Article Title: Altererythrobacter Oceanensis Sp. Nov., Isolated from the Western Pacific

Article title: Altererythrobacter oceanensis sp. nov., isolated from the Western Pacific

Journal name: Antonie van Leeuwenhoek

Author names: Yanliu Yang1, Gaiyun Zhang1*, Zhilei Sun2, Man Kit Cheung3, Cheney Huang3

Affiliation: 1 Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, Xiamen 361005, P.R. China

2 Key Laboratory of Ministry of Land and Resources for Marine oil gas Resources and Environmental Geology, Qingdao Institute of Marine Geology, Qingdao 266071, P.R.China

3 School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong SAR 852000, China

* Corresponding author: Gaiyun Zhang.

Tel: +86-592-2195833

Fax: +86-592-2085376

E-mail:

Fig.S1 Maximum-likelihood phylogenetic tree based on the secondary structure information of 16S rRNA gene sequences showing relationships between strain Y2T and E. jejuensis CNU001T and related representatives of the family Erythrobacteraceae. Rhodospirillum rubrum ATCC 11170T and Acetobacter aceti DSM 3508T served as outgroups. Bootstrap values >50 % based on 1000 replications are shown at nods. Bar, 0.02 substitutions per nucleotide position.

Fig.S2 Minimum-evolution phylogenetic tree based on the secondary structure information of 16S rRNA gene sequences showing relationships between strain Y2T and E. jejuensis CNU001T and related representatives of the family Erythrobacteraceae. Rhodospirillum rubrum ATCC 11170T and Acetobacter aceti DSM 3508T served as outgroups. Bootstrap values >50 % based on 1000 replications are shown at nods. Bar, 0.02 substitutions per nucleotide position.

Fig. S3 Transmission electron micrographs of the cells of strain Y2T

A, Cells occur singly or in pairs; B, Cells occur in irregular clumps. Cells are cultured on MA medium at 37°C for 3days. Bar, 1 µm.

Fig.S4 The polar lipid profiles of strain Y2T as revealed by two-dimensional thin-layer chromatograms and detection with 10% ethanolic molybdophosphoric acid for all lipids (A), ninhydrin for aminolipids (B), molybdenum blue for phospholipids (C), α-naphthol-sulphuric acid reagent for glycolipids (D), periodate-Schiff reagent for α-glycols (E) and Dragendoff reagent for the detection of PC (F). DPG, diphosphatidylglycerol; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PC, phosphatidylcholine; SGL, sphingoglycolipid;PL1-PL3,unknown phospholipid; GL1-GL2, unknown glycolipid; AL1-AL2, unidentified aminolipid; L1–L3, unidentified lipids.

Table S1. Fatty acid compositions of strain Y2T, E. jejuensis CNU001T and type strains of closely related Altererythrobacter species

Strains: 1, strain Y2T; 2, E. jejuensis CNU001T; 3, A. epoxidivorans JCS350T; 4, A. ishigakiensis JPCCMB0017T; 5, A. marinus H32T; 6, A. marensis MSW-14T.All data were obtained in this study using cells grown on MA for 3 days at 30°C. Values are percentages of total fatty acids; fatty acids amounting to<0.5 % in all strains are not shown. –, not present；tr, trace amount (<0.5 %).

Fatty acid / 1 / 2 / 3 / 4 / 5 / 6
straight-chain
C12:0 / 0.6 / - / 4.0 / - / 2.9 / -
C16:0 / 3.1 / 6.1 / 4.2 / 8.6 / 7.2 / 9.1
C17:0 / 2.3 / 3.1 / tr / 0.9 / 0.5 / tr
C18:0 / 0.8 / 1.2 / 0.7 / 0.9 / tr / 0.5
Unsaturated
C15:1ω6c / tr / 0.6 / 0.6 / - / - / tr
C16:1ω5c / 0.8 / tr / 1.4 / - / 1.5 / 2.1
C17:1ω6c / 41.5 / 29.7 / 16.4 / - / 6.6 / 6.8
C17:1ω8c / 11.0 / 6.4 / 1.2 / - / 0.5 / 0.6
C18:1ω5c / 0.8 / 0.9 / 0.9 / tr / 2.0 / 2.0
11-methyl C18:1ω7c / 4.4 / 4.5 / 8.0 / - / 12.2 / 12.9
Hydroxy
C14:0 2OH / 1.0 / 1.0 / 4.5 / 4.6 / 4.3 / 2.6
C15:0 2OH / 8.1 / 5.4 / 1.9 / 1.0 / 1.1 / tr
C16:0 2OH / 1.4 / 1.7 / tr / 1.5 / 2.1 / 1.5
Branched
iso-C17:0 / - / 0.9 / - / - / - / -
iso-C16:0 3OH / - / - / 1.2 / - / 0.6 / tr
Cyclic
C19:0 ω8c cyclo / - / - / - / - / tr / 0.5
C17:0 cyclo / - / - / - / 2.6 / - / -
Summed Feature*
Summed Feature 8 / 17.2 / 34 / 35.9 / 76.5 / 51.6 / 48.7
Summed Feature 3 / 4.3 / 3.6 / 15.2 / 1.7 / 4.3 / 9.5

