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Pricia Antarctica Gen. Nov., Sp. Nov., a Member of the Family Flavobacteriaceae, Isolated from Antarctic Intertidal Sediment

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Pricia Antarctica Gen. Nov., Sp. Nov., a Member of the Family Flavobacteriaceae, Isolated from Antarctic Intertidal Sediment International Journal of Systematic and Evolutionary Microbiology (2012), 62, 2218–2223 DOI 10.1099/ijs.0.037515-0 Pricia antarctica gen. nov., sp. nov., a member of the family Flavobacteriaceae, isolated from Antarctic intertidal sediment Yong Yu, Hui-Rong Li, Yin-Xin Zeng, Kun Sun and Bo Chen Correspondence SOA Key Laboratory for Polar Science, Polar Research Institute of China, Shanghai 200136, Yong Yu PR China [email protected] A yellow-coloured, rod-shaped, Gram-reaction- and Gram-staining-negative, non-motile and aerobic bacterium, designated strain ZS1-8T, was isolated from a sample of sandy intertidal sediment collected from the Antarctic coast. Flexirubin-type pigments were absent. In phylogenetic analyses based on 16S rRNA gene sequences, strain ZS1-8T formed a distinct phyletic line and the results indicated that the novel strain should be placed in a new genus within the family Flavobacteriaceae. In pairwise comparisons between strain ZS1-8T and recognized species, the levels of 16S rRNA gene sequence similarity were all ,93.3 %. The strain required + + Ca2 and K ions as well as NaCl for growth. Optimal growth was observed at pH 7.5–8.0, 17–19 6C and with 2–3 % (w/v) NaCl. The major fatty acids were iso-C15 : 1 G, iso-C15 : 0, summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1v7c), an unknown acid with an equivalent chain-length of 13.565 and iso-C17 : 0 3-OH. The major respiratory quinone was MK-6. The predominant polar lipid was phosphatidylethanolamine. The genomic DNA G+C content was 43.9 mol%. Based on the phylogenetic, phenotypic and chemotaxonomic data, strain ZS1-8T represents a novel species in a new genus in the family Flavobacteriaceae for which the name Pricia antarctica gen. nov., sp. nov. is proposed. The type strain of the type species is ZS1-8T (5JCM 17291T5DSM 23421T). The family Flavobacteriaceae, one of the major branches of sandy intertidal sediments in the Antarctic, a Flavo- the phylum Bacteroidetes (Garrity & Holt, 2001), was first bacteriaceae-like bacterium, designated strain ZS1-8T, was proposed by Jooste (1985). The name of the family was isolated (Yu et al., 2010). In the present study, the taxonomic validly published by Reichenbach (1992) before the position of the novel strain was investigated using a description of the taxon was emended by Bernardet et al. polyphasic approach. The results indicate that the strain (1996, 2002). At the time of writing, the family Flavo- represents a novel species in a new genus within the family bacteriaceae comprises 94 genera representing bacteria in Flavobacteriaceae. various terrestrial, marine and freshwater environments. In March 2007, during the 23rd Chinese National Antarctic Members of 13 of the genera, namely Aequorivita (Bowman Research Expedition, a sample of sandy sediment was & Nichols, 2002), Gelidibacter (Bowman et al., 1997), Gilli- collected from the intertidal zone on the coast close to the sia (Van Trappen et al., 2004), Lacinutrix (Bowman & Chinese Antarctic Zhongshan Station, which lies on the Nichols, 2005), Leeuwenhoekiella (Nedashkovskaya et al., Larsemann Hills of Princess Elizabeth Land (69u 229 220 S 