| The western region of China owns rich alkaliphilic Bacillus-like samples with the unique geographical climate and the natural rescources,providing unmatched advantages for developing the isolation and systematic research of alkaliphilic Bacillus-like species.In this study,120 soil samples were collected from Sichuan,Yunnan,Xizang,Qinghai,Xinjiang,Gansu Province of China,aiming to isolate alkaliphilic Bacillus-like resources,identified by polyphasic method.Using cultural method,751 strains of alkaliphilic Bacillus-like species were isolated from soil samples.Based on the result of 16S rRNA gene similarities and phylogeny analysis,it was indicated that 751 strains belonged to 12 genera including 108 species.35 strains were considered as new potential species of alkaliphilic Bacillus-like species,which 16S rRNA similarities were less than 98%.The distribution characteristics of alkaliphilic Bacillus-like species among regions were quite different,but their dominant genus were Bacillus.Based on analysis of geographical distribution diversity index,it was found that the Shannon(H')index order was Xinjiang>Ningxia>Qinghai>Sichuan>Yunnan>Tibet>Gansu respectively.5 new potential species of alkaliphilic Bacillus were identified by polyphasic method.The results were as followed.Cells of strain FJAT-45037T were aerobic,endospore-forming and motile.Temperature range for growth was 10 to 35?(optimum at 30 ?).pH range for growth was 7.5?12.0(optimum at 9.0?10.0).NaCl concentration range for growth was 0?12%(w/v)(optimum at 4%).16S rRNA gene sequence-based phylogenetic analysis revealed strain FJAT-45037T most closely related to Bacillus marmarensis GMBE 72T(97.67%)and their DNA-DNA hybridization(isDDH)value was 18.6%and their average nucleotide identity(ANI)value was 84.46%.The DNA G+C content was 39.79 mol%.The major cellular fatty acids of strain FJAT-45037T were anteiso-C15:0 and anteiso-C17:0.The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid(meso-DAP).MK-6 and MK-7 were the predominant quinones.Major polar lipids were diphosphatidyl glycerol(DPG),phosphatidyl glycerol(PG)and phosphatidyl ethanolamine(PE).On the basis of the polyphasic evidence presented,strain FJAT-45037T was considered to represent a novel member of the genus Bacillus,for which the name Bacillus alkalisalsus sp.nov.was proposed.The type strain was FJAT-45037T(= DSM 104053T= CCTCC AB 20162291).Cells of strain FJAT-45086T were aerobic,endospore-forming and motile with a single polar flagellum.Temperature range for growth was 15 to 35?(optimum at 30?).pH range for growth was 6?12.0(optimum at 9.0).NaCl concentration range for growth was 0-10%(w/v)(optimum at 4%).The major cellular fatty acids of strain FJAT-45086T were anteiso-C15:0 and C16:0.16S rRNA gene sequence-based phylogenetic analysis revealed strain FJAT-450861 most closely related to Bacillus hemicellulosilyticus JCM 9152T(96.68%)and their is DDH value was 22.7%and their ANI value was 84.79%.The DNA G+C content was 37.81 mol%.The diagnostic diamino acid of the cell-wall peptidoglycan was meso-DAP.MK-7 was the predominant quinone.Major polar lipids were DPG,PE,PG and phosphatidyl choline(PC).On the basis of the polyphasic evidence presented,strain FJAT-45086T was considered to represent a novel member of the genus Bacillus,for which the name Bacillus alkalisoli sp.nov.was proposed.The type strain was FJAT-45086T(=DSM 104056T= CCTCC AB 2016232T).Cells of strain FJAT-451221 were facultative anaerobe,endospore-forming and motile with a single polar flagellum.Temperature range for growth was 10 to 40?(optimum at 30?).pH range for growth was 7.5?12.0(optimum at 9.0).NaCl concentration range for growth was 0-10%(w/v)(optimum at 4%).16S rRNA gene sequence-based phylogenetic analysis revealed strain FJAT-451221 most closely related to Bacillus abyssalis DSM 25875T(95.87%)and their is DDH value was 27.5%and their ANI value was 68.69%.The DNA G+C content was 36.11 mol%.The major cellular fatty acids of strain FJAT-45122T were iso-C15:0,anteiso-C15:0,iso-C17:0,anteiso-Cl7:0 and iso-C17:1?10c.The diagnostic diamino acid of the cell-wall peptidoglycan was meso-DAP.MK-7 was the predominant quinone.Major polar lipids are DPG,phosphatidyl methyl ethanolamine(PME)and PG.On the basis of the polyphasic evidence presented,strain FJAT-45122T was considered to represent a novel member of the genus Bacillus,for which the name Bacillus solitudinis sp.nov.was proposed.The type strain was FJAT-45122T(= DSM 104631T= CCTCC AB 20162541).Cells of strain FJAT-453471 were aerobic,endospore-forming and non-motile.Temperature range for growth was 15 to 35 ?(optimum at 20 ?).pH range for growth was 7.5?12.0(optimum at 8.0).NaCl concentration range for growth was 0?20%(w/v)(optimum at 8%).16S rRNA gene sequence-based phylogenetic analysis revealed strain FJAT-45347T most closely related to Bacillus clarkii DSM 8720T(96.1%)and their isDDH value was 26.2%and their ANI value was 68.47%.The DNA G+C content was 40.62 mol%.The major cellular fatty acids of the strain FJAT-45347T were anteiso-C15:0 and anteiso-C17:0.The diagnostic diamino acid of the cell-wall peptidoglycan was meso-DAP.MK-7 was the predominant quinone.Major polar lipids were DPG,PE and PG;On the basis of the polyphasic evidence presented,strain FJAT-45347T was considered to represent a novel member of the genus Bacillus,for which the name Bacillus populi sp.nov.was proposed.The type strain was FJAT-45347T(= DSM 104632T= CCTCC AB 2016257T).Cells of strain FJAT-45385T were facultative anaerobe,endospore-forming and motile with a single polar flagellum.Temperature range for growth was 20 to 40?(optimum at 30 ?).pH range for growth was 7.0?11.0(optimum at 9.0).NaCl concentration range for growth was 0?10%(w/v)(optimum at 4?6%).16S rRNA gene sequence-based phylogenetic analysis revealed strain FJAT-45385T most closely related to Bacillus wakoensis N-1T(96.01%)and their isDDH value was 18.9%and their ANI value was 70.3%.The DNA G+C content was 42.4 mol%.The major cellular fatty acids of the strain FJAT-45385T were anteiso-C15:0,iso-C15:0 and C16:0.The diagnostic diamino acid of the cell-wall peptidoglycan was meso-DAP.MK-7 was the predominant quinone.Major polar lipids were DPG,PE and PG.On the basis of the polyphasic evidence presented,strain FJAT-45385T was considered to represent a novel member of the genus Bacillus,for which the name Bacillus urbisdiaboli sp.nov.was proposed.The type strain was FJAT-45385T(=DSM 104651T= CCTCC AB 2016263T). |
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