| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 Korea Research Institute of Bioscience and Biotechnology, 52 Oeundong, Yusong, Daejeon 305-333, Republic of Korea
2 Environmental Biotechnology National Core Research Center, Division of Environmental Biotechnology, Gyeongsang National University, 660-701, Korea
3 Yunnan Institute of Microbiology, Yunnan University, Kunming, Yunnan, 650091, P.R. China
Correspondence
Chang-Jin Kim
changjin{at}kribb.re.kr
 |
ABSTRACT |
|---|
 TOP ABSTRACT MAIN TEXTREFERENCES |
|---|
-meso-diaminopimelic acid as the cell-wall type; phosphatidylglycerol and diphosphatidylglycerol as the major polar lipids; anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0 as the major fatty acids) also supported assignment of the isolate to the genus Lentibacillus. On the basis of physiological and molecular properties, the isolate represents a novel species within the genus Lentibacillus, for which the name Lentibacillus lacisalsi sp. nov. is proposed. The type strain is BH260T (=KCTC 3915T=DSM 16462T).
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strains BH260T is AY667497.
A transmission electron micrograph of a cell of strain BH260T is available as a supplementary figure in IJSEM Online.
|
MAIN TEXT |
|---|
|
TOPABSTRACTMAIN TEXTREFERENCES |
|---|
). They were originally assigned to the genus Bacillus (Ash et al., 1991; Stackebrandt & Liesack, 1993; Nielsen et al., 1994; Ventosa et al., 1998), but molecular and chemical analyses have shown that they can be divided into several phylogenetically distinct lineages, such as the new genera Halobacillus (Spring et al., 1996), Gracilibacillus (Wainø et al., 1999), Virgibacillus (Heyndrickx et al., 1999), Filobacillus (Schlesner et al., 2001), Oceanobacillus (Lu et al., 2001), Tenuibacillus (Ren & Zhou, 2005) and Pontibacillus (Lim et al., 2005).
Recently, novel species isolated from salty habitats, such as Lentibacillus juripiscarius and Lentibacillus salarius, have been added to the genus since the genus Lentibacillus was proposed for novel moderately halophilic Gram-positive rods (Yoon et al., 2002; Namwong et al., 2005; Jeon et al., 2005). In this study, polyphasic taxonomy demonstrated that another novel Gram-positive bacterium, strain BH260T, isolated from a salt lake in Xin-Jiang Province in China, represents a novel species of the genus Lentibacillus.
Strain BH260T was isolated on marine agar 2216 (MA; Difco) with the addition of 20 % (w/v) NaCl [final concentration, 21·94 % (w/v) NaCl] at 28 °C for 3 days from soil sediment of a salt lake in China. Requirements for, and tolerance of, NaCl were determined in trypticase soy broth [TSB, containing (l–1) 17·0 g casein, 3·0 g soy-bean meal, 2·5 g glucose, 5·0 g NaCl, 2·5 g dipotassium phosphate] and supplemented with modified artificial sea water [containing (l–1) 0–30 % (w/v) NaCl, 5·94 g MgSO4.7H2O, 4·53 g MgCl2.6H2O, 0·64 g KCl, 1·3 g CaCl2]. Optimum growth was tested at different temperatures (4–55 °C) on MA containing 12 % (w/v) NaCl and at different pH values (5·0–11·0) in TSB supplemented with artificial sea water containing 12 % (w/v) NaCl. Anaerobic growth was determined by using incubation in an anaerobic chamber at 30 °C for 5 days on MA containing 12 % (w/v) NaCl. Cell morphology and flagellum type were studied using light microscopy (LABOPHOT apparatus; Nikon) and transmission electron microscopy (JEM-1010 apparatus; JEOL) as described by Lee et al. (2005). Motility was observed at 24 and 36 h in wet mounts by using a light microscope (E600; Nikon). Endospores were stained using Schaeffer–Fulton stain (Smibert & Krieg, 1981).