2005b), Olleya (Nichols et al., 2005), Polaribacter (Gosink T 76u 229 340 E). A novel bacterium, designated strain ZS1-8 , et al., 1998), Psychroflexus (Bowman et al., 1998), Psychro- was isolated, by the dilution plating technique, on 1/10- serpens (Bowman et al., 1997), Salegentibacter (McCammon strength marine agar (MA; Difco) after incubation in the & Bowman, 2000), Sejongia (Yi et al., 2005), Subsaxi- dark at 4 uC for up to 8 weeks (Yu et al., 2010). Costertonia bacter (Bowman & Nichols, 2005) and Subsaximicrobium aggregata JCM 13411T was obtained from the Japan (Bowman & Nichols, 2005), have been isolated from marine Collection of Microorganisms (JCM) and used as a reference and coastal zones in the Antarctic. During an investigation strain. Strains were stored at 280 uC in marine 2216 broth on the diversity of the culturable bacteria associated with (MB; Difco) supplemented with 30 % (v/v) glycerol. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene Cell morphology was examined by phase-contrast micro- sequence of strain ZS1-8T is FJ889677. scopy (Eclipse 80i; Nikon) and transmission electron Three supplementary figures are available with the online version of this microscopy (JEM-100CX II; JEOL). Colonial morphology paper. was observed on MA after incubation at 18 uC for 7 days. 2218 037515 G 2012 IUMS Printed in Great Britain Pricia antarctica gen. nov., sp. nov. Table 1. Phenotypic characteristics that distinguish strain ZS1-8T from Costertonia aggregata JCM 13411T Strains: 1, ZS1-8T;2,Costertonia aggregata JCM 13411T. All data are from this study. Both strains give a positive result for alkaline phosphatase and leucine arylamidase activities but negative for ornithine decarboxylase, lipase (C14), a-chymotrypsin, acid phosphatase, cystine arylamidase, a- galactosidase, b-glucuronidase and a-glucosidase activities. Both produce acid from aesculin and utilize D-galactose, acetic acid and citric acid but not a-cyclodextrin, D-arabitol, D-psicose, succinic acid monomethyl ester, formic acid, D-glucosaminic acid, a-hydroxybutyric acid, itaconic acid, a- ketobutyric acid, a-ketoglutaric acid, sebacic acid, succinamic acid, L-phenylalanine, thymidine or 2,3-butanediol. +, Positive; 2, negative. Characteristic 12 Enzyme activity: Arginine dihydrolase, lysine decarboxylase, urease, trypsin, b-galactosidase, b-glucosidase, N-acetyl-b-glucosaminidase, 2 + a-mannosidase and a-fucosidase Acid production from: Glycerol, L-arabinose, D-ribose, D-xylose, D-galactose, D-glucose, D-mannitol, melezitose and D-fucose + 2 Utilization of: Dextrin, glycogen, Tweens 40 and 80, N-acetyl-D-galactosamine, N-acetyl-D-glucosamine, adonitol, L-arabinose, cellobiose, 2 + L-erythritol, D-fructose, L-fucose, gentiobiose, a-D-glucose, myo-inositol, a-lactose, lactulose, maltose, D-mannitol, D-mannose, melibiose, methyl b-D-glucoside, raffinose, L-rhamnose, D-sorbitol, sucrose, trehalose, turanose, xylitol, methyl pyruvate, acetic acid, cis-aconitic acid, D-galactonic acid lactone, D-galacturonic acid, D-gluconic acid, D-glucuronic acid, b-hydroxybutyric acid, c-hydroxybutyric acid, p-hydroxyphenylacetic acid, a-ketoglutaric acid, a-ketovaleric acid, DL-lactic acid, malonic acid, propionic acid, quinic acid, D-saccharic acid, succinic acid, bromosuccinic acid, glucuronamide, L-alaninamide, D-alanine, L-alanine, L-alanyl glycine, L-asparagine, L-aspartic acid, L-glutamic acid, glycyl L-aspartic acid, glycyl L-glutamic acid, L-histidine, hydroxy L-proline, L-leucine, L-ornithine, L-proline, L-pyroglutamic