IJSEM Papers in Press. Published May 5, 2015 as doi:10.1099/ijs.0.000296
International Journal of Systematic and Evolutionary Microbiology Bacillus crescens sp. nov., isolated from soil --Manuscript Draft-Manuscript Number:
IJS-D-15-00064R2
Full Title:
Bacillus crescens sp. nov., isolated from soil
Short Title:
Bacillus crescens sp. nov.
Article Type:
Note
Section/Category:
New taxa - Firmicutes and related organisms
Corresponding Author:
Ramana Ch.V., Ph.D University of Hyderabad HYDERABAD, AP INDIA
First Author:
Shivani Y
Order of Authors:
Shivani Y Subhash Y Bharti P Dave Sasikala Ch. Ramana Ch.V., Ph.D
Manuscript Region of Origin:
INDIA
Abstract:
Two bacterial strains (JC247T and JC248) were isolated from soil samples collected from Rann of Kutch, Gujarat, India. Colonies of both strains were creamy white and cells stain Gram-positive. Cells are rods-to-curved rods (crescent shape), produce centrally located oval shaped endospores, positive for catalase and negative for oxidase. Both strains hydrolyze starch and tween-80. Both strains are negative for H2S production, indole production and nitrate reduction. Major fatty acids of both strains (>5%) are iso-C16:0, iso-C14:0, iso-C15:0, C16:1ω11c, C16:0, ¬ with minor (1%) amounts of anteiso-C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0, iso-C18:0, C14:0, C17:0,¬ C18:0, C18:1ω9c, iso-C17:1ω10c and anteiso-C17:0B/isoI. Diphosphatydilglycerol, phosphatidylethanolamine and phosphatidylglycerol are the major polar lipids of both strains. Cell wall amino acids are L-alanine, D-alanine, Dglutamic acid and meso-diaminopimelic acid. Genomic DNA G+C content was 48.2 and 46.2 mol%, respectively for strain JC247T and JC248. Both strains are closely related with mean DNA-DNA hybridization >90%. 16S rRNA gene sequence comparison of both strains indicated that they represent members of the genus Bacillus within the family Bacillaceae of the phylum Firmicutes. Both strains have sequence similarity of 96.93% with Bacillus firmus NCIMB 9366T and 5%) fatty acids of both strains are iso-
26
C16:0, iso-C14:0, iso-C15:0, C16:1ω11c, C16:0, with minor (1%) amounts of anteiso-
27
C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0,
28
C17:1ω10c and anteiso-C17:0B/isoI. Diphosphatidylglycerol, phosphatidylethanolamine
29
and phosphatidylglycerol are the major polar lipids of both strains. Cell wall amino acids
30
are L-alanine, D-alanine, D-glutamic acid and meso-diaminopimelic acid. Genomic DNA
31
G+C content was 48.2 and 48.1 mol%, respectively for strains JC247T and JC248. Both
32
strains are closely related with mean DNA-DNA hybridization >90%. 16S rRNA gene
33
sequence analysis of both strains indicated that they aremembers of the genus Bacillus
34
within the family Bacillaceae of the phylum Firmicutes. Both strains have a sequence
35
similarity of 96.93% with Bacillus firmus NCIMB 9366T and 5%) and anteiso-C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0, iso-C18:0, C14:0, C17:0,
175
C18:0, C18:1ω9c, iso-C17:1ω10c and anteiso-C17:0B/isoI are present as minor fatty acid (1%) (Supplementary Table 1).
177
Polar lipids and quinones of strains JC247T and Bacillus firmus LMG 7125T were
178
analyzed as described previously (Subhash et al., 2013; Subhash et al., 2014).
179
Phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and diphosphatidylglycerol
180
(DPG) are the major polar lipids along with five unidentified lipids (L1-5) and two
181
unidentified aminolipids (AL1-2) in the strain JC247T (JC248) (Supplementary Fig. 2A).
