Download Reclassification of Koreibacter algae as a later heterotypic synonym

International Journal of Systematic and Evolutionary Microbiology (2013), 63, 219–223
DOI 10.1099/ijs.0.040600-0
Reclassification of Koreibacter algae as a later
heterotypic synonym of Paraoerskovia marina and
emended descriptions of the genus Paraoerskovia
Khan et al. 2009 and of Paraoerskovia marina Khan
et al. 2009
P. Schumann, R. Pukall, C. Spröer and E. Stackebrandt
Correspondence
P. Schumann
Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH,
Inhoffenstraße 7B, D-38124 Braunschweig, Germany
[email protected]
16S rRNA gene sequences deposited for the type strains of Paraoerskovia marina (CTT-37T;
GenBank accession no. AB445007) and Koreibacter algae (DSW-2T; FM995611) show a
similarity of 100 %. Consequently, the type strains were subjected to a polyphasic
recharacterization under direct comparison in order to clarify their taxonomic position. PvuII
RiboPrint patterns and quantitative ratios of cellular fatty acids revealed strain-specific differences
between P. marina DSM 21750T (5CTT-37T) and K. algae DSM 22126T (5DSW-2T). The
percentage of DNA–DNA binding of 94 % indicated that the two type strains belong to the same
genomospecies. Agreement in the peptidoglycan structure and polar lipid pattern, highly similar
fatty acid profiles and MALDI-TOF mass spectra, the ability to produce acid from the same carbon
sources, corresponding enzymic activities and DNA G+C contents of 70.8±0.3 mol%, in
addition to the consistent characteristics reported in the original descriptions, support the view
that the two strains should be affiliated to the same species. According to Rules 38 and 42 of the
Bacteriological Code, Koreibacter algae should be reclassified as later heterotypic synonym of
Paraoerskovia marina, and the descriptions of the genus Paraoerskovia Khan et al. 2009 and of
Paraoerskovia marina Khan et al. 2009 are emended accordingly.
The detection of representatives of novel bacterial taxa
among isolated strains has become unbiased and rapid by
comparison of 16S rRNA gene sequences with those of type
strains deposited in comprehensive public databases. As a
consequence, a multitude of novel taxa has been described
during the last decade, and the probability of concurrent
submissions of manuscripts proposing novel taxa for
highly similar organisms is increasing.
Strains of rod-shaped actinobacteria representing a distinct
clade within the family Cellulomonadaceae (suborder
Micrococcineae) were isolated independently from marine
habitats by two research groups without being aware that
the two organisms were taxonomically highly similar. One
group submitted the proposal for a new genus and species
Paraoerskovia marina (Khan et al., 2009), whereas the other
named the novel organism Koreibacter algae (Lee & Lee,
2010). Though the description of K. algae appeared in print
8 months after the proposal of P. marina, the type strains
of the two taxa had not been compared (Schumann &
Gvozdyak, 2012). The present study is aimed at the
A supplementary figure and two supplementary tables are available with
the online version of this paper.
040600 G 2013 IUMS
reconsideration of the taxonomic status of K. algae. The
type strains K. algae DSM 22126T and P. marina DSM
21750T were therefore subjected to a re-examination of
published physiological and chemotaxonomic characteristics as well as to MALDI-TOF mass spectrometric
analysis, comparison of PvuII RiboPrint patterns and
DNA–DNA hybridization.