acid, D-serine, L-serine, L-threonine, c-aminobutyric acid, urocanic acid, inosine, uridine, phenylethylamine, putrescine, 2-aminoethanol, glycerol, DL-a-glycerol phosphate, glucose 1-phosphate and glucose 6-phosphate Table 2. Phenotypic characteristics that differentiate strain ZS1-8T from closely related members of the family Flavobacteriaceae Taxa: 1, strain ZS1-8T (data from this study); 2, Costertonia aggregata KOPRI 13342T (Kwon et al., 2006; this study); 3, Flagellimonas eckloniae DOKDO 007T (Bae et al., 2007); 4, Croceitalea (two species; Lee et al., 2008); 5, Muricauda (six species; Arun et al., 2009; Bruns et al., 2001; Hwang et al., 2009; Yoon et al., 2005b, 2008); 6, Maribacter (10 species; Barbeyron et al., 2008; Cho et al., 2008; Nedashkovskaya et al., 2004a, 2007; Yoon et al., 2005a; Zhang et al., 2009); 7, Zobellia (five species; Barbeyron et al., 2001; Nedashkovskaya et al., 2004b); 8, Pibocella ponti KMM 6031T + (Nedashkovskaya et al., 2005a). All the taxa are positive for catalase, require Na for growth and have MK-6 as their major respiratory quinone. +, Positive; 2, negative; 2/+ or +/2, variable reaction between species (where the first sign indicates the result for the type species). Characteristic 1 2 3 4 5 6 7 8 Flexirubin-type pigment 222222+ 2 Gliding motility 22222+/2 ++ Growth above 25 uC 2 +++++/2 ++ Growth at 0 uC + 22222 22 Optimum temperature for growth (uC) 17–19 28–30* 26–29 29–35 20–37 21–24D 21–35 21–24 ‘Seawater’ requirementd +++22 2 22 Facultative anaerobe 2222+/22 22 Nitrate reduction 2 + 222+/2 + 2 Oxidase activity ++22+/2 +++ Hydrolysis of: Agar 22222+/2 + 2 Casein 22+ 22 2+/2 + Gelatin 2 + 2 +/22 +/2 ++ Starch 222222/++/2 + DNA G+C content (mol%) 43.9 35.8 56.3 59-67 41–51 35–39 36–43 35.5 *Data from this study. DOptimum temperature for growth of Maribacter antarcticus CL-AP4T is recorded as 10 uC. + + + + dNa alone does not support growth because additional cations that are present in seawater, such as Mg2 ,Ca2 and/or K , are also required. http://ijs.sgmjournals.org 2219 Y. Yu and others Gram staining was determined by the Hucker staining Table 3. Cellular fatty acid contents (%) of strain ZS1-8T and method (Murray et al., 1994) and the Gram reaction was Costertonia aggregata JCM 13411T investigated by the KOH lysis method (Buck, 1982). T T Gliding motility and flexirubin-type pigment production Strains: 1, ZS1-8 ;2,Costertonia aggregata JCM 13411 . All data are from this study. 2, Not detected; ECL, equivalent chain-length. were determined as described by Bowman (2000). Oxidase activity was determined using commercial oxidase test Fatty acid 1 2 strips (bioMe´rieux). Catalase activity was detected by bubble production in 3 % (v/v) H2O2. Growth at different Straight-chain temperatures was investigated in MB for 5–30 days. The C12 : 0 2 2.1 pH range for growth was determined using MB broth at C13 : 0 2 4.2 pH 5.0–10.5 (in increments of 0.5 pH unit) and 18 uC, C15 : 0 3.6 7.0 after the pH of the medium had been adjusted by the C16 : 0 0.7 2 addition of MES, MOPS, Tris or CHES buffers (each at Branched 50 mM). Growth with various concentrations of NaCl iso-C13 : 0 2 9.5 (0–10 %, w/v, in increments of 1 %) was tested in a me- iso-C15 : 0 17.0 12.2 dium containing (l21)5gMgCl, 2 g MgSO , 0.5 g CaCl , anteiso-C15 : 0 0.6 3.8 2 4 2 2 1 g KCl, 5 g peptone and sufficient KOH to give a pH of iso-C16 : 0 1.45 + + + Unsaturated 7.5 (Smibert & Krieg, 1994).
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