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 ...... " .T•••••• . ...c ..... " ••.•A .•• T Saccharothrb: australiensis NRRL 11239 ••••• • CAC. ••••• • GTG • ...... c ..... •••••• TCC • T Saccharothrix coentleofilsca NRRL B-16115 ••••• • CAC. •••••• GTG...... c ..... • •••••T.C • T Saccharothrix espanaensis NRRL 15764 ••••• • CAC. ••••• • GTG • ...... c ..... ••••••T.C • T Saccharothrix longispora NRRL B-16116 ••••• • CAC. •..•••GTG. ••• ••••A •• • ••• C ••••• • ••••• A ••• T Saccharothri:r: mutabilis subsp. capreoills DSM 40225 ••••• • CAC. •••N •• GTG...... ••• •C ••••• • ••••• TCC • T Saccharothri:r: mlltabilis subsp. mutabilis DSM 43853 ••••• • CAC. ••N ••• GTG...... c ..... • ••••• TCC • T Saccharothrix syringae NRRL B-16468 ••••• • CAC. •••••• GTG...... c ..... • •••••T.C • T Saccharothrix te:r:asensis NRRL B-16134 ••••• • CAC. •••••• GTG. •••••••A •• • •.• C ••••• • •••••T.C •

Fig. 2. Signatures in the 16S rDNA sequence for the genera Lechevalieria, Lentzea and Saccharothrix. ses using the maximum-parsimony and maximum­ genera of the Actinosvnnel1lataceae. Although S. aero­ likelihood algorithms (trees not shown). The phylo­ ~olonigenes and S..f/ava appear to be phylogenetically genetic position of Saccharothrix cryophilis NRRL B­ close to the genus Actinosvnnel1la. neither has been 162381' indicates that this species does not belong in observed to produce corerilia with chains of motile the genus Saccharothrix and that it probably does not spores (this feature being characteristic of this genus). belong w'ithin the family Actinosynnemataceae, but It is therefore proposed that the S. aerocolonigenes-S. this will be the subject of another detailed paper. .f/ava lineage represents a new genus, to be called S. aerocolonigenes NRRL B-16l37, Sacc/zarot/zrix Lec/zeva/ieria gen. nov. violacea IMNSU 503881'. S. wavwavandensis NRRL B-16l591' and 'A. albidum' IFO"16102 form a lineage Analysis of the chemotaxonomic characteristics of that contains Lentzea albidocapi/lata DSM 440731' Lentzea albidocapi/lata DSM 440731' in the present (97'8 % mean intrageneric nucleotide similarity), the study revealed the presence ofgalactose, mannose and type species of the genus, which is also clearly ribose as diagnostic sugars and menaquinone MK­ separated phylogenetically from Sacc/zarot/zrix sensu 9(H~) as the predominant menaquinone and the stricto, with a mean nucleotide similarity of 95·8 % absence ofany significant amount of 10-methyl C18: 0 betw"een all species of both genera. These strains also fatty acids (tuberculostearic acid). This is largely in share similar chemotaxonomic characteristics. and it is agreement with the observations of Lee et al. (2000) proposed that they be transferred to the revived and regarding this strain. Galactose and mannose were fo~nd as~ emended genus Lentzea. diagnostic sugars in whole-cell hydrolysates of all strains examined. but' A. albidwn' IFO 16102 The type strains of S. aerocolonigenes and S. f/ava, and S. aerocolonigenes NRRL B-16l37 contained 1' 1' respectively NRRL B-3298 and NRRL B-1613l , ribose. while S. wavwavandensis NRRL B-16l591'. S. consistently formed a monophyletic lineage distinct aerocolonigenes NRRL-B-32981' and S ..f/ava NRRL B­ from Sacc/zarot/zrix (mean nucleotide similarity of 16131 l' did not. Whilst the genera Lec/zevalieria and 96·8 % to species of this genus) and intermediate Lentzea (including' A. albidum', S. aerocolonigenes between Lentzea (mean nucleotide similarity of96· 7 % NRRL B-16l37 and S. waVlvavandensis NRRL B­ to species of this genus) and Actinosynnel1la (mean 161591') share many diagnostic chemotaxonomic nucleotide similarity of97·3 % to species ofthis genus) characteristics with Sacc/zarot/zrix, they can be dis­ for all of the algorithms tested. The phylogenetic tinguished from this genus by the lack of hydroxy­ validity of the genera Actinosynnel1la, Lentzea and substituted fatty acids in the phosphatidyl ethanola­ Sacc/zarot/zrix \vas consistently well supported stat­ mine component of their phospholipids (Table 1). istically by all algorithms used: the S. aerocolonigenes Sacc/zarot/zrix and Lec/zevalieria strains tend to contain and S. flava lineage was consistently found between varying amounts of rhamnose in whole-cell hydro­ Actinosvnnema and Lentzea. regardless of which al­ lysates, whereas Lentzea strains were observed to lack goritlml was used. An examin'ation of the aligned (or contain only trace amounts of) rhamnose, and sequences for the l6S rDNA gene (Fig. 2) revealed that might also contain ribose. diagnostic nucleotide signatures can be used to differentiate the S. aerocolonigenes-S. flava lineage All species of Lentzea produce branching substrate from Lentzea and Actinosynnel1la and that they can mycelium, and the aerial mycelium fragments into rod­ also be used to differentiate them from the other shaped elements, although Lentzea waywayandensis