8
182
Polar lipids of strain JC247T differ from those of the type strain, Bacillus firmus LMG
183
7125T (Supplementary fig. 2B) by the presence of unidentified lipid (L1,3) and absence
184
of unidentified lipid (L6-8). Polar lipids of strain JC247T (JC248) are in concurrence with
185
the type species of the genus Bacillus subtilis sub sp. subtilis DSM10T (Kämpfer et al.,
186
2006) while absence of glycolipid in the strain JC247T (JC248) reflect the phylogenetic
187
distance. MK-7 was the major quinone of strain JC247T, JC248 and Bacillus firmus LMG
188
7125T.
189
Strain JC247T and JC248 were distinct (Table 1) from their closest validly
190
described phylogenetic neighbor, Bacillus firmus LMG 7125T (=NCIMB 9366T) with
191
respect to NaCl tolerance range, growth temperature range, pH range, antibiotic
192
susceptibility, organic substrate utilization, presence of significant levels of various fatty
193
acids (iso-C16:0 and C16:1ω7c alcohol), presence of unidentified lipid (L2,5) and absence
194
of unidentified lipids (L6-11) and higher genomic G+C mol%. Based on the phenotypic,
195
genotypic and phylogenetic analyeis, strain JC247T is designated as a as the type strain of
196
a new species, with the proposed name as Bacillus crescens sp. nov.
197
Description of Bacillus crescens sp. nov.
198 199
Bacillus crescens N.L. adj. crescens (cres'cens. L. part. adj. crescens, arising, becoming visible, appearing), of the moon in its first quarter, crescent.
200
Colonies appears creamy white. Texture of the colonies are butyrous for initial 4
201
days which turns in to friable after longer incubation (>4 days). Colony edges are entire
202
with undulate margins. Elevations of the colonies are flat. Diameter of the colonies range
203
between 2-3 mm after 5 days of growth. Cells are rod-to-curved rods (crescent shape),
204
Gram-stain-positive, 0.9-1.0 m wide and 3.0-10.0 m long. Cells are non-motile and
9
205
multiply by binary fission. Forms sub-terminal/central endospores of cylindrical to oval
206
shape (size 0.5-0.6x0.9-2.0) in a swollen sporangium. Mesophilic and halotolerant. NaCl
207
is not required for growth and tolerates up to 5% (w/v). Optimal growth occurs at pH 7-8
208
(range: 5.8-9.0). Temperature range for growth is 18-45 with optima at 30-37°C. Casein
209
and gelatin are not hydrolysed. Hydrolyses tween-80 and starch. Catalase positive and
210
oxidase negative. H2S production, indole production, ONPG test, urease and
211
denitrification negative. Growth occurs with D-galactose, D-glucose, D-fructose, D-
212
cellobiose, D-mannitol, D-lactose, D-mannose, D-sorbitol, sucrose, pyruvate, L-aspartic
213
acid and citrate. Growth does not happen with L-rhamnose, L-fucose, D-sorbitol and L-
214
glutamate. Acids are produced from D-glucose and sucrose and not from D-cellobiose,
215
D-galactose, L-arabinose, D-mannitol, D-fructose and D-mannose. Ammonium salts are
216
used as sole nitrogen source. Vitamins are not required for growth. Sensitive to
217
fosfomycin, penicillin-G, chloramphenicol, gentamicin, kanamycin, streptomycin and
218
vancomycin but resistant to rifampicin, polymyxin-B, sulfamethoxazole, nalidixic acid,
219
and tetracycline. meso-Diaminopimelic acid is the diagnostic diamino acid. Major (>5%)
220
fatty acids are iso-C16:0, iso-C14:0, iso-C15:0, C16:1ω11c, C16:0, with minor (1%)
221
amounts of anteiso-C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0, iso-C18:0, C14:0, C17:0, C18:0,
222
C18:1ω9c,
223
phosphatidylglycerol, diphosphatidylglycerol, unidentified lipids (L1-5) and unidentified
224
aminolipids (AL1-2) are present. G+C of genomic DNA is 48.2 % (by HPLC). Type
225
strain is JC247T (=KCTC 33627T= LMG 28608T), isolated from a soil at Rann of Kutch,
226
Gujarat, India. Strain JC248 is having similar characters of type strain and represents an
227
additional strain of the new species described.
iso-C17:1ω10c
and
anteiso-C17:0B/isoI.
