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Actinopolyspora Righensis Sp. Nov., a Novel Halophilic Actinomycete Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Algeria Atika Meklat, Noureddine Bouras, Abdelghani Zitouni, Florence Mathieu, Ahmed Lebrihi, Peter Schumann, Cathrin Spröer, Hans-Peter Klenk, Nasserdine Sabaou To cite this version: Atika Meklat, Noureddine Bouras, Abdelghani Zitouni, Florence Mathieu, Ahmed Lebrihi, et al.. Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Al- geria. Antonie van Leeuwenhoek, Springer Verlag, 2013, vol. 104 (n° 3), pp. 301-307. 10.1007/s10482- 013-9948-7. hal-01177618 HAL Id: hal-01177618 https://hal.archives-ouvertes.fr/hal-01177618 Submitted on 17 Jul 2015 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Open Archive Toulouse Archive Ouverte (OATAO) OATAO is an open access repository that collects the work of Toulouse researchers and makes it freely available over the web where possible. This is an author-deposited version published in: http://oatao.univ-toulouse.fr/ Eprints ID: 9759 DOI:10.1007/s10482-013-9948-7 Official URL: http://dx.doi.org/10.1007/s10482-013-9948-7 To cite this version: Meklat, Atika and Bouras, Noureddine and Zitouni, Abdelghani and Mathieu, Florence and Lebrihi, Ahmed and Schumann, Peter and Spröer, Cathrin and Klenk, Hans-Peter and Sabaou, Nasserdine Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Algeria. (2013) Antonie van Leeuwenhoek, vol. 104 (n° 3). pp. 301-307. ISSN 0003-6072 Any correspondence concerning this service should be sent to the repository administrator: [email protected] DOI 10.1007/s10482-013-9948-7 Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Algeria Atika Meklat • Noureddine Bouras • Abdelghani Zitouni • Florence Mathieu • Ahmed Lebrihi • Peter Schumann • Cathrin Spro¨er • Hans-Peter Klenk • Nasserdine Sabaou Abstract A novel halophilic actinomycete strain, chains with non-motile, smooth-surfaced and rod-shaped T T H23 , was isolated from a Saharan soil sample collected spores. Strain H23 had MK-10 (H4) and MK-9 (H4) as in Djamaˆa (Oued Righ region), El-Oued province, South the predominant menaquinones. The whole micro- Algeria. Strain H23T was identified as a member of the organism hydrolysates mainly consisted of meso-diami- genus Actinopolyspora by a polyphasic approach. Phy- nopimelic acid, galactose and arabinose. The diagnostic logenetic analysis showed that strain H23T had 16S phospholipid detected was phosphatidylcholine. The rRNA gene sequence similarities ranging from 97.8 % major cellular fatty acids were anteiso-C17:0 (37.4 %), T (Actinopolyspora xinjiangensis TRM 40136 ) to 94.8 % iso-C17:0 (14.8 %), iso-C15:0 (14.2 %), and iso-C16:0 (Actinopolyspora mortivallis DSM 44261T). The strain (13.9 %). The genotypic and phenotypic data show that grew optimally at pH 6.0–7.0, 28–32 °C and in the the strain represents a novel species of the genus presence of 15–25 % (w/v) NaCl. The substrate myce- Actinopolyspora, for which the name Actinopolyspora lium was well developed and fragmented with age. The righensis sp. nov. is proposed, with the type strain H23T aerial mycelium produced long, straight or flexuous spore (=DSM 45501T = CCUG 63368T = MTCC 11562T). Keywords Actinopolyspora righensis sp. nov. halophilic actinomycete Á Algerian Sahara Á Polyphasic taxonomy A. Meklat Á N. Bouras Á A. Zitouni Á N. Sabaou (&) A. Lebrihi Laboratoire de Biologie des Syste`mes Microbiens Universite´ Moulay Ismail, Marjane 2, BP 298, Meknes, (LBSM), Ecole Normale Supe´rieure de Kouba, Alger, Morocco Algeria e-mail: [email protected] P. Schumann Á C. Spro¨er Á H.-P. Klenk Leibniz Institute DSMZ—German Collection of A. Meklat Microorganisms and Cell Cultures, Inhoffenstraße 7B, De´partement de Biologie, Faculte´ des Sciences 38124 Braunschweig, Germany Agronomiques, Ve´te´rinaires et Biologiques, Universite´ Saaˆd Dahleb, Blida, Algeria F. Mathieu Á A. Lebrihi Laboratoire de Ge´nie Chimique, UMR 5503 (CNRS/ INPT/UPS), INPT-ENSAT, Universite´ de Toulouse, 1 Avenue de l’Agrobiopole, Auzeville Tolosane, BP 32607, 31326 Castanet-Tolosan, France Introduction Microorganisms and Cell Cultures as strain DSM 45501T, in the Culture Collection, University of The genus Actinopolyspora, which belongs to the Go¨teborg as strain CCUG 63368T, and in the Micro- family Actinopolysporaceae (Zhi et al. 2009), was bial Type Culture Collection as strain MTCC 11562T. originally established by Gochnauer et al. (1975) and the description was later emended by Tang et al. Phenotypic