Fodinicurvata halophila sp. nov., a moderately halophilic bacterium from a marine saltern

Depósi o de in es igación de la Uni e sidad de Se illa h ps://idus.us.es/ “This is an Accep ed Manusc ip o an a icle published in In e na ional Jou nal o Sys ema ic and E olu iona y Mic obiology on 01 Ma ch 2015, a ailable a : h ps://doi.o g/10.1099/ijs.0.000010 .” 1 Fodinicu a a halophila sp. no ., a mode a ely halophilic bac e ium om a ma ine 1 sal e n 2 3 Ca men In an e-Dominguez1, Paul A. Lawson2, C ys al N. Johnson2, C is ina Sánchez- 4 Po o1 and An onio Ven osa1 5 6 1Depa men o Mic obiology and Pa asi ology, Facul y o Pha macy, Uni e si y o 7 Se illa, 41012 Se illa, Spain. 8 2Depa men o Mic obiology and Plan Biology, Uni e si y o Oklahoma, No man, 9 Oklahoma, 73019 USA. 10 11 Running i le: Fodinicu a a halophila sp. no . 12 Subjec ca ego y: New axa, P o eobac e ia 13 14 Au ho o co espondence: An onio Ven osa, Depa men o Mic obiology and 15 Pa asi ology, Facul y o Pha macy, Uni e si y o Se illa, 41012 Se illa, Spain. E.-mail: 16 en [email p o ec ed]s 17 18 The GenBank/EMBL/DDBJ accession numbe o he 16S RNA gene sequence o 19 s ain BA45ALT is HG764424. 20 21 Two supplemen a y igu es and a supplemen a y able a e a ailable wi h he online 22 e sion o his pape . 23 24 Manusc ip Including Re e ences (Wo d documen ) Click he e o download Manusc ip Including Re e ences (Wo d documen ): Fodinicu a a halophila- e .doc 2 A G am-s ain-nega i e, od-shaped, acul a i ely anae obic, mode a ely halophilic 25 bac e ium, designa ed s ain BA45ALT, was isola ed om wa e o a sal e n o 26 San a Pola, Alican e, Spain. Cells we e mo ile, ca alase and oxidase posi i e. S ain 27 BA45ALT g ew a empe a u es in he ange 14-45 ºC (op imally a 37 ºC) and a 28 pH 5.0-9.0 (op imally a pH 7.5). I g ew in media con aining 5-20 % (w/ ) sal s 29 and op imally in media con aining 10 % (w/ ) sal s. Phylogene ic analysis based 30 on he compa ison o 16S RNA gene sequences e ealed ha s ain BA45ALT is a 31 membe o he genus Fodinicu a a. The closes ela i es o his s ain we e 32 Fodinicu a a enggangensis YIM D812T and Fodinicu a a sediminis YIM D82T 33 wi h sequence simila i ies o 98.2 % and 97.4 %, espec i ely. DNA-DNA 34 hyb idiza ion be ween he no el isola e and hese phylogene ically ela ed species 35 e ealed ela edness o 15 % and 30 %, espec i ely, wi h espec o he 36 a o emen ioned species. The majo cellula a y acids o s ain BA45ALT we e 37 C18:1ω7c, C16:0 and iso-C15:0. The G+C con en o he genomic DNA o his s ain 38 was 58.0 mol%. I s pola lipid pa e n consis ed o diphospha idylglyce ol, 39 phospha idylme hyle hanolamine and a numbe o unknown phospholipids and 40 lipids. Based on he pheno ypic, chemo axonomic and phylogene ic da a p esen ed 41 in his s udy s ain BA45ALT cons i u ed a no el species o he genus 42 Fodinicu a a, o which he name Fodinicu a a halophila sp. no . is sugges ed. 43 The ype s ain o Fodinicu a a halophila is s ain BA45ALT (= CCM 8504T = 44 CECT 8472T = JCM 19075T = LMG 27945T). 