*Summed features represent groups of two or three fatty acids that could not be separated by GLC with the MIDI system. Summed feature 3 contains C16:1ω6c and/or C16:1 ω7c; Summed feature 8 contains:C18:1ω7c and/or C18:1ω6c.

Table S2.Comparison of 16S rRNA gene signature nucleotide positions of the genera Altererythrobacter, Erythrobacter, Porphyrobacter, Croceicoccus and Erythromicrobiumin the family Erythrobacteraceae

Taxa; 1, Altererythrobacter; 2, Erythrobacter; 3, Porphyrobacter; 4, Croceicoccus; 5, Erythromicrobium. Nucleotide positions of bases or base pairs are given according to Escherichia coli numbering (Brosius et al. 1978). /, no signature residue.

Site / 1 / 2 / 3 / 4 / 5
78:91 / /-C / /-U / /-U / /-C / /-C
416:427 / G-U / G-C / G-U / G-U / G-U
1004 / C / C / A / A / A
1014 / A / A / A / G / G
1015 / G / G / G / A / A
1016 / A / A / A / G / G
1444:1458 / C-G / C-A / /-C / C-G / C-G

Table S3. Phenotypic characteristics of strain Y2T, E. jejuensis CNU001Tand other related members of the family Erythrobacteraceae

Strains/taxa: 1, strain Y2T(this study); 2, E. jejuensis CNU001T(this study); 3,Altererythrobacter(Wu et al. 2014; Fan et al. 2011; Kwon et al. 2007; Jeong et al. 2013; Kumar et al. 2008; Matsumoto et al. 2011; Seo and Lee 2010; Lai et al. 2009; Park et al. 2011; Nedashkovskaya et al. 2013; Lei et al. 2014; Xue et al. 2012)); 4, Erythrobacter(Yurkov et al. 1994; Yoon et al. 2004a; Denner et al. 2002; Yoon et al. 2003; Lee et al. 2010; Yoon et al. 2013; Shiba and Simidu 1982; Jung et al. 2012; Xu et al. 2010; Subhash et al. 2013; Wu et al. 2012; Yoon et al. 2005; Ivanova et al. 2005); 5, Porphyrobacter(Hiraishi et al. 2002; Hanada et al. 1997; Fuerst et al. 1993; Yoon et al. 2004b; Yoon et al. 2006; Rainey et al. 2003; Furuhata et al. 2013); 6,Croceicoccus(Xu et al. 2009);7, Erythromicrobium(Yurkov et al. 1994). Data refer to all specieswithin the genera. +, Positive; -, negative; (+) and (-), positive or negative results, respectively, reported for half or more species, but no dataavailable for the others; NA, no data available; NA (+),Positive results reported for some species, but no data available for most species; v, variablereaction; v (+) and v (-), variable, but more than half of the species display positive or negative results, respectively.