The 16S rRNA gene sequences of K. algae DSM 22126T
and P. marina DSM 21750T agree by 100 %, and the two
strains represent a common lineage within the family
Cellulomonadaceae, with the type strains of the genus
Oerskovia as the closest relatives [96.2–96.3 % sequence
similarity determined by the EzTaxon server (http://www.
eztaxon.org; Kim et al., 2012)]. The phylogenetic positions
of K. algae and P. marina among the type strains of type
species classified in the suborder Micrococcineae are shown
in a comprehensive neighbour-joining tree (Fig. S1,
available in IJSEM Online). The clustering of K. algae
DSM 22126T and P. marina DSM 21750T with the most
related type strains belonging to the families Cellulomonadaceae and Sanguibacteraceae is demonstrated by a
neighbour-joining tree (Fig. 1) reconstructed from 16S
rRNA gene sequences aligned manually by using the
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219
P. Schumann and others
Brevibacterium linens DSM 20425T (X77451)
Tropheryma whipplei Twist-MarseilleT (AF251035)
0.01
Actinotalea fermentans DSM 3133T (X83805)
Cellulomonas bogoriensis 69B4T (X92152)
Sanguibacter antarcticus KOPRI 21702T (EF211071)
94
Sanguibacter keddieii ST-74T (X79450)
98
Sanguibacter inulinus ST-50T (X79451)
88
97
Sanguibacter suarezii ST-26T (X79452)
Sanguibacter soli DCY22T (EF547937)
Sanguibacter marinus 1-19T (AJ783958)
Oerskovia enterophila DSM 43852T (X83807)
100
Oerskovia turbata DSM 20577T (X83806)
Oerskovia paurometabola DSM 14281T (AJ314851)
90
99 Oerskovia jenensis DSM 46000T (AJ314848)
T
100 Paraoerskovia marina CTT-37 (AB445007)
Koreibacter algae DSW-2T (FM995611)
Cellulomonas denverensis ATCC BAA-788T (AY501362)
99
Cellulomonas hominis CE40T (X82598)
Cellulomonas persica IT (AF064701)
Cellulomonas flavigena DSM 20109T (X83799)
81
Cellulomonas phragmiteti KB23T (AM902253)
Cellulomonas composti TR7-06T (AB166887)
Cellulomonas iranensis OT (AF064702)
Cellulomonas uda DSM 20107T (X83801)
95
Cellulomonas gelida DSM 20111T (X83800)
Cellulomonas terrae DB5T (AY884570)
Cellulomonas xylanilytica XIL 11T (AY303668)
100
Cellulomonas humilata ATCC 25174T (X82449)
Cellulomonas chitinilytica X.bub-bT (AB268586)
Cellulomonas aerilata 5420S-23T (EU560979)
Cellulomonas cellasea DSM 20118T (X83804)
Cellulomonas biazotea DSM 20112T (X83802)
100
Cellulomonas fimi DSM 20113T ( X79460)
Fig. 1. Neighbour-joining tree reconstructed from 16S rRNA gene
sequences of type strains of the genera Paraoerskovia,
Koreibacter, Cellulomonas, Oerskovia, Actinotalea and Tropheryma (family Cellulomonadaceae) as well as of the genus
Sanguibacter (family Sanguibacteraceae). The tree is based on a
1369 bp alignment. Brevibacterium linens DSM 20425T was used
as an outgroup. Bar, 0.01 substitutions per nucleotide position.
Bootstrap values (expressed as percentages of 1000 replications)
are given at branch points; only values .80 % are shown.
BioEdit program (Hall, 1999; Kimura, 1980; Saitou & Nei,
1987; Thompson et al., 1997; Felsenstein, 1985) and was
confirmed by a maximum-likelihood analysis (Felsenstein,
1981) (not shown). In order to examine the DNA–DNA
binding of K. algae DSM 22126T and P. marina DSM
21750T, cells of the two type strains were disrupted by
using a French pressure cell (Thermo Spectronic) and the
DNA in the crude lysates was purified by chromatography
on hydroxyapatite as described by Cashion et al. (1977).