45 46 47 3 The genus Fodinicu a a belongs o he amily Rhodospi illaceae, wi hin he o de 48 Rhodospi illales (P ening & T üpe , 1971) o he class Alphap o eobac e ia. This genus 49 was o iginally p oposed by Wang e al. (2009) and o da e i comp ises wo species, 50 Fodinicu a a sediminis (Wang e al., 2009) and Fodinicu a a enggangensis (Wang e 51 al., 2009). Bo h species we e isola ed om a sedimen sample collec ed om a sal 52 mine in Yunnan, sou h-wes China. The species o he genus Fodinicu a a s ain G am- 53 nega i e, a e acul a i ely anae obic, non-mo ile, ib ioid o od-shaped. Ca alase and 54 oxidase a e p oduced and polyhyd oxybu y a e (PHB) g anules a e obse ed. The 55 p edominan pola lipids consis o diphospha idylglyce ol, 56 phosphai idylme hyle hanolamine and phospha idylcholine. Phospa idylinosi ol is 57 a iable be ween species. Thei DNA G+C con en anges om 61.5 – 62.3 mol% 58 (Wang e al., 2009). 59 Recen me agenomic s udies ha e p o ided new insigh s in o he mic obial di e si y 60 along he salini y g adien o sal e ns loca ed in San a Pola and Isla C is ina in Spain 61 (Ghai e al., 2011; Fe nández e al., 2014 a,b; León e al., 2014; Ven osa e al., 2014). 62 Du ing he cou se o a s udy o sea ch o he mos abundan mic oo ganisms in hese 63 sal e ns, a no el, mode a ely halophilic, yellow o c eam pigmen ed, G am-s ain- 64 nega i e bac e ium, s ain BA45ALT, was isola ed om a wa e sample. The pH o he 65 wa e was 7.4 and i s salini y was 12 % (w/ ) sal s. A he ime o sampling, he 66 empe a u e o he wa e was 28 ºC. In his pape we desc ibe he axonomic ea u es o 67 his no el bac e ium and p opose i as a new species o he genus Fodinicu a a, as 68 Fodinicu a a halophila sp. no . 69 Aliquo s o he wa e sample we e dilu ed wi h s e ile sal solu ion, whe e 100 70 mic oli e s was pla ed on saline media and incuba ed ae obically a 37º C. The isola ion 71 4 medium used was p epa ed wi h a 15% (g l-1) sal s mix u e: NaCl, 117; MgCl2.6H2O, 72 19.5; MgSO4.7H2O, 30.5; CaCl2.2H2O, 0.3; KCl, 3; NaHCO3, 0.06; NaB , 0.23 (Subow, 73 1931), supplemen ed wi h 0.25 % (w/ ) yeas ex ac (Di co) and 1. 8 % (w/ ) aga 74 (Di co). The pH o he medium was adjus ed o 7.5 wi h 1 M KOH. A pu e cul u e o 75 s ain BA45ALT was ob ained a e se e al ans e s on he same medium. Fo ou ine 76 g ow h, he s ain was cul i a ed in he same medium bu p epa ed wi h 10 % (w/ ) 77 o al sal s. The cul u e was main ained a -80 ºC in he ou ine medium con aining 50 % 78 ( / ) glyce ol. Fodinicu a a enggangensis DSM 21160T and Fodinicu a a sediminis 79 DSM 21159T ob ained om he DSMZ (Ge man Collec ion o Mic oo ganisms and Cell 80 Cul u es) we e used as e e ence s ains and we e g own ollowing he 81 ecommenda ions o he cul u e collec ion. 82 Cell mo phology and mo ili y we e examined using an Olympus CX41 mic oscope 83 equipped wi h phase-con as op ics. Cells we e mo ile, bo h od- and ib ioid-shaped 84 (Supplemen a y Fig. S1, a ailable a IJSEM online). Colony mo phology was obse ed 85 on ou ine medium unde op imal g ow h condi ions a e incuba ion a 37º C o 5 86 days. Colonies o s ain BA45ALT ha o med on aga pla es we e ci cula , smoo h, 87 en i e, opaque, yellow-c eam pigmen ed, wi h a size o 0.3-0.5 mm. The s ain was able 88 o accumula e PHB, as was obse ed by using an Olympus CX41 mic oscope equipped 89 wi h phase-con as op ics. 90 Genomic DNA om s ain BA45ALT was ob ained using he me hod desc ibed by 91 Ma mu (1961). The 16S RNA gene o he isola e we e ampli ied by PCR using 92 o wa d p ime 16F27 and e e se p ime 16R1488 (Mellado e al., 1995). An almos 93 comple e 16S RNA gene sequence (1392 n ) o s ain BA45ALT was ob ained. The 94 iden i ica ion o he closes phylogene ic neighbou s and calcula ion o pai wise 16S 95 5 RNA gene sequence simila i y we e achie ed using he