International Journal of Systematic and Evolutionary Microbiology (2014), 64, 3496–3502

DOI 10.1099/ijs.0.063909-0

Phaeodactylibacter xiamenensis gen. nov., sp. nov., a member of the family Saprospiraceae isolated from the marine alga Phaeodactylum tricornutum Zhangran Chen,13 Xueqian Lei,13 Qiliang Lai,1,2 Yi Li,1 Bangzhou Zhang,1 Jingyan Zhang,1 Huajun Zhang,1 Luxi Yang,1 Wei Zheng,1 Yun Tian,1 Zhiming Yu,3 Hong Xu1 and Tianling Zheng1 Correspondence Tianling Zheng [email protected] Hong Xu [email protected]

1

Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystem, School of Life Sciences, Xiamen University, Xiamen 361005, PR China

2

Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, PR China

3

Key Laboratory of Marine Ecology and Environmental Science, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, PR China

A novel Gram-staining-negative, aerobic, rod-shaped, non-motile, reddish-orange and chemoheterotrophic bacteria, designated strain KD52T, was isolated from a culture of the alga Phaeodactylum tricornutum from Xiamen, Fujian Province, China. 16S rRNA gene sequence comparison showed that strain KD52T was a member of the family Saprospiraceae, forming a distinct lineage with ‘Portibacter lacus’ KCTC 23747. The 16S rRNA gene sequence similarity between strain KD52T and the type strains of species of the family Saprospiraceae ranged from 86 % to 89 %. Growth occurred at 20–37 6C (optimum, 28 6C), in the presence of 1–9 % (w/v) NaCl (optimum, 2.5 %) and at pH 5–8.5 (optimum, pH 6.0). The dominant fatty acids (.10 %) of strain KD52T were iso-C15 : 0 (33.1 %), iso-C15 : 1 G (14.8 %) and summed feature 3 (comprising C16 : 1v7c and/or C16 : 1v6c, 13.8 %). The major polar lipids were diphosphatidylglycerol, three unidentified phospholipids, four unknown lipids and one unidentified aminolipid. The DNA G+C content was 51 mol% and the major respiratory quinone was menaquinone-7 (MK-7). On the basis of phenotypic data and phylogenetic inference, strain KD52T represents a novel species of a new genus, for which the name Phaeodactylibacter xiamenensis gen. nov., sp. nov., is proposed. The type strain is KD52T (5MCCC 1F01213T5KCTC 32575T).

Members of the family Saprospiraceae, are considered to be important members of the bacterial community involved in ecophysiological activities in a variety of natural environments (Xia et al., 2008). At the time of writing, the family Saprospiraceae includes the genera Saprospira, Haliscomenobacter, Lewinella, ‘Portibacter’, Aureispira and ‘Rubidimonas’. The genera Saprospira, Haliscomenobacter, ‘Portibacter’ and ‘Rubidimonas’ each accommodate one species, Saprospira grandis (Reichenbach, 1989), Haliscomenobacter hydrossis (van Veen et al., 1973), ‘Portibacter lacus’ (Yoon et al., 2012b) and ‘Rubidimonas crustatorum’ (Yoon et al., 2012a), and the genus Lewinella accommodates seven species, Lewinella agarilytica (Lee, 2007), 3These authors contributed equally to this work. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain KD52T is KF986715. One supplementary figure is available with the online Supplementary Material.

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Lewinella antarctica (Oh et al., 2009), and Lewinella cohaerens, Lewinella lutea, Lewinella marina, Lewinella nigricans and Lewinella persica (Khan et al., 2007) The genus Aureispira comprises two species, Aureispira marina (Hosoya et al., 2006) and Aureispira maritima (Hosoya et al., 2007). Accordingly, the aim of the present work was to determine the exact taxonomic position of strain KD52T by using polyphasic characterization including the determination of phenotypic properties and a detailed phylogenetic analysis based on 16S rRNA gene sequences. Strain KD52T was isolated from a culture of the alga Phaeodactylum tricornutum from Xiamen, Fujian Province, China. The samples were serially diluted (10-fold dilution) using sterile seawater and 0.1 ml aliquots of each dilution were spread onto marine agar 2216 (MA; Difco) followed by incubation for 7 days at 28 uC. Individual colonies of distinct morphology were further purified three times and stored at 280 uC in marine broth 2216 (MB; Difco) supplemented with 10 % (v/v) glycerol. 063909 G 2014 IUMS Printed in Great Britain

Phaeodactylibacter xiamenensis gen. nov., sp. nov.

