.] similarity) [GenBank accession No.] (>97%) 1. EEC09_1183 30992 Clone SGYH1395.FRAG.MO.500m Dinophyceae Gyrodinium spirale (97%) [AB120001] All (97%) [JX842244] 2. EEC09_1001 4382 Clone BF9_3B1D (99%) [JN048120] Cercozoa Cryothecomonas sp. (99%) [GQ144679] 30_Mar, 07_Apr, 27_Apr, 20_May, 09_Jun 3. EEC09_1141 4373 Phaeocystis globosa (99%) [JX660994] Haptophyceae Phaeocystis globosa (99%) [JX660994] 25_Feb, 11_Mar, 16_Mar, 30_Mar, 07_Apr, 27_Apr, 20_May, 09_Jun 4. EEC09_1168 3133 Clone SGUH400.FRAG.MO.5m (99%) Dinophyceae Karlodinium micrum (96%) [JF791049] 25_Feb, 02_Mar, 11_Mar, 16_Mar, [JX841624] 07_Apr, 27_Apr, 20_May, 09_Jun 5. EEC09_1174 1170 Engyodontium sp. (100%) [JX896662] Fungi Engyodontium sp. (100%) [JX896662] All 6. EEC09_1179 1112 Clone TW-A1-2-6b (99%) [EU860701] Fungi Ascomycota sp. (99%) [EU723494] All 7. EEC09_1003 1011 Clone SGUH758.FRAG.MO.5m (99%) Dinophyceae Amphidinium semilunatum (95%) 02_Mar, 11_Mar, 16_Mar, 30_Mar, [JX841861] [JQ179860] 27_Apr, 20_May, 09_Jan 8. EEC09_1109 1008 Clone 897st19-43 (100%) [JQ782085] - - All 9. EEC09_1101 888 Clone 3b-G3 (99%) [FN690411] Cercozoa Protaspis sp. (98%) [FJ824125] All 10. EEC09_1151 780 Clone RS1E4A08 (99%) [HM227931] Ichthyosporea Ichthyosporea sp. (96%) [GU810145] All 11. EEC09_1052 695 Sphaeroforma arctica (99%) Ichthyosporea Sphaeroforma arctica (99%) All [JX992717] [JX992717] 12. EEC09_0476 598 Clone H3S3Be44w (98%) [JQ781892] Oomycetes Olpidiopsis porphyrae (91%) 20_May, 09_Jun [AB287418] 13. EEC09_0980 579 Clone op23 (91%) [EU567215] Cercozoa Protaspis obliqua (90%) [FJ824122] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 20_May 14. EEC09_1037 462 Clone E_MO_3H07 (100%) [JN832708] - - 25_Feb, 11_Mar, 16_Mar, 27_Apr, 20_May, 09_Jun 15. EEC09_0487 402 Clone TAGIRI-6 (99%) [AB191414] Dinophyceae Prorocentrum gracile (96%) 20_May, 09_Jun [EF517264] 16. EEC09_0914 397 Clone SGUH1454.FRAG.MO.5m (99%) Ciliophora Strombidium biarmatum (99%) 02_Mar, 11_Mar, 16_Mar, 30_Mar, [JX841568] [JX512970] 07_Apr, 27_Apr, 20_May 17. EEC09_0892 373 Hymenoscyphus sp. (99%) [KC164672] Fungi Hymenoscyphus sp. (99%) [KC164672] 25_Feb, 16_Mar, 30_Mar, 07_Apr, 27_Apr, 20_May, 09_Jun 18. EEC09_0489 360 Clone BS_DGGE_Euk-13 (99%) Dinophyceae Gymnodinium sp. (95%) [JQ639761] 20_May, 09_Jun [DQ234294]</p><p>2 No. of Closest relative, (% similarity) Closest cultured relative (% OTU similar tags Putative affiliation Sample [GenBank accession No.] similarity) [GenBank accession No.] (>97%) 19. EEC09_1158 341 Clone CS618-90 (98%) [HM369645] Dinophyceae Euduboscquella crenulata (91%) 25_Feb, 02_Mar, 11_Mar, 16_Mar, [JN606065] 30_Mar, 07_Apr, 27_Apr, 20_May 20. EEC09_1004 341 Clone 