The low level of 16S rRNA gene sequence similarity, ANI, AAI and DDH values, the independent phylogenetic position, and the differences in phenotypic properties and chemotaxonomic characteristics between strain HD8-83T, LYG-36T, DLLS-82, RC-68T and its closest phylogenetic described species of Halorussus indicated that these strains diverged from current members. Therefore, we suggest that the strains HD8-83T, LYG-36T, DLLS-82, and RC-68T represents three novel species of the genus Halorussus for which the names Halorussus halobius sp. nov. (type strain HD8-83T = CGMCC 1.15334T = JCM 31110T), Halorussus marinus sp. nov. (type strain LYG-36T = CGMCC 1.13606T = JCM 32952T, reference strain DLLS-82 = CGMCC 1.13604 = JCM 32951) and Halorussus pelagicus sp. nov. (type strain RC-68T = CGMCC 1.13609T = JCM 32953T) are proposed.
Description of Halorussus halobius sp. nov.
Halorussus halobius (ha.lo'bi.us. Gr. n. hals, halos salt; Gr. n. bius life; M.L. masc. adj. halobius living on salt).
Cells are Gram-stain-negative, motile, rod (cell size 0.8–1.0×1.0–3.5 µm) under optimal growth conditions. Colonies on agar plates are red, elevated and round. Optimal growth is obtained at 4.3 M NaCl (range: 0.9–4.8 M), 0.05 M MgCl2 (range: 0–1.0 M), 35°C (range: 25–50°C) and pH 7.5 (range: 6.0–9.5). Cells lyse in distilled water and the minimal NaCl concentration to prevent cell lysis is 0.85 M. The catalase activity and oxidase activity are positive. Anaerobic growth is observed in the presence of nitrate, with nitrate reduction to nitrite but no gas formation from nitrate. Anaerobic growth with DMSO and L-arginine is not detected. Indole formation and H2S formation are negative. The type strain can not hydrolyze gelatin, hydrolyze casein, starch or Tween 80. The following substrates are utilized as single carbon and energy sources for growth: D-glucose, D-galactose, maltose, sucrose, glycerol, D-mannitol, D-sorbitol, pyruvate, DL-lactate, succinate, L-malate and citrate. The following substrates are utilized as single carbon, nitrogen or energy sources for growth: glycine, L-arginine, L-aspartate, L-glutamate and L-ornithine. D-mannose, D-fructose, L-sorbose, D-ribose, D-xylose, lactose, starch, acetate or fumarate cannot be utilized as single carbon and energy sources for growth. L-alanine and L-lysine cannot be utilized as single carbon, nitrogen or energy sources for growth.
The major polar lipids are phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and six glycolipids, sulfated mannosyl glucosyl diether (S-DGD-1), galactosyl mannosyl glucosyl diether (TGD-1), mannosyl glucosyl diether (DGD-1), diglycosyl diether (DGD-2) and two unknown glycolipids (GL0 and GL1b). The major respiratory quinones were menaquinone MK-8 and MK-8(H2).
The DNA G + C content of strain HD8-83T is 67.3 mol%. The type strain is HD8-83T (= CGMCC 1.15334T = JCM 31110T) and was isolated from salted brown alga Laminaria produced at Dalian, Liaoning Province, China.
Description of Halorussus marinus sp. nov.
Halorussus marinus (ma.ri'nus. L. masc. adj. marinus, of the sea, marine).
Cells are Gram-stain-negative, motile, pleomorphic rod (cell size 1.0–1.5×1.2–3.6 µm) under optimal growth conditions. Colonies on agar plates are white or red, elevated and round. Optimal growth is obtained at 3.1 M NaCl (range: 0.9–4.8 M), 0.05–0.1 M MgCl2 (range: 0–1.0 M), 30–40°C (range: 20–55°C) and pH 7.0-7.5 (range: 5.0–9.5). Cells lyse in distilled water and the minimal NaCl concentration to prevent cell lysis is 0.85 M. The catalase activity is positive and the oxidase activity is negative. Nitrate reduction to nitrite is observed but gas formation from nitrate is not detected. Anaerobic growth is observed in the presence of nitrate, DMSO and L-arginine. Indole formation and H2S formation are positive. The type strain cannot hydrolyze gelatin, hydrolyze casein, starch or Tween 80. The following substrates are utilized as single carbon and energy sources for growth: D-glucose, D-mannose, D-galactose, maltose, sucrose, lactose, glycerol, D-mannitol, D-sorbitol, acetate, pyruvate, DL-lactate, succinate, L-malate, fumarate and citrate. The following substrates are utilized as single carbon, nitrogen or energy sources for growth: glycine, L-alanine, L-arginine, L-aspartate, L-glutamate, L-lysine and L-ornithine. D-fructose, L-sorbose, D-ribose, D-xylose or starch cannot be utilized as single carbon and energy sources for growth.
The major polar lipids are phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and three glycolipids, sulfated mannosyl glucosyl diether (S-DGD-1), galactosyl mannosyl glucosyl diether (TGD-1) and diglycosyl diether (DGD-2). The DNA G + C content of strain LYG-36T is 66.9 mol%. The major respiratory quinones were menaquinone MK-8 and MK-8(H2).
The type strain is LYG-36T (= CGMCC 1.13606T = JCM 32952T), reference strain is DLLS-82 (= CGMCC 1.13604 = JCM 32951), and they were isolated from salted brown alga Laminaria produced at Lianyungang, Jiangsu Province, China and Dalian, Liaoning Province, China, respectively.
Description of Halorussus pelagicus sp. nov.
Halorussus pelagicus (pe.la'gi.cus. L. masc. adj. pelagicus, belonging to the sea).
Cells are Gram-stain-negative, motile, pleomorphic rod (cell size 0.8–1.0×1.0–5.0 µm) under optimal growth conditions. Colonies on agar plates are red, elevated and round. Optimal growth is obtained at 2.6 M NaCl (range: 2.0–4.8 M), 0.3 M MgCl2 (range: 0–1.0 M), 35°C (range: 20–50°C) and pH 7.0 (range: 5.0–9.5). Cells lyse in distilled water and the minimal NaCl concentration to prevent cell lysis is 0.85 M. The catalase activity is positive and the oxidase activity is negative. Anaerobic growth is not observed in the presence of nitrate, DMSO and L-arginine, and not detected nitrate reduction to nitrite or gas formation from nitrate. Indole formation and H2S formation are positive. The type strain can hydrolyze gelatin and casein, but not hydrolyze starch or Tween 80. The following substrates are utilized as single carbon and energy sources for growth: D-glucose, D-mannose, D-galactose, sucrose, lactose, glycerol, D-mannitol, D-sorbitol, acetate, pyruvate, DL-lactate, succinate, L-malate, fumarate and citrate. The following substrates are utilized as single carbon, nitrogen or energy sources for growth: L-alanine, L-arginine, L-aspartate, L-glutamate, L-lysine and L-ornithine. D-fructose, L-sorbose, D-ribose, D-xylose,maltose or starch cannot be utilized as single carbon and energy sources for growth. Glycine cannot be utilized as single carbon, nitrogen or energy sources for growth.
The major polar lipids are phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and three glycolipids, sulfated mannosyl glucosyl diether (S-DGD-1), galactosyl mannosyl glucosyl diether (TGD-1) and diglycosyl diether (DGD-2). The DNA G + C content of strain RC-68T is 63.8 mol%. The major respiratory quinones were menaquinone MK-8 and MK-8(H2).
The type strain is RC-68T (= CGMCC 1.13609T = JCM 32953T) and was isolated from salted brown alga Laminaria produced at Rongcheng, Shandong Province, China.