International Journal ofSystematic and Evolutionary Microbiology 51 1047 D. P. Labeda and others

Table 1. Chemotaxonomic characteristics of Lecheva/ieria compared with other genera of the family Actinosynnemataceae

All of the genera have meso-diaminopimelic acid as the cell wall diamino acid, are of cell wall chemotype III and contain straight­ chain, mono-unsaturated, iso and anteiso fatty acids. Abbreviations: DPG, diphosphatidyl glycerol: PE, phosphatidyl ethanolamine: PG, phosphatidyl glycerol: PI. phosphatidyl inositol: PIMs, phosphatidyl inositol mannosides; PM£, phosphatidyl methylethanolamine.

Characteristic Acrinokilleospora Actillmynnema Asimporungiu11l Lt.'chcl'afieria Lellt:ea Sacclzarothrix

\Vhole-cdl sugar pattern Galactose. mannosc. Galactose. mannose Galactose. mannose. Galactose. m'mnose. G"l!actose. mannose. Galactose. rhamnose. rhamnose ribose rhamnose ribose mannose (trace) Phospholipid type PII PII PII PII PII PII. PIV Phospholipids PE.OH·PE PE. OH·PE. PI. PIMs. DPG PE PE PE. DPG. PG. PI PE. OH-PE. PI. PIMs. DPG.PG Predominant menaquinones MK-9(H,1 MK-9(H,). MK-9(H,;I MK-9(H,) MK-9(H,1 MK-9(H,1 MK-IO(H,'. MK-9(H,1

Table 2. Differential physiological properties of Lentzea species w. Weak positive reaction. Data for Lell1:::ea 1'io/acea were taken from Lee el al. (2000).

Property Lelltzea Lelltzea albida Lelltzea Lelltzea j'iolacea Lelltzea albidocapillata NRRL B-24073T californiellsis IMSNU 50388T lI'ayll'ayalldellsis NRRL B-24057T NRRL B-16137T NRRL B-16159T

Hydrolysis of urea w w + + + Nitrate reductase production + + Assimilation of: Acetate + + + + Citrate + + + Lactate + + Malate + + + + Production of acid from: Adonitol + + + Cellobiose + + + + Inositol + + + + Maltose + + + + Mannitol + + + + Raffinose + + + Rhamnose + + + + Sucrose + + + + Trehalose + + + + Xylose + + + + Growth at: lOoC + + + + 37°C + + + + w 42 °C + 45°C +

