International Journal of Systematic and Evolutionary Microbiology (2014), 64, 631–637 DOI 10.1099/ijs.0.057257-0

Altererythrobacter xiamenensis sp. nov., an algicidal bacterium isolated from red tide seawater

Xueqian Lei,1,23 Yi Li,1,23 Zhangran Chen,1 Wei Zheng,1 Qiliang Lai,1,3 Huajun Zhang,1 Chengwei Guan,1 Guanjing Cai,1 Xujun Yang,1 Yun Tian1 and Tianling Zheng1,2

Correspondence 1State Key Laboratory of Marine Environmental Science and Key Laboratory of MOE for Coast and Tianling Zheng Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen 361005, PR China [email protected] 2ShenZhen Research Institute of Xiamen University, ShenZhen, 518057, PR China 3Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, PR China

A Gram-stain-negative, yellow-pigmented, aerobic bacterial strain, designated LY02T, was isolated from red tide seawater in Xiamen, Fujian Province, China. Growth was observed at temperatures from 4 to 44 6C, at salinities from 0 to 9 % and at pH from 6 to 10. Phylogenetic analysis based on 16S rRNA gene sequencing revealed that the isolate was a member of the genus Altererythrobacter, which belongs to the family Erythrobacteraceae. Strain LY02T was related most closely to Altererythrobacter marensis MSW-14T (97.2 % 16S rRNA gene sequence similarity), followed by Altererythrobacter ishigakiensis JPCCMB0017T (97.1 %), Altererythrobacter epoxidivorans JCS350T (97.1 %) and Altererythrobacter luteolus SW-109T

(97.0 %). The dominant fatty acids were C18 : 1v7c,C17 : 1v6c and summed feature 3 (comprising T C16 : 1v7c and/or C16 : 1v6c). DNA–DNA hybridization showed that strain LY02 possessed low DNA–DNA relatedness to A. marensis MSW-14T, A. ishigakiensis JPCCMB0017T, A. T T epoxidivorans JCS350 and A. luteolus SW-109 (mean±SD of 33.2±1.3, 32.1±1.0, 26.7±0.7 and 25.2±1.1 %, respectively). The G+C content of the chromosomal DNA was 61.2 mol%. The predominant respiratory quinone was ubiquinone-10 (Q-10). According to its morphology, physiology, fatty acid composition and 16S rRNA gene sequence data, the novel strain most appropriately belongs to the genus Altererythrobacter, but can readily be distinguished from recognized species. The name Altererythrobacter xiamenensis sp. nov. is proposed (type strain LY02T5CGMCC 1.12494T5KCTC 32398T5NBRC 109638T).

We have studied algicidal bacterial diversity associated with (Kumar et al., 2008), Altererythrobacter marinus (Lai et al., 2009), red tide algal blooms along the Xiamen coast of China and Altererythrobacter marensis (Seo & Lee, 2010), Altererythrobacter the research has revealed novel algicidial strains (Li et al., dongtanensis (Fan et al., 2011), Altererythrobacter namhicola and 2013). This study focused on one of these isolates, designated Altererythrobacter aestuarii (Park et al., 2011), Altererythrobacter LY02T, with algicidal activity against the harmful algal ishigakiensis (Matsumoto et al., 2011), Altererythrobacter species Alexandrium tamarense. Comparative 16S rRNA gene xinjiangensis (Xue et al., 2012), Altererythrobacter gangjinensis sequence analysis indicated that strain LY02T formed a clade (Jeong et al., 2013) and Altererythrobacter troitsensis within the genus Altererythrobacter.ThegenusAltererythrobacter (Nedashkovskaya et al., 2013). Accordingly, the aim of the (family Erythrobacteraceae, order , class present work was to determine the exact taxonomic position , phylum ) was created of strain LY02T by using polyphasic characterization by Kwon et al. (2007). At the time of writing, this including the determination of phenotypic properties and genus includes 12 species with validly published names: a detailed phylogenetic analysis based on 16S rRNA gene Altererythrobacter luteolus (Yoon et al., 2005), Altererythrobacter sequences. epoxidivorans (Kwon et al., 2007), Altererythrobacter indicus Strain LY02T was isolated from surface water samples collected in October 2012 at a depth of 1–2 m from red 3These authors contributed equally to this work. tide seawater in Xiamen (24u 339 N 118u 99 E), Fujian The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene Province, China. The samples were serially diluted (10-fold sequence of strain LY02T is KC520828. dilution) in sterile seawater and 0.1 ml aliquots of each