Lewinella persica NBRC 102663T (AB301616)

100

0.02

86

Lewinella persica ATCC 23167T (AF039295) Lewinella agarilytica SST-19T (AM286229)

79

Lewinella lutea FYK2402M69T (AB301494)

99

Lewinella antarctica IMCC3223T (EF554367)

79

Lewinella marina MKG-38T (AB301495)

95

Lewinella cohaerens NBRC 102661T (AB301614) Lewinella nigricans NBRC 102662T (AB301615)

66

Phaeodactylbacter xiamenensis KD52 (KF986715) ‘Portibacter lacus’ YM8-076 (AB675658) ‘Candidatus Aquirestis calciphila’ MS-Falk1-L (AJ786341)

93

Haliscomenobacter hydrossis DSM 1100T (CP002691)

100

‘Candidatus Haliscomenobacter calcifugiens’ MS-oKlaff1-G (AJ786327)

100

Saprospira grandis JCM 21750T (AB088636) Aureispira maritima 59SAT (AB278130)

100

Aureispira marina 24T (NR_041389)

100

Pedobacter lentus DS-40T (EF446146)

Fig. 1. Neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showing the relationship of strain KD52T and representative members of the family Saprospiraceae. Bootstrap values above 70 % (expressed as percentages of 1000 replications) are given at nodes. Pedobacter lentus DS-40T was used as an outgroup. Bar, 0.02 substitutions per nucleotide position.

The genomic DNA of strain KD52T was extracted according to the method of Ausubel et al. (1995) and the 16S rRNA gene was amplified by PCR using the primer pair P27F and P1492R (DeLong, 1992). Purification of the PCR product was carried out using a TIANquick Midi purification kit (TIANGEN) and the purified PCR product was cloned into vector pMD19-T and sequenced. Sequences of related taxa were downloaded from the GenBank database and the EzTaxon-e server (http:// eztaxon-e.ezbiocloud.net/; Kim et al., 2012). Phylogenetic analysis was performed using MEGA version 4 (Tamura et al., 2007) after multiple alignment of data by DNAMAN (version 5.1). Evolutionary distances and clustering were performed by using the neighbour-joining method (Saitou & Nei, 1987). The resulting tree topology was evaluated by using bootstrap analysis based on 1000 replicates. Preliminary comparison of the 16S rRNA gene sequence of strain KD52T (1445 bp) with other sequences indicated that the new isolate showed 89 % similarity to that of ‘Portibacter lacus’ KCTC 23747 (GenBank no. AB675658), which was isolated from water (salinity 34 %, pH 6.9) collected at a harbour on Lake Notoro, Hokkaido, Japan (Yoon et al., 2012b). Sequence similarities to other members of the family Saprospiraceae were ,88 %. As shown in Fig. 1, strain KD52T clustered together with ‘Portibacter lacus’ KCTC 23747. Consequently, ‘Portibacter lacus’ KCTC 23747, http://ijs.sgmjournals.org

S. grandis JCM 21750T, L. nigricans NBRC 102662T and H. hydrossis DSM 1100T were used as reference strains and grown under the same conditions as strain KD52T, except H. hydrossis DSM 1100T for which DSMZ medium 134 was used. All these strains were used as reference strains for a number of phenotypic tests. Cell morphology and motility were observed by using transmission electron microscopy (model JEM-2100HC; JEOL) and phase-contrast light microscopy (model 50i; Nikon), with cells from the early exponential phase grown

(a)

(b)

Fig. 2. Transmission electron micrographs of cells of strain KD52 grown on MA for 5 days at 28 6C. Bars, 1 mm (a) and 2 mm (b). 3497