Phosphatidylethanolamine,
10
228
ACKNOWLEDGEMENTS
229
MOES is greatly acknowledged for funding. SY acknowledges CSIR, Government of
230
India for the award of Senior Research Fellowship.
231
acknowledged for providing the Bacillus firmus LMG 7125T in exchange.
LMG, Belgium is highly
232 233
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234
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Kumar, A. & Mayilraj, S. (2014) Bacillus mesophilum sp. nov., strain IITR‑54T,
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acid diastereomers in cell walls of Gram-positive bacteria. Lett Appl Microbiol
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Mesbah, M., Premachandran, U. & Whitman, W. B. (1989). Precise measurement of
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chromatography. Int J Syst Bacteriol 39, 159-167.
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Priest, F. G., Goodfellow, M. & Todd, C. (1988). A numerical classification of the genus Bacillus. J Gen Microbiol 134, 1847-1882.
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Reddy, S.V., Thirumala, M., Farooq, M., Sasikala, C. & Ramana, C.V. (2015)
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Bacillus lonarensis sp. nov., an alkalitolerant bacterium isolated from a soda lake.
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Arch Microbiol 197, 27-34.
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Sasser, M. (1990). Identification of bacteria by gas chromatography of cellular fatty acids. Newark, DE: MIDI Inc. Schleifer, K.H. & Kandler, O. (1972). Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36, 407-477. Schleifer, K.H. (1985). Analysis of the chemical composition and primary structure of murein. Methods Microbiol 18, 123-156.
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Seldin, L. & Dubnau, D. (1985). Deoxyribonucleic acid homology among Bacillus
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polymyxa, Bacillus macerans, Bacillus azotofixans, and other nitrogen-fixing
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Bacillus strains. Int J Syst Bacteriol 35, 151-154.
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Sonalkar, V.V., Mawlankar, R., Ramana, V.V., Joseph, N., Shouche, Y.S. &
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Dastager, S.G. (2015) Bacillus filamentosus sp. nov., isolated from sediment
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sample. Antonie van Leeuwenhoek. 107, 433-441.
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Subhash, Y., Sasikala, Ch. & Ramana Ch.V. (2014). Bacillus luteus sp. nov. isolated from a soil. Int J Syst Evol Microbiol 64, 1580-1586.
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Subhash, Y., Tushar, L., Sasikala, Ch. & Ramana, Ch.V. (2013). Falsirhodobacter
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halotolerans gen. nov. sp. nov. isolated from a solar saltern. Int J Syst Evol
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Tamura, K., Stecher, G., Peterson, D., Filipski, A. & Kumar, S. (2013). MEGA6:
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Täubel, M., Kämpfer, P., Buczolits, S., Lubitz, W. & Busse, H-J. (2003). Bacillus
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barbericus sp. nov., isolated from an experimental wall painting. Int J Syst Evol
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Microbiol 53, 725-730.
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Zhai, L., Liao, T., Xue, Y. & Ma, Y. (2012). Bacillus daliensis sp. nov., an alkaliphilic,
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Gram-positive bacterium isolated from a soda lake. Int J Syst Evol Microbiol 62,
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949-953.
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Zhang, J., Wang, J., Fang, C., Song, F., Xin, Y., Qu, L. & Ding, K. (2010).
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Bacillus oceanisediminis sp. nov., isolated from marine sediment. Int J Syst Evol
325
Microbiol 60, 2924-2929.
