Molecular signatures for Bacillus species: demarcation of the Bacillus subtilis and Bacillus cereus clades in molecular terms and proposal to limit the placement of new species into the genus Bacillus

2013 ◽  
Vol 63 (Pt_7) ◽  
pp. 2712-2726 ◽  
Author(s):  
Vaibhav Bhandari ◽  
Nadia Z. Ahmod ◽  
Haroun N. Shah ◽  
Radhey S. Gupta

The genus Bacillus is a phylogenetically incoherent taxon with members of the group lacking a common evolutionary history. Comprising aerobic and anaerobic spore-forming bacteria, no characteristics are known that can distinguish species of this genus from other similar endospore-forming genera. With the availability of complete genomic data from over 30 different species from this group, we have constructed detailed phylogenetic trees to determine the relationships among Bacillus and other closely related taxa. Additionally, we have performed comparative genomic analysis for the determination of molecular markers, in the form of conserved signature indels (CSIs), to assist in the understanding of relationships among species of the genus Bacillus in molecular terms. Based on the analysis, we report here the identification of 11 and 6 CSIs that clearly differentiate a ‘ Bacillus subtilis clade’ and a ‘ Bacillus cereus clade’, respectively, from all other species of the genus Bacillus . No molecular markers were identified that supported a larger clade within this genus. The subtilis and the cereus clades were also the largest observed monophyletic groupings among species from the genus Bacillus in the phylogenetic trees based on 16S rRNA gene sequences and those based upon concatenated sequences for 20 conserved proteins. Thus, the relationships observed among these groups of species through CSIs are independently well supported by phylogenetic analysis. The molecular markers identified in this study provide a reliable means for the reorganization of the currently polyphyletic genus Bacillus into a more evolutionarily consistent set of groups. It is recommended that the genus Bacillus sensu stricto should comprise only the monophyletic subtilis clade that is demarcated by the identified CSIs, with B. subtilis as its type species. Members of the adjoining cereus clade (referred to as the Cereus clade of bacilli), although they are distinct from the subtilis clade, will also retain the Bacillus genus name as they contain several clinically important species, and their transfer into a new genus could have serious consequences. However, all other species that are currently part of the genus Bacillus and not part of these two clades should be eventually transferred to other genera. We also propose that all novel species of the genus Bacillus must meet minimal requirements, foremost among which is that the branching of the prospective species with the Bacillus sensu stricto clade or the Cereus clade of bacilli should be strongly supported by 16S rRNA gene sequence trees or trees based upon concatenated protein sequences. Additionally, the presence of one or more of the CSIs that are specific for these clades may be used to confirm molecularly the placement of the species into these clades. The identified CSIs, in addition to their usefulness for taxonomic and diagnostic purposes, also provide novel probes for genetic and biochemical studies of these bacteria.

2015 ◽  
Vol 65 (Pt_6) ◽  
pp. 1819-1824 ◽  
Author(s):  
Sooyeon Park ◽  
Ji-Min Park ◽  
Chul-Hyung Kang ◽  
Song-Gun Kim ◽  
Jung-Hoon Yoon

A Gram-stain-negative, non-motile, aerobic and pleomorphic bacterium, designated BS-W13T, was isolated from a tidal flat on the South Sea, South Korea, and its taxonomic position was investigated using a polyphasic approach. Strain BS-W13T grew optimally at 25 °C, at pH 7.0–8.0 and in the presence of 1.0–2.0 % (w/v) NaCl. Neighbour-joining and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences showed that strain BS-W13T clustered with the type strain of Seohaeicola saemankumensis , showing the highest sequence similarity (95.96 %) to this strain. Strain BS-W13T exhibited 16S rRNA gene sequence similarity values of 95.95, 95.91, 95.72 and 95.68 % to the type strains of Sulfitobacter donghicola , Sulfitobacter porphyrae , Sulfitobacter mediterraneus and Roseobacter litoralis , respectively. Strain BS-W13T contained Q-10 as the predominant ubiquinone and C18 : 1ω7c as the major fatty acid. The polar lipid profile of strain BS-W13T, containing phosphatidylcholine, phosphatidylglycerol, phosphatidylethanolamine, one unidentified aminolipid and one unidentified lipid as major components, was distinguishable from those of some phylogenetically related taxa. The DNA G+C content of strain BS-W13T was 58.1 mol%. The phylogenetic data and differential chemotaxonomic and other phenotypic properties revealed that strain BS-W13T constitutes a novel genus and species within family Rhodobacteraceae of the class Alphaproteobacteria , for which the name Pseudoseohaeicola caenipelagi gen. nov., sp. nov. is proposed. The type strain is BS-W13T ( = KCTC 42349T = CECT 8724T).


