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Transfer of Pimelobacter Tumescens to Terrabacter Gen. Nov. As Terrabacter Tumescens Comb. Nov. and of Pimelobacter Jensenii To

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Transfer of Pimelobacter Tumescens to Terrabacter Gen. Nov. As Terrabacter Tumescens Comb. Nov. and of Pimelobacter Jensenii To INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, Jan. 1989, p. 1-6 Vol. 39, No. 1 0020-7713/89/010001-06$02.OO/O Copyright 0 1989, International Union of Microbiological Societies Transfer of Pimelobacter tumescens to Terrabacter gen. nov. as Terrabacter tumescens comb. nov. and of Pimelobacter jensenii to Nocardioides as Nocardioides jensenii comb. nov. M. D. COLLINS,l M. DORSCH,’ AND E. STACKEBRANDT’” Department of Microbiology, Agricultural Food Research Council, Institute of Food Research, Reading Laboratory, Shinjield, Reading RG2 9AT, United Kingdom,’ and Institut fur Allgemeine Mikrobiologie, Christian-Albrechts- Universitat, 2300 Kiel, Federal Republic of Germany’ The phylogenetic interrelationship of members of the genera Nocardioides and Pimelobacter were examined by using reverse transcriptase sequencing of 16s ribosomal ribonucleic acid. The sequence studies demon- strated that Nocardioides albus, Nocardioides luteus, Pimelobacter jensenii, and Pimelobacter simplex represent a coherent phylogenetic group at the genus level, whereas Pimelobacter tumescens occupies a separate line of descent. On the basis of sequence data and the chemotaxonomic distinctiveness of the latter organism, we propose that Pimelobacter tumescens be reclassified in a new genus, Terrabacter, as Terrabacter tumescens comb. nov. Arthrobacter simplex and Arthrobacter tumescens are mycelium), and its placement within the genus Nocardioides among the few named species of coryneform bacteria which has been questioned (14). contain cell wall peptidoglycans based on ~~-2,6-diaminopi- 16s ribosomal ribonucleic acid (rRNA) cataloging shows melic acid (5, 16). The two species thus differ in peptidogly- that P. simplex is well removed from the arthrobacters sensu can composition from Arthrobacter globiformis, the type strict0 and indicates that this species occupies a line of species of the genus, which contains lysine as the dibasic descent the branching point of which is as low as that of amino acid (16). Although there is now general agreement other major lines within the actinomycetes (18, 19). To date that these species should be removed from the genus Arthro- comparative sequencing studies with other members of bacter, their taxonomic position remains controversial (10). Pimelobacter and Nocardioides have not been made. There- Suzuki and Komagata (19) proposed the creation of a new fore, in an attempt to clarify the relationship of P. simplex to genus, Pimelobacter, to accommodate these species. A third the genus Nocardioides and to resolve the position of P. species, Pimelobacter jensenii, was also proposed for an tumescens, we analyzed and compared long stretches of the atypical A. simplex strain (19). However, there is evidence 16s rRNA primary structures of these taxa. of considerable chemical heterogeneity within the genus Pimelobacter (3, 12, 19). Pimelobacter (Arthrobacter) sim- plex and P. jensenii are unusual among coryneform and MATERIALS AND METHODS related bacteria in producing complex mixtures of straight- Cultures and cultivation. Type strains Nocardioides albus chain saturated, monounsaturated, and anteiso-, and iso-, DSM 43109, Nocardioides luteus NCIB 11455, P. jensenii 10-methyl branched fatty acids (3, 12, 19-21). However, DSM 20641, P. simplex NCIB 8929, and P. tumescens NCIB Pimelobacter tumescens differs from these species in lacking 8914 were grown in shake flasks (casein peptone, 10 g; yeast 10-methyl branched acids and in containing substantial lev- extract, 5 g; glucose, 5 g; NaCl, 5 g; water, 1 liter; pH 7.4) to els (ca. 20%) of monounsaturated terminally branched acids late exponential phase at 25°C. (3). P. simplex and P. jensenii also differ from P. tumescens Sequence determination and analysis. Crude ribonucleic in synthesizing 2-hydroxylated long-chain fatty acids (3, 12). acid was isolated as described previously (7). Reverse tran- The polar lipid compositions of these taxa are also quite scriptase sequencing of ribonucleic acid was achieved by different. P. simplex and P. jensenii possess diphospha- using the method of Lane et al. (11). The sequence of tidylglycerol, phosphatidylglycerol, and hydroxyphospha- oligonucleotide primers, their target sites, and the electro- tidylglycerol (3, 12, 21). However, P. tumescens lacks phoretic separation conditions used for complementary de- hydroxyphosphatidylglycerol and in addition synthesizes oxyribonucleic acid have been described by Embley et al. two unknown aminophosphoglycolipids (3, 12). Such major (7). Some sequence ambiguities were resolved by using differences in lipid composition between P. simplex and P. terminal transferase (6). Sequence data were aligned and jensenii on the one hand and P. tumescens on the other hand homology values were determined with the Beckman Micro- indicate that these taxa should probably not be accommo- genie program (15). Calculation of evolutionary distance dated in the same genus. Interestingly, the lipid composition values (Knuc) and construction of phylogenetic trees fol- of P. simplex and P. jensenii is indistinguishable from that of lowed previously published procedures (8, 9). members of the genus Nocardioides (3, 12). Indeed, primar- ily on the basis of similarity in lipid composition, O’Donnell et al. (12) proposed that P. simplex be transferred to the RESULTS AND DISCUSSION genus Nocardioides. However, P. simplex does not possess a nocardioform morphology (extended primary and aerial The partial reverse transcriptase sequences of the five strains investigated are shown aligned in Fig. 1. The homol- ogy and derived K,,, values from 1,449 bases used for * Corresponding author. determining intragroup relationships are shown in Tables 1 1 2 COLLINS ET AL. INT. J. 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