326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351
15
352 353
Table. 1. Differentiating characteristics between 1, Strain JC247T and 2, B. firmus LMG 7125T. *&1
*2
Creamy white
Creamy Yellow
Colony texture
Butyrous/Friable
Butyrous
Colony margins
Irregular undulate
Entire/Rhizoidal
Cell size
0.9-1.0x3.0-10.0
0.8-0.9x2.0-4.0
0-5 (0-2) 5.8-9.0 (7.0-8.0) 18-45 (30-37) -
0-7 (1-2) 6.0-9.4 (7.0-8.5) 20-50 (30-40) + + +
+ -
+
-
+
+ +
-
-
+ + + + +
-
+
48.1-48.2
+ 41.3
Characteristics Colony color
NaCl tolerance range (optimum %) pH range (optimum) Temperature range (optimum) (°C) Nitrate reduction Indole production Oxidase Hydrolysis of Tween-80 Casein Gelatin Organic carbon source utilization for growth D-cellobiose L-aspartic acid Acid production from D-galactose D-mannitol D-fructose Starch D-mannose Antibiotic susceptibility Rifampicin Nalidixic acid G+C content (mol %)
354 355 356 357 358 359 360 361 362 363
of the strains was done at author’s laboratory. &Strain JC248 has very similar characters as strain JC247T. Both strains have phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine as major polar lipids; meso-diaminopimelic acid as diagnostic diamino acid; non motile; both strains utilized D-mannose, D-galactose, D-glucose, D-fructose, Dmannitol, and D-lactose; not utilized L-glutamate; produced acid from D-glucose and sucrose; acid is not produced from D-cellobiose; hydrolyses starch; Sensitive to fosfomycin, penicillin-G, chloramphenicol, gentamicin, kanamycin, streptomycin and vancomycin and resistant to polymyxin-B, sulfamethoxazole and tetracycline; MK-7 is the major quinone of both strains; positive for endospore formation. *Characterization
16
364
Fig.1.Phylogenetic tree based on 16S rRNA gene sequences (1413 bp) showing the relationship of
365
strain JC247T along with the closest phylogenetic neighbors within the genus Bacillus. The
366
trees were constructed by the neighbor joining method using the MEGA6 software and
367
rooted by using Clostridium butyricum VPI3266T (AJ458420) as the out-group. Numbers at
368
nodes represent bootstrap values (based on 1000 resamplings). Bootstrap percentages refer to
369
NJ/ML/MP analysis. The GenBank accession numbers for 16S rRNA gene sequences are
370
shown in parentheses. Taxa names under “” are not validated yet, Bar 2 nucleotide
371
substitutions per 100 nucleotides.
372
17
Figure Click here to download Figure: Figure 1.pptx
Bacillus benzoevorans DSM 5391T (D78311)
Fig. 1
“Bacillus kyonggiensis NB22T ( JF896450)”
83/54/68 75/71/87
Bacillus siralis 171544T (AF071856) “Bacillus massiliosenegalensis JC6T (JF824800)” Bacillus andreesenii 8-4-E13T (HF952774)
93/81/69
Bacillus crescens JC247T (LN625239) 100/100/100
95/80/85 98/93/89
Bacillus sp. JC248 (LN625240)
Bacillus firmus NCIMB 9366T ( X60616) Bacillus oceanisediminis H2T (GQ292772)
54/-/68
“Bacillus mesophilum IITR-54T (JN210567)” Bacillus sporothermodurans 215T (U49079)
98/99/91 96/89/94 54/-/56
Bacillus acidicola 105-2T (AF547209) Bacillus shackletonii LMG 18435T (AJ250318) Bacillus subtilis ATCC 6051T (AJ276351) Bacillus neizhouensis JSM 071004T(EU925618)
100/100/100
Bacillus selenitireducens MLS10T(AF064704) Clostridium butyricum VPI3266T (AJ458420)
0.02
Click here to download Supplementary Material Files: Supplementary fig+table 5-5-15.pdf
International Journal of Systematic and Evolutionary Microbiology Bacillus crescens sp. nov., isolated from soil --Manuscript Draft-Manuscript Number:
IJS-D-15-00064R2
Full Title:
Bacillus crescens sp. nov., isolated from soil
Short Title:
Bacillus crescens sp. nov.
