Janibacter Terrae Sp. Nov., a Bacterium Isolated from Soil Around a Wastewater Treatment Plant

International Journal of Systematic and Evolutionary Microbiology (2000), 50, 1821–1827 Printed in Great Britain

Janibacter terrae sp. nov., a bacterium isolated from soil around a wastewater treatment plant

Jung-Hoon Yoon,1 Keun-Chul Lee,1 Seok-Sung Kang,1,2 Yung Hee Kho,1 Kook Hee Kang2 and Yong-Ha Park1

Author for correspondence: Yong-Ha Park. Tel: j82 42 860 4620. Fax: j82 42 860 4598. e-mail: yhpark!kribb4680.kribb.re.kr

1 Korea Research Institute of A bacterial strain, CS12T, which was isolated from soil around a wastewater Bioscience and treatment plant, was subjected to a polyphasic taxonomic study using Biotechnology (KRIBB), T PO Box 115, Yusong, phenotypic characterizations and genetic methods. The cell wall of strain CS12 Taejon, Korea contains meso-diaminopimelic acid as the diamino acid but no arabinose and

2 Department of Food and galactose. The predominant menaquinone is MK-8(H4). Mycolic acids are Life Science, absent. Strain CS12T has a cellular fatty acid proﬁle containing saturated, Sungkyunkwan University, unsaturated, branched and 10-methyl fatty acids. The major fatty acids are iso- Chunchun-dong 300, Jangan-gu, Suwon, Korea C16:0,C18:1 ω9c and anteiso-C17:0. The GMC content is 69 mol%. A phylogenetic tree based on 16S rDNA sequences showed that strain CS12T forms an evolutionary lineage within the radiation enclosing the members of the family Intrasporangiaceae and, in particular, a coherent cluster with Janibacter limosus DSM 11140T. The level of 16S rDNA similarity between strain CS12T and J. limosus DSM 11140T is 987%. The phenotypic characteristics and DNA–DNA relatedness data indicate that strain CS12T should be distinguished from J. limosus DSM 11140T. Therefore, on the basis of the data presented, a new species of the genus Janibacter, Janibacter terrae, is proposed. The type strain of the new species is strain CS12T (l KCCM 80001T l JCM 10705T).

Keywords: Janibacter terrae sp. nov., actinomycete, polyphasic taxonomy

INTRODUCTION many strains from soil around a wastewater treatment plant. Among them, one isolate (strain CS12T)was The genus Janibacter was recently proposed by Martin considered to be a Janibacter-like strain from the et al. (1997) with one species, Janibacter limosus. This results of 16S rDNA sequence comparisons. Accord- species was isolated from a 1-year-old sludge sample ingly, strain CS12T has been subjected to a polyphasic collected from a wastewater treatment plant. The characterization to investigate the possibility that it is genus Janibacter has been placed in the family Intra- a second species of the genus Janibacter. Here, we sporangiaceae, together with the genera Intraspor- describe the morphological, phenotypic, phylogenetic angium, Terrabacter, Terracoccus and Sanguibacter and genomic characteristics of this strain. On the basis (Stackebrandt et al., 1997; Prauser et al., 1997). Since of the results described, strain CS12T is considered to there is currently only one validly described species in be a new species of the genus Janibacter, for which we the genus Janibacter, the ﬁnding of new species may be propose the name Janibacter terrae sp. nov. very meaningful from the point of view of biological diversity of this genus. Recently, we have isolated many strains that are important for the bioremediation METHODS of toxic aromatic compounds from a wastewater Bacterial strain and culture conditions. Strain CS12T was treatment plant in Korea and some of these strains isolated from a soil sample taken from around a wastewater have been found to be new actinomycete species (Cho treatment plant in Korea by dilution plating on trypticase et al., 1998; Yoon et al., 1999). We have also isolated soy agar (BBL). For investigating its morphological and physiological characteristics, strain CS12T was grown at ...... 28 mC on solid or in liquid R medium (Martin et al., 1997). The GenBank accession number for the 16S rDNA sequence of strain CS12T Cell mass for analyses of its cell wall, menaquinones, mycolic is AF176948. acids and polar lipids was obtained from growth in liquid R

01369 # 2000 IUMS 1821 J.-H. Yoon and others

Terrabacter tumescens KCTC 9133T (AF005023) 83·7 T 99·5 Terracoccus luteus DSM 44267 (Y11928)