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86 aquimaris SW-110T (AY461441) 90 0.01 DSM 6997T (AF465835) Erythrobacter nanhaisediminis T30T (FJ654473) Erythrobacter vulgaris 022-2 10T (AY706935) 58 Erythrobacter flavus SW-46T (AF500004) Erythrobacter gaetbuli SW-161T (AY562220) DSM 8509T (AB013354) 78 61 Erythromicrobium ramosum DSM 8510T (AF465837) UST081027-248T (HQ203045) 55 Erythrobacter pelagi Erythrobacter citreus RE35F/1T (AF118020) Erythrobacter seohaensis SW-135T (AY562219) Erythrobacter gangjinensis K7-2T (EU428782) Altererythrobacter aestuarii KYW147T (FJ997597) Altererythrobacter namhicola KYW48T (FJ935793) indicum H25T (EF549586) Altererythrobacter indicus MSSRF26T (DQ399262) Novosphingobium tardaugens ARI-1T (AB070237) Altererythrobacter luteolus SW-109T (AY739662) 70 Altererythrobacter gangjinensis KJ7T (JF751048) Altererythrobacter xiamenensis LY02T (KC520828) Altererythrobacter ishigakiensis JPCCMB0017T (AB363004) Altererythrobacter epoxidivorans JCS350T (DQ304436) Altererythrobacter marensis MSW-14T (FM177586) Altererythrobacter marinus H32T (EU726272) Altererythrobacter xinjiangensis S3-63T (HM028673) 76 Altererythrobacter dongtanensis JM27T (GU166344) 98 Altererythrobacter troitsensis JCM 17037T (AY676115) Oceanibaculum indicum P24T (EU656113)

Fig. 1. Neighbour-joining tree showing the phylogenetic positions of strain LY02T and representatives of some other related taxa, based on 16S rRNA gene sequences. Bootstrap values (expressed as percentages of 1000 replications) are shown at branch points; only values .50 % are shown. Oceanibaculum indicum P24T was used as the outgroup. Bar, 0.01 nt

substitution rate (Knuc) units. dilution were spread onto marine agar 2216 (MA; Difco) The nearly full-length 16S rRNA gene sequence (1411 nt) of followed by incubation for 7 days at 28 uC. Individual strain LY02T was determined. Phylogenetic analysis of strain colonies of distinct morphology were further purified three LY02T based on the 16S rRNA gene sequence indicated that times and stored at 280 uC in marine broth 2216 (MB; this strain belonged to the family Erythrobacteraceae, Difco) supplemented with 10 % (v/v) glycerol. forming a robust clade within the genus Altererythrobacter (Fig. 1). Its closest relatives were A. marensis MSW-14T Genomic DNA was extracted according to the method of (97.2 % 16S rRNA gene sequence similarity), followed by A. Ausubel et al. (1995). The 16S rRNA gene sequence was ishigakiensis JPCCMB0017T (97.1 %), A. epoxidivorans amplified by PCR using primers 27F and 1492R (DeLong, JCS350T (97.1 %) and A. luteolus SW-109T (97.0 %). Strain 1992). Purification of the PCR product was carried out LY02T formed a separate phylogenetic clade with A. according to the protocol of the TIANquick midi purifica- ishigakiensis JPCCMB0017T. tion kit (Tiangen). The purified DNA was cloned into vector pMD19-T and sequenced. Sequences of related taxa were Cell morphology and motility were observed by using obtained from the GenBank database and EzTaxon-e server transmission electron microscopy (model JEM-2100HC; (http://eztaxon-e.ezbiocloud.net/) (Kim et al., 2012). JEOL) and phase-contrast light microscopy (model 50i; Phylogenetic analysis was performed using MEGA version 4 Nikon), with cells from the early exponential phase grown (Tamura et al., 2007) after multiple alignment of the data on MA at 28 uC. Colony morphology was examined from using DNAMAN (version 5.1). Evolutionary distances and cultures grown on MA for 2 days. The presence of clustering were determined by using the neighbour-joining flexirubin-type pigments was assessed using the bath- method (Saitou & Nei, 1987), and were evaluated by using ochromic shift test with 20 % KOH, as described by bootstrap values based on 1000 replications. Bernardet et al. (2002). The Gram reaction was determined

