BG61 – EGU2011-8665

Chemosymbiotic from cold seeps around NE Atlantic

Clara F. Rodrigues 1,2 , Sébastien Duperron 2, Ana Hilário 1 and Marina R. Cunha 1

1 Centro de Estudos Ambientais e do Mar, Departamento de Biologia, Universidade de Aveiro, 3810-193 Aveiro, Portugal 2 Université Pierre et Marie Curie, UMR7138 (UPMC CNRS IRD MNHN), Systématique Adaptation, Evolution, 75052 Paris, France

 [email protected]

S Western Mediterranean - Alboran Sea ib o O g a li th n 2 : Gulf of Cadiz e id r a s e • Lucinoma sp. ? ? 13 Bivalvia (see map for distribution): • Myrtea sp. • Acharax nov. sp. (1) 2 Siboglinidae: • (Petrasma) nov sp. (2) • Lamellibrachia sp. (a) The Alboran tubeworm is essentially identical to • Lucinoma nov sp. (3) • Siboglinum sp. Lamellibrachia sp. found in the eastern Mediterranean • Thyasira vulcolutre (4) • Spinaxus sentosus (5) 1 Bacteria associated: • Isorropodon nov. sp. (6) ► Not yet studied • Isorropodon sp. indet (7) 2

• Calyptogena cf. regab (8) 3 • Bathymodiolus 4

mauritanicus (9) Sag 12 CR 5 • Idas sp. (10) 1 4 10 CA 6 1 4 JB Por 5 6 7 1 Bon 1 Yum ae • Laubericoncha chuni (11) 8 11 Kid inid 1 2 ogl 13 PDE 2 Mer ib 2 9 1 S 1 2 Dar 1 2 2 3 rs ? 2 the • Callogonia cyrili (12) Gin 7 O Mek Gem 9 • Pliocardia sp. (13) 2

15 Siboglinidae: 8 • Siboglinum spp. (6 species) Mud volcanoes (MV) with chemosymbiotic species. Bon, Bonjardim MV; CA: Captain Arutyunov MV; • Siboglinum cf. poseidoni CR, Carlos Ribeiro MV; Dar, Darwin MV; Gem, Gemini MV; Gin, Ginsburg MV; JB, Jesus Baraza MV; Kid, Kidd MV; Mek, Mèknes MV; Mer, Mercator MV; PDE, Pen Duick Escarpment; Por, Porto e • Cyclobrachia sp. lv MV; Sag, Sagres MV; Yum, Yuma MV. a ? iv rs • Bobmarleya gadensis b e 1 th • Spirobrachia tripeira O • Lamellisabela denticulata Eastern Mediterranean • Lamellisabela sp. • Polybrachia spp. 8 Bivalvia:

Lamellisabella mm 1 • NID Cf. Cyclobrachia denticulata Siboglinum sp • Lucinoma kazani Bobmarleyia gadensis Juvenil NID Spirobrachia tripeira • Myrtea sp.(a) • Myrtea amorpha • Thyasira striata

Bathymodiolus mauritanicus associated clone Darwin MV_1_2 (FM213421) 8/10 clones • Idas modiolaeformis Bacteria associated: (a) 88/- Idas sp. symbiont T1 (AM402956) 85/58 Bathymodiolus mauritanicus associated clone Darwin MV_1_1 (FM213422) 2/10 clones 50/- Bathymodiolus mauritanicus associated clone Darwin MV TTR17_1_1 (????) • Idas sp. EM a b 52/- Bathymodiolus puteoserpentis symbiont (DQ321712) 98/ Bathymodiolus azoricus symbiont (AY235676) ►Phylogenetic analysis of bacterial 16S rRNA gene sequences 51 Thiotrophic 51/64 Bathymodiolus brooksi symbiont (AM236331) • Thyasira sp. EM (b) 100/99 Vesicomya gigas symbiont (AF035726) demonstrated that most bacteria were related to known sulfide- Calyptogena elongata symbiont (AF035719) 100/97 50/ Bathymodiolus sp. symbiont (AJ745718) 64 oxidizing endosymbionts found in other deep-sea chemosynthetic ; 56/- Maorithyas hadalis symbiont I (AB042413) 1 Siboglinidae: Conchocele sp. symbiont (AJ441190) 50/- 59/- Methylobacter marinus A45 (AF304197) ►Thyasira vulcolutre: symbiont sequence clusters with vesicomyids 79/76 Methylomicrobium agile ATCC35068 (X72767) • Lamellibrachia sp. Crenothrix polyspora clone 23 (DQ295890) symbionts; 82/59 Bathymodiolus puteoserpentis symbiont (U29164) 96/75 Methanotrophic Bathymodiolus brooksi symbiont (AM236330)