2013 ◽  
Vol 63 (Pt_1) ◽  
pp. 72-79 ◽  
Author(s):  
Adelfia Talà ◽  
Marcello Lenucci ◽  
Antonio Gaballo ◽  
Miriana Durante ◽  
Salvatore M. Tredici ◽  
...  

Strain SPC-1T was isolated from the phyllosphere of Cynara cardunculus L. var. sylvestris (Lamk) Fiori (wild cardoon), a Mediterranean native plant considered to be the wild ancestor of the globe artichoke and cultivated cardoon. This Gram-stain-negative, catalase-positive, oxidase-negative, non-spore-forming, rod-shaped and non-motile strain secreted copious amounts of an exopolysaccharide, formed slimy, viscous, orange-pigmented colonies and grew optimally at around pH 6.0–6.5 and 26–30 °C in the presence of 0–0.5 % NaCl. Phylogenetic analysis based on comparisons of 16S rRNA gene sequences demonstrated that SPC-1T clustered together with species of the genus Sphingomonas sensu stricto. The G+C content of the DNA (66.1 mol%), the presence of Q-10 as the predominant ubiquinone, sym-homospermidine as the predominant polyamine, 2-hydroxymyristic acid (C14 : 0 2-OH) as the major hydroxylated fatty acid, the absence of 3-hydroxy fatty acids and the presence of sphingoglycolipid supported this taxonomic position. 16S rRNA gene sequence analysis showed that SPC-1T was most closely related to Sphingomonas hankookensis ODN7T, Sphingomonas insulae DS-28T and Sphingomonas panni C52T (98.19, 97.91 and 97.11 % sequence similarities, respectively). However, DNA–DNA hybridization analysis did not reveal any relatedness at the species level. Further differences were apparent in biochemical traits, and fatty acid, quinone and polyamine profiles leading us to conclude that strain SPC-1T represents a novel species of the genus Sphingomonas , for which the name Sphingomonas cynarae sp. nov. is proposed; the type strain is SPC-1T ( = JCM 17498T = ITEM 13494T). A component analysis of the exopolysaccharide suggested that it represents a novel type of sphingan containing glucose, rhamnose, mannose and galactose, while glucuronic acid, which is commonly found in sphingans, was not detected.


2013 ◽  
Vol 63 (Pt_3) ◽  
pp. 1001-1006 ◽  
Author(s):  
Soo-Young Lee ◽  
Sooyeon Park ◽  
Tae-Kwang Oh ◽  
Jung-Hoon Yoon

A Gram-stain-negative, non-motile, non-spore-forming, rod-shaped bacterial strain, BB-Mw22T, was isolated from a tidal flat sediment of the South Sea in South Korea. It grew optimally at 30–37 °C, at pH 7.0–7.5 and in the presence of 2–3 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences revealed that strain BB-Mw22T belonged to the genus Kangiella and the cluster comprising Kangiella species and strain BB-Mw22T was clearly separated from other taxa. Strain BB-Mw22T exhibited 95.3–98.7 % 16S rRNA gene sequence similarity to the type strains of recognized Kangiella species. Strain BB-Mw22T contained Q-8 as the predominant ubiquionone and iso-C15 : 0 and iso-C11 : 0 3-OH as the major fatty acids. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine and one unidentified aminolipid. The DNA G+C content of strain BB-Mw22T was 48.9 mol%, and its mean DNA–DNA hybridization values with Kangiella geojedonensis YCS-5T, Kangiella japonica JCM 16211T and Kangiella taiwanensis JCM 17727T were 14–28 %. Phylogenetic and genetic distinctiveness and differential phenotypic properties revealed that strain BB-Mw22T is distinguishable from all recognized Kangiella species. On the basis of the data presented, strain BB-Mw22T is considered to represent a novel species of the genus Kangiella , for which the name Kangiella sediminilitoris sp. nov. is proposed. The type strain is BB-Mw22T ( = KCTC 23892T  = CCUG 62217T).