Z. Chen and others

Table 1. Differential characteristics among strain KD52T and other members of the family Saprospiraceae Strains: 1, KD52T; 2, H. hydrossis DSM 1100T; 3, S. grandis JCM 21750T; 4, ‘Portibacter lacus’ KCTC 23747; 5, L. nigricans NBRC 102662T. All data were from this study. Data for catalase, oxidase, API 20NE, API 20E and API ZYM for the five strains were obtained at the same time in this study. All strains are non-motile rods and have no flexirubin-type pigments. All of the strains are positive for oxidase activity but negative for hydrolysis of tyrosine, chitin and gelatin. In the API ZYM strip, all of the strains are positive for alkaline phosphatase, esterase (C4), leucine aminopeptidase, valine aminopeptidase, acid phosphatase and naphthol-AS-BI-phosphoamidase. In the API 20E strip, all of the strains are positive for arginine dihydrolase, lysine decarboxylase, ornithine decarboxylase, urease and gelatinase; negative for citrate utilization, H2S production, tryptophan deaminase, and acid production from mannitol, inositol, sorbitol, rhamnose, melibiose, amygdalin and arabinose. In the API 20NE strip, all strains negative for gelatin hydrolysis, denitrification, utilization of D-glucose, L-arabinose, D-mannose, D-mannitol, N-acetylglucosamine, maltose, potassium gluconate, capric acid, adipic acid, malic acid, trisodium citrate and phenylacetic acid. All strains are susceptible to chloramphenicol, cefalexin, clindamycin, vibramycin, erythromycin, minomycin, ofloxacin, piperacillin, streptomycin, ampicillin and novobiocin. +, Positive; 2, negative; W, weakly positive. Characteristic Colony colour NaCl concentration for growth (%) Range Optimum Growth temperature (uC) Range Optimum Catalase API ZYM Esterase lipase (C8) Lipase (C14) Cystine aminopeptidase Trypsin a-Chymotrypsin a-Galactosidase b-Galactosidase b-Glucuronidase a-Glucosidase b-Glucosidase N-Acetyl-b-glucosaminidase a-Mannosidase a-Fucosidase API 20E b-Galactosidase Indole production Acetoin production (Voges–Proskauer) Fermentation/oxidation (glucose) Fermentation/oxidation (sucrose) API 20NE Reduction of nitrate to nitrite Indole production D-Glucose fermentation Arginine dihydrolase Urease b-Glucosidase (aesculin hydrolysis) b-Galactosidase Susceptibility to: Carbenicillin Cefoperazone Ciprofloxacin Kanamycin Cefazolin Metronidazole Norfloxacin

3498

1

2

3

4

5

Reddish orange

Pink

Orange–red

Orange

Yellow

1–9 2.5

3–7 4

2–7 3

1–8 3

2–4 2.5

20–37 28 2

16–30 28 +

28–37 30 +

20–37 30 +

20–37 28 +

+

+

W

W

+ + + + + + + + + 2 +

+ + + 2 + 2 2 2 + 2 2

+ +

2 2 + + + 2

+ 2 + 2 2

W

W

W

W

W

2 + 2 + + 2

2 + + + + + 2 + + + + 2

2 2 + +

2 2 + 2 2

2 2 + 2 2

+ + 2 2

+ 2 2 2 2 + +

2 2 + + + + +

2 2 2 2 2 2 2

+ 2 2 + + + +

2 + 2 + + + +

+ + + 2 +

2 2

+ + + 2 + 2 +

+

+

W

2

W

W

+ + 2 +

W

2 2 2

2 2 W

2

W

W

W

+ + + + +

2 2 2 W

+

International Journal of Systematic and Evolutionary Microbiology 64

Phaeodactylibacter xiamenensis gen. nov., sp. nov.

Table 1. cont. Characteristic

1

2

3

4

5

Oxacillin Penicillin G Polymyxin B Rifampicin Co-trimoxazole Tetracycline Vancomycin Sulfafurazole Trimethoprim Neomycin Degradation of: Tween 20 Tween 40 Tween 60 Tween 80 Starch Casein Urea DNA G+C content (mol%)