EzTaxon-e ool (Kim e al., 96 2012). The 16S RNA gene sequence simila i ies be ween BA45ALT and he mos 97 closely ela ed Fodinicu a a ype s ains, Fodinicu a a enggangensis YIM D812T 98 and Fodinicu a a sediminis YIM D82T we e 98.2 and 97.4 %, espec i ely. The 16S 99 RNA gene sequence was aligned and checked agains bo h p ima y and seconda y 100 s uc u es o he 16S RNA molecule using he alignmen ool o he ARB so wa e 101 package. Phylogene ic ees we e cons uc ed using h ee di e en me hods, namely he 102 maximum-likelihood (Felsens ein, 1981), maximum-pa simony (Fi ch, 1971) and 103 neighbou -joining (Sai ou & Nei, 1987) algo i hms in eg a ed in he ARB so wa e. The 104 maximum-likelihood phylogene ic ee is p esen ed in Fig. 1. Simila ee opologies 105 we e ob ained using o he eeing me hods: neighbou -joining and maximum-pa simony 106 (Fig. S2) 107 Al hough no p ecise co ela ion exis s be ween he pe cen age o 16S RNA sequence 108 di e gence and species delinea ion, i is gene ally ecognized ha di e gence alues o 109 1.3% o mo e a e signi ican (S ackeb and & Goebel, 1994). Howe e , due o he high 110 sequence simila i y alues o i s close ela i es, DNA-DNA hyb idiza ion s udies we e 111 pe o med. These s udies we e ca ied ou using he compe i ion p ocedu e o he 112 memb ane me hod (Johnson, 1994) as desc ibed by Ven osa e al. (2004). The le els o 113 DNA-DNA ela edness o s ain BA45ALT wi h espec o Fodinicu a a 114 enggangensis DSM 21160T and Fodinicu a a sediminis DSM 21159T we e 30 and 15 115 %, espec i ely. These da a con i med ha s ain BA45ALT ep esen s a no el species 116 o he genus Fodinicu a a, ha ing DNA-DNA hyb idiza ion alues <70 % wi h espec 117 o he ype s ains o he mos closely ela ed species o he genus Fodinicu a a (Waine 118 e al., 1987; S ackeb and & Goebel, 1994). 119 6 Op imal condi ions o g ow h and ange we e de e mined by g owing he s ain in he 120 ou ine medium p epa ed a he ollowing salini y alues: 0.5, 3, 5, 7, 7.5, 8, 10, 12, 15, 121 20 and 25% (w/ ) o al sal s and a empe a u es om 5 o 45 ºC (in inc emen s o 5 ºC) 122 and om 35 o 40 ºC in inc emen s o one uni . The pH ange o isola e was 123 de e mined in he ou ine medium a op imal salini y adjus ed o pH alues anging 124 om 4 o 10 (in inc emen s o one pH uni ). The g ow h was de e mined by moni o ing 125 he op ical densi y using a spec opho ome e a 600 nm. 126 S ain BA45ALT g ew in media con aining 5-20 % (w/ ) sal s and op imally in media 127 con aining 10 % (w/ ) sal s. No g ow h was obse ed in he absence o NaCl. I g ew a 128 empe a u es in he ange 14-45 ºC (op imally a 37 ºC) and a pH 5.0-9.0 (op imally a 129 pH 7.5). The s ain was able o g ow anae obically, de e mined a e incuba ion in an 130 anae obic chambe (GasPak Anae obic sys em, BBL). 131 All pheno ypic ea u es we e ca ied ou g owing he s ain in he ou ine medium 132 p epa ed a 10% (w/ ) o al sal s, pH 7.5 and a 37 ºC. The p oduc ion o acid om 133 di e en ca bohyd a es was es ed in a medium wi h 0.5 % (w/ ) yeas ex ac and 134 supplemen ed wi h 1% (w/ ) o he ca bohyd a e es ed (s e ilized sepa a ely) (Ven osa 135 e al., 1982). To de e mine he u iliza ion o di e en o ganic subs a es as ca bon and 136 ene gy sou ces, a medium con aining 0.05 % (w/ ) yeas ex ac and supplemen ed wi h 137 1 % (w/ ) o he es ed subs a e, was used (Ven osa e al., 1982). The esul s o he 138 u iliza ion o di e en subs a es a e included in he