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1 Applied Microbiology Division, National Institute of Agricultural Science and Technology, Rural Development Administration (RDA), Suwon 441-707, Korea
2 Korean Agricultural Culture Collection, Microbial Genetics Division, National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea
3 National Institute of Subtropical Agriculture, RDA, Jeju 690-150, Republic of Korea
4 Taean Lily Experimental Station, Chungnam Provincial Agricultural Research and Extension Services, Taean 357-952, Republic of Korea
5 National Institute of Highland Agriculture, RDA, Pyongchang 232-955, Republic of Korea
Correspondence
Soon-Wo Kwon
swkwon{at}rda.go.kr
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ABSTRACT |
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7c. The predominant isoprenoid quinone was ubiquinone 10 (Q-10). Diphosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidylethanolamine, phosphatidyldimethylethanolamine, phosphatidylglycerol, phosphatidylcholine and four unknown lipids were found as the polar lipid components. The DNA G+C content was 67.0 mol%. On the basis of the phenotypic and phylogenetic features studied, we propose that strain 5715S-12T be assigned to a novel species of the genus Aurantimonas, for which the name Aurantimonas ureilytica sp. nov. (type strain 5715S-12T =KACC 11607T =DSM 18598T) is proposed.
-hydroxybutyrateThe GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain 5715S-12T is DQ883810.
A transmission electron micrograph of a cell of strain 5715S-12T, polar lipid profiles of strain 5715S-12T and related strains and a 16S rRNA gene sequence-based maximum-parsimony tree are available as supplementary material with the online version of this paper.
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). A second species, Aurantimonas altamirensis, was proposed recently for an isolate from a subterranean environment (Jurado et al., 2006). With the genus Fulvimarina, isolated from marine environments (Cho & Giovannoni, 2003), this genus took up a distinct phylogenetic position separable from the other families within the order Rhizobiales. Thus, these two genera were classified into an eleventh family within the order Rhizobiales of the Alphaproteobacteria, the family Aurantimonadaceae. In the course of the study of bacterial strains isolated from air samples, we isolated a yellow-coloured bacterium, 5715S-12T. The air samples were collected with an MAS-100 air sampler (Merck) (single-stage multiple-hole impactor) in Suwon city, Korea, on 15 July 2005. The sampler contained Petri dishes with R2A agar (BBL) amended with 200 µg cycloheximide ml–1 (Sigma). After sampling, plates were incubated at 28 °C for 5 days.
For phenotypic characterization, R2A medium was used as the basal medium. For transmission electron microscopy, cells were grown for 48 h on R2A medium, negatively stained with 0.5 % (w/v) uranyl acetate and examined with an LEO model 912AB electron microscope. Gram staining, catalase, oxidase, poly--hydroxybutyrate (PHB) formation and hydrolysis of casein, DNA, pectin and starch were determined by the method of Smibert & Krieg (1994). Hydrolysis of CM-cellulose (0.1 %, w/v), chitin (1 %, w/v) and tyrosine (0.5 %, w/v) was tested by the appearance of clear zones around colonies. Temperature, salinity and pH ranges for growth were tested over the ranges of 5–45 °C, 0–10 % (w/v) NaCl and pH 4.0–10.0. Pigment analysis was performed as reported by Denner et al. (2003). Further biochemical characteristics were determined using the API 20NE, API ID 32GN and API ZYM systems, following the instructions of the manufacturer (bioMérieux).
Isoprenoid quinones were extracted from lyophilized cells and analysed by HPLC as described previously (Groth et al., 1996). Polar lipid profiles were determined according to the methods of Minnikin et al. (1984). Whole-cell fatty acids were analysed according to the standard protocol of the MIDI/Hewlett Packard Microbial Identification System (Sasser, 1990) after cultivation on marine agar 2216 (MA; Difco) for 3 days at 28 °C. The DNA G+C content was measured according to Mesbah et al. (1989) using a reversed-phase column (Supelcosil LC-18S; Supelco).
The 16S rRNA gene of the isolate was amplified by a PCR and sequenced directly using an ABI Prism 310 Genetic Analyzer (Applied Biosystems). The closest known relatives of the novel isolate were determined by performing GenBank/EMBL/DDBJ database searches. The software package MEGA version 3.1 (Kumar et al., 2004) was used for all analyses. Distances (using distance options according to Kimura's two-parameter model) and clustering using the neighbour-joining and maximum-parsimony methods were determined by using bootstrap values based on 1000 replicates.
Strain 5715S-12T grew on R2A, MA, nutrient agar (Difco) and tryptic soy agar (Difco), but did not grow on MacConkey agar (Difco). Colonies were yellow, round and convex with clear margins after 3 days on R2A. Cells were short rods with more than one polar flagellum (Supplementary Fig. S1 available in IJSEM Online). Strain 5715S-12T yielded carotenoid pigments with peaks in the absorption spectrum at 447 and 470–471 nm and with a slight inflexion at 424–427 nm. Detailed phenotypic properties are summarized in Table 1 and the species description.