905st23-36 (99%) [JQ782096] Labyrinthulomycetes Aplanochytrium sp. (99%) [EU851174] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 07_Apr, 27_Apr, 20_May, 09_Jun 21. EEC09_1121 336 Clone 15H3Te94T (99%) [JQ782075] Bacillariophyta Stephanodiscus sp. (99%) [AB430594] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 07_Apr, 27_Apr 22. EEC09_0966 321 Clone SGYH791.FRAG.MO.500m - - 25_Feb, 02_Mar, 11_Mar, 16_Mar, (99%) [JX842562] 30_Mar, 07_Apr, 20_May 23. EEC09_1025 299 Clone SCM28C40 (99%) [AY664893] Dinophyceae Warnowia sp. (98%) [FJ947040] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 20_May, 09_Jun 24. EEC09_0939 296 Clone T4A4_15 (99%) [HQ394058] Telonemida Telonema antarcticum (98%) 02_Mar, 11_Mar, 16_Mar, 07_Apr, [AJ564773] 27_Apr, 20_May, 25. EEC09_1066 247 Gyrodinium sp. (99%) [HE611580] Dinophyceae Gyrodinium sp. (99%) [HE611580] 11_Mar, 16_Mar, 30_Mar, 07_Apr, 27_Apr, 20_May 26. EEC09_0816 240 Clone PRTBE7302 (99%) [HM800000] Fungi Pseudozyma aphidis (99%) [JN940465] 07_Apr, 27_Apr, 09_Jun 27. EEC09_0827 239 Clone SGUH594.FRAG.MO.5m (98%) Dinophyceae Gyrodinium rubrum (96%) [AB120003] 27_Apr, 20_May [JX841751] 28. EEC09_1185 236 Clone 561 (99%) [HQ696569] Cercozoa Protaspis sp. (96%) [FJ824125] All 29. EEC09_1069 230 Clone T0_78 (99%) [HQ394148] Cercozoa Cryothecomonas sp. (99%) [GQ144679] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 07_Apr, 27_Apr, 20_May 30. EEC09_1028 230 Clone SGYH1214.FRAG.MO.500m Cryptophyta Geminigera cryophila (95%) [JF791030] 25_Feb, 02_Mar, 11_Mar, 16_Mar, (99%) [JX842127] 30_Mar, 07_Apr 31. EEC09_1091 230 Clone Ch8A2mG4 (98%) [JF730792] Fungi Spizellomyces acuminatus (94%) 25_Feb, 02_Mar, 11_Mar, 16_Mar, [JN940936] 30_Mar 32. EEC09_1175 209 Clone ANT-Roth-MECL-73 (99%) Cercozoa Cercozoa sp. (97%) [FJ824126] 25_Feb, 02_Mar, 11_Mar, 16_Mar, [FJ985906] 27_Apr, 20_May 33. EEC09_1143 208 Pseudo-nitzscia sp. (100%) [JN934671] Bacillariophyta Pseudo-nitzscia sp. (100%) [JN934671] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 07_Apr, 27_Apr, 20_May 34. EEC09_1061 208 Clone DSGM-53 (100%) [AB275053] - - 25_Feb, 02_Mar, 11_Mar, 16_Mar, 07_Apr, 27_Apr, 20_May</p><p>3 No. of Closest relative, (% similarity) Closest cultured relative (% OTU similar tags Putative affiliation Sample [GenBank accession No.] similarity) [GenBank accession No.] (>97%) 35. EEC09_0670 199 Clone DGGE band K (99%) Ciliophora Strombidium biarmatum (97%) 25_Feb, 11_Mar, 16_Mar, 30_Mar, [JQ988915] [JX512970] 07_Apr, 20_May, 09_Jun 36. EEC09_1173 195 Clone KRL01E39 (99%) [JN090899] Cercozoa Cryothecomonas aestivalis (96%) 25_Feb, 02_Mar, 11_Mar, 16_Mar, [AF290539] 27_Apr 37. EEC09_1154 178 Clone 6c-F7 (99%) [FN690390] Cercozoa Ebria tripartita (97%) [DQ303923] 25_Feb, 02_Mar, 16_Mar, 20_May 38. EEC09_1090 177 Protomyces inouyei (99%) [AY548295] Fungi Protomyces inouyei (99%) [AY548295] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 20_May 39. EEC09_1142 156 Saprolegnia sp. (89%) [FJ794909] Oomycetes Saprolegnia sp. (89%) [FJ794909] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 27_Apr 40. EEC09_0905 149 Clone 15H3Te96Z (100%) [JQ782078] Bacillariophyta Minidiscus sp. (100%) [FJ590771] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 27_Apr, 20_May 41. EEC09_1072 143 Clone 1816G03 (98%) [FR874823] Ciliophora Acineta flava (98%) [HM140400] 25_Feb, 11_Mar, 16, Mar, 20_May 42. EEC09_0491 138 Clone BT_Euk_F6 (93%) [KC306524] - - 09_Jun 43. EEC09_1064 126 Clone ANT-Roth-MECL-14 (99%) Cercozoa Protaspsis obliqua (96%) [FJ824122] All [FJ985885] 44. EEC09_1130 117 Clone SGYH1395.FRAG.MO.500m Dinophyceae Gyrodinium spirale (97%) [AB120001] 25_Feb, 02_Mar, 11_Mar, 16_Mar, (97%) [JX842244] 30_Mar, 20_May, 09_Jul 45. EEC09_0830 116 Clone CNCIII51_5 (99%) [HM581780] Labyrinthulomycetes Thraustochytriidae sp. (90%) 11_Mar, 16_Mar, 30_Mar, 07_Apr, [DQ367047] 27_Apr, 20_May 46. EEC09_0916 113 Clone SGUH612.FRAG.MO.5m (99%) Picobiliphytes Picobiliphyte sp. (99%) [JN934893] 25_Feb, 02_Mar, 11_Mar, 16, Mar, [JX841765] 07_Apr, 27_Apr, 20_May, 09_Jun 47. EEC09_0479 112 Clone Out-139 (99%) [JF971997] Dinophyceae Pelagodinium beii (99%) [JF791066] 25_Feb, 11_Mar, 07_Apr, 20_May, 09_Jun 48. EEC09_0853 111 Clone SHAA649 (99%) [JQ226560] Bacillariophyta Eunotogramma sp. (90%) [JN975245] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 30_Mar, 07_Apr, 27_Apr 49. EEC09_1068 110 Clone F5-2-24 (100%) [HM138181] Choanoflagellida Didymoeca costata (96%) [EU011923] 11_Mar, 16_Mar, 07_Apr, 27_Apr, 09_Jun 50. EEC09_0938 110 Clone EukaV4-22 (99%) [JF727608] Cercozoa Cryothecomonas longipes (99%) 25_Feb, 02_Mar, 11_Mar, 16_Mar [AF290540]</p><p>4 No. of Closest relative, (% similarity) Closest cultured relative (% similarity) OTU similar tags Putative affiliation Sample [GenBank accession No.] [GenBank accession No.] (>97%) 51. EEC09_1065 100 Clone BTQB20030806.0083 (94%) Apicomplexa Selenidium serpulae (93%) [DQ683562] 25_Feb, 02_Mar, 11_Mar, 16_Mar, [FJ221514] 07_Apr, 27_Apr, 20_May, 09_Jun 52. EEC09_0834 98 Clone T4B4_8 (99%) [HQ394102] Haptophyceae Chrysochromulina strobilus (99%) 11_Mar, 16_Mar, 27_Apr, 20_May, [FN599060] 09_Jun 53. EEC09_0504 95 Cryothecomonas aestivalis (99%) Cercozoa Cryothecomonas aestivalis (99%) 16_Mar, 20_May, 09_Jun [AF290539] [AF290539] 54. EEC09_1163 93 Clone EukaV4-12 (100%) [JF727604] Bacillariophyta Chaetoceros peruvianus (98%) 25_Feb, 02_Mar, 11_Mar, 16_Mar, [HQ912650] 30_Mar, 27_Apr 55. EEC09_1054 89 Clone JPeuk-77 (100%) [AY145111] Chlorophyta Nannochloris sp. (99%) [JQ315642] 25_Feb, 02_Mar, 11_Mar, 16_Mar, 20_May, 09_Jun 56. EEC09_1165 83 Clone NPK2_111 (94%) [EU371149] Unclassified Pirsonia verrucosa (94%) [JF791043] 25_Feb, 02_Mar, 11_Mar, 16_Mar, Stramenopiles 30_Mar, 07_Apr, 09_Jun 57. EEC09_0930 80 Clone CiliV4-240 (93%) [JF727599] Fungi Chytridiales sp. (89%) [AF164261] 25_Feb, 02_Mar 58. EEC09_0691 79 Clone DSGM-19 (99%) [AB275019] Dinophyceae Stoeckeria algicida (99%) [AJ841809] 16_Mar, 27_Apr, 09_Jun 59. EEC09_0490 76 Clone Q1-17 (99%) [JQ420086] Dinophyceae Karlodinium micrum (99%) [JF790988] 09_Jun</p><p>5 Table S2. Basic physical and chemical parameters at the 9 sampling dates (SOMLIT network, http://somlit.epoc.u- bordeaux1.fr/fr/). Water temperature (°C) and salinity (PSU) were measured using a Seabird SBE 25 Sealogger CTD, inorganic nutrients concentrations were determined with an Alliance Integral Futura Autoanalyser II [1-2]. Chlorophyll-a (Chl-a) concentrations were measured on 90% (v/v) acetone extracted particulate material isolated by filtration on GF/F glass-fibre filters (Whatman). Concentrations were determined by fluorescence using a 10-AU Turner Designs© fluorometer [3]. Biomass of micro-autrotrophs (m-auto, mainly diatoms and Phaeocystis globosa) inferred by microscopic counts [from 4]. na: not available</p><p>NO3+NO2 Silicate Phosphate N/P Temperature Salinity Chl a Biomass (µM) (µM) (µM) (°C) (µg L-1) m-auto (µg C L-1)</p><p>Feb 25th 14.72 0.70 0.45 33 5.59 33.70 11.4 225 Mar 2nd 14.01 0.40 0.25 57 6.29 33.70 22.9 351 Mar 11th 13.31 0.53 0.17 80 6.74 33.75 12.6 239 Mar 16th 11.00 0.38 0.17 71 7.70 33.51 6.8 84 Mar 30th 8.68 0.85 0.17 81 8.47 33.66 3.3 71 Apr 7th 6.31 0.90 0.19 40 8.66 34.01 2.1 66 Apr 27th 0.09 3.42 0.23 1.0 11.60 33.97 4.9 638 May 20th 0.11 0.88 0.09 1.8 13.34 33.45 5.2 na June 9th 0.12 1.05 0.08 6.3 15.15 34.19 4.9 123</p><p>6 REFERENCES</p><p>1. Aminot A, Kerouel R (2004) Dissolved organic carbon, nitrogen, and phosphorus in the N–E Atlantic and the N–W Mediterranean with particular reference to non-refractory fractions and degradation. Deep-Sea Res I 51:1975-1999 2. Strickland J, Parsons T (1972) A practical handbook of seawater analysis. Bull Fish Res Board Can 167:1- 310 3. Lorenzen CJ (1966) A method for continuous measurement of in vivo chlorophyll concentration. Deep-Sea Res I 13:223-247 4. Grattepanche J-D, Vincent D, Breton E, Christaki U (2011) Phytoplankton growth and Microzooplankton grazing during a spring bloom in the eastern English Channel. J Exp Mar Biol Ecol 404:87-97</p><p>7</p>