| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Korea Research Institute of Bioscience and Biotechnology (KRIBB), PO Box 115, Yusong, Taejon, Korea
Correspondence
Jung-Hoon Yoon
jhyoon{at}kribb.re.kr
 |
ABSTRACT |
|---|
 TOP ABSTRACT MAIN TEXTREFERENCES |
|---|
7c and/or iso-C15 : 0 2-OH and 10-methyl C16 : 0. The DNA G+C content was 58.1 mol%. A phylogenetic analysis based on 16S rRNA gene sequences showed that strain ISL-40T belongs to the genus Marinobacter. Strain ISL-40T exhibited 16S rRNA gene sequence similarity values of 93.5–96.4 % to the type strains of recognized Marinobacter species. The differential phenotypic properties and phylogenetic distinctiveness of strain ISL-40T revealed that it is separate from recognized Marinobacter species. On the basis of phenotypic, phylogenetic and genetic data, therefore, strain ISL-40T represents a novel species of the genus Marinobacter, for which the name Marinobacter salicampi sp. nov. is proposed. The type strain is ISL-40T (=KCTC 12972T=CCUG 54357T).
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain ISL-40T is EF486354.
|
MAIN TEXT |
|---|
|
TOPABSTRACTMAIN TEXTREFERENCES |
|---|
, currently comprises 16 species with validly published names including the recently described species Marinobacter algicola (Green et al., 2006), Marinobacter vinifirmus (Liebgott et al., 2006), Marinobacter koreensis (Kim et al., 2006), Marinobacter gudaonensis (Gu et al., 2007) and Marinobacter salsuginis (Antunes et al., 2007). Phylogenetic analyses based on 16S rRNA gene sequences have shown that the genus Marinobacter falls within the class Gammaproteobacteria (Anzai et al., 2000; Gu et al., 2007). Here, we report the taxonomic characterization of a Marinobacter-like strain, ISL-40T, isolated from a marine solar saltern. Sediment collected from a marine solar saltern of the Yellow Sea at Byunsan, Korea, was used as the source for the isolation of bacterial strains. Strain ISL-40T was isolated by means of the standard dilution plating technique at 30 °C on marine agar 2216 (MA; Difco) supplemented with 8 % (w/v) NaCl. The morphological, physiological and biochemical characteristics of strain ISL-40T were investigated using routine cultivation at 30 °C on MA supplemented with 6 % (w/v) NaCl. Cell morphology was examined by using light microscopy (E600; Nikon) and transmission electron microscopy (TEM). Flagellation was determined by using a TEM (CM-20; Philips) with cells from exponentially growing cultures. To visualize flagella, cells were negatively stained with 1 % (w/v) phosphotungstic acid and the grids were examined after being air-dried.
Growth under anaerobic conditions was determined after incubation in a Forma anaerobic chamber on MA supplemented with 6 % (w/v) NaCl and on MA supplemented with 6 % (w/v) NaCl supplemented with potassium nitrate (0.1 %, w/v), both of which had been prepared anaerobically using nitrogen. Growth in the absence of NaCl was investigated using trypticase soy broth prepared according to the formula of the Difco medium except that NaCl was excluded. Growth at various NaCl concentrations was investigated in marine broth 2216 (MB; Difco) or trypticase soy broth (Difco). Growth at various temperatures (4–45 °C) was measured on MA supplemented with 6 % (w/v) NaCl. Catalase and oxidase activities and hydrolysis of casein, starch and Tweens 20, 40, 60 and 80 were determined as described by Cowan & Steel (1965). Hydrolysis of hypoxanthine, tyrosine and xanthine was tested on MA supplemented with 6 % (w/v) NaCl, using the substrate concentrations described by Cowan & Steel (1965). Hydrolysis of aesculin, gelatin and urea and reduction of nitrate were investigated as described previously (Lanyi, 1987) with the modification that artificial seawater supplemented with 6 % (w/v) NaCl was used for the preparation of media. The artificial seawater contained the following (l–1 distilled water): 23.6 g NaCl, 0.64 g KCl, 4.53 g MgCl2 . 6H2O, 5.94 g MgSO4 . 7H2O and 1.3 g CaCl2 . 2H2O (Bruns et al., 2001).