Intrasporangium calvum IFO 12989T (D85486) 85·5

Strain CS12T (AF176948)

100 Janibacter limosus DSM 11140T (Y08539)

Sanguibacter inulinus NCFB 3024T (X79451) 56·2 Sanguibacter keddieii NCFB 3025T (X79450) 55·8 100 Sanguibacter suarezii NCFB 3023T (X79452)

Cellulomonas flavigena NCIMB 8073T (X79463)

81·8 Microbacterium lacticum DSM 20427T (X77441)

Brachybacterium faecium DSM 4810T (X83810) 98·9 T 56·2 Dermabacter hominis DSM 7083 (X91034)

Jonesia denitrificans DSM 20603T (X83811)

Brevibacterium linens DSM 20425T (X77451)

Dermatophilus congolensis ATCC 14637T (L40615)

88·1 Micrococcus luteus ATCC 381 (M38242)

Nocardia asteroides DSM 43757T (X80606) 95·9 T 58·2 Rhodococcus rhodochrous DSM 43241 (X79288)

Pseudonocardia halophobica DSM 43089T (Y08534)

Geodermatophilus obscurus DSM 43160T (X92356)

Microsphaera multipartita JCM 9543T (Y08541)

Nocardioides albus KCTC 9186T (AF004988)

100 Luteococcus japonicus DSM 10546T (Z78208)

Streptomyces griseus subsp. griseus KCTC 9080T (M76388)

Atopobium minutum NCFB 2751T (X67148)

0·01

...... Fig. 1. Phylogenetic tree showing the positions of strain CS12T and some other related taxa based on 16S rDNA sequences. Scale bar represents 0n01 substitution per nucleotide position. Bootstrap values (expressed as percentages of 1000 replications) greater than 50% are shown at the branch points.

medium. Biomass of strain CS12T and J. limosus DSM agar (BBL), nutrient agar and solid R medium for fatty acid 11140T for DNA extraction was also obtained from growth methyl ester (FAME) analysis. in liquid R medium. The strains were cultivated at 28 mCon Morphological and physiological tests. The morphology of a horizontal shaker at 150 r.p.m. and the broth cultures were cells was examined by light microscopy and transmission checked for purity microscopically before they were har- electron microscopy (TEM). Presence or absence of ﬂagella vested by centrifugation. Strain CS12T and J. limosus DSM was examined by TEM using cells from exponentially 11140T were also grown at 28 mC for 7 d on trypticase soy growing cultures. These were negatively stained with 1%