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7c (62.2 %), and moderate amounts of C18 : 1 2-OH (12.7 %), C16 : 0 (9.0 %) and summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 17c) (6.9 %) were also detected (Table 2). The fatty acid profile of strain 5715S-12T was similar to that of A. altamirensis S21BT except for the presence of a small amount (1.3 %) of C16 : 0 3-OH in the former strain (Table 2). The DNA G+C content was 67.0 mol%.
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) shows that strain 5715S-12T forms a cluster with the two Aurantimonas species and F. pelagi. According to the maximum-parsimony tree (Supplementary Fig. S3 available in IJSEM Online), strain 5715S-12Twas grouped with A. altamirensis S21BT within the genus Aurantimonas clade. Based on these results, it is concluded that strain 5715S-12T represents a novel species of the genus Aurantimonas, for which the name Aurantimonas ureilytica sp. nov. is proposed.
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Cells are motile, Gram-negative, non-spore-forming, short rods (1.0–1.5 µm wide and 1.5–2.5 µm long). Strictly anaerobic and catalase- and oxidase-positive. The ranges (optima) of temperature, pH and NaCl concentration for growth are 4–37 °C (25–30 °C), pH 4–9 (pH 7–8) and 0–3 %. Negative for nitrate reduction, indole production, glucose fermentation and arginine dihydrolase. Cells hydrolyse starch, Tween 80 and urea, but not aesculin, casein, chitin, CM-cellulose, DNA, gelatin, pectin or tyrosine. According to API 20NE and API ID 32GN test strips, D-glucose, L-arabinose, D-mannose, D-mannitol, malic acid, L-rhamnose, D-ribose, inositol, sucrose, sodium acetate, lactic acid, L-alanine, potassium 5-ketogluconate, glycogen, D-melibiose, L-fucose, D-sorbitol, propionic acid, valeric acid, potassium 2-ketogluconate, 3-hydroxybutyric acid and 4-hydroxybenzoic acid are assimilated. Does not assimilate N-acetylglucosamine, maltose, potassium gluconate, capric acid, adipic acid, trisodium citrate, phenylacetic acid, itaconic acid, suberic acid, sodium malonate, 3-hydroxybenzoic acid, L-serine, salicin, L-histidine or L-proline. According to API ZYM test strips, positive for activities of alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase and naphthol-AS-BI-phosphohydrolase and negative for activities of lipase (C14), valine arylamidase, cystine arylamidase, trypsin, 
-chymotrypsin, -galactosidase, -galactosidase, -glucuronidase, -glucosidase, -glucosidase, N-acetyl--glucosaminidase, -mannosidase and -fucosidase. The major fatty acid is C18 : 17c. The predominant isoprenoid quinone is Q-10. The DNA G+C content of the type strain is 67.0 mol%.
The type strain is 5715S-12T (=KACC 11607T =DSM 18598T), isolated from air in the Republic of Korea.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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Denner, E. B. M., Smith, G. W., Busse, H.-J., Schumann, P., Narzt, T., Polson, S. W., Lubitz, W. & Richardson, L. L. (2003). Aurantimonas coralicida gen. nov., sp. nov., the causative agent of white plague type II on Caribbean scleractinian corals. Int J Syst Evol Microbiol 53, 1115–1122.
Groth, I., Schumann, P., Weiss, N., Martin, K. & Rainey, F. A. (1996). Agrococcus jenensis gen. nov., sp. nov., a new genus of actinomycetes with diaminobutyric acid in the cell wall. Int J Syst Bacteriol 46, 234–239.
Jurado, V., Gonzalez, J. M., Laiz, L. & Saiz-Jimenez, C. (2006). Aurantimonas altamirensis sp. nov., a member of the order Rhizobiales isolated from Altamira Cave. Int J Syst Evol Microbiol 56, 2583–2585.
Kumar, S., Tamura, K. & Nei, M. (2004). MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5, 150–163.
Mesbah, M., Premachandran, U. & Whitman, W. B. (1989). Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. Int J Syst Bacteriol 39, 159–167.
Minnikin, D. E., O'Donnell, A. G., Goodfellow, M., Alderson, G., Athalye, M., Schaal, A. & Parlett, J. H. (1984). An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids. J Microbiol Methods 2, 233–241.[CrossRef]
Sasser, M. (1990). Identification of bacteria by gas chromatography of cellular fatty acids. Technical Note no. 101. Newark, DE: MIDI Inc.
Smibert, R. M. & Krieg, N. R. (1994). Phenotypic characterization. In Methods for General and Molecular Bacteriology, pp. 607–654. Edited by P. Gerhardt, R. G. E. Murray, W. A. Wood & N. R. Krieg. Washington, DC: American Society for Microbiology.
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