Article Type:
Note
Section/Category:
New taxa - Firmicutes and related organisms
Corresponding Author:
Ramana Ch.V., Ph.D University of Hyderabad HYDERABAD, AP INDIA
First Author:
Shivani Y
Order of Authors:
Shivani Y Subhash Y Bharti P Dave Sasikala Ch. Ramana Ch.V., Ph.D
Manuscript Region of Origin:
INDIA
Abstract:
Two bacterial strains (JC247T and JC248) were isolated from soil samples collected from Rann of Kutch, Gujarat, India. Colonies of both strains were creamy white and cells stain Gram-positive. Cells are rods-to-curved rods (crescent shape), produce centrally located oval shaped endospores, positive for catalase and negative for oxidase. Both strains hydrolyze starch and tween-80. Both strains are negative for H2S production, indole production and nitrate reduction. Major fatty acids of both strains (>5%) are iso-C16:0, iso-C14:0, iso-C15:0, C16:1ω11c, C16:0, ¬ with minor (1%) amounts of anteiso-C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0, iso-C18:0, C14:0, C17:0,¬ C18:0, C18:1ω9c, iso-C17:1ω10c and anteiso-C17:0B/isoI. Diphosphatydilglycerol, phosphatidylethanolamine and phosphatidylglycerol are the major polar lipids of both strains. Cell wall amino acids are L-alanine, D-alanine, Dglutamic acid and meso-diaminopimelic acid. Genomic DNA G+C content was 48.2 and 46.2 mol%, respectively for strain JC247T and JC248. Both strains are closely related with mean DNA-DNA hybridization >90%. 16S rRNA gene sequence comparison of both strains indicated that they represent members of the genus Bacillus within the family Bacillaceae of the phylum Firmicutes. Both strains have sequence similarity of 96.93% with Bacillus firmus NCIMB 9366T and 5%) fatty acids of both strains are iso-
26
C16:0, iso-C14:0, iso-C15:0, C16:1ω11c, C16:0, with minor (1%) amounts of anteiso-
27
C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0,
28
C17:1ω10c and anteiso-C17:0B/isoI. Diphosphatidylglycerol, phosphatidylethanolamine
29
and phosphatidylglycerol are the major polar lipids of both strains. Cell wall amino acids
30
are L-alanine, D-alanine, D-glutamic acid and meso-diaminopimelic acid. Genomic DNA
31
G+C content was 48.2 and 48.1 mol%, respectively for strains JC247T and JC248. Both
32
strains are closely related with mean DNA-DNA hybridization >90%. 16S rRNA gene
33
sequence analysis of both strains indicated that they aremembers of the genus Bacillus
34
within the family Bacillaceae of the phylum Firmicutes. Both strains have a sequence
35
similarity of 96.93% with Bacillus firmus NCIMB 9366T and 5%) and anteiso-C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0, iso-C18:0, C14:0, C17:0,
175
C18:0, C18:1ω9c, iso-C17:1ω10c and anteiso-C17:0B/isoI are present as minor fatty acid (1%) (Supplementary Table 1).
177
Polar lipids and quinones of strains JC247T and Bacillus firmus LMG 7125T were
178
analyzed as described previously (Subhash et al., 2013; Subhash et al., 2014).
179
Phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and diphosphatidylglycerol
180
(DPG) are the major polar lipids along with five unidentified lipids (L1-5) and two
181
unidentified aminolipids (AL1-2) in the strain JC247T (JC248) (Supplementary Fig. 2A).
8
182
Polar lipids of strain JC247T differ from those of the type strain, Bacillus firmus LMG
183
7125T (Supplementary fig. 2B) by the presence of unidentified lipid (L1,3) and absence
184
of unidentified lipid (L6-8). Polar lipids of strain JC247T (JC248) are in concurrence with
185
the type species of the genus Bacillus subtilis sub sp. subtilis DSM10T (Kämpfer et al.,
186
2006) while absence of glycolipid in the strain JC247T (JC248) reflect the phylogenetic
187
distance. MK-7 was the major quinone of strain JC247T, JC248 and Bacillus firmus LMG
188
7125T.
