Cellular morphology was examined after
growth on MA at 30 °C for 2 days by transmission electron microscopy. Gliding motility was assessed on the edge of a hanging drop of a fresh MB culture as recommended by Bernardet et al. (2002). Anaerobic growth was evaluated on MA in an anaerobic chamber system (Coy Laboratory Products Inc.). The pH range (4–9 at 1 pH unit intervals) for growth was determined using MB. The final pH was adjusted with NaOH and HCl solutions after autoclaving. The requirements for sea salts (0%, 1%, 3%, 5%, 10%, 20% and 30%, w/v; Sigma) were tested using R2A medium (Conda). The temperature range for growth was determined on MA at 5–50 °C at 5 °C intervals. Catalase and oxidase activities as well as hydrolysis of gelatin, starch and Tween 80 using MA as the basal medium were tested as described by Smibert & Krieg (1994). DNase test agar (Difco) supplemented with 2.5% (w/v) NaCl was used for DNase assay. PLX3397 mw Arginine Selleck CX-4945 dihydrolase and urease activities, nitrate reduction, acid production from glucose and indole production tests were performed using an API 20NE kit (bioMérieux) according to the manufacturer’s instructions, and other enzymatic activities were determined using an API ZYM kit (bioMérieux). Kits were inoculated with a heavy bacterial suspension in AUX media (bioMérieux) supplemented with 2.5% (w/v) NaCl. Carbon source utilization was tested by incubation at 37 °C
for 2 weeks on basal agar medium supplemented with yeast extract (0.64 g KCl, 23.6 g NaCl, 5.94 g MgSO4·7H2O, 4.53 g MgCl2·6H2O, 1.3 g CaCl2·2H2O, 0.2 g NH4Cl, 0.2 g NaNO3, 15 g Bacto agar, 0.05 g yeast extract, per liter distilled water; Choi & Cho, 2006) containing 0.2% of the carbon source. DNA G+C content was determined by HPLC analysis of deoxyribonucleosides
as described by Mesbah et al. (1989), using a reverse-phase column (Supelcosil LC-18-S; Supelco). Experiments were performed in triplicates. Chemotaxonomic characteristics selleck inhibitor were determined from cells grown on MA or in MB at 30 °C for 2–3 days. Fatty acid methyl ester analysis was carried out by GLC according to the instructions of the Microbial Identification system (MIDI). Isoprenoid quinones were isolated by the method of Minnikin et al. (1984) and analysed by HPLC (Varian) as described by Collins (1985). Flexirubin-type pigments were sought using the KOH test according to Bernardet et al. (2002). Polar lipids were extracted from freeze-dried cell materials by the method of Tindall (1990a, b) and separated by 2D silica-gel thin-layer chromatography. Total lipids and specific functional groups were detected using molybdophosphoric acid, molybdenum blue spray, ninhydrin and α-naphthol, as described previously (Minnikin et al., 1984). The nearly complete 16S rRNA gene sequence of strain JC2131T was obtained (1428 bp). The GenBank accession number for the 16S rRNA gene sequence of the strain JC2131T is FJ387163.