DNA–DNA hybridization was performed in duplicate as
described by De Ley et al. (1970) under consideration of
the modifications proposed by Huss et al. (1983) using a
model Cary 100 Bio UV/Vis spectrophotometer equipped
with a Peltier-thermostatted 666 multicell changer and a
temperature controller with in-situ temperature probe
(Varian). DNA of K. algae DSM 22126T and P. marina
DSM 21750T showed 94 % binding (in 26 SSC with 10 %
formamide at 71 uC; average of two results, 98 and 89 %
220
binding), indicating that the two type strains belong to the
same genomospecies (Wayne et al., 1987). According to a
MALDI-TOF mass spectrometric analysis carried out as
described by Tóth et al. (2008), the mass spectra of K. algae
DSM 22126T and P. marina DSM 21750T are almost
identical (Fig. 2). However, investigation of the type strains
of K. algae and P. marina by the RiboPrinter system, which
is capable of differentiation of strains (Bruce, 1996; Barney
et al., 2001), revealed that they were not identical but
display distinct band patterns when PvuII was used as the
restriction enzyme (Fig. 3).
When comparing the descriptions of K. algae and P.
marina, several differing phenotypic characteristics are
obvious (Khan et al., 2009; Lee & Lee, 2010). K. algae was
reported to be aerobic and oxidase- and leucine arylamidase-negative, while facultatively anaerobic growth as well
as positive oxidase and leucine arylamidase reactions were
reported for P. marina. In contrast to P. marina, K. algae is
described as incapable of starch degradation and acid
production from L-arabinose, D-fructose, D-galactose, Dglucose, D-mannose, D-xylose, lactose, cellobiose, sucrose
and salicin. According to its description, K. algae contains
phosphatidylinositol as polar lipid but lacks mannose as a
cell-wall sugar and differs in these chemotaxonomic
features from the data reported for P. marina. Khan et al.
(2009) reported the peptidoglycan type A4a L-Lys–L-Ser–DGlu and a DNA G+C content of 70 mol% for P. marina,
while Lee & Lee (2010) published the type A3a Lys–Ser and
a DNA G+C content of 68.3 mol% for K. algae. With
respect to physiological features not mentioned above,
morphology, colony appearance, motility, growth temperature, pH and NaCl tolerance as well as menaquinone
patterns, the data reported for K. algae and P. marina were
either identical or very similar or the differences were
within ranges that would not allow the unambiguous
differentiation of the two organisms (Table S1; Khan et al.,
2009; Lee & Lee, 2010).
In order to re-examine the published data, K. algae DSM
22126T and P. marina DSM 21750T were cultivated on
medium no. 92 (http://www.dsmz.de) at 28 uC and
subjected to the API 50CH and API ZYM test systems
(bioMérieux) following the manufacturer’s instructions.
Oxidase activity was tested according to the method of
Smibert & Krieg (1994). The capacity for anaerobic growth
was examined by incubating the organisms on medium no.
92 agar plates using Anaerocult A (Merck). Degradation of
starch was studied according to Gordon et al. (1973).
Both K. algae DSM 22126T and P. marina DSM 21750T
exhibited facultatively anaerobic growth in the present
study. Reinvestigation of the physiological characteristics
of K. algae DSM 22126T and P. marina DSM 21750T
revealed agreement in almost all features except for only
weak acid production from turanose and a weak esterase
reaction for K. algae DSM 22126T, while P. marina DSM
21750T was scored as positive for these two reactions
(Table S1).
