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NITE Biological Resource Center (NBRC), National Institute of Technology and Evaluation, 2-5-8 Kazusakamatari, Kisarazu, Chiba 292-0818, Japan
Correspondence
Tomohiko Tamura
tamura-tomohiko{at}nite.go.jp
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ABSTRACT |
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The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequences of Actinocorallia aurea sp. nov. NBRC 14752T and strain NBRC 15120 are AB006177 and AB006160, respectively.
A table detailing the cellular fatty acid contents of Actinocorallia aurea sp. nov. NBRC 14752T and selected members of the genus Actinocorallia, and a scanning electron micrograph of cells of the strain are available with the online version of this paper.
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. Subsequently, Zhang et al. (2001) enlarged the monotypic genus by reclassifying four species of the genus Actinomadura and emending the genus description. At the time of writing, the genus comprises six species with validly published names, including the recently described species Actinocorallia cavernae (Lee, 2006).
During a taxonomic study of strains assigned to genera with non-validly published names, it was revealed that two strains, NBRC 14752 and NBRC 15120, described as ‘Sarraceniospora aurea’ (Furihata et al., 1989) formed a phylogenetic cluster with the members of the genus Actinocorallia. In this paper, we describe the results of polyphasic taxonomic studies of the two strains and propose to assign them to a novel species of the genus Actinocorallia.
PCR-amplification and sequencing of the 16S rRNA gene and phylogenetic analysis were performed as described previously (Tamura & Hatano, 2001). Strains NBRC 14752 and NBRC 15120 had identical 16S rRNA gene sequences and formed a monophyletic cluster with members of the genus Actinocorallia (Fig. 1). 16S rRNA gene sequence similarity calculations after a neighbour-joining analysis indicated that the closest relatives of both strains were Actinocorallia herbida (98.9 % gene sequence similarity), Actinocorallia aurantiaca (98.6 %), Actinocorallia glomerata (98.3 %), Actinocorallia longicatena (98.1 %) and Actinocorallia libanotica (98.1 %). Lower sequence similarities were found with Actinocorallia cavernae (97.3 %).
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). Freeze-dried cells for chemotaxonomic analyses were obtained from a culture grown in yeast extract-glucose broth (1.0 % yeast extract; 1.0 % D-glucose; pH 7.0) on a rotary shaker at 28 °C. The major menaquinones were MK-9(H4), MK-9(H6) and MK-9(H8) and a small amount of MK-9(H2) was also present. The organisms have meso-diaminopimelic acid (meso-DAP), glutamic acid and alanine as cell-wall amino acids and glucose, galactose and small amounts of madurose and xylose as the whole-cell sugars (wall chemotype/sugar pattern III/B sensu Lechevalier & Lechevalier, 1970). The data differed from those reported by Furihata et al. (1989), in which the presence of arabinose and a trace of glycine were revealed and the cell-wall chemotype was designated as II/D. The diagnostic phospholipids phosphatidylethanolamine and phosphatidylglycerol were detected in strains NBRC 14752 and NBRC 15120, but phosphatidylcholine and some other phospholipids, including glucosamine, were not detected (phospholipid type PII sensu Lechevalier et al., 1977). The DNA G+C contents of strains NBRC 14752 and NBRC 15120 were 71.6 and 72.2 mol%, respectively. Mycolic acids were not detected. The acyl type of the muramic acid was acetyl. The fatty acid profile of strains NBRC 14752 and NBRC 15120 (see Supplementary Table S1 in IJSEM Online) contained C16 : 0 (21 % and 22 %, respectively), C18 : 1 (19 % and 18 %), 10-methyl C18 : 0 (10 % and 14 %), C17 : 1 (13 % and 9 %), C16 : 1 (8 % and 12 %), 10-methyl C16 : 0 (4 % and 10 %), 10-methyl C17 : 0 (7 % and 5 %) and iso-C16 : 0 (6 % and 5 %) (fatty acid type 3a sensu Kroppenstedt, 1985). No significant differences in the fatty acid profiles were found between strains NBRC 14752 and NBRC 15120 and A. herbida.
