Halobellus limi sp nov and Halobellus salinus sp nov., isolated from two marine solar salterns

Two halophilic archaea, strains TBN53(T) and CSW2.24.4(T), were characterized to elucidate their taxonomic status. Strain TBN53(T) was isolated from the Taibei marine solar saltern near Lianyungang city, Jiangsu province, China, whereas strain CSW2.24.4(T) was isolated from a saltern crystallizer in Victoria, Australia. Cells of the two strains were pleomorphic, stained Gram-negative and produced red-pigmented colonies. Strain TBN53(T) was able to grow at 25-55 degrees C (optimum 45 degrees C), with 1.4-5.1 M NaCl (optimum 2.6-3.9 M NaCl), with 0-1.0 M MgCl2 (optimum 0-0.1 M MgCl2) and at pH 5.5-9.5 (optimum pH 7.0), whereas strain CSW2.24.4(T) was able to grow at 25-45 degrees C (optimum 37 degrees C), with 2.6-5.1 M NaCl (optimum 3.4 M NaCl), with 0.01-0.7 M MgCl2 (optimum 0.05 M MgCl2) and at pH 5.5-9.5 (optimum pH 7.0-7.5). Cells of the two isolates lysed in distilled water. The minimum NaCl concentrations that prevented cell lysis were 8% (w/v) for strain TBN53(T) and 12% (w/v) for strain CSW2.24.4(T). The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate, with two glycolipids chromatographically identical to sulfated mannosyl glucosyl diether and mannosyl glucosyl diether, respectively. Trace amounts of other unidentified lipids were also detected. On the basis of 16S rRNA gene sequence analysis, strains TBN53(T) and CSW2.24.4(T) showed 94.1% similarity to each other and were closely related to Halobellus clavatus TNN18(T) (95.0 and 94.7% similarity, respectively). Levels of rpoB' gene sequence similarity between strains TBN53(T) and CSW2.24.4(T), and between these strains and Halobellus clavatus TNN18(T) were 88.5, 88.5 and 88.1%, respectively. The DNA G+C contents of strains TBN53(T) and CSW2.24.4(T) were 69.2 and 67.0 mol%, respectively. The level of DNA-DNA relatedness between strain TBN53(T) and strain CSW2.24.4(T) was 25%, and these two strains showed low levels of DNA-DNA relatedness with Halobellus clavatus TNN18(T) (30 and 29% relatedness, respectively). Based on these phenotypic, chemotaxonomic and phylogenetic properties, two novel species of the genus Halobellus are proposed to accommodate these two strains, Halobellus limi sp. nov. (type strain TBN53(T)=CGMCC 1.10331(T)=JCM 16811(T)) and Halobellus salinus sp. nov. (type strain CSW2.24.4(T)=DSM 18730(T)=CGMCC 1.10710(T)=JCM 14359(T)).