2014 ◽  
Vol 64 (Pt_6) ◽  
pp. 1841-1846 ◽  
Author(s):  
Sooyeon Park ◽  
Ji-Min Park ◽  
Chul-Hyung Kang ◽  
Jung-Hoon Yoon

A Gram-stain-negative, non-motile, aerobic, curved-to-spiral-rod-shaped bacterium, designated AH-MY2T, was isolated from a tidal flat on Aphae island in the sea to the south-west of South Korea, and its taxonomic position was investigated using a polyphasic taxonomic approach. Strain AH-MY2T grew optimally at 30 °C, at pH 7.0–8.0 and in the presence of 2.0 % (w/v) NaCl. Neighbour-joining, maximum-likelihood and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences showed that strain AH-MY2T clustered with the type strain of Terasakiella pusilla and that this cluster joined the clade comprising the type strains of species of the genus Thalassospira . Strain AH-MY2T exhibited 16S rRNA gene sequence similarity values of 90.6 % to the type strain of Terasakiella pusilla and of less than 91.0 % to the type strains of other species with validly published names. Strain AH-MY2T contained Q-10 as the predominant ubiquinone and C18 : 1ω7c as the major fatty acid. The major polar lipids detected in strain AH-MY2T were phosphatidylglycerol, phosphatidylethanolamine, two unidentified aminolipids and one unidentified glycolipid. The DNA G+C content of strain AH-MY2T was 56.0 mol%. The phylogenetic data and differential chemotaxonomic and other phenotypic properties revealed that strain AH-MY2T represented a novel genus and species within the family Rhodospirillaceae of the class Alphaproteobacteria , for which the name Aestuariispira insulae gen. nov., sp. nov. is proposed. The type strain of Aestuariispira insulae is AH-MY2T ( = KCTC 32577T = CECT 8488T).


Author(s):  
Xing-Kui Zhou ◽  
Ying Huang ◽  
Ti- Kun Zhang ◽  
Jian- Jin Liu ◽  
Zi- Yi Liu ◽  
...  

An aerobic, rod-shaped, Gram-stain-positive, actinobacterial strain, designated 1.0914T, was isolated from a stalactite sample collected from a cave located in Guizhou Province, southwest PR China. Based on 16S rRNA gene sequence analysis, strain 1.0914T shared highest similarities values with Nocardioides pelophilus CGMCC 4.7388T (97.7 %), Nocardioides immobilis CCTCC AB 2017083T (97.5 %) and Nocardioides silvaticus CCTCC AB 2018079T (97.3 %) and values lower than 97.0 % to other members of the genus Nocardioides . Phylogenetic trees based on 16S rRNA gene sequences indicated that strain 1.0914T formed an isolated branch with N. pelophilus CGMCC 4.7388T, N. immobilis CCTCC AB 2017083T and N. silvaticus CCTCC AB 2018079T. The polar lipids contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and one unidentified phospholipid in the cellular membrane. The major fatty acids were identified as iso-C16 : 0, C18 : 1 ω9c, C17 : 1 ω8c and C16 : 0. The predominant respiratory quinone was MK-8(H4) and ll-diaminopimelic acid was the diagnostic diamino acid in the cell-wall peptidoglycan. The genomic DNA G+C content was 71.1 mol%. The orthologous average nucleotide identiy values between N. pelophilus CGMCC 4.7388T, N. immobilis CCTCC AB 2017083T, N. silvaticus CCTCC 2018079T and strain 1.0914T were 82.3, 81.7 and 81.9 % respectively. DNA–DNA hybridization values between N. pelophilus CGMCC 4.7388T, N. immobilis CCTCC AB 2017083T, N. silvaticus CCTCC 2018079T and strain 1.0914T were 25.2, 24.6 and 24.5 % respectively. The phylogenetic, phenotypic and chemotaxonomic data supported the classification of strain 1.0914T as representing a new species of Nocardioides , for which the name Nocardioides stalactiti sp. nov. is proposed. The type strain is 1.0914T (=CCTCC AB 2018266T=KCTC 49243T).