Gram staining was determined using the bioMérieux Gram stain kit according to the manufacturer's instructions. Catalase activity was determined by assessing the production of oxygen bubbles in 3 % (v/v) aqueous hydrogen peroxide solution. Oxidase activity was tested for by assessing the oxidation of 1 % (w/v) tetramethyl-p-phenylenediamine (Merck). Hydrolysis of aesculin, casein, starch, Tween 80, urea, hypoxanthine, tyrosine and xanthine was determined on MA, according to methods described previously (Lanyi, 1987; Smibert & Krieg, 1994). Nitrate reduction was determined according to the method of Lanyi (1987). Acid production from carbohydrates was tested for as described by Leifson (1963); all suspension media were supplemented with artificial sea water containing 12 % (w/v) NaCl. GC analysis of fatty acid methyl esters was performed using cells grown at 30 °C for 3 days on MA with the addition of 10 % (w/v) NaCl according to the instructions of the Microbial Identification System (MIDI; Microbial ID). Analyses of peptidoglycan, polar lipids and isoprenoid quinones were carried out using the methods described by Komagata & Suzuki (1987). The DNA G+C content of strain BH260T was determined by reverse-phase HPLC (HP 1090 apparatus; Hewlett Packard) using the method of Tamaoka & Komagata (1984).
Sequencing and assembly of the 16S rRNA gene was carried out as described previously (Bakermans & Madsen, 2002). The resultant 16S rRNA gene sequence was compared with available 16S rRNA gene sequences from GenBank by using the BLAST program (http://www.ncbi.nlm.nih.gov/BLAST/) and was aligned with closely related 16S rRNA gene sequences with CLUSTAL W software (Thompson et al., 1994). Sequence similarity values were computed using Similarity Matrix version 1.1 (Ribosomal Database Project II, http://35.8.164.52/html/; Cole et al., 2003). Phylogenetic trees were constructed using three different methods: neighbour-joining, maximum-likelihood and maximum-parsimony algorithms available in PHYLIP software, version 3.6 (Felsenstein, 2002). Evolutionary distance matrices were calculated according to the algorithm of the Kimura two-parameter model (Kimura, 80) for the neighbour-joining method. To evaluate the stability of the phylogenetic tree, a bootstrap analysis (1000 replications) was performed with the SEQBOOT, DNADIST, NEIGHBOR and CONSENSE programs in the PHYLIP package.
Strain BH260T on MA medium supplemented with 12 % (w/v) NaCl formed cream-coloured, low-convex and circular/slightly irregular colonies and the colony morphologies were consistent at various salinities of NaCl. Strain BH260T showed halophilic properties, growing in TSB containing 5–25 % (w/v) NaCl; optimum growth occurred in TSB with 12–15 % (w/v) NaCl. Growth was not observed in the absence of NaCl. Growth was observed at temperatures between 15 and 40 °C, with optimum growth at 30–32 °C. Strain BH260T grew at pH values in the range 7·0–9·5 in 12 % (w/v) NaCl-containing TSB; optimal growth was observed at pH 8·0. Cells of strain BH260T from early and late growth phases showed obvious Gram-positive reactions. The cells of the isolate were strictly aerobic, short rods, 0·4–0·6 µm wide and 1·2–3·0 µm long (see the transmission electron micrograph available as a supplementary figure in IJSEM Online). Cells of the isolate produced single spherical terminal endospores within swollen sporangia. Cell motility was facilitated by means of flagella on the sides of the cells. Anaerobic growth was not observed under anaerobic conditions after 5 days at 30 °C on MA with 12 % (w/v) NaCl.