species desc ip ion. 139 Ca alase ac i i y was de e mined by adding a 1% (w/ ) H2O2 solu ion o colonies on 140 solid medium. The oxidase es was pe o med using a D ySlide assay (Di co). S ain 141 BA45ALT was ca alase and oxidase posi i e. The ollowing es s we e conduc ed as 142 desc ibed by Cowan and S eel (1977): hyd olysis o aesculin, casein, gela in, Tween 80, 143 7 and s a ch; he u ease, me hyl ed, and indole p oduc ion, Vogues-P oskaue es ; ni a e 144 and ni i e educ ion, and Simmons ci a e. The ollowing es s we e nega i e: me hyl 145 ed, Voges-P oskaue , indole p oduc ion om yp one and Simmons’s ci a e, casein 146 and s a ch hyd olysis. The s ain was posi i e o ni a e educ ion bu ni i e was no 147 educed. Bo h gela in and aesculin we e hyd olyzed and he u ease es was posi i e. 148 O he pheno ypic cha ac e is ics o s ain BA45ALT a e included in he species 149 desc ip ion. 150 An imic obial compounds sensi i i y es s we e pe o med by sp eading he cul u e 151 suspension on solid medium pla es and applying discs imp egna ed wi h an imic obial 152 compounds ollowing he echnique o Baue -Ki by (Baue e al., 1966). Suscep ibili y 153 o an imic obial compounds was de e mined on aga pla es using an imic obial agen 154 discs wi h he ollowing concen a ions: ampicillin (10 µg), chlo amphenicol (30 µg), 155 e y h omycin (15 µg), gen amicin (10 µg), nalidixic acid (30 µg), neomycin (10 µg), 156 no obiocin (30 µg), penicillin G (10 U) and i ampicin (30 µg). S ain BA45ALT was 157 esis an o ampicillin, e y h omycin, gen amicin, nalidixic acid, neomycin, no obiocin, 158 penicillin G, and i ampicin bu suscep ible o chlo amphenicol. 159 Se e al pheno ypic di e ences we e obse ed be ween s ain BA45ALT and he wo 160 closely ela ed Fodinicu a a species, including hose conce ning mo ili y, 161 physiological and cul u al ea u es, ni a e educ ion, hyd olysis o gela in and aesculin, 162 H2S p oduc ion, and u iliza ion o se e al ca bohyd a es (Table 1). 163 The G+C con en o he genomic DNA o s ain BA45ALT was de e mined om he 164 mid-poin alue (Tm) o he he mal dena u a ion p o ile (Ma mu & Do y, 1962), 165 ob ained wi h a Pe kin-Elme UV-Vis Lambda 20 spec opho ome e a 260 nm. The 166 8 DNA G+C con en o s ain BA45ALT was 58.0 mol%, a alue close o hose epo ed 167 o species o he genus Fodinicu a a (Wang e al., 2009). 168 Cell biomass o a y acid, pola lipid analyses and quinone composi ion o s ain 169 BA45ALT, Fodinicu a a sediminis DSM 21159T and Fodinicu a a enggangensis 170 DSM 21160T was ob ained by cul i a ion in nu ien aga (NA) supplemen ed wi h 5 % 171 NaCl, incuba ed a 28 ºC o 5 days. The whole-cell a y acid composi ion o s ain 172 BA45ALT and he e e ence s ains was de e mined on cul u es eaching he la e- 173 exponen ial s age o g ow h acco ding o he ou quad an s s eak me hod (Sasse , 174 1990). Fa y acid me hyl es e s we e p epa ed and analysed acco ding o he s anda d 175 p o ocol o he Mic obial Iden i ica ion Sys em (MIDI, Ve sion 6.1; Iden i ica ion 176 Lib a y TSBA40 4.1; Mic obial ID). Ex ac s we e analyzed using a Hewle Packa d 177 model HP6890A gas ch oma og aph equipped wi h a lame-ioniza ion de ec o as 178 desc ibed by Kämp e & K oppens ed (1996). The a y acid p o iles o s ain 179 BA45ALT included C18:1 ω7c (35.6 %), iso-C16:0 (16.3 %) and iso-C15:0 (10.1 %) as he 180 majo a y acids, ollowed by iso-C17:1 ω9c (5.8 %), C17:0 (5.5 %), C16:0 (3.3 %), C18:1 181 2OH (3 %). The a y acid p o ile o s ain BA45ALT was clea ly di e en om hose o 182 he ela ed axa. Al hough all h ee species p oduce C18:1 ω7c as a majo p oduc , he 183 no el o ganism also p oduces majo amoun s o iso-C16:0 and iso-C15:0 which a e lacking 184 in Fodinicu a a sediminis DSM 21159T and Fodinicu a a enggangensis DSM 185 21160T (Table S1). 