1887

Abstract

Two extremely halophilic archaea, strains RO2-11 and HO2-1, were isolated from two Chinese marine solar salterns, Rudong solar saltern and Haimen solar saltern, respectively. Cells of the two strains were polymorphic and Gram-stain-negative; colonies were red-pigmented. The two strains grew at NaCl concentrations of 2.6–4.3 M (optimum 3.9 M) and required at least 0.1 M Mg for growth. They were able to grow over a pH range of 6.0–8.0 and a temperature range of 20–50 °C, with optimal pH of 7.5 and optimal temperature of 37 °C. The major polar lipids of strain RO2-11 and strain HO2-1 were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and three glycolipids, two of them chromatographically identical to S-DGD-1 and DGD-1, the third unidentified. The 16S rRNA gene sequence similarity of strain RO2-11 and strain HO2-1 was 99.3 % and highest sequence similarity with the closest relative () was 91.4 %. Based on the data obtained, the two isolates could not be classified in any recognized genus of the family . Strain RO2-11 and strain HO2-1 are thus considered to represent a novel species of a new genus within the family , for which the name gen. nov., sp. nov. is proposed. The type strain is RO2-11 (=CGMCC 1.7738 =JCM 15772).

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2010-06-01
2019-10-16
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Micrographs of cells of strains RO2-11 and HO2-1 in liquid medium under optimal conditions. (a) Scanning electron micrograph of cells of strain RO2-11 . (b) Scanning electron micrograph of cells of strain HO2-1. (c) Phase-contrast micrograph of strain RO2-11 showing pleomorphic forms. For scanning electron microscopy examination, 0.5 ml samples were fixed overnight at 4 °C by adding glutaraldehyde to a final concentration of 5%. A 10 µl sample was smeared on a polylysine-coated coverslip and air-dried. The coverslip was then serially dehydrated in 40, 70, 90 and 100% ethanol solutions (10 min at each stage), critical-point dried, mounted on a specimen stub, sputter-coated with gold and viewed in a Hitachi S-4800 scanning electron microscope.

IMAGE

Thin-layer chromatograms on Merck silica gel 60 F254 aluminium-backed thin-layer plates of the phospholipids (a) and glycolipids (b, c) from strain RO2-11 and related members of the family . The plate (a) was detected by spraying with the phosphate stain reagent (Kates, 1986) to detect phospholipids. The plate (b) subjected to single development in the solvent chloroform/methanol/acetic acid/water (85:22.5:10:4, by vol.) was detected by spraying with 0.5% α-naphthol in methanol/water (1:1, v/v) and then with sulfuric acid/ethanol (1:1, v/v) followed by heating at 120 °C for 5 min to detect glycolipids only. The plate (c) subjected to double development in the same solvent as plate (b) was detected by spraying with sulfuric acid/ethanol (1:2, v/v) followed by heating at 150 °C for 3 min to detect phospholipids and glycolipids. Lanes: 1 and 7, CGMCC 1.2367 (=ATCC 33170); 2 and 8, CGMCC 1.2150 ; 3 and 9, Strain HO2-1; 4 and 10, Strain RO2-11 ; 5, CGMCC 1.1784 ; 6 and 11, CGMCC 1.5332 . Circled spots are glycolipids of strain RO2-11T. Abbreviations: PG, phosphatidylglycerol; PGP-Me, phosphatidylglycerol phosphate methyl ester; GL, glycolipid; F, first dimension of TLC; S, second dimension of TLC.

IMAGE

Phylogenetic tree derived from maximum-parsimony analysis based on 16S rRNA gene sequences showing the relationship of strain RO2-11 , strain HO2-1 and other close relatives within the family . Only bootstrap values greater than 50% are shown (1000 replications). Bar, 50 expected changes per 1000 nucleotide positions. [ PDF] (401 KB)

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