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1 Ecole Nationale Supérieure Agronomique de Toulouse, INPT, Laboratoire de Génie Chimique, UMR 5503 (CNRS/INPT/UPS), 1, avenue de l'Agrobiopôle, B. P. 107, F31 326 Castanet-Tolosan Cedex, France
2 Laboratoire de Recherche sur les Produits Bioactifs et la Valorisation de la Biomasse, Ecole Normale Supérieure de Kouba, B.P. 92, 16 050 Vieux-Kouba, Algiers, Algeria
3 Centre de Recherche Scientifique et Technique sur les Régions Arides, Front de l'Oued, B. P. 1682, 07 000 Biskra, Algeria
4 Microbial Genomics and Bioprocessing Research Unit, National Center for Agricultural Utilization Research, USDA-Agricultural Research Service, Peoria, IL 61604, USA
Correspondence
N. Sabaou
sabaou{at}yahoo.fr
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Published online ahead of print on 27 February 2004 as DOI 10.1099/ijs.0.02679-0.
The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain SA 233T is AY054972.
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). The genera Lentzea and Lechevalieria differ from Saccharothrix by the absence of rhamnose and phosphatidyl hydroxyethanolamine or only of rhamnose, respectively, and constitute a lineage phylogenetically distinct from Saccharothrix (Labeda et al., 2001). During studies of the taxonomy and antibiotic production of actinomycetes of Saharan soils from Algeria, we obtained an isolate from soil collected in the palm grove of Adrar oasis, which we have reported to produce three novel dithiolopyrrolone antibiotics (Lamari et al., 2002). Here we describe the characterization and classification of this isolate, which is proposed as the novel species Saccharothrix algeriensis sp. nov. Isolate SA 233T has been deposited in the Agricultural Research Culture Collection and in the Deutsche Sammlung von Mikroorganismen und Zellkulturen under numbers NRRL B-24137T and DSM 44581T, respectively.
Strain SA 233T was isolated from a Saharan soil sample collected at a palm grove in Adrar, Algeria, by a dilution agar plating method using humic acid/B vitamin agar medium (Hayakawa & Nonomura, 1987) supplemented with streptomycin sulphate (10 µg ml–1) and actidione (50 µg ml–1). Biomass for chemotaxonomic analysis was grown on yeast extract/malt extract broth, on a rotary shaker for 4 days at 30 °C, harvested by centrifugation and washed twice with distilled water.
Detailed observation of mycelium morphology was performed using a scanning electron microscope (Stereoscan 260; Cambridge Instruments). Cultural characteristics observed on media from the International Streptomyces Project (ISP) (Shirling & Gottlieb, 1966), nutrient agar and Bennett agar (Waksman, 1961) were recorded after 7–14 days incubation at 28 °C. Colours were determined according to the ISCC–NBS centroid colour chart (Kelly & Judd, 1976).
Tryptone/yeast extract agar (ISP medium 1), peptone/yeast extract/iron agar (ISP medium 6) and tyrosine agar (ISP medium 7) (Shirling & Gottlieb, 1966) were used to determine melanoid pigment production. Decomposition of adenine, guanine, hypoxanthine, tyrosine and xanthine were determined as described by Gordon et al. (1974), and arbutin and aesculin decomposition, gelatin liquefaction, starch hydrolysis and nitrate reductase production were determined as described by Marchal & Bourdon (1973). Utilization of Tween 80 and testosterone was determined by the methods of Sierra (1957) and Goodfellow (1971), respectively. Carbohydrate and organic acid assimilation were determined as described by Goodfellow (1971) and Gordon et al. (1974), respectively. Sensitivity to lysozyme was studied using the method of Gordon & Barnett (1977). Sensitivity to phenol, potassium tellurite, sodium azide, sodium chloride and crystal violet were determined on glucose/yeast extract agar as described by Athalye et al. (1985). Growth at different temperatures and pH and in the presence of various antibiotics was determined on the same media.
