1887

Abstract

Four halophilic archaeal strains, designated TNN18, TBN12, TNN28 and TBN19, were isolated from brines sampled from two artificial marine solar salterns in eastern China. Strains TNN18 and TNN28 were isolated from the Tainan marine solar saltern, whereas strains TBN12 and TBN19 were from the Taibei marine solar saltern. Colonies of the four strains were red-pigmented and their cells were pleomorphic, motile, Gram-reaction-negative rods. Strains TNN18 and TBN12 were able to grow at 25–50 °C (optimum 37 °C), in 10–30 % (w/v) NaCl (optimum 15 %), with 0–1.0 M MgCl (optimum 0.05 M) and at pH 5.5–9.0 (optimum pH 7.0–7.5), while strains TNN28 and TBN19 were able to grow at 20–50 °C (optimum 37 °C), in 15-30 % (w/v) NaCl (optimum 18–20 %), in 0.005–1.0 M MgCl (optimum 0.01–0.3 M) and at pH 6.0–9.0 (optimum pH 7.0–7.5). Cells of these strains lyse in distilled water; minimal NaCl concentrations to prevent cell-lysis are 10 % (w/v) for strains TNN18 and TBN12 and 12 % (w/v) for strains TNN28 and TBN19. The major polar lipids of strains TNN18 and TBN12 were phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and one major glycolipid (GL1), which was chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-1). Minor amounts of other lipids (GL0, GL2, GL3 and GL4) were also detectable. The polar lipid profiles of strains TNN28 and TBN19 contained PG, PGP-Me, GL1, which was chromatographically identical to S-DGD-1, and three to four minor unidentified glycolipids (GL2–GL5). Phylogenetic analyses revealed that strains TNN18 and TBN12 formed a distinct clade with strains of the closest related species, (91.5–91.8 % 16S rRNA gene sequence similarity) and strains TNN28 and TBN19 formed a distinct clade with strains of the species (89.9–93.3 % similarity) and two members of the genus (92.5–93.3 % similarity). The DNA G+C contents of strains TNN18, TBN12, TNN28 and TBN19 were 61.5, 62.4, 61.9 and 61.5 mol%, respectively. DNA–DNA hybridization values between strains TNN18 and TBN12, and strains TNN28 and TBN19 were 82.9 % and 88.2 %, respectively. The phenotypic, chemotaxonomic and phylogenetic properties suggest that the four strains represent two novel species of two new genera within the family , for which the names gen. nov., sp. nov. (type strain TNN18 = CGMCC 1.10118 = JCM 16424) and gen. nov., sp. nov. (type strain TNN28 = CGMCC 1.10123 = JCM 16425) are proposed.

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2011-11-01
2019-12-08
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Phase-contrast micrographs of strain TNN18 (a), strain TBN12 (b), TNN28 (c) and strain TBN19 (d) grown on liquid NOM-3 medium to late-exponential phase at 37 °C . Bar, 2 μm.

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Analysis of the lipid composition of strain TNN18 , strain TBN12, strain TNN28 and strain TBN19 using TLC. (a-d) Two-dimensional TLC of strains TNN18 (a), TBN12 (b), TNN28 (c) and TBN19 (d). (e-f) One-dimensional TLC. Lanes: 1 and 7, CGMCC 1.2367 (=ATCC 33170); 2, Strain TNN18 ; 3, Strain TBN12; 4 and 11, CGMCC 1.2150 ; 5, CGMCC 1.6168 ; 6, Halogranum rubrum RO2-11 ; 8, Halosimplex carlsbadense JCM 11222 ; 9, Strain TNN28 ; 10, Strain TBN19; 12, Haloarcula marismortui CGMCC 1.1784 . Circled spots are minor glycolipids of two strains. Abbreviations: PG, phosphatidylglycerol; PGP-Me, phosphatidylglycerol phosphate methyl ester; PGS, phosphatidylglycerol sulfate; GL, glycolipid; DGD-1, mannosyl glucosyl diether; DGD-2, an unknown diglycosyl diether; S-DGD-1, sulfated mannosyl glucosyl diether; S2-DGD, bisulfated diglycosyl diether; TGD-1, galactosyl mannosy glucosyl diether; TGD-2, glucosyl mannosyl glucosyl diether; S-TGD-1, sulfated galactosyl mannosy glucosyl diether; S-TeGD, sulfated tetraglycosyl diether; F, first dimension of TLC; S, second dimension of TLC.

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