1822 International Journal of Systematic and Evolutionary Microbiology 50 Janibacter terrae sp. nov.

(w\v) phosphotungstic acid, and after air drying, grids were Nucleotide sequence accession numbers. GenBank\EMBL examined with a model CM-20 transmission electron micro- accession numbers for reference 16S rDNA gene sequences scope (Philips). Catalase activity was determined by bubble used in the phylogenetic analysis are shown in Fig. 1. formation in a 3% hydrogen peroxide solution. Oxidase activity was determined by oxidation of 1% p-aminodi- methylaniline oxalate. DNase activity was determined as RESULTS described previously (Cowan & Steel, 1965) with DNase test Morphological and physiological characteristics agar (Difco). Nitrate reduction, indole production, methyl T red and Voges–Proskauer reactions, hydrogen sulfide pro- Strain CS12 is a Gram-positive, non-acid-fast, non- duction and hydrolysis of aesculin were tested as described spore-forming and non-motile bacterium. Its cells are previously (Lanyi, 1987). Hydrolyses of casein, gelatin, cocci, which are 0n6–1n1 µm in diameter after 24 h hypoxanthine, starch, Tween 80, tyrosine and xanthine, and culture in liquid R medium at 30 mC and occur singly, production of urease were also determined as described in pairs, in short chains or often in irregular clumps. previously (Cowan & Steel, 1965). Hydrolysis of arbutin was Cells were always cocci in all growth stages, both in determined according to the method of Kurup & Fink liquid and on solid R media. Colonies on R and (1975). Acid production from carbohydrates was detected nutrient agar were similar to those of J. limosus DSM by the method of Hugh & Leifson (1953). Tolerance of NaCl 11140T in colour and morphology. Colonies of strain and growth at various temperatures were measured on R T medium and brain heart infusion (BHI) medium. CS12 were cream-coloured on R agar and pale- cream-coloured on nutrient agar. Colonies of J. T Isolation of DNA. Chromosomal DNA was isolated and limosus DSM 11140 on nutrient agar were described purified according to the method of Yoon et al. (1996), with as white by Martin et al. (1997), but it is more the exception that ribonuclease T1 was used together with appropriate to describe these as being pale-cream- ribonuclease A. coloured. Colonies of strain CS12T were circular, Chemotaxonomic characterization. The isomer type of the opaque, glistening and convex on R and nutrient agar. The growth of strain CS12T on trypticase soy agar was diamino acid of the cell wall was analysed by the method of T Komagata & Suzuki (1987) and wall sugars were determined poor relative to J. limosus DSM 11140 on the same by the method of Saddler et al. (1991). Menaquinones were medium. Neither substrate mycelia nor primary my- analysed as described by Komagata & Suzuki (1987). celia were seen. Presence or absence of mycolic acids was determined using T the methods of Minnikin et al. (1975). Polar lipids were Strain CS12 grew optimally at pH 7n0–8n0, and growth extracted using the procedures of Minnikin et al. (1984) and was inhibited or slow below pH 5n0 and above pH 10. identified by two-dimensional TLC and by spraying with It grew optimally at 28–30 mC. Growth did not occur at appropriate detection reagents (Komagata & Suzuki, 1987). 40 mC on solid R medium or on solid or in liquid BHI Fatty acids were extracted and analysed following the media, but occurred weakly at 40 mC in liquid R T instructions of the Microbial Identification System (MIDI). medium. Both strain CS12 and J. limosus DSM 11140T grew in the presence of 8% (w\v) NaCl in Determination of GjC content. GjC content was de- liquid R and BHI media, but strain CS12T did not termined by the HPLC method of Tamaoka & Komagata grow in liquid R or BHI media containing 10% NaCl. (1984). No growth occurred under anaerobic conditions. T DNA–DNA hybridization. DNA–DNA hybridization to de- Physiological properties of strain CS12 , together with T termine genomic relatedness was performed fluorometrically those of J. limosus DSM 11140 , which were sim- by the method of Ezaki et al. (1989) using photobiotin- ultaneously determined, are given in Table 1. labelled DNA probes and microdilution wells. 16S rDNA sequencing and phylogenetic analysis. 16S rDNA Chemotaxonomic characteristics and DNA base was amplified by PCR using the two universal primers composition described previously (Yoon et al., 1998). The PCR product T was purified with a QIAquick PCR purification kit (Qiagen) Strain CS12 contained meso-diaminopimelic acid as and the 16S rDNA sequenced using an ABI PRISM BigDye the diamino acid in the cell wall. The sugars arabinose Terminator Cycle Sequencing Ready Reaction kit (Applied and galactose were not detected, although ribose and Biosystems). The purified sequencing reaction mixtures were galactose were present. The predominant isoprenoid automatically electrophoresed using an Applied Biosystems quinone was MK-8(H%) and the cellular phospholipids model 310 automatic DNA sequencer. Alignment of sequ- were diphosphatidylglycerol, phosphatidylglycerol ences was carried out with   software (Thompson and phosphatidylinositol. Mycolic acids were absent. et al., 1994). Gaps at the 5h and 3h ends of the alignment Strain CS12T had a cellular fatty acid profile con- were omitted from further analyses. Evolutionary distance taining saturated, unsaturated, branched and 10- matrices were calculated using the algorithm of Jukes & Cantor (1969) with the  program within the  methyl fatty acids and lacked hydroxy fatty acids (Table 2). The fatty acid profile of strain CS12T was package (Felsenstein, 1993). A phylogenetic tree was con- T structed by the neighbour-joining method (Saitou & Nei, different from that of J. limosus DSM 11140 in the 1987) with the  program of the same package. The composition of some fatty acids. In particular, C"(:!, one of the major fatty acids in J. limosus DSM 11140T, stability of relationships was assessed by a bootstrap analysis T of 1000 data sets by using the programs , , was a minor component in strain CS12 , and anteiso- T  and  of the  package. C"(:!, a minor component in J. limosus DSM 11140 ,

International Journal of Systematic and Evolutionary Microbiology 50 1823 J.-H. Yoon and others