189
Strain JC247T and JC248 were distinct (Table 1) from their closest validly
190
described phylogenetic neighbor, Bacillus firmus LMG 7125T (=NCIMB 9366T) with
191
respect to NaCl tolerance range, growth temperature range, pH range, antibiotic
192
susceptibility, organic substrate utilization, presence of significant levels of various fatty
193
acids (iso-C16:0 and C16:1ω7c alcohol), presence of unidentified lipid (L2,5) and absence
194
of unidentified lipids (L6-11) and higher genomic G+C mol%. Based on the phenotypic,
195
genotypic and phylogenetic analyeis, strain JC247T is designated as a as the type strain of
196
a new species, with the proposed name as Bacillus crescens sp. nov.
197
Description of Bacillus crescens sp. nov.
198 199
Bacillus crescens N.L. adj. crescens (cres'cens. L. part. adj. crescens, arising, becoming visible, appearing), of the moon in its first quarter, crescent.
200
Colonies appears creamy white. Texture of the colonies are butyrous for initial 4
201
days which turns in to friable after longer incubation (>4 days). Colony edges are entire
202
with undulate margins. Elevations of the colonies are flat. Diameter of the colonies range
203
between 2-3 mm after 5 days of growth. Cells are rod-to-curved rods (crescent shape),
204
Gram-stain-positive, 0.9-1.0 m wide and 3.0-10.0 m long. Cells are non-motile and
9
205
multiply by binary fission. Forms sub-terminal/central endospores of cylindrical to oval
206
shape (size 0.5-0.6x0.9-2.0) in a swollen sporangium. Mesophilic and halotolerant. NaCl
207
is not required for growth and tolerates up to 5% (w/v). Optimal growth occurs at pH 7-8
208
(range: 5.8-9.0). Temperature range for growth is 18-45 with optima at 30-37°C. Casein
209
and gelatin are not hydrolysed. Hydrolyses tween-80 and starch. Catalase positive and
210
oxidase negative. H2S production, indole production, ONPG test, urease and
211
denitrification negative. Growth occurs with D-galactose, D-glucose, D-fructose, D-
212
cellobiose, D-mannitol, D-lactose, D-mannose, D-sorbitol, sucrose, pyruvate, L-aspartic
213
acid and citrate. Growth does not happen with L-rhamnose, L-fucose, D-sorbitol and L-
214
glutamate. Acids are produced from D-glucose and sucrose and not from D-cellobiose,
215
D-galactose, L-arabinose, D-mannitol, D-fructose and D-mannose. Ammonium salts are
216
used as sole nitrogen source. Vitamins are not required for growth. Sensitive to
217
fosfomycin, penicillin-G, chloramphenicol, gentamicin, kanamycin, streptomycin and
218
vancomycin but resistant to rifampicin, polymyxin-B, sulfamethoxazole, nalidixic acid,
219
and tetracycline. meso-Diaminopimelic acid is the diagnostic diamino acid. Major (>5%)
220
fatty acids are iso-C16:0, iso-C14:0, iso-C15:0, C16:1ω11c, C16:0, with minor (1%)
221
amounts of anteiso-C15:0, anteiso-C17:0, iso-C16:1 H, iso-C17:0, iso-C18:0, C14:0, C17:0, C18:0,
222
C18:1ω9c,
223
phosphatidylglycerol, diphosphatidylglycerol, unidentified lipids (L1-5) and unidentified
224
aminolipids (AL1-2) are present. G+C of genomic DNA is 48.2 % (by HPLC). Type
225
strain is JC247T (=KCTC 33627T= LMG 28608T), isolated from a soil at Rann of Kutch,
226
Gujarat, India. Strain JC248 is having similar characters of type strain and represents an
227
additional strain of the new species described.
iso-C17:1ω10c
and
anteiso-C17:0B/isoI.
Phosphatidylethanolamine,
10
228
ACKNOWLEDGEMENTS
229
MOES is greatly acknowledged for funding. SY acknowledges CSIR, Government of
230
India for the award of Senior Research Fellowship.
231
acknowledged for providing the Bacillus firmus LMG 7125T in exchange.