Author(s):  
Lei Song ◽  
Hongcan Liu ◽  
Ying Huang ◽  
Xin Dai ◽  
Yuguang Zhou

A Gram-stain-negative, rod-shaped bacterial strain, designated SW123T, was isolated from a deep-sea water sample collected from the Indian Ocean. Strain SW123T was strictly aerobic, catalase- and oxidase-positive. The predominant cellular fatty acids were iso-C15 : 0, iso-C17 : 0 and summed feature 9 (comprising C16 : 0-methyl or iso-C17 : 1 ω9c). Ubiquinone-8 was the sole respiratory quinone. The major polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol. The genomic DNA G+C content was 49.4 mol%. 16S rRNA gene sequence analysis showed that strain SW123T was closely related to Aliidiomarina shirensis AIST (96.7 % sequence similarity), Aliidiomarina iranensis GBPy7T (96.3%), Aliidiomarina haloalkalitolerans AK5T (96.0%) and Aliidiomarina celeris F3105T (95.9%). Phylogenetic trees based on 16S rRNA gene sequences showed that strain SW123T represented a novel member of the genus Aliidiomarina , forming a distinct cluster with A. celeris F3105T. On the basis of phylogenetic inference and phenotypic characteristics, we propose that strain SW123T represents a novel species of the genus Aliidiomarina , with the name Aliidiomarina indica sp. nov. The type strain is SW123T (=CGMCC1.16169T=KCTC 82234T).


2013 ◽  
Vol 63 (Pt_5) ◽  
pp. 1684-1689 ◽  
Author(s):  
Sooyeon Park ◽  
Jung-Hoon Yoon

A Gram-negative, non-spore-forming, non-flagellated, coccoid-, oval- or rod-shaped strain, designated M-M23T, was isolated from seashore sediment at Geoje island, South Korea. Strain M-M23T grew optimally at 25 °C, at pH 7.0–8.0 and in the presence of 2 % (w/v) NaCl. Phylogenetic trees based on 16S rRNA gene sequences revealed that strain M-M23T clustered with the type strains of the two species of the genus Hirschia , with which it exhibited 97.6–98.1 % 16S rRNA gene sequence similarity. Sequence similarity with the type strains of other recognized species was <90.8 %. Strain M-M23T contained Q-10 as the predominant ubiquinone and C18 : 1ω7c and C16 : 0 as the major fatty acids. The major polar lipids of strain M-M23T were phosphatidylglycerol and two unidentified lipids. The DNA G+C content of strain M-M23T was 45.4 mol%. DNA–DNA relatedness between the isolate and Hirschia baltica DSM 5838T and Hirschia maritima JCM 14974T was 22±7.2 and 14±5.6 %, respectively. The differential phenotypic properties, together with the phylogenetic and genetic distinctiveness, revealed that strain M-M23T is separate from the other described members of the genus Hirschia . On the basis of the data presented, strain M-M23T is considered to represent a novel species of the genus Hirschia , for which the name Hirschia litorea sp. nov. is proposed. The type strain is M-M23T ( = KCTC 32081T  = CCUG 62793T). An emended description of the genus Hirschia is also provided.


2014 ◽  
Vol 64 (Pt_10) ◽  
pp. 3411-3416 ◽  
Author(s):  
Zhi-Ping Zhong ◽  
Ying Liu ◽  
Hong-Can Liu ◽  
Fang Wang ◽  
Lei Song ◽  
...  

A Gram-stain-negative bacterium, strain TS-T11T, was isolated from Tuosu lake, a saline lake (salinity 5.4 %, w/v) in the Qaidam basin, Qinghai province, China. Its taxonomic position was determined by using a polyphasic approach. Cells of strain TS-T11T were non-spore-forming rods, 0.6–0.8 µm wide and 0.8–2.2 µm long, and motile by means of a single polar flagellum. Strain TS-T11T was strictly heterotrophic and aerobic. Cells were positive for catalase and oxidase. Growth was observed in the presence of 0.5–11.0 % (w/v) NaCl (optimum 4.0–6.0 %), at 4–40 °C (optimum 30–35 °C) and at pH 6.0–10.5 (optimum pH 7.5–8.5). Strain TS-T11T contained iso-C15 : 0, iso-C17 : 0 and iso-C17 : 1ω9c as the predominant fatty acids (>10 %). The major respiratory quinone was Q-8. The polar lipids consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and nine uncharacterized phospholipids. The G+C content of genomic DNA was 46.8 mol% (T m). Phylogenetic trees based on 16S rRNA gene sequences showed that strain TS-T11T was associated with the genus Idiomarina , and showed highest 16S rRNA gene sequence similarity to Idiomarina aestuarii KYW314T (97.4 %) and Idiomarina salinarum ISL-52T (97.4 %). DNA–DNA relatedness of strain TS-T11T to I. aestuarii JCM 16344T and I. salinarum DSM 21900T was 22.2±2.4 and 11.5±1.6 %, respectively. Based on the data presented above, it was concluded that strain TS-T11T represents a novel species of the genus Idiomarina , for which the name Idiomarina planktonica sp. nov. is proposed. The type strain is TS-T11T ( = CGMCC 1.12458T = JCM 19263T).