NRRL B-16l59T produces only relatively sparse aerial S. violacea is transferred as Lentzea violacea comb. mycelium in comparison with the other Lentzea nov., S. wavwavandensis is transferred as Lentzea species. Both species of Lechevalieria produce very lvanvavandellsis -comb. nov. and' A. albidum' IFO sparse aerial mycelium on agar media and have l6i02 "is validly described as Lentzea albida sp. nov. not been observed to produce sporangia, coremia or Because it is proposed that the type strain of S. motile spores, which differentiates them from aerocolonigenes be transferred to the new genus Actinokineospora and Actinosynnema. Lechevalieria and as strain NRRL B-16137 is a member ofthe genus Lentzea, it is described as the new It is proposed that the formal description ofthe revived species Lentzea cal(forniensis sp. nov. The physio­ genus Lentzea be emended to include updated chemo­ logical properties that differentiate these species are taxonomic data from both the present study and that shown in Table 2 and the formal descriptions are given of Lee et al. (2000). On the basis ofthe data presented, below.

1048 International Journal ofSystematic and Evolutionary Microbiology 51 Revival of Lentzea. and Lecheva/ieria gen. nov.

T Table 3. Differential physiological properties of type strain of Lentzea albida is IFO 16102 ( = NRRL T Lechevalieria species B-24073 ).

\Y, Weak positive reaction. Description of Lentzea californiensis sp. nov. Property LeclIel'aliel'ia Lecllel'aliel'ia Lentzea cal(forniensis (caLi.for.ni.en'sis. N.L. gen. n. ael'ocololligelles flal'a NRRL T T californiensis of California, referring to the source of NRRL B-3298 B-16131 this isolate, soil from California). ~

Growth in the presence of: Substrate mycelium is yellow' to orange-brown. White 4% NaCl + aerial mycelium is produced. Orange, soluble pigments 5% NaCl + may be produced on Czapek's agar. Casein, aesculin, Utilization of lactate + hypoxanthine, starch and tyrosine are hydrolysed. Production of acid from: Does not hydrolyse adenine, allantoin or xanthine. Adonitol + w Grows in the presence of5 % NaC!. Assimilates citrate, Salicin + malate, propionate and succinate. Does not assimilate Growth at 45 DC + acetate. benzoate. lactate. mucate. oxalate or tartrate. Acid is produced from arabinose, cellobiose, dextrin, fructose, galactose, glucose, glycerol, inositoL lactose, maltose. mannitoL mannose. melibiose. raffinose. The physiological properties that differentiate the rhamnose, sucrose, trehalose and xylose; acid is not Lechevalieria species can be seen in Table 3 and the produced from adonitoL dulcitol, erythritoL methyl fJ­ formal descriptions of the genus Lechevalieria and the xyloside or sorbitoL Grows at 37°C but not at 42 dc. species Lechevalieria aerocolonigenes and Lechevalieria Isolated from a soil sample from California. Strain fiava are also given below. NRRL B-16137 was originally classified as Saccharo­ thrix aerocolonigenes. The type strain of Lentzea californiensis is NRRL B-16137T (= DSM 43393 T = T Emended description of Lentzea nom. rev., emend. IM'RU 550 ). The formal description of Yassin et al. (1995) is emended to reflect the following changes in the Description of Lentzea violacea comb. nov. chemotaxonomic characteristics. The diagnostic whole-cell sugars include galactose and mann;se and Basonym Saccharothrix violacea Lee et al. 2000. might also include ribose. The major menaquinone is The morphological and physiological properties of MK-9(HJ. Tuberculostearic acid is found only in this species have been described previously (Lee et al., trace quantities, if at alL in fatty acid profiles. The 2000). The type strain of Lentzea violacea is IMSNU sequence of the 16S rRNA gene contains a genus­ T 50388 . specific diagnostic nucleotide signature pattern, namely TCCA (617-620) and GCC (843-845). Description of Lentzea waywayandensis comb. nov.