98/- Bathymodiolus japonicus symbiont (AB036711) ► 93/76 Bathymodiolus mauritanicus: was shown to possess a dual Bathymodiolus sp. symbiont (AJ745717) 50/- Bathymodiolus childressi symbiont (U05595) symbiosis with thio- and methanotrophic symbionts; Bathymodiolus platifrons symbiont (AB036710) 55/- 52/- Bathymodiolus mauritanicus associated clone Darwin MV TTR17_1_2 (??????) ► Other bivalves: under study. 93/70 Thyasira flexuosa symbiont (L01575) Bacteria associated: 93/65 Solemya pusilla symbiont (U62130) Anodontia phillipiana symbiont (L25711) 76/- 97/58 Solemya terraeregina symbiont (U62131) Lucina pectinata symbiont (X84980) (a) Phylogenetic tree of bacterial 16S rRNA gene Lucina nassula symbiont (X95229) ►Lamellibrachia sp.: gammaproteobacterium closely related to other 68/- 89/71 sequences associated with the gills of some bivalve Codakia orbicularis symbiont (X84979) Lucinoma aequizonata symbiont (M99448) sulfide-oxidizing tubeworm symbionts. 98/55 100/ species from the Gulf of Cadiz. Minimum evolution trees 86 Lucinoma kazani symbiont (AM236336) Lucinoma annulata symbiont (M99449) 78/- ► Myrtea sp. and Thyasira sp.EM: gammaproteobacterial endosymbionts; the Myrtea- derived from LogDet distance analysis. Lucinoma sp. associated clone Mercator MV_1_1 (FM213432) 12/14 clones 68/- Lucina floridana symbiont (L25707) (b) FISH hybridization on T. vulcolutre gill filaments with the Codakia costata symbiont (L25712) associated bacterium is closely related to lucinid symbionts from both deep-sea and 99/88 Lucina nassula associated clone CK_G1_5 (EU487789) ThyGoC193 probe. No signal was observed from the Thiotrophic 51/- Beggiatoa alba ATCC33555 (AF110274) coastal species, whereas the Thyasira -associated bacterium is closely related to the ciliated zone, whereas all bacteriocytes from the lateral Thioploca ingrica (L40998) Captain Arutyunov MV sediment clone CpA BacB18 (FN397816) Acharax sp. ‘Oregon 56’ symbiont (AJ441197) symbiont of a T. flexuosa from coastal waters off the U.K. zone were fully hybridized (in green), indicating 100/100 Acharax sp. associated clone Yuma MV_1_1 (FM213423) 8/8 clones 78/- bacteriocytes filled with bacteria. Nuclei of thyasirid Acharax sp. associated clone Ginsburg MV_1_1 (FM213424) 10/10 clones ► Solemyidae: no bacteria found, but it was a small single specimen 96/- Acharax sp. ‘Pakistan’ symbiont (AJ441188) cells, which are counterstained with DAPI, appear blue Thiobacillus prosperus DSM5130 (AY034139) 100/99 51/- Solemya occidentalis symbiont (U41049) Petrasma sp. associated clone Yuma MV_8 _1 (FM213425) 2/2 clones Figure 4 Rodrigues et al 61/- 100 Solemya velum symbiont (M90415) C Siboglinum sp. 4 (Mek) associated clone W15_8 (4/9 clones) 100/100 63 Captain Arutyunov MV sediment clone CpA BacB37 (FN397817) 51/- Petrasma sp. associated clone Mercator MV_1_1 (FM213426) 13/15 clones Siboglinum sp. 5 (Gem) associated clone W16_8 (6/12 clones) Petrasma sp. associated clone Gemini MV_1_1 (FM213435) 10/10 clones 69 Spirobrachia tripeira (Por3) associated clone W4_1 (10/10 clones) Oligobrachia haakonmosbiensis symbiont clone Ohaa2 (AM883179) Thiorhodovibrio winogradskyi MBIC2776T (AB016986) Rainbow hydrothermal vent sediment clone pIR3BC07 (AY354137) Frenulate 