Table 1. Morphological and physiological characteristics of strain CS12T and J. limosus DSM 11140T ...... j, Positive reaction; –, negative reaction; W, weakly positive reaction. Both strains were positive for catalase, decomposition of gelatin, nitrate reduction, production of H#S and growth in the presence of 2, 4, 6 and 8% NaCl. Both strains were negative for decomposition of arbutin, aesculin, hypoxanthine, starch, urea and xanthine, production of indole, Voges–Proskauer and methyl red tests, and acid production from -arabinose, -cellobiose, -fructose, -galactose, glycerol, inulin, lactose, maltose, -mannitol, -mannose, -raﬃnose, -rhamnose, -ribose, salicin, -sorbitol, starch, trehalose and -xylose.

Characteristic Strain CS12T J. limosus DSM 11140T

Colour of colonies Cream or pale cream Cream or pale cream Morphology Cocci Cocci, rods Motility – – Gram staining jj Spore formation – – Acid production from -glucose – j Decomposition of:* Casein jj j Tween 80 jj j Tyrosine jj j Oxidase W – DNase j W Growth in the presence of: 10% NaCl (R medium) – j 10% NaCl (BHI medium) – – Growth on BHI medium at: 28 mC jj 37 mC jj 40 mC–– Growth on R medium at: 10 mC jj 28 mC jj 37 mC j – 40 mC j –

* jj, Positive reaction stronger than that of J. limosus DSM 11140T. was present in large amounts in strain CS12T.In lineage within the radiation enclosing the members of addition, some fatty acids were characteristically the family Intrasporangiaceae and, especially, a co- detected in only strain CS12T or J. limosus DSM herent cluster with J. limosus DSM 11140T (Fig. 1). T T 11140 (Table 2). The genomic DNA GjC content of The relationship between strain CS12 and J. limosus strain CS12T was 69 mol%. DSM 11140T is supported by a bootstrap resampling value of 100%. The level of 16S rDNA sequence similarity between strain CS12T and J. limosus DSM Phylogenetic analysis based on 16S rDNA sequence T T 11140 was 98n7%, whilst strain CS12 had levels of An almost complete 16S rDNA sequence of 1480 16S rDNA similarity of 94n3–96n2% with the members nucleotides (approx. 96% of the Escherichia coli T of the genera Terrabacter, Terracoccus, Intraspor- sequence) was determined for strain CS12 . Strain angium and Sanguibacter. CS12T has highest 16S rDNA similarity with the genera Janibacter, Terrabacter, Terracoccus, Intra- DNA–DNA relatedness sporangium and Sanguibacter, all members of the family Intrasporangiaceae (Stackebrandt et al., 1997; DNA–DNA hybridization was performed to deter- Martin et al., 1997; Prauser et al., 1997). Accordingly, mine the genomic relatedness between strain CS12T its 16S rDNA sequence was compared with known 16S and J. limosus DSM 11140T. These species exhibited rDNA sequences of members of the family Intraspor- two independent levels of DNA–DNA relatedness of angiaceae and other actinomycetes. The resulting 22n3% and 23n7%, respectively, when each of their phylogenetic tree showed that strain CS12T forms a DNAs was separately used as labelled DNA probe.

1824 International Journal of Systematic and Evolutionary Microbiology 50 Janibacter terrae sp. nov.

Table 2. Cellular fatty acid proﬁles of strain CS12T and J. limosus DSM 11140T on trypticase soy agar (TSA), nutrient agar (NA) and R agar ...... Values are percentages of total cellular fatty acids.