Susceptibility to antibiotics was investigated on MA supplemented with 6 % (w/v) NaCl using discs containing the following concentrations of antibiotic: 100 U polymyxin B; 50 µg streptomycin; 20 U penicillin G; 100 µg chloramphenicol; 10 µg ampicillin; 30 µg cephalothin; 30 µg gentamicin; 5 µg novobiocin; 30 µg tetracycline; 30 µg kanamycin; 15 µg lincomycin; 15 µg oleandomycin; 30 µg neomycin and 100 µg carbenicillin. Acid production from carbohydrates was tested as described by Leifson (1963). Utilization of various substrates for growth was determined as described by Yurkov et al. (1994). Enzyme activities were determined by using the API ZYM system (bioMérieux).
Cell biomass for DNA extraction and for isoprenoid quinone analysis was obtained from cultivation in MB supplemented with 6 % (w/v) NaCl at 30 °C. Chromosomal DNA was isolated and purified according to the method described by Yoon et al. (1996), except that RNase T1 was used in combination with RNase A to minimize contamination of the RNA. The 16S rRNA gene was amplified using a PCR with two universal primers, as described previously (Yoon et al., 1998). Sequencing of the amplified 16S rRNA gene and phylogenetic analysis were performed as described by Yoon et al. (2003). Isoprenoid quinones were analysed as described by Komagata & Suzuki (1987), using reversed-phase HPLC. For cellular fatty acid analysis, cell mass of strain ISL-40T was harvested from agar plates after cultivation for 7 days at 30 °C on MA supplemented with 6 % (w/v) NaCl. The fatty acids were extracted and the fatty acid methyl esters prepared according to the standard protocol of the MIDI/Hewlett Packard Microbial Identification System (Sasser, 1990). The DNA G+C content was determined using the method of Tamaoka & Komagata (1984), with the modification that the DNA was hydrolysed using nuclease P1 (Sigma) and the resultant nucleotides were analysed by reversed-phase HPLC.
The morphological, cultural, physiological and biochemical characteristics of strain ISL-40T are given in the species description (see below) or shown in Table 1. The almost-complete 16S rRNA gene sequence of strain ISL-40T determined in this study comprised 1496 nt, representing approximately 96 % of the Escherichia coli 16S rRNA gene sequence. 16S rRNA gene sequence analysis revealed that strain ISL-40T was phylogenetically most closely related to the genus Marinobacter. In the phylogenetic tree constructed using the neighbour-joining algorithm, strain ISL-40T fell within the clade comprising species of the genus Marinobacter (Fig. 1). Strain ISL-40T exhibited 16S rRNA gene sequence similarity values of 96.3 and 96.4 % to Marinobacter bryozoorum and M. gudaonensis, respectively, and values of 93.5–95.9 % with the type strains of the other species of the genus Marinobacter. The gene sequence similarity values with respect to other species used in the phylogenetic analysis were less than 90.4 %.
|
|
; Gu et al., 2007). The fatty acids of strain ISL-40T consisted of the following (each constituting >0.5 % of total fatty acids): straight-chain fatty acids C16 : 0 (18.4 %), C12 : 0 (6.7 %), C18 : 0 (5.2 %), C17 : 0 (3.3 %) and C16 : 0 alcohol (0.6 %); summed feature 3 (C16 : 17c and/or iso-C15 : 0 2-OH, 18.3 %); 10-methyl fatty acids C16 : 0 (16.9 %), C17 : 0 (2.0 %) and C18 : 0 (1.2 %); saturated fatty acids C18 : 19c (7.7 %), C18 : 17c (5.5 %), C17 : 18c (1.8 %), C16 : 19c (1.2 %) and C18 : 15c (0.8 %); hydroxyl fatty acids C12 : 0 3-OH (6.1 %) and C11 : 0 3-OH (0.6 %); branched fatty acid iso-C17 : 0 (1.2 %) and summed feature 2 (iso-C16 : 1, C14 : 0 3-OH and/or an unknown fatty acid with an equivalent chain-length of 10.928, 0.7 %). This fatty acid profile was similar to those of other species of the genus Marinobacter, although there were differences in the proportions of some fatty acids, probably because of differences in cultivation conditions and extraction procedures (Green et al., 2006; Kim et al., 2006; Gu et al., 2007). The DNA G+C content of strain ISL-40T was 58.1 mol%.
Strain ISL-40T is distinguishable from two phylogenetically related recognized species of the genus Marinobacter by means of differences in several phenotypic characteristics, as shown in Table 1
. The phylogenetic distinctiveness of ISL-40T, together with the differential phenotypic properties, is sufficient to allocate the strain to a separate species from the recognized Marinobacter species (Stackebrandt & Goebel, 1994). Therefore, on the basis of the data presented, strain ISL-40T should be placed within the genus Marinobacter as a novel species, for which the name Marinobacter salicampi sp. nov. is proposed.