2 + + 2 2 2 + 2 2 +

2 2 + + 2 + 2 + + +

2 + + +

+

W

+ +

2 2 + + 2 + + 2 + +

+ + + 2 2 2 2 51.0

2 2 2 2 +

2 2 2 2 2 + 2 47.1

+ + + 2 2 2 2 54.3

+ 2 2 2 + + + 52.9

on MA at 28 uC. The presence of flexirubin-type pigments was assessed using the bathochromic shift test with 20 % (w/v) KOH, as described by Bernardet et al. (2002). Colony morphology was examined from cultures grown on MA for 6 days. Gliding motility was investigated as described by Bowman (2000). The Gram reaction was determined by using the bioMe´rieux Gram stain kit according to the manufacturer’s instructions. Anaerobic growth was assessed on MA that was autoclaved and cooled to room temperature under nitrogen atmosphere (99.999 % purity). Triplicate cultures were grown in 50 ml anaerobic serum bottles sealed with thick butyl rubber stoppers and aluminium caps, and incubated statically in the dark at 28 uC for 21 days. Growth in MB was tested at 4, 20, 28, 30, 37 and 40 uC and at pH 3.0–10.0 (at 1 pH unit intervals from pH 3.0–7.0 and 0.5 pH unit intervals from pH 7.0–10.0). The pH of MB was adjusted prior to sterilization using the following buffers: citric acid/sodium citrate (pH 3.0–6.0), Na2HPO4/ citric acid (pH 7.0–8.0) and lysine/NaOH (pH 9.0–10.0). Verification of the pH after autoclaving revealed only minor changes (Su et al., 2013). The NaCl concentration range and optimum for growth were determined in NaCl-free MB (5 g tryptone, 1 g yeast extract, 1 l distilled water, pH 7.6~7.8), supplemented with 0–7 % (at 1 % intervals) and 9–13 % (at 2 % intervals) (w/v) NaCl. Catalase and oxidase activities were assessed by addition of 3 % hydrogen peroxide to exponential-phase colonies and by using oxidase reagent (bioMe´rieux), respectively. Hydrolysis of starch, chitin, tyrosine, casein, gelatin, urea and Tweens 20, 40, 60 and 80 was tested using MA supplemented with 0.5 % (w/v) starch and 1 % (w/v) of the other substrates. Results were examined twice after growth on agar plates for 3 and 5 days. Carbon source utilization tests were carried out using API http://ijs.sgmjournals.org

W

+ 47.8

W

+ +

W

W

+ + 2 +

+ + W

50CH (bioMe´rieux). The above-mentioned tests were carried out on strain KD52T and the four reference strains. Biochemical tests were carried out using API 20NE, API 20E and API ZYM strips (bioMe´rieux) according to the manufacturer’s instructions, except that the NaCl concentration in all tests was adjusted to 3.0 %. Susceptibility to antibiotics was tested on MA for 5 days by using filter paper discs (Oxoid) containing various antibiotics (La´nyı´, 1987; Smibert & Krieg, 1994). All above-mentioned tests were incubated at 28 uC. All strains were tested using MA or MB except H. hydrossis DSM 1100T (grown on DSMZ medium 134). The novel isolate displayed basic characteristics of members of the family Saprospiraceae, e.g. rod-shaped, nonmotile cells (Fig. 2). Other phenotypic properties of strain KD52T are given in the species description and in Table 1. To determine the DNA G+C content, genomic DNA was extracted from cells cultured on MA for 6 days at 28 uC and analysed by reverse-phase HPLC (Tamaoka & Komagata, 1984). The DNA G+C content of the new isolate, KD52T, was 51 mol%. For cellular fatty acid analysis, the fatty acids of strain KD52T and the four reference strains grown on MA (except H. hydrossis DSM 1100T grown on DSMZ medium 134) at 28 uC for 6 days were saponified, methylated and extracted using the standard protocol of MIDI (Sherlock Microbial Identification System, version 6.0B). The fatty acids were analysed by GC (model 6850; Agilent Technologies) and identified by using the TSBA6 database of the Microbial Identification System (Sasser, 1990). The five strains had similar growth rates at 28 uC and the same physiological age at the time they were harvested. The dominant fatty acids (.10 %) of strain KD52T were iso-C15 : 0 (33.1 %), 3499

Z. Chen and others

iso-C15 : 1 G (14.8 %) and summed feature 3 (comprising C16 : 1v7c and/or C16 : 1v6c, 13.8 %). Significant amounts (.7 %) of iso-C17 : 0 3-OH were also present. The fatty acid compositions of the four reference strains were similar, with minor differences in the respective proportions of some fatty acids. Strain KD52T mainly differed from the other four strains by the presence of C13 : 1 at 12–13 and from H. hydrossis DSM 1100T and L. nigricans NBRC 102662T by the absence of iso-C15 : 1 F (Table 2). The major polar lipids of strain KD52T were diphosphatidylglycerol, three unidentified phospholipids, four unknown lipids and one unidentified aminolipid (Fig. S1, available in the online Supplementary Material).

a reverse-phase column (2 mm6125 mm, 3 mm, RP18; Macherey-Nagel) using methanol/heptane, 9 : 1 (v/v), as the eluent. The major quinone present in strain KD52T was MK-7, in line with species of the family Saprospiraceae (Daligault et al., 2011; Khan et al., 2007; Yoon et al., 2012a).