186 The pola lipids o s ain BA45ALT and he e e ence s ains we e analyzed as desc ibed 187 by G o h e al. (1996). The pola lipids de ec ed in s ain BA45ALT we e 188 diphospha idylglyce ol, phospha idylme hyle hanolamine and a numbe o unknown 189 phospholipids and lipids (Supplemen a y Figu e S2). I is impo an o no e ha in ou 190 15 Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. & Higgins, D. J. 312 (1997). The clus al X windows in e ace: Flexible s a egies o mul iple sequence 313 alignmen aided by quali y analysis ools. Nucleic Acids Res 24, 4876–4882. 314 Tindall, B. J., Rosselló-Mó a, R., Busse, H.-J., Ludwig, W. & Kämp e , P. (2010). 315 No es on he cha ac e iza ion o p oka yo e s ains o axonomic pu poses. In J Sys 316 E ol Mic obiol 60, 249–266. 317 Tindall, B. J., Siko ski, J., Smibe , R. A. & K ieg, N. R. (2007). Pheno ypic 318 cha ac e iza ion and he p inciples o compa a i e sys ema ics. In Me hods o Gene al 319 and Molecula Mic obiology, pp. 330-393. Edi ed by C. A. Reddy, T. J. Be e idge, J. 320 A. B eznak, G. A. Ma zlu , T. M. Schmid and R. L. Snyde . Washing on, D. C.: ASM 321 P ess. 322 To eblanca, M., Rod iguez-Vale a, F., Juez, G., Ven osa, A.,Kameku a, M. & 323 Ka es, M. (1986). Classi ica ion o non-alkaliphilic halobac e ia based on nume ical 324 axonomy and pola lipid composi ion, and desc ip ion o Haloa cula gen. no . and 325 Halo e ax gen. no . Sys Appl Mic obiol 8, 89–99. 326 Ven osa, A., Quesada, E., Rod íguez-Vale a, F., Ruiz-Be aque o, F. & Ramos- 327 Co menzana, A. (1982). Nume ical axonomy o mode a ely halophilic G am-nega i e 328 ods. J Gen Mic obiol 128, 1959–1968. 329 Ven osa, A., Gu ié ez, M. C., Kameku a, M., Z yagin se a, I. S. & O en, A. 330 (2004). Taxonomic s udy o Halo ub um dis ibu um and p oposal o Halo ub um 331 e es e sp. no . In J Sys E ol Mic obiol 54, 389–392. 332 16 Ven osa, A., Fe nández, A. B., León, M. J., Sánchez-Po o, C. & Rod iguez- 333 Vale a, F. (2014). The San a Pola sal e n as a model o s udying he mic obio a o 334 hype saline en i onmen s. Ex emophiles 18, 811-824. 335 Wang, Y. X., Liu, J.-H., Zhang, X. X., Chen, Y.-G., Wang, Z. G., Chen, Y., Li, Q. 336 Y., Peng, Q. & Cui, X. L. (2009). Fodinicu a a sediminis gen. no ., sp. no . and 337 Fodinicu a a enggangensis sp. no ., poly-ß-hyd oxybu y a e-p oducing bac e ia in 338 he amily Rhodospi illaceae. In J Sys E ol Mic obiol 59, 2575–2581. 339 Wayne, L. G., B enne , D. J., Colwell, R. R., G imon , P. A. D., Kandle , O., 340 K iche sky, M. I., Moo e, L. H., Moo e, W. E. C., Mu ay, R. G. E., S ackeb and , 341 E., S a , M. P. & T üpe , H. G. (1987). Repo o he ad hoc commi ee on 342 econcilia ion o app oaches o bac e ial sys ema ics. In J Sys Bac e iol 37, 463-464. 