An extremely halophilic Archaeon belonging to the order Halobacteriales was isolated from the solar salterns of Cabo Rojo, Puerto Rico. The organism, designated strain PR5 T , is rod-shaped, non-motile and requires at least 12 % (w/v) NaCl to grow. The strain is highly thermotolerant : its temperature optimum is 50 SC and growth is possible up to 60 SC. Polar lipid analysis revealed the presence of the bis-sulfated glycolipid S 2 -DGD-1 as sole glycolipid and the absence of the glycerol diether analogue of phosphatidylglycerosulfate. Both C 20 ,C 20 and C 20 ,C 25 core lipids are present. The GMC content of the DNA is 63 3 mol %. According to 16S rDNA sequence data, strain PR5 T is closely related to the representatives of the genera Haloterrigena and Natrinema, but on the basis of its phenotypic properties, 16S rDNA sequence and DNA-DNA hybridization studies, strain PR5 T cannot be assigned to any of the recognized species within these genera. On the basis of its polar lipid composition, the isolate has been assigned to the genus Haloterrigena. The creation of a new species, Haloterrigena thermotolerans, is therefore proposed to accommodate this isolate. The type strain is strain PR5 T (l DSM 11552 T l ATCC 700275 T ).
A moderately halophilic, spore-forming, Gram-positive, short-rod-shaped bacterium, designated strain MS10 T , was isolated from the surface of leaves of the black mangrove Avicennia germinans and was subjected to a polyphasic taxonomic study. Strain MS10 T was able to grow at NaCl concentrations in the range 5-20 % (w/v) with optimum growth at 10 % (w/v) NaCl. Growth occurred at temperatures of 10-50 6C (optimal growth at 33-35 6C) and pH 6.0-9.0 (optimal growth at pH 7.0). Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain MS10 T fell within the branch encompassing members of the genus Halobacillus and was most closely related to Halobacillus dabanensis JCM 12772 T (99.2 % 16S rRNA gene sequence similarity). The DNA G+C content of strain MS10 T was 45.7 mol%, the major respiratory isoprenoid quinone was MK-7 and the cell-wall peptidoglycan was of the L-Orn-D-Asp type, characteristics consistent with its affiliation to the genus Halobacillus. Strain MS10 T showed a level of DNA-DNA hybridization with H. dabanensis JCM 12772 T of 29 % and levels below 70 % were also obtained with respect to other recognized members of the genus Halobacillus. The major fatty acids of strain MS10 T were iso-C 16 : 0 , anteiso-C 15 : 0 , iso-C 14 : 0 and iso-C 15 : 0 . Overall, the phenotypic, genotypic and phylogenetic results presented in this study demonstrate that strain MS10 T represents a novel species of the genus Halobacillus, for which the name Halobacillus mangrovi sp. nov. is proposed. The type strain is MS10 T (5CECT 7206 T 5CCM 7397 T ).The genus Halobacillus was first described by Spring et al. (1996) et al., 2005). Besides the ability to produce ellipsoidal or spherical endospores at the central or subterminal position, one of the main features of the genus Halobacillus is the cell-wall peptidoglycan type based on L-Orn-D-Asp, in contrast to the meso-diaminopimelic acid or L-lysine type cell-wall peptidoglycan found in related genera (Shida et al., 1997;Yoon et al., 2003). There have been no previous reports of members of the genus Halobacillus associated with mangroves that thrive in saline habitats.Avicennia germinans (black mangrove) is a type of mangrove that has developed mechanisms to tolerate high salt concentrations in unstable substrates, to obtain oxygen from almost anoxic sediments and to reproduce under harsh environmental conditions. One of these mechanisms is the ability to secrete salt crystals on to the surface of its leaves when the salt concentration is in excess of that normally present in the vascular system (Lugo & Snedaker, 1975). In the present study, we determined the taxonomic position of bacterial strain MS10 T , which was isolated from the surface of leaves of the black mangrove during a microbial diversity study. On the basis of the results presented, we show that strain MS10 T represents a novel species of the genus Halobacillus.The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain MS10 T is DQ888316. & Gibbons, 1960). This p...
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