Morphological characteristics were observed by using scanning electron microscopy as described previously (Tamura et al., 2000). Strains NBRC 14752 and NBRC 15120 formed a straight to flexuous spore chain on the tip of coralloid sporophore as seen in A. herbida (Iinuma et al., 1994). A structure of the sheath, similar in shape to a North American pitcher plant of the genus Sarracenia, was occasionally observed at the tip of the spore chains as previously reported by Furihata et al. (1989) (see Supplementary Fig. S1 in IJSEM Online). The tip of the sheath was similar in appearance to the operculum of a pitcher plant and the spore chain resembled the pitcher tube.
Determination of the cultural and physiological characteristics was performed as described previously (Gordon et al., 1974; Seino et al., 1985; Shirling & Gottlieb, 1966; Yokota et al., 1993). Detailed results are provided in the species description and in Table 1. The colour of the aerial spore mass of strains NBRC 14752 and NBRC 15120 was nearly identical to that of A. herbida, but the colour of the vegetative mycelium was different. The two strains did not produce a soluble pigment on tyrosine agar (ISP medium 7). Gelatin liquefaction and growth were observed at 37 °C (Table 1).
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, 1989) was applied to determine DNA–DNA relatedness. The DNA–DNA relatedness of strain NBRC 14752 with strain NBRC 15120 was 60 and 67 % (reciprocal values), 27 % and 34 % with A. herbida NBRC 15485T, and less than 18 % with A. aurantiaca NBRC 16147T, A. glomerata NBRC 15960T and A. longicatena NBRC 15961T. The two strains are distinguishable from the other members of the genus Actinocorallia by several phenotypic properties as shown in Table 1
. Therefore, on the basis of morphological, physiological and chemotaxonomic data, DNA–DNA hybridization and phylogenetic distinctiveness, ‘Sarraceniospora aurea’ NBRC 14752 and strain NBRC 15120 should be placed in the genus Actinocorallia as members of a novel species, for which the name Actinocorallia aurea sp. nov. is proposed.
Description of Actinocorallia aurea sp. nov.
Actinocorallia aurea (au're.a. L. fem. adj. aurea golden).
The description is based on the data obtained in this study and also reported by Furihata et al. (1989). Aerobic, Gram-positive, non-acid-fast actinomycete that produces branching, non-fragmented vegetative hyphae. Straight to flexuous chains of aerial spores are formed on the tip of coralloid sporophores. Spores are short, non-motile rods (0.6–0.9x1.0–1.5 µm). Vegetative mycelium is yellow on maltose-Bennett's agar, yeast extract-malt-extract agar (ISP medium 2), inorganic salts-starch agar (ISP medium 4), glycerol-asparagine agar (ISP medium 5) and peptone-yeast extract-iron agar (ISP medium 6) and greyish white on oatmeal (ISP medium 3) and tyrosine agars (ISP medium 7). The aerial spore mass is yellow on yeast extract-starch agar, maltose-Bennett's agar, 1/4 strength ISP medium 2 and ISP medium 3. Tests for starch hydrolysis, nitrate reduction and gelatin liquefaction are positive. Does not produce a soluble pigment on ISP medium 7. Starch is weakly hydrolysed. Calcium malate is not dissolved. Tests for the coagulation and peptonization of milk give negative results. Optimum temperature for growth is 20–30 °C. Grows at 37 °C, but does not grow at 45 °C. Does not grow on 4 % NaCl. D-Glucose, L-rhamnose, D-xylose and maltose are utilized. The cell-wall peptidoglycan contains meso-DAP as the diagnostic diamino acid and the whole-cell sugars contain madurose as the diagnostic sugar, indicating that the cell-wall chemotype is III/B. The major fatty acids are C16 : 0, C18 : 1, 10-methyl C18 : 0 and C17 : 1. The major menaquinones are MK-9(H4), MK-9(H6) and MK-9(H8). The major polar lipids are phosphatidylglycerol and phosphatidylethanolamine. The DNA G+C content is 71–73 mol% (determined by HPLC).
The type strain, NBRC 14752T (=DSM 44434T), was isolated from soil.
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REFERENCES |
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