Fatty acid Strain CS12T J. limosus DSM 11140T

TSA NA R agar TSA NA R agar

Saturated fatty acids C"%:! 0n10n20n20n10n3 C"&:! 0n50n10n81n90n41n2 C"':! 0n80n73n02n55n17n5 C"(:! 0n80n31n416n35n914n2 C"):! 0n40n61n02n85n97n3 C"*:! 0n70n50n9 Unsaturated fatty acids C"&:" ω6c 0n30n20n2 C"(:" ω8c 11n06n37n128n711n917n5 C"):" ω7c 0n82n11n10n51n51n0 C"):" ω9c 12n818n613n39n539n222n6 C"*:" iso I* 0n30n30n3 Branched fatty acids iso-C"#:! 0n10n20n10n1 iso-C"$:! 0n10n1 ante-C"$:! 0n10n10n2 iso-C"%:! 1n10n61n10n90n3 iso-C"&:! 4n64n19n00n60n20n5 ante-C"&:! 1n11n32n6 iso-C"':" H* 1n31n10n40n1 iso-C"':! 31n927n022n819n111n613n1 iso-C"(:! 6n24n95n80n91n21n3 iso-C"(:" ω9c 4n75n22n9 ante-C"(:" ω9c 1n11n21n1 ante-C"(:! 9n112n314n60n50n91n1 iso-C"):! 0n70n50n41n81n81n5 10-Methyl fatty acids C"':! 0n30n70n4 C"(:! 4n43n51n15n52n91n7 C"):! 0n32n80n21n24n81n1 Summed features† 34n16n68n81n92n54n0 61n80n53n82n92n5 * The double bond positions indicated by upper-case letters are unknown. † Summed features represent groups of two or three fatty acids which could not be separated by GLC with the MIDI system. Summed feature 3 contained one or more of following fatty acids: iso-C"&:! 2-OH and\or C"':" ω7c. Summed feature 6 contained one or more of following fatty acids: C"*:" ω9c and\or C"*:" ω11c.

DISCUSSION genus Janibacter. Comparative chemotaxonomic analyses confirm that strain CS12T belongs to the genus Phylogenetic inference based on 16S rDNA sequence Janibacter. Strain CS12T is clearly differentiated from comparison shows that strain CS12T is the nearest members of the genera Intrasporangium, Sanguibacter, phylogenetic neighbour of J. limosus DSM 11140T and Terrabacter and Terracoccus by the type of diamino both are closely related to the genera Intrasporangium, acid in the cell wall and by the profile of cellular fatty Sanguibacter, Terrabacter and Terracoccus, all mem- acids (Martin et al., 1997). Strain CS12T contains bers of the family Intrasporangiaceae (Stackebrandt et meso-diaminopimelic acid as the diamino acid in the al., 1997; Prauser et al., 1997) (Fig. 1). These data cell wall, whereas the genera Intrasporangium, Terra- suggest that strain CS12T may be a member of the bacter and Terracoccus all have -diaminopimelic

