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Korea Research Institute of Bioscience and Biotechnology (KRIBB), PO Box 115, Yusong, Taejon, Korea
Correspondence
Jung-Hoon Yoon
jhyoon{at}kribb.re.kr
| ABSTRACT |
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The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains KSL-17T, KSL-18 and KSL-19 are DQ121389, DQ121390 and DQ121391, respectively.
A table giving the cellular fatty acid content of strains KSL-17T, KSL-18 and KSL-19 is available as supplementary material in IJSEM Online.
| MAIN TEXT |
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Strains KSL-17T, KSL-18 and KSL-19 were isolated by using the standard dilution plating technique at 30 °C on 10x diluted nutrient agar (Difco) with the pH adjusted to 9·0 using Na2CO3. To investigate their morphological, physiological and biochemical characteristics, strains KSL-17T, KSL-18 and KSL-19 were routinely cultivated at 28 °C on 2x diluted nutrient agar with the pH adjusted to 8·0. Morphological, physiological, cultural and biochemical properties were examined as described by Yoon et al. (2005a)
. Growth in the absence of NaCl was investigated in trypticase soy broth prepared according to the formula of the Difco medium except that no NaCl was used. Growth at various NaCl concentrations (0·5 %, w/v, and 1·07·0 %, w/v, at intervals of 1·0 %) was investigated in trypticase soy broth (Difco). The pH range for growth was determined in 2x diluted nutrient broth (Difco) that was adjusted to various pH values (initial pH 4·510·5 at intervals of 0·5 pH units). The pH was adjusted, prior to sterilization, to various levels by the addition of HCl or Na2CO3. Cell biomass for DNA extraction and for the analyses of cell-wall and isoprenoid quinones was obtained by cultivation at 28 °C in 2x diluted nutrient broth (pH 8·0). Chemotaxonomic and molecular systematic studies were performed as described by Yoon et al. (2005a)
. The isomer type of the diamino acid in the cell-wall peptidoglycan was analysed using TLC according to the method described by Komagata & Suzuki (1987)
. For fatty acid methyl ester analysis, cell mass from the three strains was harvested from 2x diluted nutrient agar (pH 8·0) after incubation for 10 days at 28 °C.
The morphological, cultural, physiological and biochemical characteristics of strains KSL-17T, KSL-18 and KSL-19 are given in the species description (see later) or are shown in Table 1
. The 16S rRNA gene sequences of strains KSL-17T, KSL-18 and KSL-19 were identical and each comprised 1491 nt, representing approximately 96 % of the Escherichia coli 16S rRNA gene sequence. Comparative 16S rRNA gene sequence analyses showed that the three strains were phylogenetically affiliated to the genus Nocardioides (Fig. 1
). In the phylogenetic trees based on the neighbour-joining, maximum-parsimony and maximum-likelihood algorithms, strains KSL-17T, KSL-18 and KSL-19 formed a distinct phylogenetic lineage within the radiation of the cluster comprising Nocardioides species (Fig. 1
). Similarity values between the 16S rRNA gene sequences of the three strains and those of recognized Nocardioides species ranged from 92·6 % (Nocardioides albus KCTC 9186T) to 95·2 % (N. kribbensis KSL-2T). Values for sequence similarity to other species included in the phylogenetic analysis were below 93·3 %.
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8c (0·56·6 %), 10-methyl C17 : 0 (1·55·8 %), iso-C14 : 0 (2·36·5 %) and so on (see the supplementary table in IJSEM Online). The chemotaxonomic characteristics were consistent with the affiliation of the strains to the genus Nocardioides (Yoon et al., 1997
Phylogenetic analyses based on 16S rRNA gene sequences and chemotaxonomic data revealed that the three strains can be assigned to the genus Nocardioides (Miller et al., 1991
; Tamura & Yokota, 1994
; Park et al., 1999
; Urzì et al., 2000
; Wang et al., 2001
). The mean DNADNA relatedness levels between strains KSL-17T, KSL-18 and KSL-19 were 8590 %, indicating that the three strains represent the same genomic species (Wayne et al., 1987
). They also shared almost identical phenotypic properties. The levels of 16S rRNA gene sequence similarity are low enough to exclude the possibility of assigning the three strains to a previously described Nocardioides species (Stackebrandt & Goebel, 1994
). The three strains were distinguishable from the other Nocardioides species on the basis of some phenotypic properties, as shown in Table 1
. Therefore, on the basis of the data presented, strains KSL-17T, KSL-18 and KSL-19 should be classified in the genus Nocardioides as members of a novel species, for which the name Nocardioides lentus sp. nov. is proposed.
Description of Nocardioides lentus sp. nov.
Nocardioides lentus (len'tus. L. masc. adj. lentus slow, delayed, referring to slow growth).
Cells are aerobic, non-endospore-forming rods (0·40·7x1·04·5 µm) in the exponential phase of growth. Cells show rod-to-coccus morphogenesis from the early exponential phase to the stationary phase. Gram-positive but Gram-variable in old cultures. Colonies are circular, convex, smooth, glistening, yellow in colour and 0·51·0 mm in diameter after 10 days incubation on 2x diluted nutrient agar at 28 °C. Neither substrate mycelium nor aerial mycelium is formed. Growth occurs at 4 and 34 °C, but not at 35 °C. The optimal pH for growth is 8·0; growth occurs at pH 6·5 and 9·5, but not at pH 6·0 or 10·0. Growth occurs in the presence of 05 % (w/v) NaCl, with an optimum at 0·5 % (w/v) NaCl. Weakly positive for oxidase activity. Tweens 20, 40 and 60 are hydrolysed. D-Sorbitol is utilized, but adonitol is not. The cell-wall peptidoglycan contains LL-2,6-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinone is MK-8(H4). The major fatty acid is iso-C16 : 0; 10-methyl fatty acids are present. The DNA G+C content is 74·674·8 mol% (determined by HPLC). Other phenotypic characteristics are given in Table 1
.
The type strain, KSL-17T (=KCTC 19039T=DSM 16315T), was isolated from an alkaline soil in Kwangchun, Korea. Reference strains are KSL-18 and KSL-19.
| ACKNOWLEDGEMENTS |
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