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Table 1. Characteristics that differentiate strain LY02T from closely related species of the genus Altererythrobacter

Strains: 1, LY02T;2,A. luteolus SW-109T (Yoon et al., 2005); 3, A. marensis MSW-14T (Seo & Lee, 2010); 4, A. epoxidivorans JCS350T (Kwon et al., 2007); 5, A. ishigakiensis JPCCMB0017T (Matsumoto et al., 2011). Data for catalase, oxidase, API 20NE, API 20E and API ZYM for all five strains were done at the same time in this study. All the strains were positive for catalase activity, oxidase activity, and hydolysis of Tweens 20, 40 and 60 and tyrosine; negative for hydrolysis of chitin, gelatin and urea, and flexirubin-type pigments. In API 20NE strips, all strains were positive for b- glucosidase (aesculin hydrolysis); negative for indole production, arginine dihydrolase, gelatin hydrolysis, and utilization of L-arabinose, D- mannose, D-mannitol, N-acetylglucosamine, potassium gluconate, capric acid, trisodium citrate and phenylacetic acid. In API 20E tests, all were positive for tryptophan deaminase and acetoin production; and negative for citrate utilization, H2S production and acid production from inositol. In API ZYM strips, all strains were positive for alkaline phosphatase, leucine aminopeptidase, valine aminopeptidase, trypsin, a-chymotrypsin, acid phosphatase and naphthol-AS-BI-phosphoamidase; negative for b-glucuronidase, b-glucosidase, N-acetyl-b-glucosaminidase, a-mannosidase and a-fucosidase. +, Positive; W, weakly positive; 2, negative; ND, not determined. All data are from this study except growth temperature, salinity range, growth pH and the DNA G+C contents of A. luteolus SW-109T, A. marensis MSW-14T, A. epoxidivorans JCS350T and A. ishigakiensis JPCCMB0017T, which are from Yoon et al. (2005), Seo & Lee (2010), Kwon et al. (2007) and Matsumoto et al. (2011), respectively.

Characteristic 12345

Colony colour Yellow Yellow Yellow Yellow Orange NaCl tolerance (%, w/v) 0–9 0–9 0–9 0–6 ND NaCl optimum (%) 1.5 2 0–4 2 1.5–2 Growth temperature (uC) 4–44 4–36 4–42 20–40 25–40 Optimum temperature (uC) 28–30 30 30 35 35 Growth pH 6–10 ND 6.1–11.1 6–8.5 5.5–8.5 Optimal pH 7.0 7–8 7.1 6.5 7.5 Hydrolysis of: Starch 2 W 222 Tween 80 2 ++++ Casein 2 + 2 + 2 Aesculin +++ W + API ZYM results Esterase (C4) 22W + 2 Esterase lipase (C8) 22W + 2 Lipase (C14) 222+ 2 Cystine arylamidase W + 2 + W a-Galactosidase 2222+ b-Galactosidase 2 + 2 W + a-Glucosidase ++222 API 20NE results Reduction of nitrate + 222+ Denitrification 2 + 222 D-Glucose fermentation + 222+ Urease + 2222 b-Galactosidase 2 + 2 ++ D-Glucose ++222 Maltose 2 + 2 + 2 Adipic acid + 22+ 2 Malic acid 22+ 22 API 20E results Lysine decarboxylase W ++2 + Ornithine decarboxylase W ++2 + Gelatinase ++2 + 2 Sorbitol 222+ 2 Rhamnose 2 + 222 Sucrose ++2 + 2 Melibiose 2 + 222 Amygdalin ++222 Susceptibility to: Ampicillin ++222 Carbenicillin ++2 ++ Cephradin 2 + 22+

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Table 1. cont.

