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Revival of the Genus Lentzea and Proposal for Lechevalieria Gen International Journal ofSystematic and Evolutionary Microbiology (2001), 51, 1045-1050 Printed in Great Britain Revival of the genus Lentzea and proposal for Lechevalieria gen. nov. 1 Microbial Properties D. P. Labeda,l K. Hatano/ R. M. Kroppenstedt3 and T. Tamura2 Research Unit, National Center for Agricultural Utilization Research, 1815 N. University Street, Author for correspondence: D. P. Labeda. Tel: + 13096816397. Fax: + 1309681 6672. e-mail: labedadp«(lmail.ncauLusda.gov Agricultural Research Service, US Department of Agriculture, Peoria, IL 61604, USA The genus Saccharothrix is phylogenetically heterogeneous on the basis of analysis of almost complete 165 rONA sequences. An evaluation of 2 Institute for Fermentation Osaka, Osaka, Japan chemotaxonomic, morphological and physiological properties in the light of the molecular phylogeny data revealed that several species are misclassified. 3 DSMZ-Germany Collection of Microorganisms and Cell Saccharothrix aerocolonigenes NRRL 8·3298T and Saccharothrix flava NRRL 8­ Cultures, Braunschweig, 16131T constitute a lineage distinct from Saccharothrix and separate from Germany Lentzea. The genus Lechevalieria gen. nov. is proposed for these species. Lechevalieria aerocolonigenes comb. nov. is the type species and S. flava is transferred as Lechevalieria flava comb. nov. Although Lentzea albidocapillata, the type species of the genus Lentzea, was transferred recently to the genus Saccharothrix, the revival of Lentzea is clearly supported by molecular phylogenetic and chemotaxonomic data. The description of the revived genus is emended to include galactose, mannose and traces of ribose as diagnostic whole-cell sugars and MK-9(H4) as the principal menaquinone and elimination of tuberculostearic acid as a diagnostic component in the fatty acid profile. T Saccharothrix waywayandensis NRRL 8·16159 , S. aerocolonigenes NRRL 8­ 16137 and'Asiosporangium albidum' IFO 16102 are members of the amended genus Lentzea on the basis of phylogenetic and chemotaxonomic properties. S. waywayandensis is transferred to Lentzea as Lentzea waywayandensis comb. nov., while the new species Lentzea californiensis sp. nov. and Lentzea albida sp. nov. are described for S. aerocolonigenes NRRL 8-16137 and I A. albidum' IFO 16102, respectively. Nucleotide signatures in the 165 rONA sequences are defined that are diagnostic for the genera Lechevalieria, Lentzea and Saccharothrix. Keywords: Pseudonocardineae. Actinosynnemataceae. Saccharothrix INTRODUCTION B-3298T and Saccharothrix {lava NRRL B-16l31 T form a lineage distinct from' Saccharothrix and sep­ A phylogenetic analysis of species of the genus arate from Lentzea. The previous study of Tamura Saccharothrix and allied taxa within the family Actino­ & Hatano (1998) also demonstrated that . Asio­ synnemataceae, based on partial l6S rDNA sequences sporangium albiduIII' IFO 16102 is phylogenetically (Labeda & Kroppenstedt, 2000), demonstrated that related to Saccharolhrix and Lentzea. A recent pro­ Saccharothrix is phylogenetically heterogeneous and posal (Lee et al., 2000) suggested that Lentzea albido­ that several ofthe described species are misclassified. It capillata, the type species of the genus Lentzea, be was noted that Saccharothrix wavwavandensis NRRL transferred into the genus Saccharothrix, thus effec­ B-165l59T is more closely related phylogenetically to tively abolishing thi; genus under the rules of the Lentzea albidocapillata than to Saccharothrix sensu Bacteriological Code of Nomenclature (Lapage et al., stricto and that Saccharothrix aerocolonigenes NRRL 1992), but the phylogenetic and chemotaxonomic data in the present study do not support this proposal. An The GenBank accession number for the almost complete sequence of the evaluation of the chemotaxonomic, morphological 165 rONA gene of Saccharothrix aeroco/onigenes NRRL B-16137 is and physiological properties of these actinomycetes AF174435. was undertaken to clarify their taxonomic position. 01477 1045 D. P. Labeda and others METHODS procedures described previously (Labeda & KroppenstedL 2000). Strains, cultivation and maintenance. The strains evaluated in the present study are listed in Fig. I. Primary storage of Phylogenetic analysis. The 16S rONA sequences obtained in strains was as lyophilized ampoules of mycelial and spore this study were aligned manually with actinomycete ref­ suspensions in sterile beefserum held at 4°C. Working stock erence sequences obtained from the Ribosomal Database cultures were maintained on slants of ATCC medium no. Project (Maidak el al., 1994) and GenBank in the ARB 172 (Cote el al.. 1984) and stored at 4°C until needed. software environment for sequence data that was developed Biomass for extraction of DNA was