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1 Microbial Genomics and Bioprocessing Research Unit, National Center for Agricultural Utilization Research, Agricultural Research Service, 1815 N. University Street, US Department of Agriculture, Peoria, IL 61604, USA
2 DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
Correspondence
D. P. Labeda
labedadp{at}mail.ncaur.usda.gov
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Published online ahead of print on 1 April 2005 as DOI 10.1099/ijs.0.63496-0.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of Stackebrandtia nassauensis sp. nov. NRRL B-16338T is AY650268.
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), strain LLR-40K-21T (=NRRL B-16338T=DSM 44728T), isolated by Mary P. Lechevalier from a soil sample collected on the road side in Nassau, Bahamas, was found to be phylogenetically most closely related to the genus Glycomyces, but clearly not a member this genus. The strain was subjected to a polyphasic study using standard methods for chemotaxonomic and morphological evaluation. In addition to strain NRRL B-16338T, the type strains of all described Glycomyces species were included in this study, i.e. Glycomyces rutgersensis NRRL B-16106T, G. harbinensis NRRL 15337T, G. tenuis NRRL B-16895T, G. algeriensis NRRL B-16327T, G. arizonensis NRRL B-16153T and G. lechevalieriae NRRL B-16149T.
The strain was cultivated on NZ-amine starch agar (DSMZ medium no. 554) at 28 °C. Morphological observations were made on yeast extract-malt extract agar (ISP-2), inorganic salts-starch agar (ISP-4), glycerol-asparagine agar (ISP-5) (Shirling & Gottlieb, 1966), Czapek's sucrose agar (Pridham & Lyons, 1980) and DSMZ medium no. 554 (DSMZ, 2001).
For the analyses of fatty acids, about 40 mg of cells was scraped from agar plates. For the other chemical analyses, cells were grown in liquid medium and harvested by centrifugation. Chemotaxonomic analyses of strains for polar lipids, menaquinones and fatty acids were performed using previously described methods (Grund & Kroppenstedt, 1989; Minnikin et al., 1984; Sasser, 1990). Cell-wall diamino acid isomer was determined by the method of Staneck & Roberts (1974) and whole-cell sugar content was determined as alditol acetates by the method of Saddler et al. (1991). The acyl type of the muramic acid in the cell wall was determined by the method of Uchida & Aida (1977).
Genomic DNA for sequencing of the 16S rRNA gene was isolated from growth on DSMZ medium no. 554 plates using UltraClean microbial DNA isolation kits (Mo Bio Laboratories) and then amplified and sequenced following previously described procedures (Labeda & Kroppenstedt, 2000) The 16S rRNA gene sequence was deposited in GenBank with the accession number AY650268. A phylogenetic tree was constructed according to the neighbour-joining method of Saitou & Nei (1987) and the stability of the groupings was estimated by bootstrap analysis (Felsenstein, 1989). The G+C content of the DNA was determined by HPLC according to Mesbah et al. (1989).
Physiological tests, including production of acid from carbohydrates, utilization of organic acids and hydrolysis and decomposition of adenine, guanine, hypoxanthine, tyrosine, xanthine, casein, aesculin, urea and hippurate, were evaluated using the media of Gordon et al. (1974). Phosphatase activity was measured using the method of Kurup & Schmitt (1973). The temperature range for growth was determined on slants of DSMZ Medium No. 554.
Phylogenetic analysis clearly demonstrated that strain NRRL B-16338T belongs to the order Actinomycetales (Stackebrandt et al., 1997) and is well separated from all Glycomyces type strains (16S rRNA gene sequence similarity to the Glycomyces species studied is only 92 %) that represent the nearest phylogenetic neighbours (Fig. 1). Evaluation of the data with several other phylogenetic algorithms, including maximum-parsimony and maximum-likelihood (Felsenstein, 1989), provided dendrograms that exhibited a stable relationship between NRRL B-16338T and the species within the genus Glycomyces. The exact placement of the suborder Glycomycineae within the class Actinobacteria is somewhat different for each computational method, possibly reflecting the small number of taxa available and sequenced within this suborder.
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. The whole-cell sugar pattern for strain NRRL B-16338T consists of ribose and inositol, with traces of arabinose and mannose, whereas in all Glycomyces strains, ribose, xylose mannose and galactose are found as diagnostic sugars. NRRL B-16338T and all of the Glycomyces type strains were found to contain N-glycolylmuramic acid. Strain NRRL B-16338T and the Glycomyces strains synthesize phosphatidylglycerol and diphosphatidylglycerol but NRRL B-16338T exhibits two additional unknown phospholipids; all Glycomyces strains are rich in sugar-containing phospholipids such as phosphatidylinositolmannosides and two phosphoglycolipids of unknown structure. The fatty acid pattern clearly separates NRRL B-16338T from the Glycomyces strains by the occurrence of 10-methyl-branched heptadecanoic acid and iso-branched 2-hydroxy fatty acids which are missing in Glycomyces (Table 2). The menaquinone pattern of NRRL B-16338T, where the predominant menaquinones are MK-10(H4), MK-10(H6), MK-11(H4) and MK-11(H6), is also different from that of the genus Glycomyces, where the pattern appears almost species-specific but consists of menaquinones with 9, 10, 11 or 12 isoprene units with various degrees of hydrogenation (Table 3).