Description of Lentzea albida sp. nov. Basonym Saccharothrix waywayandensis Labeda and Lyons 1989. Lentzea albida (al'bi.da. L. fem. adj. albida whitish. referring to the colour of the aerial mycelium). The morphological and physiological properties of this species have been described previously (Labeda & Substrate mycelium is a yellowish-orange in colour on Lyons, 1989). The type strain of Lentzea wayway­ most media. Copious white aerial mycelium is pro­ andensis is NRRL B-16159T (= DSM 44232T = IFO T duced. Soluble pigments are not produced. Casein, 14970 ). aesculin, hypoxanthine, starch and tyrosine are hydro­ lysed. No hydrolysis ofadenine, allantoin or xanthine. Growth occurs in the presence of 4 and 5 % NaC!. Description of Lechevalieria gen. nov. Assimilates acetate, citrate, malate, propionate and Leclzevalieria (Le.che.vaLi.er'i.a. N.L. fem. n. Lech­ succinate. Does not assimilate benzoate. lactate. evalieria of Lechevalier. named after the American mucate, oxalate or tartrate. Acid is produced from microbiologists Hubert and Mary Lechevalier. who adonitoL arabinose, cellobiose, dextrin, fructose, ga­ contributed substantially to the field of actinomycete lactose, glucose, glyceroL inositol, maltose, mannitol, biology during their careers at the Waksman Institute mannose. melibiose. rhamnose. sucrose. trehalose and of Microbiology). xylose. Acid is weakly produced from lactose. No acid is produced from dulcitoL erythritol, methyl fJ-xy­ Branching vegetative mycelium (approx. 0·5 11m in loside. raffinose or sorbitoL Grows at 37 and 42°C but diameter) is produced. Very scant aerial mycelium is not at 45 dc. Isolated from a soil sample collected in produced on some media. Gram-positive. Lysozyme­ Jianbxi Province, People's Republic of China. The resistant. Catalase-positive and aerobic. The cell wall

