Abstract
Two novel (18CT and 6C) Gram-stain-positive, rod shaped, motile and endospore-forming bacterial strains were isolated from Lonar soda lake, India. Based on 16S rRNA gene sequence analysis, strains 18CT and 6C were identified as belonging to the class Firmibacteria, and were most closely related to Bacillus cohnii KCTC 3572T (99.3 and 99.9%, respectively), Bacillus zhanjiangensis KCTC 13713T (97.4 and 98.0%, respectively), Bacillus halmapalus LMG 17950T (97.0 and 97.6%, respectively) and other members in the genus Bacillus (<97.0%). However, the DNA–DNA relatedness between 18CT and 6C and B. cohnii KCTC 3572T (49.6 ± 0.9 and 51.6 ± 0.7, respectively), B. zhanjiangensis KCTC 13713T (42.9 ± 0.8 and 47.1 ± 0.3, respectively) and B. halmapalus LMG 17950T (39.9 ± 0.8 and 40.8 ± 0.3, respectively) indicated that the novel strains were distantly related to these strains. Further, the high 16S rRNA gene sequence similarity (100%) and DNA–DNA relatedness (90 ± 5%) suggested that strains 18CT and 6C were members of a genomospecies. The strains grew optimally at a pH of 7.5 with 2–3% (w/v) NaCl and temperature of 37 °C. Strains 18CT and 6C were catalase and oxidase negative. The cell wall of strain 18CT contained meso-diaminopimelic acid as the diagnostic diamino acid, which was in contrast with its nearest neighbour B. cohnii KCTC 3572T, which contained ornithine and aspartic acid. Polar lipids include diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), an unknown phospholipid (PL) and three unknown lipids (L1-3). The predominant isoprenoid quinone was MK-7. iso-C15:0 (32.5%) was the predominant fatty acid and significant proportions of anteiso-C15:0 (19.5%), C16:0 (11.5%), iso-C17:0 (9.5%) and anteiso-C17:0 (6.3%) were also detected. The DNA G + C content of strains 18CT and 6C were 39.3 and 39.2 mol%, respectively. The results of molecular, biochemical and chemotaxonomic tests showed a clear differentiation of strains 18CT and 6C from all other members of the genus Bacillus, for which the name Bacillus catenulatus sp. nov. is proposed. The type strain is 18CT (=KCTC 33781T = CGMCC 1.15475T).
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SVR and MT thank Department of Science and Technology, Government of India, for providing FTSYS project Grants (SB/FT/LS-115/2012 and SB/FT/LS-320/2012, respectively).
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Communicated by Erko Stackebrandt.
The EMBL accession number for the 16S rRNA gene sequences of strains 18CT and 6C are LT617055 and LT837689, respectively. The DPD Taxonumber of the type strain 18CT is TA00143.
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Supplementary Fig. 1. Phase contrast photograph of strain 18CT; A, growth at pH 7.5; B, growth at pH 9.5. Bar, 0.5 μm. Supplementary Fig. 2. Two-dimensional thin-layer chromatogram of polar lipid extracts of strain 18CT and phylogenetically related Bacillus species. The first direction was developed in CHCl3:CH3OH:H2O (75: 32: 4 v/v) and the second in CHCl3: CH3OH: CH3COOH: H2O (86: 16: 15: 4 v/v). PG, phosphatidylglycerol; DPG, diphosphatidylglycerol; PE, phosphatidylethanolamine; PL, phospholipid; L1-8, Unknown lipids. Supplementary Fig. 3. Thin-layer chromatogram of cell wall extracts of 18CT and phylogenetically related Bacillus species. This was developed in methanol: pyridine: water: 12 N HCl (32: 4: 7: 1 v/v) (PPTX 838 kb)
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Sultanpuram, V.R., Mothe, T., Chintalapati, S. et al. Bacillus catenulatus sp. nov., an alkalitolerant bacterium isolated from a soda lake. Arch Microbiol 199, 1391–1397 (2017). https://doi.org/10.1007/s00203-017-1413-y
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DOI: https://doi.org/10.1007/s00203-017-1413-y