The isomeric form of diaminopimelic acid and predominant whole-cell sugars were detected following standard procedures described by Becker et al. (1964) and Lechevalier & Lechevalier (1970). Phospholipids and mycolic acids were analysed using the procedure of Minnikin et al. (1977, 1980). The fatty acid profile was determined by the method of Grund & Kroppenstedt (1989).
Genomic DNA for sequencing was isolated, purified and sequenced following the procedures described by Labeda & Kroppenstedt (2000). The 16S rDNA sequence was manually aligned with actinomycete reference sequences obtained from the Ribosomal Database Project (Maidak et al., 1994) and GenBank in the ARB software environment for sequence data developed by W. Ludwig and O. Strunk (Lehrstuhl für Mikrobiologie, University of Munich, Germany). Evolutionary distances were calculated within ARB by the method of Kimura (1980) and linkages by the neighbour-joining method of Saitou & Nei (1987); maximum-parsimony and maximum-likelihood analyses were also performed in ARB. The topologies of the trees resulting from neighbour-joining and maximum-parsimony analyses were evaluated by bootstrap analysis of the data with 500 resamplings.
Genomic DNA was isolated as described by Labeda (1998) and DNA–DNA relatedness between strains was determined spectrophotometrically in 5x SSC (1x SSC is 0·15 M sodium chloride and 0·015 M sodium citrate) and 20 % dimethyl sulphoxide at 66 °C (melting point–23 °C) by the method of De Ley et al. (1970).
Strain SA 233T exhibited good growth on ISP-2, ISP-5 and Bennett agar, with well-developed, yellow orange aerial mycelium that fragmented into rod-shaped spores. The spores had a smooth surface (Fig. 1) and were non-motile. No endospores, sporangia, sclerotia or synnemata were observed. Growth was moderate on ISP-3 and nutrient agar and poor on ISP-4 agar, with poorly developed aerial mycelium. The substrate mycelium exhibited little or no fragmentation on either solid or liquid media. The substrate mycelium was vivid yellow, orange yellow or yellow brown. A bright yellow soluble pigment was produced on ISP-2 and ISP-3 agar, whereas a yellow brown soluble pigment was produced on Bennett agar. The isolate did not produce melanoid pigments on ISP-1, ISP-6 or ISP-7 agar.
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). Diagnostic phospholipids detected were phosphatidyl ethanolamine (with phosphatidyl hydroxy-ethanolamine) and glucosamine-containing phospholipids, corresponding to phospholipids type PIV (Lechevalier et al., 1977). Mycolic acids were not detected. The predominant fatty acid was iso-branched hexadecanoate (iso-C16 : 0), and significant amounts of iso-C16 : 1 (position of the unsaturation unknown), iso-2-hydroxy-C16 : 0 and iso-C15 : 0 fatty acids were also present. The cellular fatty acid composition of the strain SA 233T is given in Table 1. The morphological and chemical characteristics described above clearly support the placement of strain SA 233T within the genus Saccharothrix.
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. In addition, a comparison with the physiologically nearest species, Saccharothrix australiensis NRRL 11239T, showed differences in characteristics of the cultures (colour of aerial and substrate mycelia and colour of diffusible pigments) and in the utilization of cellobiose, dextrin, mannose, sorbitol, erythritol, sodium acetate, sodium citrate and sodium propionate. Complete physiological characteristics of strain SA 233T are given in the formal description below.
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. Maximum-parsimony calculation resulted in a similar tree topology. The similarity of the 16S rDNA sequence of strain SA 233T to those of the other species of the genus Saccharothrix ranged from 97·5 to 98·8 %, with Saccharothrix australiensis NRRL 11239T having the closest match. Other species within the genus Saccharothrix that have been determined to be distinct based on DNA–DNA relatedness have 16S rRNA gene sequence similarities in this range of values. Strain SA 233T was determined to have only 55·9 % genomic DNA–DNA relatedness (based on a mean of duplicate determinations) to S. australiensis NRRL 11239T, the phylogenetically closest species of the genus Saccharothrix, which is clearly below the 70 % relatedness guideline proposed by Wayne et al. (1987) for the delineation of separate species. All of the data support the designation of strain SA 233T as representing a novel species of the genus Saccharothrix, for which we propose the name Saccharothrix algeriensis sp. nov.
