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Biological Resource Center, National Institute of Technology and Evaluation, 2-5-8 Kazusakamatari, Kisarazu, Chiba, 292-0812, Japan
Correspondence
Tomohiko Tamura
tamura-tomohiko{at}nite.go.jp
| ABSTRACT |
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The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of Planotetraspora silvatica TT 00-51T is AB112082.
A scanning electron micrograph of strain TT 00-51T is available as supplementary material in IJSEM Online.
| MAIN TEXT |
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During a study of the distribution of actinomycetes in the subtropical field, strain TT 00-51T was isolated from soil of Amami Island, Japan. The isolate was judged to belong to the genus Planotetraspora by phylogenetic analysis and chemotaxonomic characteristics, and to represent a novel species of the genus Planotetraspora by DNADNA relatedness and physiological characteristics. Therefore, we propose the name Planotetraspora silvatica sp. nov. for strain TT 00-51T.
Strain TT 00-51T (=NBRC 100141T=DSM 44746T) was isolated from a sample of forest soil in Amami Island, Kagoshima prefecture, Japan. This organism was isolated on humic acid/vitamin (HV) agar (Hayakawa & Nonomura, 1987
) by using the yeast extract/SDS method (Hayakawa & Nonomura, 1989
). Freeze-dried cells for chemotaxonomic analyses were grown in yeast extract/glucose broth [10 g yeast extract, 10 g D-glucose (l distilled water)1, pH 7·0] on a rotary shaker at 28 °C. P. mira NBRC 15435T was also analysed to compare whole-cell sugar patterns.
Cultural and physiological characteristics were determined as described previously (Gordon et al., 1974
; Seino et al., 1985
; Shirling & Gottlieb, 1966
; Yokota et al., 1993
). Morphology was observed by scanning electron microscopy as described previously (Tamura et al., 2000
).
Strain TT 00-51T developed sporangia at the ends of short sporangiophores on branched aerial mycelium. Each sporangium was seen to contain four spores. Globose bodies were not observed. Spores were oval to short rods (0·60·9x0·81·5 µm); motile spores were not observed. A micrograph is available as supplementary material in IJSEM Online.
The strain exhibited good growth on yeast extract/malt extract agar, glycerol/asparagine agar and tyrosine agar, and developed white colonies. In contrast, P. mira NBRC 15435T developed yellow colonies on yeast extract/malt extract agar. The new strain developed white aerial mycelium on yeast extract/malt extract agar, inorganic salts/starch agar, glycerol/asparagine agar, tyrosine agar and HV agar.
The strain utilized mannose, lactose, galactose, methyl
-D-glucoside, maltose, rhamnose, melibiose and raffinose. Decomposition of hypoxanthine and L-tyrosine was positive, but resistance to 4 % NaCl, hydrolysis of starch, utilization of lactate, malate, succinate, citrate and oxalate and decomposition of urea, adenine, cellulose and calcium malate were negative. The strain showed good growth at 2530 °C. In contrast, P. mira NBRC 15435T did not utilize maltose or galactose or decompose L-tyrosine.
Analyses of whole-cell sugar pattern, cell-wall amino acids, menaquinones, cellular fatty acids, isomers of A2pm, acyl type of peptidoglycan, mycolic acid and DNA G+C content were performed as described previously (Tamura et al., 1994
).
The new strain contained glucose, madurose, galactose, rhamnose and 3-O-methylmannose as whole-cell sugars. In this study, consistent with the results of Kudo (2001)
, P. mira NBRC 15435T was found to contain madurose and rhamnose, but not xylose or arabinose. The amino acids in the cell wall of the new strain were meso-A2pm, alanine and glutamic acid; this corresponds to murein type A1
, according to Schleifer & Kandler (1972)
. The predominant isoprenoid quinones of the new strain were MK-9(H4), MK-9(H2) and MK-9. Phosphatidylethanolamine and an unidentified phospholipid containing glucosamine were detected as diagnostic phospholipids, but phosphatidylglycerol and phosphatidylcholine were not detected. The major isoprenoid quinone and phospholipid pattern of strain TT 00-51 T were consistent with those of P. mira (Kudo, 2001
). Mycolic acids were not detected. The glycan moiety of the murein contained acetyl residues. The cellular fatty acids consisted of iso-branched, anteiso-branched, saturated, unsaturated and 10-methylated fatty acids, corresponding to fatty acid pattern 3d of Kroppenstedt (1985)
. The strain contained diagnostic amounts of 10-methylated C18 : 0 and iso-C16 : 0 (>14 %). The DNA G+C content of the isolate ranged from 70 to 71 mol%.
The microplate hybridization method developed by Ezaki et al. (1988
, 1989)
was applied with minor modifications to determine DNADNA relatedness (Tamura et al., 1999
). The strain exhibited DNADNA relatedness levels of 3842 % with P. mira NBRC 15435T.
PCR amplification and sequencing of the 16S rRNA gene were performed as described previously (Tamura & Hatano, 2001
) with a model ABI PRISM 3100 genetic analyser (Applied Biosystems) according to the manufacturer's protocol. Phylogenetic analysis of 16S rRNA gene sequences was performed as described previously (Tamura & Hatano, 2001
). This phylogenetic analysis revealed that the isolate fell within the cluster of the family Streptosporangiaceae and, with P. mira NBRC 15435T, formed a line of descent that was distinct from other actinomycetes of this family (Fig. 1
). The signature nucleotides of the 16S rRNA gene at positions 502543 (A-U) and 11161184 (U-G) of P. mira NBRC 15435T and the isolate were different from those of other members of the family Streptosporangiaceae [positions 502543 (G-C), 11161184 (C-G)]. The closest neighbours are members of the genera Acrocarpospora (similarity values 95·897·1 %) and Herbidospora (similarity values 95·897·1 %).
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On the basis of morphological, chemotaxonomic and phylogenetic criteria, strain TT 00-51T is considered to belong to the genus Planotetraspora and to represent a distinct species, based on DNADNA relatedness and physiological characteristics (Table 1
). We propose that the isolate should be classified as the type strain of a novel species, Planotetraspora silvatica sp. nov. (type strain, TT 00-51T=NBRC 100141T=DSM 44746T).
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, according to Schleifer & Kandler (1972)
Description of Planotetraspora silvatica sp. nov.
Planotetraspora silvatica (sil.va'ti.ca. L. fem. adj. silvatica of the forest).
Morphological, chemotaxonomic and general characteristics are as given above for the genus. Brownish, soluble pigment is produced on tyrosine agar (ISP medium 7). Starch is not hydrolysed. Hydrolysis of gelatin is negative or weakly positive. Calcium malate is not decomposed. Coagulation and peptonization of milk are positive. Optimum temperature for growth is 2530 °C. Does not grow at 37 °C. Does not grow on 4 % NaCl. Glucose, mannose, lactose, galactose, methyl
-D-glucoside, maltose, rhamnose and melibiose are utilized, but dulcitol, erythritol, adonitol and arabinose are not. As major cellular fatty acids, 10-methylated C18 : 0 and iso-C16 : 0 are present. DNA G+C content is 71 mol%.
The type strain is TT 00-51T (=NBRC 100141T=DSM 44746T). Habitat is soil.
| ACKNOWLEDGEMENTS |
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