Characteristic 12345

Ciprofloxacin 2 +++2 Cefalexin 2 + 2 ++ Gentamicin ++2 ++ Vibramycin 2 + 2 ++ Erythromycin ++++2 Norfloxacin 2 + 222 Oxacillin 2 + 222 Penicillin G + 22+ 2 Polymyxin B 2 + 222 Piperacillin ++2 ++ Sulphafurazole + 2 + 22 Neomycin ++2 ++ DNA G+C content (mol%) 61.2 60.3 63.1 54.5 59.1 by using the bioMe´rieux Gram stain kit according to the (10 mg), carbenicillin (100 mg), cefalexin (30 mg), cefazolin manufacturer’s instructions. Gliding motility was investi- (30 mg), cefobid (30 mg), cephradin (30 mg), chlorampheni- gated as described by Bowman (2000). Anaerobic growth col (30 mg), ciprofloxacin (5 mg), clindamycin (2 mg), was assessed in MB that was autoclaved and cooled to cotrimoxazole (25 mg), erythromycin (15 mg), gentamicin room temperature under nitrogen atmosphere (99.999 % (10 mg), kanamycin (30 mg), metronidazole (5 mg), mino- purity). Triplicate cultures were grown in 50 ml anaerobic mycin (30 mg), neomycin (30 mg), norfloxacin (10 mg), serum bottles sealed with thick butyl rubber stoppers and novobiocin (5 mg), ofloxacin (5 mg), oxacillin (1 mg), aluminium caps, and incubated statically in the dark at penicillin G (10 mg), piperacillin (100 mg), polymyxin B 30 uC for 21 days. The optimal growth temperature was (30 IU), rifampicin (5 mg), streptomycin (10 mg), sulpha- determined over the range 0–45 uC (0, 4, 10, 15, 20, 25, 28, furazole (300 mg), tetracycline (30 mg), trimethoprim 30, 32, 35, 38, 40, 42, 43, 44 and 45 uC) in MB in triplicate (5 mg), vancomycin (30 mg) and vibramycin (30 mg). The and measured by spectrophotometry based on OD600 physiological and biochemical characteristics of strain values every 12 h for 3 days. The NaCl concentration range LY02T are given in the species descriptions and in Table 1. and optimum for growth were determined in NaCl-free DNA–DNA hybridization experiments were performed at 2216 medium (per litre distilled water: 5.0 g tryptone, 1.0 g 42 uC in triplicate with genomic DNA of strain LY02T and yeast extract, pH 7.6–7.8), supplemented with various A. marensis MSW-14T, A. ishigakiensis JPCCMB0017T, A. concentrations of NaCl [0–5.0 % (w/v) at 0.5 % intervals epoxidivorans JCS350T and A. luteolus SW-109T according and 5.0–12.0 % at 1 % intervals]. The pH range for growth to the method of Liu & Shao (2005). The results showed was determined in MB that was adjusted to pH 3.0–12.0 that strain LY02T possessed a low level of DNA–DNA (at 1 pH unit intervals). The pH of MB was adjusted prior relatedness (mean±SD of 33.2±1.3, 32.1±1.0, 26.7±0.7 to sterilization using the following buffers: citric acid/ and 25.2±1.1 %, respectively) with them. According to sodium citrate (pH 3.0–6.0), Na HPO /citric acid (pH 7.0– 2 4 Wayne et al. (1987), a strain can be discriminated as 8.0) and lysine/NaOH (pH 9.0–10.0). Verification of the pH representing a different bacterial species if the cut-off value values after autoclaving revealed only minor changes (Su is less than 70 %. Therefore, strain LY02T should be et al., 2013). Catalase activity was determined by addition of considered to represent a novel species of the genus 3 % (v/v) hydrogen peroxide to exponential-phase colonies, Altererythrobacter. and the oxidase reaction was tested by using oxidase reagent (bioMe´rieux). Hydrolysis of starch, chitin, tyrosine, casein, Fatty acids of strain LY02T and A. marensis MSW-14T, A. gelatin, urea and Tweens 20, 40, 60 and 80 was tested using ishigakiensis JPCCMB0017T, A. epoxidivorans JCS350T and MA supplemented with 0.5 % (w/v) starch and 1 % (w/v) of A. luteolus SW-109T in whole cells grown on MA at 28 uC the other substrates. Results were examined twice after for 48 h were extracted, saponified and esterified using growth on agar plates for 3 and 5 days. Further biochemical the standard protocol of MIDI (Sherlock Microbial tests of strain LY02T and the reference strains were Identification System, version 6.0B). The five strains had conducted using the API 20E and API 20NE test kits similar growth rates and had the same physiological age (bioMe´rieux) at 30 uC for 3 days and the API ZYM test kit after growth on MA plates at 28 uC for 48 h. The fatty acids (bioMe´rieux) at 30 uC for 24 h according to the manufac- were analysed by GC (Agilent Technologies 6850) and turer’s instructions. All the commercial kits were inoculated identified by using the TSBA 6.0 database of the Microbial with bacterial suspensions prepared in 3 % (w/v) NaCl. Identification System (Sasser, 1990). The four reference Susceptibility to antibiotics was tested on MA at 30 uC for type strains were tested under the same conditions in this 2 days by using the following discs (Oxoid): ampicillin study with strain LY02T. As shown in Table 2, the major