Characteristic / 1 / 2 / 3 / 4 / 5 / 6 / 7
Motility / - / - / v(-) / v(-) / v(+) / + / +
Presence of BChl a / - / - / (-) / v(-) / v(+) / - / +
oxidase / + / + / v(+) / + / v(+) / - / +
Growth in NaCl (%):
Range / 0-5 / 1-5 / 0-12 / 0-14 / 0-7 / 0-10 / NA
Optimum / 2 / 2 / 0-4 / 2-5 / 0-2 / 0-1 / NA
Growth pH:
Range / 6-10 / 6-10 / 5-11.1 / 4-11 / 6-9 / 6-9 / NA
Optimum / 7-8 / 7-8 / 6-9 / 6.5-9 / 6.5-9 / 7 / 7.0-8.5
Growth temperature (°C):
Range / 4-40 / 4-37 / 4-45 / 4-45 / 10-55 / 4-42 / NA
Optimum / 35-37 / 30-35 / 25-37 / 25-37 / 30-50 / 25 / 25-30
Hydrolysis of:
Aesculin / + / + / v(+) / v / v(+) / + / NA
Casein / + / - / v(-) / v(-) / v(-) / + / NA
Gelatin / + / - / v(-) / v(-) / v(-) / + / -
Starch / - / - / v(-) / v(-) / v(+) / + / -
Tween 20 / - / + / v(+) / v / NA(+) / + / NA
Tween 80 / + / + / v(+) / v(+) / v(+) / + / -
Utilization of:
Citrate / + / - / (-) / v(-) / (-) / - / +
D-fructose / - / - / v / v(-) / v(-) / - / +
D-glucose / + / - / v(-) / v(+) / + / + / +
D-mannose / + / - / - / v / v(-) / + / NA
L-arabinose / - / - / v(-) / v(-) / v(-) / - / -
L-Malate / - / - / v / v(-) / v(-) / + / +
Maltose / - / - / v(+) / v / (+) / + / +
D-Mannitol / - / - / v(-) / v / v(-) / - / -
DNA G+C content (mol%) / 60 / 58.9 / 54.5-67.2 / 59.5-67# / 63.8-66.9 / 71.5 / 62.5-68.5
major cellular fatty acids / C17:1 ω6c , C18:1ω7c/C18:1ω6c and C17:1ω8c / C17:1ω6c, C18:1 ω7c /C18:1ω6c and C17:1ω8c / C18:1ω7c, C16:1ω7c,
C17:1ω6c / C18:1ω7c, C16:1ω6c/C16:1ω7c,C17:1ω6c and C16:0 / C18:1ω7c and C17:1ω6c / anteiso-C15:0 / NA
Major polarlipid* / DPG,PE, PC, PG and SGL / DPG, PE, PC, PG and SGL / PE, PG, DPG and SGL / DPG, PG, PE and PC / NA / PG and PC / NA

*DPG, Diphosphatidylglycerol; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; SGL, sphingoglycolipid.

# Range of DNA G+C contentreported for all the species in the genus Erythrobacter exceptE. jejuensis CNU001T

Brosius J, Palmer ML, Kennedy PJ, Noller HF (1978) Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 75 (10):4801-4805

Denner EB, Vybiral D, Koblizek M, Kampfer P, Busse HJ, Velimirov B (2002) Erythrobacter citreus sp. nov., a yellow-pigmented bacterium that lacks bacteriochlorophyll a, isolated from the western Mediterranean Sea. International journal of systematic and evolutionary microbiology 52 (Pt 5):1655-1661

Fan ZY, Xiao YP, Hui W, Tian GR, Lee JS, Lee KC, Quan ZX (2011) Altererythrobacter dongtanensis sp. nov., isolated from a tidal flat. International journal of systematic and evolutionary microbiology 61 (Pt 9):2035-2039. doi:10.1099/ijs.0.024380-0

Fuerst JA, Hawkins JA, Holmes A, Sly LI, Moore CJ, Stackebrandt E (1993) Porphyrobacter neustonensis gen. nov., sp. nov., an aerobic bacteriochlorophyll-synthesizing budding bacterium from fresh water. Int J Syst Bacteriol 43 (1):125-134

Furuhata K, Edagawa A, Miyamoto H, Kawakami Y, Fukuyama M (2013) Porphyrobacter colymbi sp. nov. isolated from swimming pool water in Tokyo, Japan. The Journal of general and applied microbiology 59 (3):245-250

Hanada S, Kawase Y, Hiraishi A, Takaichi S, Matsuura K, Shimada K, Nagashima KV (1997) Porphyrobacter tepidarius sp. nov., a moderately thermophilic aerobic photosynthetic bacterium isolated from a hot spring. Int J Syst Bacteriol 47 (2):408-413

Hiraishi A, Yonemitsu Y, Matsushita M, Shin YK, Kuraishi H, Kawahara K (2002) Characterization of Porphyrobacter sanguineus sp. nov., an aerobic bacteriochlorophyll-containing bacterium capable of degrading biphenyl and dibenzofuran. Archives of microbiology 178 (1):45-52. doi:10.1007/s00203-002-0423-5