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Aerobic, Gram-positive, catalase-positive. Forms a copious yellow orange aerial mycelium that fragments into rod-shaped spores. Spore surface is smooth. Substrate mycelium is vivid yellow, orange yellow or yellowish brown. A bright yellow soluble pigment is produced but no melanoid pigment. Growth occurs between 18 and 45 °C but not at 48 °C. Utilizes D-fructose, D-galactose, D-glucose, maltose, D-trehalose, glycerol, acetate, citrate, pyruvate and succinate as carbon sources, but not L-arabinose, D-cellobiose, dextrin, D-lactose, D-mannose, D-melezitose, melibiose, methyl 
-D-glucoside, D-raffinose, L-rhamnose, D-ribose, sucrose, D-xylose, adonitol, dulcitol, meso-erythritol, meso-inositol, D-mannitol, D-sorbitol, benzoate, butyrate, oxalate, propionate and tartrate. Aesculin, casein, gelatin, Tween 80 and tyrosine are hydrolysed. No hydrolysis of adenine, arbutin, guanine, hypoxanthine, starch, testosterone or xanthine. Nitrate reductase is produced. Growth occurs at pH 5 and 9 and in the presence of 0·005 % lysozyme, 0·01 % potassium tellurite, 0·001 % crystal violet and 0·05 % phenol. Growth does not occur in the presence of 5 % NaCl, 0·001 % sodium azide and 0·1 % phenol. Susceptible to chloramphenicol (25 µg ml–1), erythromycin (10 µg ml–1), kanamycin (25 µg ml–1), novobiocin (10 µg ml–1) and penicillin (10 µg ml–1) but resistant to cycloserin (10 µg ml–1), gentamicin (5 µg ml–1), oxytetracycline (25 µg ml–1), rifampicin (5 µg ml–1), streptomycin (10 µg ml–1) and vancomycin (5 µg ml–1). Type III cell wall (meso-diaminopimelic acid, galactose, mannose and rhamnose in whole-cell hydrolysates). Phospholipids type PIV (phosphatidyl ethanolamine and glucosamine-containing phospholipids). The predominant fatty acid is iso-C16 : 0 (31·26 %), followed by iso-H-C16 : 0 (14·00 %), iso-2-hydroxy-C16 : 0 (10·44 %) and iso-C15 : 0 (10·06 %). Mycolic acids are absent.
The type strain is SA 233T (=NRRL B-24137T=DSM 44581T), isolated from a Saharan soil sample collected at a palm grove in Adrar, Algeria. The species description is based on a single strain and hence serves as the strain description.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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Becker, B., Lechevalier, M. P., Gordon, R. E. & Lechevalier, H. A. (1964). Rapid differentiation between Nocardia and Streptomyces by paper chromatography of whole-cell hydrolysates. Appl Microbiol 12, 421–423.[Medline]
De Ley, J., Cattoir, H. & Reynaerts, A. (1970). The quantitative measurement of DNA hybridization from renaturation rates. Eur J Biochem 12, 133–142.[Medline]
Goodfellow, M. (1971). Numerical taxonomy of some nocardioform bacteria. J Gen Microbiol 69, 33–80.[Medline]
Gordon, R. E. & Barnett, D. A. (1977). Resistance to rifampin and lysozyme of strains of some species of Mycobacterium and Nocardia as a taxonomic tool. Int J Syst Bacteriol 27, 176–178.
Gordon, R. E., Barnett, D. A., Handerhan, J. E. & Pang, C.-H. N. (1974). Nocardia coeliaca, Nocardia autotrophica, and the nocardin strain. Int J Syst Bacteriol 24, 54–63.