The major respiratory lipoquinone of strain BH260T was MK-7. The fatty acid profile of the strain was characterized by the presence of branched and saturated fatty acids such as anteiso-C15 : 0 (50·78 %), anteiso-C17 : 0 (18·20 %) and iso-C16 : 0 (11·95 %) as the major fatty acids, in common with the type strains of the genus Lentibacillus; their fatty acid compositions were somewhat different (Table 1). Cell-wall peptidoglycan analysis indicated A1 type, meso-diaminopimelic acid as the diagnostic diamino acid. This peptidoglycan type is characteristic of Lentibacillus species (Yoon et al., 2002). The major polar lipids of the test strain were diphosphatidylglycerol and phosphatidylglycerol. The genomic DNA G+C content of strain BH139T was 44 mol%, a value that falls within the defined range for the genus Lentibacillus (Jeon et al., 2005). The major fatty acid profile, the major lipoquinone, the cell-wall type and the DNA G+C content of strain BH260T were typical of the genus Lentibacillus (Table 2).
|
|
). The topologies of phylogenetic trees obtained using the maximum-likelihood and maximum-parsimony algorithms were similar to that of the tree constructed using neighbour-joining analysis (data not shown). The levels of 16S rRNA gene sequence similarity for strain BH260T with respect to Lentibacillus salsus KCTC 3911T, L. juripiscarius JCM 12147T and L. salicampi KCCM 41560T were 96·0, 95·5 and 95·3 %, respectively – values that are generally sufficient to define a novel species (Rosselló-Mora & Amann, 2001; Stackebrandt et al., 2002).
|
Description of Lentibacillus lacisalsi sp. nov.
Lentibacillus lacisalsi (la.ci.sal'si. L. masc. n. lacus lake; L. adj. salsus -a -um salted, salt; N.L. gen. n. lacisalsi of a salt lake).
Cells are approximately 0·4–0·6 µm wide and 1·2–3·0 µm long, strictly aerobic, motile rods with peritrichous flagella. Colonies are cream-coloured, low-convex and circular/slightly irregular on MA supplemented with 10 % (w/v) NaCl. Growth occurs at 15–40 °C (optimum, 30–32 °C), pH 7·0–9·5 (optimum, pH 8·0) and 5–25 % (w/v) NaCl (optimum, 12–15 %). Nitrate is reduced to nitrite. Hydrolysis of urea, L-tyrosine, hypoxanthine, casein, starch, Tween 80, aesculin and xanthine is not observed. Acids produced from D-fructose, D-ribose, D-xylose and L-arabinose but not from glycerol, D-glucose, maltose, D-trehalose, 
-D-lactose, D-mannitol, D-mannose, rhamnose, adonitol, raffinose, arbutin, D-salicin or D-melibiose. The major isoprenoid quinone is MK-7. Cell wall contains meso-diaminopimelic acid (A1 type). The predominant polar lipids are phosphatidylglycerol and diphosphatidylglycerol. The major fatty acids are anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The DNA G+C content is 44 mol% (by HPLC).
The type strain, BH260T (=KCTC 3915T=DSM 16462T), was isolated from a salt lake in China.
|
ACKNOWLEDGEMENTS |
|---|
|
REFERENCES |
|---|
|
TOPABSTRACTMAIN TEXTREFERENCES |
|---|
Arahal, D. R., Marquez, M. C., Volcani, B. E., Schleifer, K. H. & Ventosa, A. (2000). Reclassification of Bacillus marismortui as Salibacillus marismortui comb. nov. Int J Syst Evol Microbiol 50, 1501–1503.[Abstract]
Ash, C., Farrow, J. A. E., Wallbanks, S. & Collins, M. D. (1991). Phylogenetic heterogeneity of the genus Bacillus as revealed by comparative analysis of small-subunit ribosomal-RNA sequences. Lett Appl Microbiol 13, 202–206.
Bakermans, C. & Madsen, E. L. (2002). Diversity of 16S rDNA and naphthalene dioxygenase genes from coal-tar-waste-contaminated aquifer waters. Microb Ecol 44, 95–106.[Medline]
Cole, J. R., Chai, B., Marsh, T. L. & 8 other authors (2003). The ribosomal database project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy. Nucleic Acids Res 31, 442–443.
Felsenstein, J. (2002). PHYLIP – Phylogeny Inference Package, version 3.6a. Seattle, WA: University of Washington.
Heyndrickx, M., Lebbe, L., Kersters, K., De Vos, P., Forsyth, G. & Logan, N. A. (1999). Virgibacillus: a new genus to accommodate Bacillus pantothenticus (Proom and Knight 1950). Emended description of Virgibacillus pantothenticus. Int J Syst Bacteriol 48, 99–106.