Ivanova EP, Bowman JP, Lysenko AM, Zhukova NV, Gorshkova NM, Kuznetsova TA, Kalinovskaya NI, Shevchenko LS, Mikhailov VV (2005) Erythrobacter vulgaris sp. nov., a novel organism isolated from the marine invertebrates. Systematic and applied microbiology 28 (2):123-130. doi:10.1016/j.syapm.2004.11.001

Jeong SH, Jin HM, Lee HJ, Jeon CO (2013) Altererythrobacter gangjinensis sp. nov., a marine bacterium isolated from a tidal flat. International journal of systematic and evolutionary microbiology 63 (Pt 3):971-976. doi:10.1099/ijs.0.039024-0

Jung YT, Park S, Oh TK, Yoon JH (2012) Erythrobacter marinus sp. nov., isolated from seawater. International journal of systematic and evolutionary microbiology 62 (Pt 9):2050-2055. doi:10.1099/ijs.0.034702-0

Kumar NR, Nair S, Langer S, Busse HJ, Kampfer P (2008) Altererythrobacter indicus sp. nov., isolated from wild rice (Porteresia coarctata Tateoka). International journal of systematic and evolutionary microbiology 58 (Pt 4):839-844. doi:10.1099/ijs.0.65523-0

Kwon KK, Woo JH, Yang SH, Kang JH, Kang SG, Kim SJ, Sato T, Kato C (2007) Altererythrobacter epoxidivorans gen. nov., sp. nov., an epoxide hydrolase-active, mesophilic marine bacterium isolated from cold-seep sediment, and reclassification of Erythrobacter luteolus Yoon et al. 2005 as Altererythrobacter luteolus comb. nov. International journal of systematic and evolutionary microbiology 57 (Pt 10):2207-2211. doi:10.1099/ijs.0.64863-0

Lai Q, Yuan J, Shao Z (2009) Altererythrobacter marinus sp. nov., isolated from deep seawater. International journal of systematic and evolutionary microbiology 59 (Pt 12):2973-2976. doi:10.1099/ijs.0.008193-0

Lee YS, Lee DH, Kahng HY, Kim EM, Jung JS (2010) Erythrobacter gangjinensis sp. nov., a marine bacterium isolated from seawater. International journal of systematic and evolutionary microbiology 60 (Pt 6):1413-1417. doi:10.1099/ijs.0.015743-0

Lei X, Li Y, Chen Z, Zheng W, Lai Q, Zhang H, Guan C, Cai G, Yang X, Tian Y, Zheng T (2014) Altererythrobacter xiamenensis sp. nov., an algicidal bacterium isolated from red tide seawater. International journal of systematic and evolutionary microbiology 64 (Pt 2):631-637. doi:10.1099/ijs.0.057257-0

Matsumoto M, Iwama D, Arakaki A, Tanaka A, Tanaka T, Miyashita H, Matsunaga T (2011) Altererythrobacter ishigakiensis sp. nov., an astaxanthin-producing bacterium isolated from a marine sediment. International journal of systematic and evolutionary microbiology 61 (Pt 12):2956-2961. doi:10.1099/ijs.0.024729-0

Nedashkovskaya OI, Cho SH, Joung Y, Joh K, Kim MN, Shin KS, Oh HW, Bae KS, Mikhailov VV, Kim SB (2013) Altererythrobacter troitsensis sp. nov., isolated from the sea urchin Strongylocentrotus intermedius. International journal of systematic and evolutionary microbiology 63 (Pt 1):93-97. doi:10.1099/ijs.0.038836-0

Park SC, Baik KS, Choe HN, Lim CH, Kim HJ, Ka JO, Seong CN (2011) Altererythrobacter namhicola sp. nov. and Altererythrobacter aestuarii sp. nov., isolated from seawater. International journal of systematic and evolutionary microbiology 61 (Pt 4):709-715. doi:10.1099/ijs.0.021196-0

Rainey FA, Silva J, Nobre MF, Silva MT, da Costa MS (2003) Porphyrobacter cryptus sp. nov., a novel slightly thermophilic, aerobic, bacteriochlorophyll a-containing species. International journal of systematic and evolutionary microbiology 53 (Pt 1):35-41

Seo SH, Lee SD (2010) Altererythrobacter marensis sp. nov., isolated from seawater. International journal of systematic and evolutionary microbiology 60 (Pt 2):307-311. doi:10.1099/ijs.0.011031-0

Shiba T, Simidu U (1982) Erythrobacter longus gen. nov., sp. nov., an aerobic bacterium which contains bacteriochlorophyll a. International Journal of Systematic Bacteriology 32 (2):211-217