Respiratory quinones were analysed at the Chinese Center for Industrial Culture Collection (CICC) on an LDC Analytical (Thermo Separation Products) HPLC fitted with

Description of Phaeodactylibacter gen. nov.

Table 2. Cellular fatty acid contents of strain KD52T and closely related members of the family Saprospiraceae

Cells are Gram-staining-negative, non-motile, strictly aerobic and chemoheterotrophic rods. The dominant fatty acids are iso-C15 : 0, iso-C15 : 1 G and summed feature 3 (comprising C16 : 1v7c and/or C16 : 1v6c). The major respiratory quinone is menaquinone-7 (MK-7). A member of the family Saprospiraceae, phylum Bacteroidetes, according to 16S rRNA gene sequence studies. In addition to diphosphatidylglycerol, the polar lipids consist of three unidentified phospholipids, four unknown lipids and one unidentified aminolipid. The type species is Phaeodactylibacter xiamenensis.

Strains: 1, KD52T; 2, H. hydrossis DSM 1100T; 3, S. grandis JCM 21750T; 4, ‘Portibacter lacus’ KCTC 23747; 5, L. nigricans NBRC 102662T. All data are from this study. Results are presented as percentages of the total fatty acids. Fatty acids making up less than 1 % are marked as tr (trace); ND, not detected. Fatty acid iso-C13 : 0 C13 : 1 at 12–13 iso-C14 : 0 C14 : 0 iso-C15 : 0 iso-C15 : 1 G iso-C15 : 1 F anteiso-C15 : 0 C15 : 1v6c C15 : 0 3-OH iso-C15 : 0 3-OH iso-C16 : 0 C16 : 0 C16 : 0 3-OH iso-C16 : 1 G iso-C16 : 0 3-OH iso-C17 : 0 C17 : 0 C17 : 1v6c C17 : 0 3-OH iso-C17 : 0 3-OH C18 : 0 anteiso-C19 : 0 Summed feature 3* Summed feature 9*

1 tr 1.4 tr 1.2 33.1 14.8 ND

tr 4.7 tr tr 3.0 4.4 3.1 2.8 tr tr tr tr 1.3 8.9 tr

2

3

4

1.6

2.9 tr tr tr 32.8 3.9

tr tr tr tr 47.4 10.9

ND

ND

tr

tr 7.2 1.3 2.4 tr tr 1.0 1.3 4.2

ND

2.7 tr 27.5 12.6 10.1 4.1 ND

ND

ND

ND

4.0 tr 1.9 tr tr tr 1.8

ND

4.5 2.4 2.7 3.3 3.2 tr

ND

ND

ND

ND

tr

tr 1.0 32.3 tr

1.9 1.2 12.0 tr

ND

ND

4.5

3.6 tr

5 tr

On the basis of morphological, physiological and chemotaxonomic characteristics, as well as phylogenetic inference (Tables 1 and 2, Fig. 1), strain KD52T should be assigned to a novel species of a new genus in the family Saprospiraceae, for which the name Phaeodactylibacter xiamenensis gen. nov., sp. nov. is proposed.

Phaeodactylibacter (Phae.o.dac.ty.li.bac9ter. N.L. neut. n. Phaeodactylum generic name of an alga; N.L. masc. n. bacter a rod; N.L. masc. n. Phaeodactylibacter rod isolated from a culture of Phaeodactylum).

ND

1.7 tr 32.9 ND

5.0 tr ND

tr 2.6 1.4 5.5 tr

Description of Phaeodactylibacter xiamenensis sp. nov. Phaeodactylibacter xiamenensis (xi.a.men.en9sis. N.L. masc. adj. xiamenensis of Xiamen, a city in Fujian, China, where the type strain was first isolated).

*Summed features represent groups of two or three fatty acids that could not be separated by GLC with the MIDI system. Summed feature 3: C16 : 1v7c and/or C16 : 1v6c; summed feature 9: iso-C17 : 1v9c and/or 10-methyl C16 : 0.