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), while observations of growth on Czapek's sucrose agar (not shown) have revealed regions of aerial mycelia that appear almost ready to fragment into spores, although the latter process has never been found.
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On the basis of phylogenetic position and novel chemotaxonomy, strain NRRL B-16338T is clearly not a member of the genus Glycomyces and therefore a new genus is proposed for this strain, to be named Stackebrandtia. The type species of Stackebrandtia is designated Stackebrandtia nassauensis. This genus may well represent a new family within the suborder Glycomycineae Rainey et al. 1997, but it is premature to base this proposal on a genus containing one species. The description of the family Glycomycetaceae is therefore emended to permit inclusion of the genus Stackebrandtia.
Description of Stackebrandtia gen. nov.
Stackebrandtia (Stack.e.brandt'i.a. N.L. fem. n. Stackebrandtia named for Erko Stackebrandt, a German microbiologist who has contributed significantly to the molecular systematics of prokaryotes, including actinobacteria).
Aerobic. Gram-positive, non-motile actinomycetes. Branched substrate mycelium (approx. 0·5 µm in diameter) and, on some media, aerial mycelia are produced. Mycolic acids are absent. Catalase-positive. Contains meso-diaminopimelic acid as the diamino acid. Cell walls contain N-glycolylmuramic acid. The whole-cell sugar pattern consists of ribose and inositol, with traces of arabinose and mannose. The phospholipid pattern consists of phosphatidylglycerol, diphosphatidylglycerol and two additional unknown phospholipids. The predominant menaquinones are MK-10(H4), MK-10(H6), MK-11(H4) and MK-11(H6). Fatty-acid profile is rich in branched-chain and saturated components including 10-methyl-branched heptadecanoic acid and iso-branched 2-hydroxy fatty acids. Phylogenetically, the nearest neighbour is the genus Glycomyces. Type species is Stackebrandtia nassauensis.
Description of Stackebrandtia nassauensis sp. nov.
Stackebrandtia nassauensis (nas.sau.en'sis. N.L. fem. adj. nassauensis from Nassau, named after the place of origin of the type strain, Nassau, Providence, Bahamas).
Displays the following properties in addition to those given in the genus description. Pale yellow to pale tan vegetative mycelium; white to yellowish-white aerial hyphae produced on most media, especially ISP-2. Degrades or hydrolyses allantoin, casein, aesculin, gelatin, hypoxanthine, starch and tyrosine. No degradation of adenine or xanthine. Produces phosphatase. Weakly reduces nitrates. Weakly assimilates acetate and malate; does not assimilate benzoate, citrate, lactate, mucate, oxalate, propionate, succinate or tartrate. Acid is produced from arabinose, cellobiose, dextrin, fructose, galactose, glucose, glycerol, lactose, maltose, mannose, melibiose, methyl 
-D-glucoside, raffinose, rhamnose, salicin, sorbitol, sucrose, trehalose and xylose; no acid produced from adonitol, dulcitol, erythritol, inositol, mannitol, melezitose or methyl 
-xyloside. Temperature range for growth is 15–37 °C. Grows in the presence of 4–9 % NaCl. G+C content of the DNA is 72·4 mol% (HPLC).
The type strain, NRRL B-16338T (LLR-40K-21T=DSM 44728T), was isolated by Mary Lechevalier from a soil sample from Nassau, Bahamas.
Emended description of the family Glycomycetaceae Rainey et al. 1997
Glycomycetaceae (Gly.co.my.ce.ta'ceae. N.L. masc. n. Glycomyces type genus of the family; -taceae ending to denote a family; N.L. fem. pl. n. Glycomycetaceae the Glycomyces family).
Aerobic. Gram-positive, non-acid-fast, non-motile actinomycetes comprising the genera Glycomyces and Stackebrandtia. Branched substrate mycelium (approx. 0·35–0·5 µm in diameter) and, on some media, aerial mycelia are produced. Mycolic acids are absent. Catalase-positive. Contains meso-diaminopimelic acid as the diamino acid. Cell walls contain N-glycolylmuramic acid. Whole-cell sugar pattern contains ribose as a diagnostic sugar along with other genus-specific sugars. Phospholipid content includes phosphatidylglycerol and diphosphatidylglycerol in addition to other genus-specific phospholipids. The 16S rRNA gene sequence pattern of 16S rRNA signature nucleotides contains 70–98 (A–U), 415 (C), 449 (C), 534 (G), 681–709 (A–U), 825–875 (G–C), 999–1041 (C–G), 1059–1198 (C–G), 1064–1192 (G–G), 1117–1183 (A–U) and 1309–1328 (C–G). The type genus is Glycomyces.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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Evtushenko, L. I., Taptykova, S. D., Akimov, V. N., Semyonova, S. A. & Kalakoutskii, L. V. (1991). Glycomyces tenuis sp. nov. Int J Syst Bacteriol 41, 154–157.
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Labeda, D. P. & Kroppenstedt, R. M. (2004). Emended description of the genus Glycomyces and description of Glycomyces algeriensis sp. nov., Glycomyces arizonensis sp. nov. and Glycomyces lechevalierae sp. nov. Int J Syst Evol Microbiol 54, 2343–2346.
Labeda, D. P., Testa, R. T., Lechevalier, M. P. & Lechevalier, H. A. (1985). Glycomyces, a new genus of the Actinomycetales. Int J Syst Evol Microbiol 35, 417–421.
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