98 Siboglinum sp. 2 (Dar1) associated clone W13_1 (11/16 clones) 55/60 Solemya reidi symbiont (L07864) 70 Gulf of Cadiz sediment clone GoC_Bac_97_D1_C0_M0 (FJ813578) associated Oligobrachia haakonmosbiensis symbiont clone Ohaa1(AM883178) Bobmarleya gadensis (Car) associated clone W17_4 (11/11 clones) bacteria 0.02 Gulf of Guinea 100 90 94 Lamellisabella denitculata (Por9) associated clone W2_1 (10/10 clones) 64 Barents Sea sediment clone 4 2B 50 (FJ800278) Siboglinum fiordicum symbiont clone 12_2 (EU086761) 70 100 Siboglinum fiordicum clone 16_3 (EU086765) 73 Oligobrachia mashikoi symbiont E (AB271123) 100 Oligobrachia mashikoi symbiont (AB057751) 88 Kazan MV sediment clone KZNMV-10-B65 (FJ712538) 100 Lucina floridana symbiont clone BJS72-017 (AB238979) Thiotrophic Leucothrix mucor DSM2157 (X87277) 68 14 Bivalvia: Bivalvia (cont.): Hydrothermal vent gastropod associated clone SF_C7-H11 (AY531559) ► Siboglinidae: Phylogenetic analyses indicate that the 93 99 Rimicaris exoculata ectosymbiont clone LOG_272/6-9_D1( FN393024) Hydrothermal Rimicaris sp. ectosymbiont clone SMAR_146/6-23_A10 (FM203403) 100 Japan Trench seep sediment clone CS5.30 (AB069800c) 100 vent primary symbiont in most cases belongs to the Undetermined genus sp. (Por5) associated clone W11_2 (10/10 clones) • Acharax sp. • Wareniconcha guinensis 98 Shinkaia crosnieri associated clone HAT0_581 (AB476178) associated 100 Hydrothermal vent chimney clone Ba9 (FJ640825) 54 Shinkaia crosnieri associated clone 1232 (AB440173) bacteria Gammaproteobacteria and were related to thiotrophic and 76 Shinkaia crosnieri associated clone B16 (EU107481) 100 Solemya velum symbiont (M90415) • Elenaconcha guiness 82 • Lucinoma myriamae Petrasma sp. associated clone Yuma MV_8 _1 (FM213425) Thiobacillus prosperus DSM5130 (AY034139) methanotrophic symbionts from other marine invertebrates, 100 Escarpia spicata symbiont Alvin #2837 (AF165909) 55 Sclerolinum contortum symbiont clone Scon (AM883183) 91 Tevnia jerichonana symbiont (AY129118) • Graecina karinae • Laubericoncha chuni 100 whereas members of the microflora were related to multiple Riftia pachyptila symbiont (AY129115) 92 Ridgeia piscesae symbiont (U77480) 100 Lucinoma sp. associated clone Mercator MV_1_1 (FM213432) 80 Lucinoma kazani symbiont (AM236336) bacterial phyla. 95 • Joellina dosiniformis • Bathymodiolus aff. boomerang Loripes lacteus symbiont clone 2C (FJ752447) Codakia orbicularis symbiont (X84979) Thiotrophic Thyasira flexuosa symbiont (L01575) Lamellibrachia cf. luymesi symbiont (AY129100) ► This is the first molecular evidence of methanotrophic Lamellibrachia sp. 1 symbiont (AY129112)1 100 • Thyasira sp. nov sp. 2 Siboglinidae: Lamellibrachia sp. symbiont clone V11.1 (FM165438) Lamellibrachia columna symbiont (U77481) bacteria in at least one frenulate species. Lamellibrachia sp. symbiont (U77479) 75 Lamellibrachia sp. symbiont (DQ232902) Escarpia laminata symbiont (AY129107) 54 • Isorropodon bigoti • Lamellibrachia spp. siboglinid associated clone 1 (AM888202) ► No selection for specific symbionts - environmental 99 Bathymodiolus japonicus symbiont (AB036711) Bathymodiolus puteoserpentis symbiont (U29164) 89 Methylobacter marinus A45 (AF304197) acquisition as