Displays the following characteristics in addition to those listed in the genus description. Cells are 5.0–10.7 mm in length and 0.5–0.6 mm in diameter. Colonies grown for 7 days on MA are reddish orange, circular with entire margins, 1–3 mm in diameter, smooth, translucent and raised. Flexirubin pigments are not produced. Growth occurs at 20–37 uC (optimum, 28 uC), with 1–9 % (w/v) NaCl (optimum, 2.5 %) and at pH 5–8.5 (optimum, pH 6.0). Tweens 20, 40 and 60 are hydrolysed; Tween 80, starch, tyrosine, chitin, casein, urea, and gelatin are not hydrolysed. Catalase-negative and oxidase-positive. In the API ZYM strip, alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, valine arylamidase, cystine aminopeptidase, trypsin, a-chymotrypsin, acid phosphatase, naphthol-AS-BI-phosphoamidase, a-galactosidase, b-galactosidase, b-glucuronidase, a-glucosidase, b-glucosidase, Nacetyl-b-glucosaminidase and a-fucosidase activities are present; weak lipase (C14) activity is present; a-mannosidase activity is absent. In API 20NE strips, positive for reduction of nitrate to nitrite, b-glucosidase (aesculin hydrolysis),

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International Journal of Systematic and Evolutionary Microbiology 64

ND

13.8 1.6

ND

13.0 tr 1.0 12.6 tr

ND

ND

tr 1.2 1.3 ND

3.6 17.0 6.0 ND

10.8 1.8

Phaeodactylibacter xiamenensis gen. nov., sp. nov.

gelatin hydrolysis and b-galactosidase; negative for denitrification, indole production, D-glucose fermentation, arginine dihydrolase, urease activity, and utilization of Dglucose, L-arabinose, D-mannose, D-mannitol, N-acetylglucosamine, maltose, potassium gluconate, capric acid, adipic acid, malic acid, trisodium citrate and phenylacetic acid. In the API 20E strip, positive for b-galactosidase, arginine dihydrolase, lysine decarboxylase, ornithine decarboxylase, urease activity, acetoin production and gelatinase; negative for citrate utilization, H2S production, tryptophan deaminase, indole production and acid production from glucose, mannitol, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin and arabinose. In the API 50CH strip, positive for utilization of arbutin, aesculin ferric citrate, potassium 2ketogluconate and potassium 5-ketogluconate; negative for the other carbon sources. Susceptible to ampicillin, chloramphenicol, carbenicillin, cefoperazone, ciprofloxacin, clindamycin, vibramycin, erythromycin, cefazolin, minomycin, metronidazole, ofloxacin, penicillin G, polymyxin B, piperacillin, streptomycin, vancomycin, novobiocin, neomycin, cefalexin and gentamicin; resistant to kanamycin, norfloxacin, oxacillin, rifampicin, co-trimoxazole, tetracycline, sulfafurazole and trimethoprim. The major fatty acids are iso-C15 : 0, iso-C15 : 1 G and summed feature 3 (comprising C16 : 1v7c and/or C16 : 1v6c). The complete fatty acid composition is given in Table 2. The type strain is KD52T (5MCCC 1F01213T5KCTC 32575T), isolated from a culture of the marine alga Phacodactylum tricornutum in Xiamen, China. The DNA G+C content of the type strain is 51.0 mol%.

Acknowledgements This work was supported by the National Nature Science Foundation of China (41376119), the Joint project of National Natural Science Foundation of China (NSFC) and Shandong province, Marine Ecology and Environmental Sciences (grant no. U1406403) and Public Science and Technology Research Funds Projects of Ocean (201305016, 201305022). We would like to thank Professor I. J. Hodgkiss from Hongkong University for help with English.

Daligault, H., Lapidus, A., Zeytun, A., Nolan, M., Lucas, S., Del Rio, T. G., Tice, H., Cheng, J.-F., Tapia, R. & other authors (2011).

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arachidonic acid-containing bacterium isolated from the southern coastline of Thailand. Int J Syst Evol Microbiol 56, 2931–2935. Hosoya, S., Arunpairojana, V., Suwannachart, C., Kanjana-Opas, A. & Yokota, A. (2007). Aureispira maritima sp. nov., isolated

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Emended descriptions of the genus Lewinella and of Lewinella cohaerens, Lewinella nigricans and Lewinella persica, and description of Lewinella lutea sp. nov. and Lewinella marina sp. nov. Int J Syst Evol Microbiol 57, 2946–2951. Kim, O.-S., Cho, Y.-J., Lee, K., Yoon, S.-H., Kim, M., Na, H., Park, S.-C., Jeon, Y. S., Lee, J.-H. & other authors (2012). Introducing EzTaxon-

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A novel Gram-staining-negative, aerobic, rod-shaped, non-motile, reddish-orange and chemoheterotrophic bacteria, designated strain KD52(T), was isolat...
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