grown as 7 d streak by Wolfgang Ludwig and Oliver Strunk (Lehrstuhl flir cultures on ATCC medium no. 172 age'll' plates. Mikrobiologie, University of Munich, Germany). The program PHYLa_WIN (Galtier el al., 1996) was used to Chemotaxonomy. Chemotaxonomic analysis of strains for calculate evolutionary distances (using the method of menaquinones, fatty acids and whole-cell sugars was per­ Kimura, 1980) and linkages (using the neighbour-joining formed using methods described previously (Grund & method of Saitou & Nei. 1987) and to perform maximum­ KroppenstedL 1989). parsimony and maximum-likelihood analyses. The topo­ Physiological tests. Physiological tests, including those for graphies of the trees resulting from neighbour-joining and the production ofacid from carbohydrates, the utilization of maximum-parsimony analyses were evaluated by bootstrap organic acids and the hydrolysis and decomposition of analysis of the data (with 500 resamplings). adenine, guanine, hypoxanthine, tyrosine, xanthine, casein, aesculin, urea and hippurate, \vere evaluated by using the RESULTS AND DISCUSSION media of Gordon el al. (1974). Allantoin hvdrolvsis was evaluated in the basal medium suggested bv Gordon el al. The neighbour-joining dendrogram for the family (1974) for aesculin hydrolysis. Pl{'osphatase activity was Actinosynnemataceae, resulting from phylogenetic evaluated by using the method of Kurup & Schmitt (1973). analysis of 16S rDNA sequences, is shown in Fig. 1; it The temperature range for growth was determined on slants can be clearly observed that the genus Saccharothrix of ATCC medium no. 172 agar (Cote el al., 1984). sensll lata exhibits phylogenetic heterogeneity upon DNA isolation, 16S rONA gene amplification and sequencing. analysis based on almost complete sequences of 16S Genomic DNA was isolated, purified and sequenced using rDNA. Similar tree topographies resulted from analy- .-----Lentzea violacea IMNSU 50388T/AJ242633 Lentzea albidocapillata DSM 44073T (= NRRL B-24057T)1X84321 '-----Lentzea californiensis NRRL B-16137T/AF174435 Lentzea albida IFO 16102T (= NRRL B-24073T)/AB006176 Lentzea waywayandensis NRRL B-16159T/AFl14813 r---Lechevalieria aerocolonigenes NRRL B-3298T/AFl14804 '----Lechevalieria flava NRRL B-16131 T/AFl14808 r-----Actinosynnema mirum DSM 43827TIX84447 74 Actinosynnema pretiosum subsp. pretiosum NRRL B-16060T/AFl14800 98 Saccharothrix mutabilis subsp. capreolus DSM 40225TIX76965 Saccharothrix mutabilis subsp. mutabilis DSM 43853TIX76966 Saccharothrix espanaensis NRRL 15764T/AFl14807 Saccharothrix syringae NRRL B-16468T/AFl14812 Saccharothrix coeruleofusca NRRL B-16115T/AFl14805 82 Saccharothrix australiensis NRRL 11239T/AFl14803 '-----Saccharothrix texasensis NRRL B-16134T/AFl14814 '----Saccharothrix longispora NRRL B-16116T/AFl14809 r------Actinokineospora riparia NRRL B-16432T/AFl14802 '------Actinokineospora inagensis NRRL B-24050T/AFl14699 r---Actinokineospora diospyrosa NRRL B-24047T/AFl14797 Actinokineospora globicatena NRRL B-24048T/AFl14798 r------Kutzneria kofuensis NRRL B-24061 T/AFl14801 '---------Saccharothrix cryophilis NRRL B-16238T/AFl14806 '-------Streptoalloteichus hindustanus IFO 15115T/D85497 '----------Actinoalloteichus cyanogriseus IFO 14455T/AB006178 '----------- Amycolatopsis orientalis IFO 12806T/D86935.1 r-------------Saccharomonospora viridis ATCC 15386TIX54286 '-----------Saccharopolyspora hirsuta ATCC 27875T1X53196 '-----------Kibdelosporangium aridum subsp. aridum ATCC 39323TIX53191 '------------------Arthrobacter globiformis DSM 20124T/M23411 0·10 Fig. 1. Phylogenetic dendrogram reconstructed from evolutionary distances (Kimura, 1980) by the neighbour-joining method (Saitou & Nei, 1987), indicating the positions of species of the genera within the family Aetinosynnemataceae. Bar, 0·1 nucleotide substitutions per site. 1046 International Journal ofSystematic and Evolutionary Microbiology 51 Revival of Lentzea. and Lechevalieria gen. nov. 60~ H~ 62~ 84~ ~oo~ I I I I T Lechevalieria aerocolonigenes NRRL B-3298 AAACTTGGGG CTTAACCCCG AGCCTGCGGT ACGTTCTCCG GAAACCGGTA Lechevalieriaj/ava NRRL B-16131 T .......... .......... .......... ......... .. .......... T Lentzea albida IFO 16102 ........... ••••• •T •• A ••• T •••••• .. .cc... T• ••••••A ••• T Lentzea albidocapillata DSM 44073 .......... ••••• •T •• A •• •T .•.••• ...cc... T• •••• G.TC •• T Lentzea californiensis NRRL B-16137 .......... ••••• •T •• A .•• T ••• C •• .. .cc... T• •••••• A ••• T Lentzea violacea IMSNU 50388 .......... •••••• T •• A •• •T •••••• . ..cc...T• •••• G.TC •• T Lentzea waywayandensis NRRL B-16159 .......... •••••• T ••A •• •T •••••• ••• CC ••• T• •••••• TC •• T Actinosylllzema mirum DSM 43827 ........... ........... ............ '" .c..... ••••••A ••• T Actinosynnema pretiosllm subsp. pretiosum :NRRL B-16060 ........
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