Grund, E. & Kroppenstedt, R. M. (1989). Transfer of five Nocardiopsis species to the genus Saccharothrix Labeda et al., 1984. Syst Appl Microbiol 12, 267–274.
Hayakawa, M. & Nonomura, H. (1987). Humic acid–vitamin agar, a new medium for the selective isolation of soil actinomycetes. J Ferment Technol 65, 501–509.[CrossRef]
Kelly, K. L. & Judd, D. B. (1976). Color. Universal Language and Dictionary of Names (National Bureau of Standards Special Publication 440). Washington, DC: US Department of Commerce.
Kimura, M. (1980). A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16, 111–120.[CrossRef][Medline]
Labeda, D. P. (1998). DNA relatedness among the Streptomyces fulvissimus and Streptomyces griseoviridis phenotypic cluster groups. Int J Syst Bacteriol 48, 829–832.
Labeda, D. P. (2002). The family Actinosynnemataceae. In The Prokaryotes: an Evolving Electronic Resource for the Microbiological Community, 3rd edn, release 3.9, 1 April 2002. Edited by M. Dworkin et al. New York: Springer. http://www.prokaryotes.com
Labeda, D. P. & Kroppenstedt, R. M. (2000). Phylogenetic analysis of Saccharothrix and related taxa: proposal for Actinosynnemataceae fam. nov. Int J Syst Evol Microbiol 50, 331–336.[Abstract]
Labeda, D. P., Hatano, K., Kroppenstedt, R. M. & Tamura, T. (2001). Revival of the genus Lentzea and proposal for Lechevalieria gen. nov. Int J Syst Evol Microbiol 51, 1045–1050.[Abstract]
Lamari, L., Zitouni, A., Boudjella, H., Badji, B., Sabaou, N., Lebrihi, A., Lefebvre, G., Seguin, E. & Tillequin, F. (2002). New dithiolopyrrolone antibiotics from Saccharothrix sp. SA 233. I. Taxonomy, fermentation, isolation, and biological activities. J Antibiot 55, 696–701.[Medline]
Lechevalier, M. P. & Lechevalier, H. A. (1970). Chemical composition as a criterion in the classification of aerobic actinomycetes. Int J Syst Bacteriol 34, 435–444.
Lechevalier, M. P., de Bièvre, C. & Lechevalier, H. A. (1977). Chemotaxonomy of aerobic actinomycetes: phospholipid composition. Biochem Syst Ecol 5, 249–260.[CrossRef]
Maidak, B. L., Larsen, N., McCaughey, M. J., Overbeek, R., Olsen, G. J., Fogel, K., Blandy, J. & Woese, C. R. (1994). The Ribosomal Database Project. Nucleic Acids Res 22, 3485–3487.
Marchal, N. & Bourdon, J. L. (1973). Milieux de Culture et Identification Biochimique des Bactéries. Paris: Doin (in French).
Minnikin, D. E., Patel, P. V., Alshamaony, L. & Goodfellow, M. (1977). Polar lipid composition in the classification of Nocardia and related bacteria. Int J Syst Bacteriol 27, 104–107.
Minnikin, D. E., Hutchinson, I. G., Caldicott, A. B. & Goodfellow, M. (1980). Thin-layer chromatography of methanolysates of mycolic acid-containing bacteria. J Chromatogr 188, 221–233.[CrossRef]
Saitou, N. & Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425.[Abstract]
Shirling, E. B. & Gottlieb, D. (1966). Methods for characterization of Streptomyces species. Int J Syst Bacteriol 16, 313–340.
Sierra, G. (1957). A simple method for the detection of lipolytic activity of micro-organisms and some observations on the influence of contact between the cells and fatty substrates. Antonie van Leeuwenhoek 23, 15–22.[Medline]
Waksman, S. A. (1961). Classification, identification, and descriptions of genera and species. In The Actinomycetes, vol. 2, pp. 331–332. Baltimore: Williams & Wilkins.
Wayne, L. G., Brenner, D. J., Colwell, R. R. & 9 other authors (1987). Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. Int J Syst Bacteriol 37, 463–464.
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