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T T Table 2. Cellular fatty acid contents of strain LY02 and fatty acids of strain LY02 were C18 : 1v7c (61.6 %), closely related species of the genus Altererythrobacter C17 : 1v6c (10.0 %) and summed feature 3 (comprising C v7c and/or C v6c, 9.6 %), which accounted for Strains: 1, LY02T;2,A. luteolus SW-109T;3,A. marensis MSW-14T;4, 16 : 1 16 : 1 81.3 % of the total. The fatty acid profile of strain LY02T A. epoxidivorans JCS350T;5,A. ishigakiensis JPCCMB0017T. All data was in good agreement with those of members of the genus were generated in this study. Values are percentages of total fatty Altererythrobacter, but it can be differentiated from other acids; fatty acids amounting to ,0.5 % in all strains are not shown. species based on the percentage of C16 : 1v5c and 11-methyl TR, Trace amount (,0.5 %); ND, not detected. C18 : 1v7c. Fatty acid 1 2 3 4 5 The G+C content of the chromosomal DNA was Saturated determined according to the methods of Mesbah & Whitman (1989) using reversed-phase HPLC. The DNA C11 : 0 ND ND ND ND 0.6 G+C content of strain LY02T was 61.2 mol% and was in C12 : 0 1.0 ND 0.5 ND ND C13 : 0 TR ND ND ND ND the range previously reported for species of the genus C14 : 0 0.6 0.5 0.8 0.7 0.5 Altererythrobacter (54.5–71.5 mol%). C16 : 0 3.5 6.6 7.4 3.8 9.6 The major respiratory quinone of strain LY02T was C17 : 0 TR ND TR TR 2.4 ubiquinone-10 (Q-10), analysis of which was carried out C 0.7 0.5 2.4 0.6 1.3 18 : 0 by the Identification Service of the DSMZ, Braunschweig, Branched Germany. This trait was in accordance with the properties iso-C ND TR ND ND ND 12 : 0 of the genus Altererythrobacter. iso-C15 : 1 G ND TR ND TR TR anteiso-C17 : 1 A ND ND ND TR ND On the basis of morphological, physiological and chemo- Cyclic taxonomic characteristics, together with data from 16S rRNA C19 : 0v8c cyclo ND 0.9 TR ND ND gene sequence comparisons and DNA–DNA hybridization Hydroxy experiments, strain LY02T should be assigned to a novel C10 : 0 3-OH ND TR 0.5 ND ND species within the genus Altererythrobacter, for which the C 2-OH ND ND TR ND ND 11 : 0 name Altererythrobacter xiamenensis sp. nov. is proposed. C12 : 0 2-OH ND TR ND ND ND C12 : 0 3-OH TR ND ND ND ND Description of Altererythrobacter xiamenensis C13 : 0 2-OH ND ND ND TR TR sp. nov. C14 : 0 2-OH 1.6 2.3 0.6 