Heyrman, J., Logan, N. A., Busse, H. J., Balcaen, A., Lebbe, L., Rodriguez-Diaz, M., Swings, J. & De Vos, P. (2003). Virgibacillus carmonensis sp. nov., Virgibacillus necropolis sp. nov. and Virgibacillus picturae sp. nov., three novel species isolated from deteriorated mural paintings, transfer of the species of the genus Salibacillus to Virgibacillus, as Virgibacillus marismortui comb. nov. and Virgibacillus salexigens comb. nov., and emended description of the genus Virgibacillus. Int J Syst Evol Microbiol 53, 501–511.
Jeon, C. O., Lim, J.-M., Lee, J. C., Lee, G. S., Lee, J.-M., Xu, L.-H., Jiang, C.-L. & Kim, C.-J. (2005). Lentibacillus salarius sp. nov., isolated from saline sediment in China, and emended description of the genus Lentibacillus. Int J Syst Evol Microbiol 55, 1339–1343.
Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16, 111–120.[CrossRef][Medline]
Komagata, K. & Suzuki, K. (1987). Lipid and cell-wall analysis in bacterial systematics. Methods Microbiol 19, 161–208.
Lanyi, B. (1987). Classical and rapid identification methods for medically important bacteria. Methods Microbiol 19, 1–67.
Lee, E. M., Jeon, C. O., Choi, I., Chang, K. S. & Kim, C. J. (2005). Silanimonas lenta gen. nov., sp. nov., a slightly thermophilic and alkaliphilic gammaproteobacterium isolated from a hot spring. Int J Syst Evol Microbiol 55, 385–389.
Leifson, E. (1963). Determination of carbohydrate metabolism of marine bacteria. J Bacteriol 85, 1183–1184.
Lim, J.-M., Jeon, C. O., Song, S. M. & Kim, C.-J. (2005). Pontibacillus chungwhensis gen. nov., sp. nov., a moderately halophilic Gram-positive bacterium from a solar saltern in Korea. Int J Syst Evol Microbiol 55, 165–170.
Lu, J., Nogi, Y. & Takami, H. (2001). Oceanobacillus iheyensis gen. nov., sp. nov., a deep-sea extremely halotolerant and alkaliphilic species isolated from a depth of 1050 m on the Iheya Ridge. FEMS Microbiol Lett 205, 291–297.[CrossRef][Medline]
Namwong, S., Tanasupawat, S., Smitinont, T., Visessanguan, W., Kudo, T. & Itoh, T. (2005). Characterization of Lentibacillus salicampi strains and Lentibacillus juripiscarius sp. nov. isolated from fish sauce in Thailand. Int J Syst Evol Microbiol 55, 315–320.
Nielsen, P., Rainey, F. A., Outtrup, H., Priest, F. G. & Fritze, D. (1994). Comparative 16S rDNA sequence analysis of some alkaliphilic bacilli and the establishment of a sixth rRNA group within the genus Bacillus. FEMS Microbiol Lett 117, 61–66.[CrossRef]
Oren, A. (2002). Diversity of halophilic microorganisms: environments, phylogeny, physiology, and applications. J Ind Microbiol Biotechnol 28, 56–63.[CrossRef][Medline]
Ren, P. G. & Zhou, P. J. (2005). Tenuibacillus multivorans gen. nov., sp. nov., a moderately halophilic bacterium isolated from saline soil in Xin-Jiang, China. Int J Syst Evol Microbiol 55, 95–99.
Rosselló-Mora, R. & Amann, R. (2001). The species concept for prokaryotes. FEMS Microbiol Rev 25, 39–67.[Medline]
Schlesner, H., Lawson, P. A., Collins, M. D., Weiss, N., Wehmeyer, U., Völker, H. & Thomm, M. (2001). Filobacillus milensis gen. nov., sp. nov., a new halophilic spore-forming bacterium with Orn-D-Glu-type peptidoglycan. Int J Syst Evol Microbiol 51, 425–431.[Abstract]
Smibert, R. M. & Krieg, N. R. (1981). General characterization. In Manual of Methods for General Microbiology, pp. 409–443. Edited by P. Gerhardt, R. G. E. Murray, R. N. Costilow, E. W. Nester, W. A. Wood, N. R. Krieg & G. B. Phillips. Washington, DC: American Society for Microbiology.