Subhash Y, Tushar L, Sasikala C, Ramana Ch V (2013) Erythrobacter odishensis sp. nov. and Pontibacter odishensis sp. nov. isolated from dry soil of a solar saltern. International journal of systematic and evolutionary microbiology 63 (Pt 12):4524-4532. doi:10.1099/ijs.0.052183-0

Wu HX, Lai PY, Lee OO, Zhou XJ, Miao L, Wang H, Qian PY (2012) Erythrobacter pelagi sp. nov., a member of the family Erythrobacteraceae isolated from the Red Sea. International journal of systematic and evolutionary microbiology 62 (Pt 6):1348-1353. doi:10.1099/ijs.0.029561-0

Wu YH, Xu L, Meng FX, Zhang DS, Wang CS, Oren A, Xu XW (2014) Altererythrobacter atlanticus sp. nov., isolated from deep-sea sediment. International journal of systematic and evolutionary microbiology 64 (Pt 1):116-121. doi:10.1099/ijs.0.052951-0

Xu M, Xin Y, Yu Y, Zhang J, Zhou Y, Liu H, Tian J, Li Y (2010) Erythrobacter nanhaisediminis sp. nov., isolated from marine sediment of the South China Sea. International journal of systematic and evolutionary microbiology 60 (Pt 9):2215-2220. doi:10.1099/ijs.0.014027-0

Xu XW, Wu YH, Wang CS, Wang XG, Oren A, Wu M (2009) Croceicoccus marinus gen. nov., sp. nov., a yellow-pigmented bacterium from deep-sea sediment, and emended description of the family Erythrobacteraceae. International journal of systematic and evolutionary microbiology 59 (Pt 9):2247-2253. doi:10.1099/ijs.0.004267-0

Xue X, Zhang K, Cai F, Dai J, Wang Y, Rahman E, Peng F, Fang C (2012) Altererythrobacter xinjiangensis sp. nov., isolated from desert sand, and emended description of the genus Altererythrobacter. International journal of systematic and evolutionary microbiology 62 (Pt 1):28-32. doi:10.1099/ijs.0.025437-0

Yoon BJ, Lee DH, Oh DC (2013) Erythrobacter jejuensis sp. nov., isolated from seawater. International journal of systematic and evolutionary microbiology 63 (Pt 4):1421-1426. doi:10.1099/ijs.0.038349-0

Yoon JH, Kang KH, Oh TK, Park YH (2004a) Erythrobacter aquimaris sp. nov., isolated from sea water of a tidal flat of the Yellow Sea in Korea. International journal of systematic and evolutionary microbiology 54 (Pt 6):1981-1985. doi:10.1099/ijs.0.63100-0

Yoon JH, Kang SJ, Lee MH, Oh HW, Oh TK (2006) Porphyrobacter dokdonensis sp. nov., isolated from sea water. International journal of systematic and evolutionary microbiology 56 (Pt 5):1079-1083. doi:10.1099/ijs.0.63840-0

Yoon JH, Kim H, Kim IG, Kang KH, Park YH (2003) Erythrobacter flavus sp. nov., a slight halophile from the East Sea in Korea. International journal of systematic and evolutionary microbiology 53 (Pt 4):1169-1174

Yoon JH, Lee MH, Oh TK (2004b) Porphyrobacter donghaensis sp. nov., isolated from sea water of the East Sea in Korea. International journal of systematic and evolutionary microbiology 54 (Pt 6):2231-2235. doi:10.1099/ijs.0.63226-0

Yoon JH, Oh TK, Park YH (2005) Erythrobacter seohaensis sp. nov. and Erythrobacter gaetbuli sp. nov., isolated from a tidal flat of the Yellow Sea in Korea. International journal of systematic and evolutionary microbiology 55 (Pt 1):71-75. doi:10.1099/ijs.0.63233-0

Yurkov V, Stackebrandt E, Holmes A, Fuerst JA, Hugenholtz P, Golecki J, Gad'on N, Gorlenko VM, Kompantseva EI, Drews G (1994) Phylogenetic positions of novel aerobic, bacteriochlorophyll a-containing bacteria and description of Roseococcus thiosulfatophilus gen. nov., sp. nov., Erythromicrobium ramosum gen. nov., sp. nov., and Erythrobacter litoralis sp. nov. Int J Syst Bacteriol 44 (3):427-434