International Journal of Systematic and Evolutionary Microbiology 50 1825 J.-H. Yoon and others acid (Prauser et al., 1997), and the genus Sanguibacter Colonies are circular, opaque, glistening and convex, has -lysine (Fernandez-Garayzabal et al., 1995). and cream-coloured on R agar and pale-cream- Differences in major fatty acids are also valuable in coloured on nutrient agar. Neither substrate mycelia distinguishing strain CS12T from the other four genera. nor primary mycelia are seen. The optimal growth Both the genera Intrasporangium and Terrabacter have temperature is 28–30 mC. Growth does not occur at iso-C"&:! (Prauser et al., 1997), the genus Terracoccus 40 mC on solid R medium or on solid or in liquid BHI has iso-C"&:! and anteiso-C"&:! (Prauser et al., 1997) media, but occurred weakly at 40 mC in liquid R and the genus Sanguibacter has anteiso-C"&:! and C"':! medium. The optimal pH for growth is 7n0–8n0, and (Fernandez-Garayzabal et al., 1995; Martin et al., growth is inhibited below pH 5n0 and above pH 10. 1997). Although strain CS12T and J. limosus DSM Growth occurs in the presence of 8% NaCl in liquid R 11140T show a few differences in their fatty acid and BHI media. No growth occurs under anaerobic compositions, both contain iso-C"':! as a major fatty conditions. Catalase- and DNase-positive. Urease- acid (Table 2). The predominant menaquinones found negative. Oxidase is weakly positive only on R agar. in the genera Intrasporangium and Sanguibacter are Casein, gelatin, Tween 80 and tyrosine are hydrolysed. different from that of strain CS12T. The genera Arbutin, aesculin, hypoxanthine, starch and xanthine Intrasporangium and Sanguibacter have MK-8 and are not hydrolysed. Nitrate is reduced to nitrite. H#Sis T MK-9(H%), respectively, whilst strain CS12 has MK- produced. Indole is not produced. Voges–Proskauer 8(H%). However, the predominant menaquinones of and methyl red reactions are negative. No acid is Terrabacter and Terracoccus are the same as that of produced from -arabinose, -cellobiose, -fructose, strain CS12T (Martin et al., 1997; Prauser et al., 1997). -galactose, -glucose, glycerol, inulin, lactose, malt- Polar lipid profiles also distinguish strain CS12T and ose, -mannitol, -mannose, -raffinose, -rhamnose, the genus Janibacter from the genera Intrasporangium, -ribose, salicin, -sorbitol, starch, trehalose or - Terrabacter and Terracoccus (Martin et al., 1997; xylose. The cell wall contains meso-daminopimelic Prauser et al., 1997). The genus Intrasporangium is also acid. The predominant menaquinone is MK-8(H%). morphologically distinctive in having hyphae, a prop- The major fatty acids are iso-C"' !,C") " ω9c and T : : erty not found in strain CS12 (Kalakoutskii et al., anteiso-C"(:! or C"(:" ω8c. The polar lipids are 1967; Schumann et al., 1997). Accordingly, the mor- diphosphatidylglycerol, phosphatidylglycerol and phological, chemotaxonomic and phylogenetic data phosphatidylinositol. The GjC content is 69 mol% all indicate that strain CS12T is a member of the genus (determined by HPLC). Isolated from soil around a Janibacter. wastewater treatment plant, Korea. The type strain is T T T Strain CS12T is similar to J. limosus DSM 11140T in its CS12 (l KCCM 80001 l JCM 10705 ). morphological and in most of its physiological characteristics. However, there are some differences between ACKNOWLEDGEMENTS strain CS12T and J. limosus DSM 11140T, including those in their ability to degrade some substrates, their This work was supported by grants HS2321 and HS2701 tolerance of 10% NaCl and their growth at 37 mC and from the Ministry of Science and Technology (MOST) of the 40 mC (Table 1). It is noteworthy that the fatty acid Republic of Korea. profile of strain CS12T also differs from that of J. T limosus DSM 11140 in the composition of some fatty REFERENCES acids (Table 2). The 16S rDNA sequences of strain CS12T and J. limosus DSM 11140T have 19 bp se- Cho, Y.-G., Yoon, J.-H., Park, Y.-H. & Lee, S.-T. (1998). Simul- quence differences in the region compared. Levels of taneous degradation of p-nitrophenol and phenol by a newly DNA–DNA relatedness provide decisive evidence that isolated Nocardioides sp. J Gen Appl Microbiol 44, 303–309. T T strain CS12 and J. limosus DSM 11140 are members Cowan, S. T. & Steel, K. J. (1965). Manual for the Identification of of different genomic species (Wayne et al., 1987). Medical Bacteria. London: Cambridge University Press. Therefore, differences in some phenotypic character- Ezaki, T., Hashimoto, Y. & Yabuuchi, E. (1989). Fluorometric istics and genetic distinctiveness together indicate that deoxyribonucleic acid-deoxyribonucleic acid hybridization in strain CS12T is a new species of the genus Janibacter. microdilution wells as an alternative to membrane filter T hybridization in which radioisotopes are used to determine On the basis of the data described above, strain CS12 genetic relatedness among bacterial strains. Int J Syst Bacteriol should be placed in the genus Janibacter as a new 39, 224–229. species, for which the name Janibacter terrae sp. nov. Felsenstein, J. (1993). : phylogenetic inference package, is proposed. version 3.5. Seattle: University of Washington. Fernandez-Garayzabal, J. F., Dominguez, L., Pascual, C., Jones, D. Description of Janibacter terrae sp. nov. & Collins, M. (1995). Phenotypic and phylogenetic character- Janibacter terrae (ter rae. M.L. gen. n. terrae of the ization of some unknown coryneform bacteria isolated from h bovine blood and milk: description of Sanguibacter gen. nov. earth). Lett Appl Microbiol 20, 69–75. Cells are non-spore-forming and non-motile cocci, Hugh, R. & Leifson, E. (1953). The taxonomic significance of which occur singly, in pairs, in short chains or often in fermentative versus oxidative metabolism of carbohydrates by irregular clumps. Gram-positive and non-acid-fast. various Gram negative bacteria. J Bacteriol 66, 24–26.