Smibert, R. M. & Krieg, N. R. (1994). Phenotypic characterization. In Methods for General and Molecular Bacteriology, pp. 607–654. Edited by P. Gerhardt. Washington, DC: American Society for Microbiology.
Spring, S., Ludwig, W., Marquez, M. C., Ventosa, A. & Schleifer, K.-H. (1996). Halobacillus gen. nov., with description of Halobacillus litoralis sp. nov. and Halobacillus trueperi sp. nov., and transfer of Sporosarcina halophila to Halobacillus halophila comb. nov. Int J Syst Bacteriol 46, 492–496.
Stackebrandt, E. & Liesack, W. (1993). Nucleic acids and classification. In Handbook of New Bacterial Systematics, pp. 152–189. Edited by M. Goodfellow & A. G. O'Donnell. London: Academic Press.
Stackebrandt, E., Frederiksen, W., Garrity, G. M. & 10 other authors (2002). Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology. Int J Syst Evol Microbiol 52, 1043–1047.[Abstract]
Tamaoka, J. & Komagata, K. (1984). Determination of DNA base composition by reverse-phase high-performance liquid chromatography. FEMS Microbiol Lett 25, 125–128.
Thompson, J. D., Higgins, D. G. & Gibson, T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22, 4673–4680.
Ventosa, A., Nieto, J. J. & Oren, A. (1998). Biology of moderately halophilic aerobic bacteria. Microbiol Mol Biol Rev 62, 504–544.
Wainø, M., Tindall, B. J., Schumann, P. & Ingvorsen, K. (1999). Gracilibacillus gen. nov., with description of Gracilibacillus halotolerans gen. nov., sp. nov.; transfer of Bacillus dipsosauri to Gracilibacillus dipsosauri comb. nov., and Bacillus salexigens to the genus Salibacillus gen. nov., as Salibacillus salexigens comb. nov. Int J Syst Bacteriol 49, 821–831.
Yoon, J. H., Kang, K. H. & Park, Y. H. (2002). Lentibacillus salicampi gen. nov., sp. nov., a moderately halophilic bacterium isolated from a salt field in Korea. Int J Syst Evol Microbiol 52, 2043–2048.[Abstract]
This article has been cited by other articles:
|
|
 |
|
  J.-C. Lee, W.-J. Li, L.-H. Xu, C.-L. Jiang, and C.-J. Kim Lentibacillus salis sp. nov., a moderately halophilic bacterium isolated from a salt lake Int J Syst Evol Microbiol, August 1, 2008; 58(8): 1838 - 1843. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S.-Y. Lee, W.-Y. Choi, T.-K. Oh, and J.-H. Yoon Lentibacillus salinarum sp. nov., isolated from a marine solar saltern in Korea Int J Syst Evol Microbiol, January 1, 2008; 58(1): 45 - 49. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Yuan, P. Ren, J. Liu, Y. Xue, Y. Ma, and P. Zhou Lentibacillus halodurans sp. nov., a moderately halophilic bacterium isolated from a salt lake in Xin-Jiang, China Int J Syst Evol Microbiol, March 1, 2007; 57(3): 485 - 488. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Pakdeeto, S. Tanasupawat, C. Thawai, S. Moonmangmee, T. Kudo, and T. Itoh Lentibacillus kapialis sp. nov., from fermented shrimp paste in Thailand Int J Syst Evol Microbiol, February 1, 2007; 57(2): 364 - 369. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Tanasupawat, A. Pakdeeto, S. Namwong, C. Thawai, T. Kudo, and T. Itoh Lentibacillus halophilus sp. nov., from fish sauce in Thailand. Int J Syst Evol Microbiol, August 1, 2006; 56(Pt 8): 1859 - 1863. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