LMG, Belgium is highly
232 233
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326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351
15
352 353
Table. 1. Differentiating characteristics between 1, Strain JC247T and 2, B. firmus LMG 7125T. *&1
*2
Creamy white
Creamy Yellow
Colony texture
Butyrous/Friable
Butyrous
Colony margins
Irregular undulate
Entire/Rhizoidal
Cell size
0.9-1.0x3.0-10.0
0.8-0.9x2.0-4.0
0-5 (0-2) 5.8-9.0 (7.0-8.0) 18-45 (30-37) -
0-7 (1-2) 6.0-9.4 (7.0-8.5) 20-50 (30-40) + + +
+ -
+
-
+
+ +
-
-
+ + + + +
-
+
48.1-48.2
+ 41.3
Characteristics Colony color
NaCl tolerance range (optimum %) pH range (optimum) Temperature range (optimum) (°C) Nitrate reduction Indole production Oxidase Hydrolysis of Tween-80 Casein Gelatin Organic carbon source utilization for growth D-cellobiose L-aspartic acid Acid production from D-galactose D-mannitol D-fructose Starch D-mannose Antibiotic susceptibility Rifampicin Nalidixic acid G+C content (mol %)
354 355 356 357 358 359 360 361 362 363
of the strains was done at author’s laboratory. &Strain JC248 has very similar characters as strain JC247T. Both strains have phosphatidylglycerol, diphosphatidylglycerol and phosphatidylethanolamine as major polar lipids; meso-diaminopimelic acid as diagnostic diamino acid; non motile; both strains utilized D-mannose, D-galactose, D-glucose, D-fructose, Dmannitol, and D-lactose; not utilized L-glutamate; produced acid from D-glucose and sucrose; acid is not produced from D-cellobiose; hydrolyses starch; Sensitive to fosfomycin, penicillin-G, chloramphenicol, gentamicin, kanamycin, streptomycin and vancomycin and resistant to polymyxin-B, sulfamethoxazole and tetracycline; MK-7 is the major quinone of both strains; positive for endospore formation. *Characterization
16
364
Fig.1.Phylogenetic tree based on 16S rRNA gene sequences (1413 bp) showing the relationship of
365
strain JC247T along with the closest phylogenetic neighbors within the genus Bacillus. The
366
trees were constructed by the neighbor joining method using the MEGA6 software and
367
rooted by using Clostridium butyricum VPI3266T (AJ458420) as the out-group. Numbers at
368
nodes represent bootstrap values (based on 1000 resamplings). Bootstrap percentages refer to
369
NJ/ML/MP analysis. The GenBank accession numbers for 16S rRNA gene sequences are
370
shown in parentheses. Taxa names under “” are not validated yet, Bar 2 nucleotide
371
substitutions per 100 nucleotides.
372
17
Figure Click here to download Figure: Figure 1.pptx
Bacillus benzoevorans DSM 5391T (D78311)
Fig. 1
“Bacillus kyonggiensis NB22T ( JF896450)”
83/54/68 75/71/87
Bacillus siralis 171544T (AF071856) “Bacillus massiliosenegalensis JC6T (JF824800)” Bacillus andreesenii 8-4-E13T (HF952774)
93/81/69
Bacillus crescens JC247T (LN625239) 100/100/100
95/80/85 98/93/89
Bacillus sp. JC248 (LN625240)
Bacillus firmus NCIMB 9366T ( X60616) Bacillus oceanisediminis H2T (GQ292772)
54/-/68
“Bacillus mesophilum IITR-54T (JN210567)” Bacillus sporothermodurans 215T (U49079)
98/99/91 96/89/94 54/-/56
Bacillus acidicola 105-2T (AF547209) Bacillus shackletonii LMG 18435T (AJ250318) Bacillus subtilis ATCC 6051T (AJ276351) Bacillus neizhouensis JSM 071004T(EU925618)
100/100/100
Bacillus selenitireducens MLS10T(AF064704) Clostridium butyricum VPI3266T (AJ458420)
0.02
Click here to download Supplementary Material Files: Supplementary fig+table 5-5-15.pdf