1826 International Journal of Systematic and Evolutionary Microbiology 50 Janibacter terrae sp. nov.

Jukes, T. H. & Cantor, C. R. (1969). Evolution of protein mol- Saitou, N. & Nei, M. (1987). The neighbor-joining method: a new ecules. In Mammalian Protein Metabolism, vol. 3, pp. 21–132. method for reconstructing phylogenetic trees. Mol Biol Evol 4, Edited by H. N. Munro. New York: Academic Press. 406–425. Kalakoutskii, L. V., Kirillova, I. P. & Krassilnikov, N. A. (1967). A Schumann, P., Prauser, H., Rainey, F. A., Stackebrandt, E. & new genus of the Actinomycetales – Intrasporangium gen. nov. Hirsch, P. (1997). Friedmanniella antarctica gen. nov., sp. nov., J Gen Microbiol 48, 79–85. an -diaminopimelic acid-containing actinomycete from ant- Komagata, K. & Suzuki, K.-I. (1987). Lipids and cell-wall analysis arctic sandstone. Int J Syst Bacteriol 47, 278–283. in bacterial systematics. Methods Microbiol 19, 161–203. Stackebrandt, E., Rainey, F. A. & Ward-Rainey, N. L. (1997). Kurup, V. P. & Fink, J. N. (1975). A scheme for the identification Proposal for a new hierarchic classification system, Actino- of thermophilic actinomycetes associated with hypersensitivity bacteria classis nov. Int J Syst Bacteriol 47, 479–491. pneumonitis. J Clin Microbiol 2, 55–61. Tamaoka, J. & Komagata, K. (1984). Determination of DNA base Lanyl, B. (1987). Classical and rapid identification methods for composition by reversed-phase high-performance liquid medically important bacteria. Methods Microbiol 19, 1–67. chromatography. FEMS Microbiol Lett 25, 125–128. Martin, K., Schumann, P., Rainey, F. A., Schuetze, B. & Groth, I. Thompson, J. D., Higgins, D. G. & Gibson, T. J. (1994).  (1997). Janibacter limosus gen. nov., sp. nov., a new actino- : improving the sensitivity of progressive multiple sequence mycete with meso-diaminopimelic acid in the cell wall. Int J Syst alignment through sequence weighting, position-specific gap Bacteriol 47, 529–534. penalties and weight matrix choice. Nucleic Acids Res 22, 4673–4680. Minnikin, D. E., Alshamaony, L. & Goodfellow, M. (1975). Differentiation of Mycobacterium, Nocardia, and related taxa Wayne, L. G., Brenner, D. J., Colwell, R. R. & 9 other authors by thin-layer chromatographic analysis of whole-organism (1987). International Committee on Systematic Bacteriology. methanolysates. J Gen Microbiol 88, 200–204. Report of the ad hoc committee on reconciliation of approaches 37, Minnikin, D. E., O’Donnell, A. G., Goodfellow, M., Alderson, G., to bacterial systematics. Int J Syst Bacteriol 463–464. Athalye, M., Schaal, A. & Parlett, J. H. (1984). An integrated Yoon, J.-H., Kim, H., Kim, S.-B., Kim, H.-J., Kim, W. Y., Lee, S. T., procedure for the extraction of bacterial isoprenoid quinones Goodfellow, M. & Park, Y.-H. (1996). Identification of Saccharo- and polar lipids. J Microbiol Methods 2, 233–241. monospora strains by the use of genomic DNA fragments and Prauser, H., Schumann, P., Rainey, F. A., Kroppenstedt, R. M. & rRNA gene probes. Int J Syst Bacteriol 46, 502–505. Stackebrandt, E. (1997). Terracoccus luteus gen. nov., sp. nov., Yoon, J.-H., Lee, S. T. & Park, Y.-H. (1998). Inter- and intraspecific an -diaminopimelic acid-containing coccoid actinomycete phylogenetic analysis of the genus Nocardioides and related from soil. Int J Syst Bacteriol 47, 1218–1224. taxa based on 16S rDNA sequences. Int J Syst Bacteriol 48, Saddler, G. S., Tavecchia, P., Lociuro, S., Zanol, M., Colombo, L. & 187–194. Selva, E. (1991). Analysis of madurose and other actinomycete Yoon, J.-H., Cho, Y.-G., Lee, S. T., Suzuki, K.-I., Nakase, T. & Park, whole cell sugars by gas chromatography. J Microbiol Methods Y.-H. (1999). Nocardioides nitrophenolicus sp. nov., a p-nitro- 14, 185–191. phenol-degrading bacterium. Int J Syst Bacteriol 49, 675–680.

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