A polyphasic study was undertaken to determine the taxonomic position of two halophilic archaeal strains, TH32T and YPL4T, isolated from saline soil and a salt mine in PR China, respectively. Strains TH32T and YPL4T both have two dissimilar 16S rRNA genes. The two strains exhibited sequence similarities of 91.5-95.5 % for 16S rRNA genes and 90.9 % for the rpoB' gene. Sequence similarities of 16S rRNA genes and the rpoB' gene between the two strains and the current four members of Halosimplex were 90.6-97.4 % and 91.4-93.5 %, respectively. Phylogenetic analysis revealed that the two strains formed different branches separating them from the current Halosimplex members. Several phenotypic characteristics differentiate strains TH32T and YPL4T from current Halosimplex members. The polar lipids of the two strains are phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and four glycolipids. Two of the glycolipids are chromatographically identical to disulfated mannosyl glucosyl diether and sulfated mannosyl glucosyl diether, respectively, and the remaining two glycolipids are unidentified. The average nucleotide identity (ANI) and in silico DNA-DNA hybridization (DDH) values between the two strains and the current members of Halosimplex (ANI 80.4-89.2 % and in silico DDH 24.0-41.8 %) were much lower than the threshold values proposed as a species boundary, suggesting that the two strains represent novel species of Halosimplex. The values between the two strains (ANI 81.3 % and in silico DDH 24.9 %) were also much lower than the recommended threshold values, which revealed that the two strains represent two genomically different species of Halosimplex. These results showed that strains TH32T (=CGMCC 1.15190T=JCM 30840T) and YPL4T (=CGMCC 1.15329T=JCM 31108T) represent two novel species of Halosimplex, for which the names Halosimplex halophilum sp. nov. and Halosimplex salinum sp. nov. are proposed.
Keywords: Halosimplex halophilum sp. nov.; Halosimplex salinum sp. nov.; halophilic archaea; saline soil; salt mine.