Abstract
Two alkaliphilic and halotolerant Gram-stain positive, rod-shaped and endospore-forming bacteria, designated strains 12-3T and 12-4, were isolated from saline and alkaline soils collected in Lindian county, Heilongjiang province, China. Both strains were observed to grow well at a wide range of temperature and pH values, 10–45 °C and pH 8–12, with optimal growth at 37 °C and pH 9.0, respectively. Growth of the two strains was found to occur at total salt concentrations of 0–12 % (w/v), with an optimum at 4 % (w/v). The G+C contents of the genomic DNA of strains 12-3T and 12-4 were determined to be 42.7 and 42.4 mol%, respectively, and the major cellular fatty acids were identified as anteiso-C15:0 and anteiso-C17:0. In isolate 12-3T, meso-diaminopimelic acid was found to be the diagnostic diamino acid of the cell wall peptidoglycan; diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were identified as the major cellular polar lipids; and menaquinone-7 was identified as the predominant isoprenoid quinone. Strains 12-3T and 12-4 share very close 16S rRNA gene sequence similarity (99.74 %) and their DNA–DNA relatedness was 95.3 ± 0.63 %, meaning that the two strains can be considered to belong to the same species. 16S rRNA gene sequence-based phylogenetic analysis revealed strains 12-3T and 12-4 exhibit high similarities to Bacillus pseudofirmus DSM 8715T (98.7 %), Bacillus marmarensis DSM 21297T (97.2 %) and Bacillus nanhaiisediminis CGMCC 1.10116T (97.1 and 97.0 %, respectively). DNA–DNA hybridization values between isolate 12-3T and the type strains of closely related Bacillus species were below 30 %. On the basis of the polyphasic evidence presented, strains 12-3T and 12-4 are considered to represent a novel species of the genus Bacillus, for which the name Bacillus lindianensis sp. nov. is proposed. The type strain is 12-3T (DSM 26864T = CGMCC 1.12717T).
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Acknowledgments
We thank Yuqin Zhang of the Institute of Medicinal Biotechnology, Academy of Medical Sciences for the respiratory menaquinone analysis. This work was supported by the Beijing Nova Program (2011033), the Forestry Industry Research Special Funds for Public Welfare Projects (201304409), the Fundamental Research Funds for the Central Universities (YX2014-15), National Science and Technology Ministry (2012BAC09B03) and the National Science Foundation of China (J1103516).
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The GenBank accession numbers for the 16S rRNA gene sequences of strains 12-3T and 12-4 are KP954690 and KP954691 respectively.
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10482_2015_616_MOESM1_ESM.pptx
Supplementary material 1 (PPTX 47 kb). Fig. S1. Neighbour-joining tree based on 16S rRNA gene sequences showing the phylogenetic position of strains 12-3T and 12-4 compared to the most closely related members of the genus Bacillus. Numbers at branching points represent bootstrap values from 1000 replicates; only values ≥ 70 % are shown. Accession numbers are given in parentheses. Bar 0.005 substitutions per nucleotide position
10482_2015_616_MOESM2_ESM.pptx
Supplementary material 2 (PPTX 46 kb). Fig. S2. Minimum-evolution tree based on 16S rRNA gene sequences showing the phylogenetic position of strains 12-3T and 12-4 compared to the most closely related members of the genus Bacillus. Numbers at branching points represent bootstrap values from 1000 replicates; only values ≥70 % are shown. Accession numbers are given in parentheses. Bar 0.005 substitutions per nucleotide position
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Dou, G., Liu, H., He, W. et al. Bacillus lindianensis sp. nov., a novel alkaliphilic and moderately halotolerant bacterium isolated from saline and alkaline soils. Antonie van Leeuwenhoek 109, 149–158 (2016). https://doi.org/10.1007/s10482-015-0616-y
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DOI: https://doi.org/10.1007/s10482-015-0616-y