1.5 TR C15 : 0 2-OH TR ND TR 0.6 TR Altererythrobacter xiamenensis (xia.men.en9sis. N.L. masc. C16 : 0 2-OH 0.5 1.1 1.3 TR TR adj. xiamenensis of Xiamen, a city in Fujian, China, where C16 : 1 2-OH TR 0.6 ND ND ND the type strain was first isolated). iso-C11 : 0 3-OH ND ND ND 0.5 ND iso-C16 : 0 3-OH TR 0.6 TR 1.4 ND Cells are long and rod-shaped (0.3–0.5 mm in diameter and C18 : 1 2-OH ND TR ND ND ND 1.4–3.5 mm in length, Fig. 2.). Growth is visible after 24 h Unsaturated of incubation on MA at 28 uC. Colonies on MA are shiny, C15 : 1v6c 0.4 TR TR 0.6 ND yellow and circular with regular, smooth edges, and are 1– C16 : 1v5c 0.8 3.8 1.4 1.2 ND 2 mm in diameter after 48 h incubation at 28 uC, opaque C17 : 1v6c 10.0 ND 11.5 21.3 ND and raised in the centre. Growth occurs at 4–44 uC C17 : 1v8c 0.6 ND ND 1.7 ND (optimum 28–30 uC), at pH 6–10 (optimum pH 7) and C18 : 1v5c 2.2 3.1 2.5 1.0 0.79 at NaCl concentrations of 0–9 % (w/v), with optimal C v7c 61.6 44.5 47.1 36.6 81.7 18 : 1 growth at 1.5 % (w/v); growth does not occur in the C v9c TR TR TR 0.5 TR 18 : 1 absence of NaCl. Growth does not occur under anaerobic 11-methyl 4.6 9.6 14.7 12.0 ND conditions on MB. Flexirubin-type pigments are not C18 : 1v7c Summed produced. Positive for catalase, oxidase, reduction of features* nitrate, and hydrolysis of Tweens 20, 40 and 60 and 3 9.6 23.5 5.9 13.1 1.0 tyrosine. Negative for hydrolysis of casein, chitin, starch, 4 ND ND ND TR ND Tween 80, urea and gelatin. In API 20NE strips, positive for 9 ND ND ND TR ND reduction of nitrate to nitrite, D-glucose fermentation, urease activity, b-glucosidase (aesculin hydrolysis), and *Summed features represent groups of two or three fatty acids that utilization of D-glucose and adipic acid; negative for could not be separated by GLC with the MIDI system. Summed denitrification, indole production, arginine dihydrolase, feature 3 was listed as C16 : 1v7c and/or C16 : 1v6c; summed feature 4 gelatin hydrolysis, b-galactosidase, and utilization of L- was listed as iso-C17 : 1 I and/or anteiso-C17 : 1 B; summed feature 9 was arabinose, D-mannose, D-mannitol, maltose, N-acetylglu- listed as iso-C17 : 1v9c and/or C16 : 0 10-methyl. cosamine, potassium gluconate, capric acid, malic acid, trisodium citrate and phenylacetic acid. In API 20E strips, positive for urease activity, tryptophan deaminase, acetoin