International Journal ofSystematic and Evolutionary Microbiology 51 1049 D. P. Labeda and others is of type III (meso-diaminopimelic acid). The whole­ REFERENCES cell sugar pattern consists of galactose, mannose and Cote, R., Daggett, P.-M., Gantt, M. J., Hay, R., Jong, S.-c. & Pienta, traces of rhamnose. Possesses the type PII phospho­ P. (1984). ATCC Media Handbook. Manassas. VA: American lipid pattern, with significant quantities of phospha­ Type Culture Collection. tidyl ethanolamine lacking hydroxylated fatty acids. Galtier, N., Gouy, M. & Gautier, C. (1996). SEA VIEW and PHYLO_ The predominant menaquinone is MK-9(H ). The q WIN: two graphic tools for sequence alignment and molecular fatty acid profile consists of saturated and mono­ phylogeny. Comput Appl Biosci 12, 543-548. unsaturated iso and anteiso fatty acids. Phylo­ Gordon, R. E., Barnett, D. A., Handerhan, J. E. & Pang, C. H. genetically, the genus represents a line ofdescent in the (1974). Nocardia coeliaca, Nocardia autotrophica. and the Actinosynnemataceae adjacent to the genus Saccharo­ nocardin strain. Int J Syst Bacteriol24. 54-63. thrix and close to the genera Actinosynnema and Grund, E. & Kroppenstedt, R. M. (1989). Transfer of five Lentzea. The sequence of the 16S rRNA gene contains Nocardiopsis species to the genus Saccharothri.\ Labeda et al. a genus-specific diagnostic nucleotide signature pat­ 1984. Syst Appl Microbio/12. 267-274. tern, namely TT (844-845) and GGT (1107-1109). The Kimura, M. (1980). A simple method for estimating evolutionary type species is Lechevalieria aerocolonigenes. rates of base substitutions through comparative studies of nucleotide sequences. J Mol Ero/16. 111-120. Kurup, P. V. & Schmitt, J. A. (1973). Numerical taxonomy of Description of Lechevalieria aeroco/onigenes comb. Nocardia. Can J Microbio/19. 1035-1048. nov. Labeda, D. P. (1986). Transfer of" Nocardia aerocololligenes" (Shinobu and Kawato 1960) Pridham 1970 into the genus Basonym Saccharothrix aerocolonigenes (ex Shinobu Sacclwrothrix Labeda. Testa. Lechevalier. and Lechevalier 1984 and Kawato 1960) Labeda 1986. as Saccharothrix aerocololligenes sp. nov. lilt J Syst Bacteriol 36.109-110. The morphological and physiological properties of Labeda, D. P. & Kroppenstedt, R. M. (2000). Phylogenetic analysis this species have been described previously (Labeda, of Saccharothrix and related taxa: proposal for Actino­ 1986). The type strain of Lechevalieria aerocolonigenes sVllllemataceae fam. nov. lilt J Syst Evol Alicrobiol50. 331-336. is NRRL B-3298T (= ATCC 23870T = CCRC 13661 T Labeda, D. P. & Lyons, A. J. (1989). Saccharothrix texasellsis sp. T T T = DSM 40034 = IFO 13195 = JCM 4150 ). nov. and Saccharothrix \l'aylmyalldellsis sp. nov. lilt J Syst BacterioI39.355-358. Lapage, S. P., Sneath, P. H. A., Lesser, E. F., Skerman, V. B. D., Seeliger, H. P. R. & Clark, W. A. (editors). (1992). International Description of Lechevalieria flava comb. nov. Code of Nomellclature of Bacteria (l990 Revision). Bacterio­ logical Code. Washington. DC: American Society for Micro­ Basonym Saccharothrix flava (Gauze et al. 1974) biology. Grund and Kroppenstedt 1990. Lee, S. D., Kim, E. 5., Roe, J.-H., Kim, J., Kang, 5.-0. & Hah, Y. C. (2000). Saccharothrix violacea sp. nov., isolated from a gold The morphological and physiological properties of mine cave, and Saccharothrix albidocapillata comb. nov. 1111 J this species have been described previously (Grund & Syst Evol Microbiol50. 1315-1323. KroppenstedL 1989). The type strain of Lechevalieria L., N., T Maidak, B. Larsen, McCaughey, M. J., Overbeek, R., Olsen, {lava is NRRL B-16131 T (= A TCC 29533 = CCRC G. J., Fogel, K., Blandy, J. & Woese, C. R. (1994). The Ribosomal T T T T 13328 = DSM 43885 = IFO 14521 = INA 2171 Database Project. Nucleic Acids Res 22.3485-3487. T = JCM 3296 ). Saitou, N. & Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. !VIol BioI Evol 4, 406-425. ACKNOWLEDGEMENTS Tamura, T. & Hatano, K. (1998). Phvlogenetic analvses on the strains belonging to invalidated ge'ner:l of the order Actino­ The able technical assistance of E. N. Hoekstra during the mycetales. Actinomycetologica 12. 15-28. analysis of physiological properties, the isolation and puri­ Yassin, A. F., Rainey, F. A., Brzezinka, H., Jahnke, K.-D., fication of DNA, the peR amplification of 165 rDNA and Weissbrodt, H., Budzikiewicz, H., Stackebrandt, E. & Schaal, K. P. the sequence determinations for this study is gratefully (1995). Lellt::ea gen. nov., a new genus of the order Actino­ acknowledged. mycetales. 1111 J Syst Bacteriol45. 357-363.

Suppllea by U.S. DeDt of Agriculture National Center tor Agncultural UtilizatIon Research, Peona, Illinois

1050 International Journal ofSystematic and Evolutionary Microbiology 51