Description of Marinobacter salicampi sp. nov.
Marinobacter salicampi (sa.li.cam'pi. L. n. sal salt; L. n. campus field; N.L. gen. n. salicampi of a salt field).
Cells are Gram-negative, aerobic, straight or curved rods (0.4–0.8x1.0–7.0 µm). Motile by means of a single flagellum. Colonies on MA supplemented with 6 % (w/v) NaCl are circular to slightly irregular, raised, smooth, yellowish white in colour and 0.8–1.2 mm in diameter after 7 days incubation at 30 °C. Optimal growth occurs at 30 °C. Optimal pH for growth is between 7.0 and 8.0; growth occurs at pH 5.5, but not at pH 5.0. Optimal growth occurs in the presence of approximately 8 % (w/v) NaCl. Growth occurs in the presence of 15 % (w/v) NaCl, but not in the absence of NaCl or in the presence of more than 16 % (w/v) NaCl. Tweens 20, 40 and 60 are hydrolysed, but aesculin, casein, L-tyrosine, hypoxanthine and xanthine are not. Acid is not produced from the following substrates: L-arabinose, D-cellobiose, D-fructose, D-galactose, D-glucose, lactose, maltose, D-mannose, D-melezitose, melibiose, D-raffinose, L-rhamnose, D-ribose, sucrose, trehalose, D-xylose, D-mannitol, D-sorbitol and myo-inositol. Benzoate and salicin are not utilized. Susceptible to ampicillin, carbenicillin, cephalothin, chloramphenicol, penicillin G, polymyxin B and streptomycin, but not to lincomycin, novobiocin, neomycin or oleandomycin. In assays with the API ZYM system, alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, naphthol-AS-BI-phosphohydrolase and N-acetyl-
-glucosaminidase are present, but lipase (C14), valine arylamidase, cystine arylamidase, trypsin, 
-chymotrypsin, acid phosphatase, -galactosidase, -galactosidase, -glucuronidase, -glucosidase, -glucosidase, -mannosidase and -fucosidase are absent. The predominant ubiquinone is Q-9. The major fatty acids (each constituting >10 % of total fatty acids) are C16 : 0, summed feature 3 (C16 : 17c and/or iso-C15 : 0 2-OH) and 10-methyl C16 : 0. The DNA G+C content is 58.1 mol% (determined by HPLC). Other phenotypic characteristics are given in Table 1.
The type strain, ISL-40T (=KCTC 12972T=CCUG 54357T), was isolated from a marine solar saltern of the Yellow Sea at Byunsan, Korea.
|
ACKNOWLEDGEMENTS |
|---|
|
REFERENCES |
|---|
|
TOPABSTRACTMAIN TEXTREFERENCES |
|---|
Anzai, Y., Kim, H., Park, J.-Y., Wakabayashi, H. & Oyaizu, H. (2000). Phylogenetic affiliation of the pseudomonads based on 16S rRNA sequence. Int J Syst Evol Microbiol 50, 1563–1589.[Abstract]
Bruns, A., Rohde, M. & Berthe-Corti, L. (2001). Muricauda ruestringensis gen. nov., sp. nov., a facultatively anaerobic, appendaged bacterium from German North Sea intertidal sediment. Int J Syst Evol Microbiol 51, 1997–2006.[Abstract]
Cowan, S. T. & Steel, K. J. (1965). Manual for the Identification of Medical Bacteria. London: Cambridge University Press.
Gauthier, M. J., Lafay, B., Christen, R., Fernandez, L., Acquaviva, M., Bonin, P. & Bertrand, J.-C. (1992). Marinobacter hydrocarbonoclasticus gen. nov., sp. nov., a new, extremely halotolerant, hydrocarbon-degrading marine bacterium. Int J Syst Bacteriol 42, 568–576.
Green, D. H., Bowman, J. P., Smith, E. A., Gutierrez, T. & Bolch, C. J. S. (2006). Marinobacter algicola sp. nov., isolated from laboratory cultures of paralytic shellfish toxin-producing dinoflagellates. Int J Syst Evol Microbiol 56, 523–527.
Gu, J., Cai, H., Yu, S.-L., Qu, R., Yin, B., Guo, Y.-F., Zhao, J.-Y. & Wu, X.-L. (2007). Marinobacter gudaonensis sp. nov., isolated from an oil-polluted saline soil in a Chinese oilfield. Int J Syst Evol Microbiol 57, 250–254.