previously proposed for this group of 97 Methylomicrobium agile ATCC35068 (X72767) • Isorropodon striatum 100 Methylomonas methanica (AF150806) Methanotrophic 74 Methylomonas rubra (AF150807) 75 Guaymas Basin hydrothermal plume clone1E-037 (FJ980791) siboglinids 75 Siboglinum cf. poseidoni (Cap2) associated clone W10 _91 (9/9 clones) 96 Kazan MV sediment clone KZNMV-0-B15 (FJ712410) • Isorropodon atalante 96 Acharax sp. ‘Pakistan’ symbiont (AJ441188) Acharax sp. associated clone Ginsburg MV_1_1 (FM213424) 100 Acharax sp. associated clone Yuma MV_1_1 (FM213423) Captain Arutyunov MV sediment clone CpA BacB18 (FN397816) Thiotrophic 99 Siboglinum sp. 1 (Cap1) associated clone W9_14 (2/11 clones) • Calyptogena valdiviae Thioploca ingrica (L40998) 81 Pogonophora sp. JT-1 symbiont (AB070215) Olavius crassitunicatus symbiont clone g2-P13-I-41 (AJ620508) 100 Methylophaga marina DSM 5989 (X95459) • Calyptogena regab Methylophaga thalassica DSM5690 (X95460) 99 Arctic sediment clone S11-106 (EU287289) 98 71 13 C-methane enrichment clone Methane SIP2-4-23 (GU584312) Siboglinum sp. 1 (Cap1) associated clone W9_1 (8/11 clones) Methylotrophic 100 Idas sp. symbiont clone M2.41 (AM402960) 74 Bathymodiolus hirtus symbiont (AB250699) 54 Bathymodiolus heckerae symbiont clone M12.2 (AM236326) 71 Acinetobacter lwoffii HJ7 (AY176770) 96 100 Daqing oilfield water clone DQB-T10 (GQ415369 ) 100 Siboglinum sp. 4 (Mek) associated clone W15_10 (1/9 clone) Heterotrophic Contaminated groundwater clone 300A-B09 (AY661997) Lamellibrachia sp. symbiont TW-2 (AB042404) Beggiatoa sp. 402 (AY583996) After Krylova & Sahling 2010; Oliver et al . submitted 100 Beggiatoa alba ATCC33555 (AF110274) Thiotrophic Thiovirga sulfuroxydans S007 (AB118236) 100 Photobacterium profundum 3TCK (DQ027054) 100 Siboglinum sp. 3 (Gin) associated clone W12_3 (1/10 clones) Photobacterium indicum (AY771742) (c) Phylogenetic tree showing the relationship of the Photobacterium phosphoreum ATCC11040 (X74687) Siboglinum sp. 3 (Gin) associated clone W12_4 (1/10 clones) Heterotrophic Vestimentiferan symbiont TW-6 (AB042418) Gammaproteobacteria 16S rRNA gene sequences recovered from 100 Bacteria associated: 97 Pseudomonas fluorescens ATCC17386 (AF094726) 79 Pseudomonas congelans DSM14939 (AJ492828) several species of frenulates from the Gulf of Cadiz. The tree was Inanidrilus leukodermatus symbiont 1 (AJ890100) Allochromatium vinosum (M26629) 0.02 91 80 Thiorhodovibrio winogradskyi MBIC2776T (AB016986) Thiotrophic obtained using Mimimum Evolution and LogDet. s ►Thyasira nov sp.: gammaproteobacterium related to Maorithyas hadalis symbiont II, but e lv ? both are related to environmental sequences and branch far from any other known a v rs i e symbionts; b h 2 t ► Calyptogena regab and Laubericoncha chuni : see Decker et al . (Poster BG66 ACKNOWLEDGMENTS O •Thanks are due to Dr. Graham Oliver (National Museum of Wales, Cardiff), Dr. Gordon Webster and Prof. Andrew J. Weightman EGU2011-11570); (Cardiff University), with whom some of the work as been done. ► Other taxa: under study. * We also thank Carole Decker (IFREMER) for carry this poster. * CFR is supported by the FCT grant (SFRH/BPD/64154/2009) and AH is supported by the FCT grant SFRH/BPD/22383/2005