characterization (2011). The genus currently contains 8 recognized species: Actinopolyspora halophila (Gochnauer Cultural characteristics of the new isolated strain were et al. 1975), A. mortivallis (Yoshida et al. 1991), determined after 3 weeks incubation at 30 °C on the A. xinjiangensis (Guan et al. 2010), A. egyptensis International Streptomyces Project media (Shirling (Hozzein and Goodfellow 2011), A. alba and A. and Gottlieb 1966), CM agar (Chun et al. 2000) and erythraea (Tang et al. 2011), A. algeriensis (Meklat nutrient agar (Waksman 1961). The ISCC–NBS et al. 2012) and A. saharensis (Meklat et al. 2013). colour name charts (Kelly and Judd 1976) were used The two species ‘A. egyptensis’ and ‘A. saharensis’ to determine colony colours. Morphological charac- were described recently and have not yet been teristics were observed under light microscopy (B1, validated. Motic) and scanning electron microscopy (Hitachi, The genus Actinopolyspora was shown to exhibit S450) using cultures grown on CM agar at 30 °C for distinct chemotaxonomic characteristics: cell-wall 4 weeks. Temperature range (10–45 °C), pH range chemotype IVA (meso-diaminopimelic acid, arabinose (5.0–9.0) and NaCl (0–35 %, w/v) tolerance for and galactose in whole-cell hydrolysates; Lechevalier growth were determined on nutrient agar after incu- and Lechevalier 1970), phospholipid type PIII (phos- bating for 21 days at 30 °C. Utilization of carbohy- phatidylcholine as characteristic phospholipid; Leche- drates, milk coagulation and peptonization, and valier et al. 1977), MK-9 (H4) and MK-10 (H4), MK-9 decarboxylation of organic acids were evaluated using (H4) and MK-9 (H2) or MK-6 as the predominant the method of Gordon et al. (1974). Degradation of menaquinones, iso-C16:0 and anteiso-C17:0 as the major other organic compounds was studied using protocol fatty acids and absence of mycolic acids (Gochnauer as described by Goodfellow (1971). Lysozyme sensi- et al. 1989; Guan et al. 2010; Tang et al. 2011). During tivity, methyl red and Voges–Proskauer tests, H2S an investigation of the soil actinobacterial community production and reduction of nitrate were determined in the Sahara desert of Algeria, an Actinopolyspora- according to the methods of Gordon and Barnett like strain was isolated. The aim of the present study (1977) and Marchal et al. (1987), respectively. was to determine the taxonomic position of this strain using polyphasic taxonomic approach. Chemotaxonomy Biomass for chemotaxonomy studies was obtained by Materials and methods cultivating the cell in shake flasks (250 rpm, 30 °C, 10 days) using CM broth containing 15 % NaCl (w/v). Strain and culture conditions Analysis of cell-wall amino acids and sugars in whole- cell hydrolysates were carried out according to the Strain H23T was isolated from moderately saline soil methods described by Becker et al. (1964) and (electrical conductivity = 2.3 mS cm-1) of Oued Righ Lechevalier and Lechevalier (1970). Phospholipids region (El-Oued) during an investigation of actinobac- were extracted, examined and identified by using the teria diversity in Saharan soils (Meklat et al. 2011). procedure developed by Minnikin et al. (1977). The Serially diluted sample was plated on complex medium cellular fatty acid composition was determined as (CM) agar (Chun et al. 2000) supplemented with 20 % described by Sasser (1990) using the microbial (w/v) NaCl and incubated for 5 weeks at 30 °C. identification system (MIDI). Cellular menaquinones After primary isolation and purification, the isolate were extracted and purified as described by Minnikin was preserved both on slants of CM agar at 4 °C and as et al. (1984) and were analyzed by HPLC (Kroppen- 20 % (v/v) glycerol suspensions at -20 °C. Strain stedt 1982, 1985). Analysis of mycolic acids was H23T was deposited in the German Collection of performed using the method of Minnikin et al. (1980). Phylogenetic analyses Extraction of chromosomal DNA of H23T strain was carried out as described by Liu et al. (2000). Amplifi- cation of the 16S rRNA gene by PCR and the sequencing of the purified PCR products were performed as described previously by Rainey et al. (1996). The resulting 16S rRNA gene sequence was compared with sequences obtained from public databases to determine the most closely related species using EzTaxon-e server (http://eztaxon-e.ezbiocloud.net/; Kim et al. 2012). Multiple alignments were performed using the CLUS- TAL W program (Larkin et al. 2007).
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