Kim, B.-Y., Weon, H.-Y., Yoo, S.-H., Kim, J.-S., Kwon, S.-W., Stackebrandt, E. & Go, S.-J. (2006). Marinobacter koreensis sp. nov., isolated from sea sand in Korea. Int J Syst Evol Microbiol 56, 2653–2656.
Komagata, K. & Suzuki, K. (1987). Lipid and cell wall analysis in bacterial systematics. Methods Microbiol 19, 161–207.
Lanyi, B. (1987). Classical and rapid identification methods for medically important bacteria. Methods Microbiol 19, 1–67.
Leifson, E. (1963). Determination of carbohydrate metabolism of marine bacteria. J Bacteriol 85, 1183–1184.
Liebgott, P.-P., Casalot, L., Paillard, S., Lorquin, J. & Labat, M. (2006). Marinobacter vinifirmus sp. nov., a moderately halophilic bacterium isolated from a wine-barrel-decalcification wastewater. Int J Syst Evol Microbiol 56, 2511–2516.
Romanenko, L. A., Schumann, P., Rohde, M., Zhukova, N. V., Mikhailov, V. V. & Stackebrandt, E. (2005). Marinobacter bryozoorum sp. nov. and Marinobacter sediminum sp. nov., novel bacteria from the marine environment. Int J Syst Evol Microbiol 55, 143–148.
Sasser, M. (1990). Identification of bacteria by gas chromatography of cellular fatty acids, MIDI Technical Note 101. Newark, DE: MIDI Inc.
Stackebrandt, E. & Goebel, B. M. (1994). Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44, 846–849.
Tamaoka, J. & Komagata, K. (1984). Determination of DNA base composition by reversed-phase high-performance liquid chromatography. FEMS Microbiol Lett 25, 125–128.[CrossRef]
Yoon, J.-H., Kim, H., Kim, S.-B., Kim, H.-J., Kim, W. Y., Lee, S. T., Goodfellow, M. & Park, Y.-H. (1996). Identification of Saccharomonospora strains by the use of genomic DNA fragments and rRNA gene probes. Int J Syst Bacteriol 46, 502–505.
Yoon, J.-H., Lee, S. T. & Park, Y.-H. (1998). Inter- and intraspecific phylogenetic analysis of the genus Nocardioides and related taxa based on 16S rDNA sequences. Int J Syst Bacteriol 48, 187–194.
Yoon, J.-H., Shin, D.-Y., Kim, I.-G., Kang, K. H. & Park, Y.-H. (2003). Marinobacter litoralis sp. nov., a moderately halophilic bacterium isolated from sea water from the East Sea in Korea. Int J Syst Evol Microbiol 53, 563–568.
Yurkov, V., Stackebrandt, E., Holmes, A., Fuerst, J. A., Hugenholtz, P., Golecki, J., Gad'on, N., Gorlenko, V. M., Kompantseva, E. I. & Drews, G. (1994). Phylogenetic positions of novel aerobic, bacteriochlorophyll a-containing bacteria and description of Roseococcus thiosulfatophilus gen. nov., sp. nov., Erythromicrobium ramosum gen. nov., sp. nov., and Erythrobacter litoralis sp. nov. Int J Syst Bacteriol 44, 427–434.
This article has been cited by other articles:
|
|
 |
|
  C.-Y. Wang, C.-C. Ng, W.-S. Tzeng, and Y.-T. Shyu Marinobacter szutsaonensis sp. nov., isolated from a solar saltern Int J Syst Evol Microbiol, October 1, 2009; 59(10): 2605 - 2609. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Aguilera, M. L. Jimenez-Pranteda, K. Kharroub, A. Gonzalez-Paredes, J. J. Durban, N. J. Russell, A. Ramos-Cormenzana, and M. Monteoliva-Sanchez Marinobacter lacisalsi sp. nov., a moderately halophilic bacterium isolated from the saline-wetland wildfowl reserve Fuente de Piedra in southern Spain Int J Syst Evol Microbiol, July 1, 2009; 59(7): 1691 - 1695. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. M. Handley, M. Hery, and J. R. Lloyd Marinobacter santoriniensis sp. nov., an arsenate-respiring and arsenite-oxidizing bacterium isolated from hydrothermal sediment Int J Syst Evol Microbiol, April 1, 2009; 59(4): 886 - 892. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D.-C. Zhang, H.-R. Li, Y.-H. Xin, Z.-M. Chi, P.-J. Zhou, and Y. Yu Marinobacter psychrophilus sp. nov., a psychrophilic bacterium isolated from the Arctic Int J Syst Evol Microbiol, June 1, 2008; 58(6): 1463 - 1466. [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 | |