2014 ◽  
Vol 64 (Pt_8) ◽  
pp. 2812-2818 ◽  
Author(s):  
Zhi-Ping Zhong ◽  
Ying Liu ◽  
Hong-Can Liu ◽  
Fang Wang ◽  
Yu-Guang Zhou ◽  
...  

A Gram-staining-negative bacterium, strain DSG-S4-2T, was isolated from Dasugan Lake, a saline lake (salinity 3.1 %, w/v) in Qaidam basin, Qinghai, China and its taxonomic position was determined by using a polyphasic approach. Cells of strain DSG-S4-2T were non-spore-forming rods, 0.5–0.8 µm wide and 1.2–3.8 µm long and motile by means of a single polar flagellum. Strain DSG-S4-2T was strictly heterotrophic and aerobic, catalase-positive and oxidase-negative. PufLM and coxL genes were present, bacteriochlorophyll a (BChl a) and a carotenoid pigment were produced. Growth was observed in the presence of 0–8.0 % (w/v) NaCl (optimum, 1.0–2.0 %), at 20–40 °C (optimum, 35 °C) and pH 6.5–10.5 (optimum, pH 7.5–8.0). Strain DSG-S4-2T contained Q-10 as the sole respiratory quinone. The polar lipids contained two aminolipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, sulphoquinovosyldiacylglyceride, phosphatidylcholine and some unknown phospholipids, like the other members of the genus Roseibium . The predominant fatty acid (>70 %) was summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The DNA G+C content was 61.4 mol% (determined from melting temperature). Phylogenetic trees (neighbour-joining, maximum-likelihood and maximum-parsimony) based on 16S rRNA gene sequences showed that strain DSG-S4-2T was associated with the members of the genus Roseibium , with highest 16S rRNA gene sequence similarity to Roseibium denhamense OCh 254T (96.3 %) and Roseibium hamelinense OCh 368T (96.3 %). Based on the data presented above, it is concluded that strain DSG-S4-2T represents a novel species of the genus Roseibium , for which the name Roseibium aquae sp. nov. is proposed. The type strain is DSG-S4-2T ( = CGMCC 1.12426T = JCM 19310T).


2014 ◽  
Vol 64 (Pt_8) ◽  
pp. 2551-2557 ◽  
Author(s):  
Ja-Min Park ◽  
Sooyeon Park ◽  
Yong-Taek Jung ◽  
Hyangmi Kim ◽  
Jung-Sook Lee ◽  
...  

A Gram-stain-negative, strictly aerobic, non-flagellated and rod-shaped bacterial strain, designated GJR-7T, was isolated from coastal sand of the South Sea of South Korea. Strain GJR-7T grew optimally at 30 °C, at pH 7.0–7.5 and without NaCl. Phylogenetic trees based on 16S rRNA gene sequences revealed that strain GJR-7T clustered with the type strains of Sphingopyxis wooponensis , Sphingopyxis rigui and Sphingorhabdus planktonica , with which it exhibited 16S rRNA gene sequence similarity values of 96.0–96.3 %. Sequence similarities to the type strains of other recognized species were less than 95.5 %. Strain GJR-7T contained Q-10 as the predominant ubiquinone and C18 : 1ω7c, 11-methyl C18 : 1ω7c and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) or C14 : 0 2-OH. The major polar lipids were sphingoglycolipid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and one unidentified glycolipid. The DNA G+C content of strain GJR-7T was 55.5 mol%. Differential phenotypic properties, together with phylogenetic distinctiveness, revealed that strain GJR-7T is separated from the type strains of Sphingopyxis wooponensis , Sphingopyxis rigui and Sphingorhabdus planktonica . On the basis of the data presented, strain GJR-7T is considered to represent a novel species of the genus Sphingorhabdus , for which the name Sphingorhabdus arenilitoris sp. nov. is proposed. The type strain is GJR-7T ( = KCTC 42051T = CECT 8531T). It is also proposed that Sphingopyxis wooponensis and Sphingopyxis rigui should be reclassified as members of the genus Sphingorhabdus .


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