343 17 Table 1. Cha ac e is ics ha di e en ia e s ain BA45ALT om ela ed species o he 344 genus Fodinicu a a. 345 Taxa: 1, s ain BA45ALT; 2, Fodinicu a a enggangensis DSM 21160T; 3, 346 Fodinicu a a sediminis DSM 21159T. All da a a e om he p esen s udy, excep 347 whe e indica ed. +, Posi i e eac ion; -, nega i e eac ion. 348 Cha ac e is ic 1 2 3 Colony pigmen a ion Yellow-c eam C eam-whi e C eam-whi e Cell size (wid h x leng h; µm) 0.05-0.1 x 0.2-1.5 0.2-04 x 0.5-1.3 0.3-0.5 x 0.7-1.5 Mo ili y + - - Tempe a u e ange (ºC) 14-45 15-42 15-42 NaCl ange (%, w/ ) 5-20 1.5-20 1.5-20 pH ange 5-9 6.5-8.5 6.5-8.5 Ni a e educ ion + - - Hyd olysis o : Aesculin Gela in + + - - + - U iliza ion o ca bon sou ces: F uc ose Lac ose Fuma a e Hippu a e L-o ni hine + + + + + - - - - - - - - - - Simmons ci a e - + + H2S p oduc ion + - - Majo a y acids C18:1 ω7c, iso-C16:0 and iso- C15:0 C18:1 ω7c, 2-OH-C18:1 and C16:0 C18:1 ω7c, 2-OH-C18:1 and C16:0 Pola lipids Diphospha idylglyce ol, phospha idylme hyle hanola mine and unknown phospholipids and lipids Diphospha idylglyce ol, phospha idylme hyle hanola mine and ou unknown phospholipids and wo unknown lipids Diphospha idylglyce ol, phospha idylme hyle hanol a mine and ou unknown phospholipids and wo unknown lipids DNA G+C con en (mol%) 58.0 62.3 61.5 Sou ce Sal wa e Deposi o sal mine* Deposi o sal mine* *Da a om Wang e al. (2009). 349 350 351 18 352 Legend o igu e 353 354 Fig. 1. Maximum-likelihood phylogene ic ee based on 16S RNA gene sequences, 355 showing he ela ionships be ween s ain BA45ALT and ep esen a i es o he amily 356 Rhodospi illaceae. Boo s ap pe cen ages (based on 1000 eplica ions) ≥ 70 % a e 357 shown a b anch poin s. Ba ep esen s 0.1 subs i u ions pe nucleo ide posi ion. 358 Azospi illum melinis TMCY 0552T (DQ022958) Azospi illum la gimobile ACM 2041T (X90759) Azospi illum o yzae COC8T (AB185396) Azospi illum zeae N7T (DQ682470) Azospi illum lipo e um ATCC 29707T (M59061) Azospi illum hiophilum BV-sT ( EU678791) Azospi illum canadense DS2T (DQ393891) Azospi illum ugosum IMMIB AFH-6T (AM419042) Azospi illum b asilense NCIMB 11860T (Z29617) Azospi illum halop ae e ens Au4T (X79731) Rhodocis a cen ena ia ATCC 43720T (D12701) Rhodocis a pekingensis 3-pT (AF523824) Ske manella pa ooensis ACM 2042T (X90760) Inquilinus ginsengisoli Gsoil 080T (AB245352) Inquilinus limosus AU476T (AY043374) Thalassobaculum li o eum CL-GR58T (EF203900) Phaeospi illum molischianum ATCC 14031T (M59067) Phaeospi illum ul um NCIMB 11762T (D14433) Phaeospi illum chand amohanii JA145T (AM779061) Magne ospi illum g yphiswaldense MSR-1T (Y10109) Magne ospi illum magne o ac icum DSM 3856T (Y10110) Telma ospi illum sibe iense 26-4b1T (AF524863) De lu iicoccus anus Ben 114T (AF179678) Tis ella mobilis IAM 14872T (AB071665) Fodinicu a a sediminis YIM D82T (FJ357426) Fodinicu a a enggangensis YIM D812T (FJ357427) Rhodo ib io salina um NCIMB 2243T (D14432) Rhodo ib io sodomensis DS1T (M59072) Caenispi illum bisanense K92T (EF100694) Roseospi illum pa um 930IT (AJ011919) Roseospi a mediosalina BN 280T (AJ000989) Roseospi a ma ina CE2105T (AJ298879) Roseospi a goensis JA135T (AM283537) Roseospi a isakhapa namensis JA131T (AM282560) Rhodospi a uepe i ATCC 70022T (AJ001276) Rhodospi illum pho ome icum DSM 122T (AJ222662) Rhodospi illum ub um ATCC 11170T (CP000230) Thalassospi a lucen ensis QMT2T (AF358664) Esche ichia coli ATCC 11775T (X80725) 0.10 Fodinicu a a halophila BA45ALT (HG764424) 97 100 97 100 100 100 99 96 98 100 99 100 93 100 96 100 100 77 78 88 72 81 78 Figu e 1 Click he e o download Figu e: Fig. 1.pp Supplemen a y Ma e ial File Click he e o download Supplemen a y Ma e ial Files: Suppl Ma e ial Fodinicu a a halophila.pd