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Fig. 2. Transmission electron micrographs of cells of strain LY02T grown on MA for 24 h at 28 6C. Bars, 0.5 mm (left), 1 mm (right). production, gelatinase, acid production from glucose, Program in University (20120121130001) and Science and Technology sucrose and amygdalin, lysine decarboxylase and ornithine Innovation Funds of Shenzhen (JCYJ20120615161239998). We thank decarboxylase; negative for b-galactosidase, citrate utiliza- Professor I. J. Hodgkiss from Hong Kong University for help with the English text. tion, H2S production, indole production, and acid production from mannitol, inositol, sorbitol, rhamnose, melibiose and arabinose. In API ZYM tests, positive for References alkaline phosphatase, leucine aminopeptidase, valine ami- nopeptidase, trypsin, a-chymotrypsin, acid phosphatase, Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J., naphthol-AS-BI-phosphoamidase and a-glucosidase; weakly Smith, J. A. & Struhl, K. (editors) (1995). Short Protocols in Molecular positive for lipase (C14), cystine aminopeptidase; negative Biology: a Compendium of Methods from Current Protocols in for esterase (C4), esterase lipase (C8), a-galactosidase, b- Molecular Biology, 3rd edn. New York: Wiley. galactosidase, b-glucuronidase, b-glucosidase, N-acetyl-b- Bernardet, J.-F., Nakagawa, Y., Holmes, B. & Subcommittee on the glucosaminidase, a-mannosidase and a-fucosidase. Sensitive taxonomy of Flavobacterium and Cytophaga-like of the International Committee on Systematics of Prokaryotes (2002). m to ( g per disc unless otherwise noted; Oxoid): ampicillin Proposed minimal standards for describing new taxa of the family (10), chloramphenicol (30), carbenicillin (100), gentamicin Flavobacteriaceae and emended description of the family. Int J Syst (10), erythromycin (15), kanamycin (30), penicillin G (10), Evol Microbiol 52, 1049–1070. piperacillin (100), rifampicin (5), vancomycin (30), novo- Bowman, J. P. (2000). Description of Cellulophaga algicola sp. nov., biocin (5), sulphafurazole (300) and neomycin (30); isolated from the surfaces of Antarctic algae, and reclassification of resistant to cephradin (30), cefobid (30), ciprofloxacin (5), Cytophaga uliginosa (ZoBell and Upham 1944) Reichenbach 1989 as cefalexin (30), clindamycin (2), vibramycin (30), cefazolin Cellulophaga uliginosa comb. nov. Int J Syst Evol Microbiol 50, 1861–1868. (30), minomycin (30), metronidazole (5), norfloxacin (10), DeLong, E. F. (1992). Archaea in coastal marine environments. Proc ofloxacin (5), oxacillin (1), polymyxin B (30 IU), strep- Natl Acad Sci U S A 89, 5685–5689. tomycin (10), cotrimoxazole (25), tetracycline (30) and Fan, Z.-Y., Xiao, Y.-P., Hui, W., Tian, G.-R., Lee, J.-S., Lee, K. C. & trimethoprim (5). The predominant fatty acids are Quan, Z.-X. (2011). Altererythrobacter dongtanensis sp. nov., isolated C18 : 1v7c,C17 : 1v6c and summed feature 3 (comprising from a tidal flat. Int J Syst Evol Microbiol 61, 2035–2039. C16 : 1v7c and/or C16 : 1v6c). The predominant respiratory Jeong, S. H., Jin, H. M., Lee, H. J. & Jeon, C. O. (2013). quinone is ubiquinone-10 (Q-10). Altererythrobacter gangjinensis sp. nov., a marine bacterium isolated from a tidal flat. Int J Syst Evol Microbiol 63, 971–976. The type strain, LY02T (5CGMCC 1.12494T5KCTC T T Kim, O. S., Cho, Y. J., Lee, K., Yoon, S. H., Kim, M., Na, H., Park, S. C., 32398 5NBRC 109638 ), was isolated from red tide Jeon, Y. S., Lee, J. H. & other authors (2012). Introducing EzTaxon-e: seawater in Xiamen, Fujian Province, China. The DNA a prokaryotic 16S rRNA gene sequence database with phylotypes that G+C content of the type strain is 61.2 mol%. represent uncultured species. Int J Syst Evol Microbiol 62, 716–721. Kumar, N. R., Nair, S., Langer, S., Busse, H.-J. & Ka¨ mpfer, P. (2008). Altererythrobacter indicus sp. nov., isolated from wild rice (Porteresia Acknowledgements coarctata Tateoka). Int J Syst Evol Microbiol 58, 839–844. This work was supported by the National Nature Science Foundation Kwon, K. K., Woo, J.-H., Yang, S.-H., Kang, J.-H., Kang, S. G., Kim, of China (40930847), Public Science and Technology Research Funds S.-J., Sato, T. & Kato, C. (2007). Altererythrobacter epoxidivorans gen. Projects of Ocean (201305016, 201305022), Special Fund for PhD nov., sp. nov., an epoxide hydrolase-active, mesophilic marine

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