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Koreibacter algae is a synonym of Paraoerskovia marina
MSP dendrogram
Cellulomonas hominis DSM 9581T
Cellulomonas denverensis DSM 15764T
Cellulomonas cellasea DSM 20118T
Cellulomonas chitinilytica DSM 17922T
Cellulomonas xylanilytica DSM 16933T
Cellulomonas terrae DSM 17791T
Cellulomonas biazotea DSM 20112T
Cellulomonas fimi DSM 20113T
Cellulomonas uda DSM 20107T
Cellulomonas persica DSM 14784T
Cellulomonas gelida DSM 20111T
Cellulomonas bogoriensis DSM 16987T
Oerskovia jenensis DSM 46000T
Oerskovia turbata DSM 20577T
Oerskovia paurometabola DSM 14281T
Oerskovia enterophila DSM 43852T
Cellulomonas phragmiteti DSM 22512T
Cellulomonas flavigena DSM 20109T
Cellulomonas iranensis DSM 14785T
Actinotalea fermentans DSM 3133T
Cellulomonas aerilata DSM 18649T
Paraoerskovia marina DSM 21750T
Koreibacter algae DSM 22126T
1000 900
800
700
600 500
Distance
400
300
200
100
0
Fig. 2. Score-orientated dendrogram generated by the BioTyper software (version 2.0; Bruker Daltonics) showing the similarity
of MALDI-TOF mass spectra of cell extracts of P. marina DSM 21750T, K. algae DSM 22126T and selected type strains of the
family Cellulomonadaceae.
Purified cell walls were isolated after disruption of cells by
shaking with glass beads and subsequent trypsin digestion,
according to the method described by Schumann (2011).
Amino acids and peptides in cell-wall hydrolysates were
analysed by two-dimensional TLC on cellulose plates by
using previously described solvent systems (Schleifer,
1985). The molar ratios of amino acids were determined
by GC/MS (320-MS Quadrupole GC/MS; Varian) of Nheptafluorobutyryl amino acid isobutyl esters (Schumann,
2011). Total hydrolysates of the peptidoglycan (4 M HCl,
16 h at 100 uC) of P. marina DSM 21750T and K. algae
DSM 22126T contained the amino acids lysine, alanine,
serine and glutamic acid in molar ratios of 1.0 : 1.8 : 0.7 : 2.1
and 1.0 : 1.6 : 0.6 : 2.0, respectively. The peptides L-Ala–DGlu, L-Lys–L-Ser and L-Lys–D-Ala were detected by twodimensional TLC of partial hydrolysates of the peptidoglycan
15.00
30.00
60.00
8.00
6.00
2.00
3.00
4.00
1.00
kb
P. marina DSM 21750T
K. algae DSM 22126T
Fig. 3. PvuII RiboPrint patterns of P. marina DSM 21750T and K.
algae DSM 22126T normalized by using the BioNumerics software
(Applied Maths).
http://ijs.sgmjournals.org
(4 M HCl, 0.75 h at 100 uC). Dinitrophenylation according
to Schumann (2011) revealed that glutamic acid represented
the N terminus of the interpeptide bridge. On the basis
of these results, it was concluded that K. algae DSM 22126T
and P. marina DSM 21750T correspond in showing
the peptidoglycan type A4a L-Lys–L-Ser–D-Glu, A11.48
(Schleifer & Kandler, 1972; Schumann, 2011). The data
from the present study confirm the peptidoglycan structure
reported for P. marina DSM 21750T by Khan et al. (2009),
but disagree with the result of Lee & Lee (2010). Galactose
and traces of xylose were the sugars of purified cell walls of
the two type strains, as determined according to the
procedure of Staneck & Roberts (1974). Galactose as a major
cell-wall sugar in K. algae was also reported by Lee & Lee
(2010), but the occurrence of mannose in the cell wall of P.
marina (Khan et al., 2009) could not be confirmed in the
present study.
Polar lipids were extracted, separated by two-dimensional
TLC and identified by spraying with ninhydrin, anaphthol, molybdophosphoric acid and Zinzadze reagent
according to Tindall et al. (2007). Phosphatidylglycerol,
diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides were the polar lipids found in
both K. algae DSM 22126T and P. marina DSM 21750T
(Fig. 4) in the present study, in disagreement with the
differing lipid patterns reported previously (Khan et al.,
2009; Lee & Lee, 2010).
P. marina DSM 21750T and K. algae DSM 22126T agreed in
their growth behaviour, and sufficient cells of comparable
physiological age could be harvested for fatty acid analysis
from the third streak quadrant of plates after cultivation on
tryptic soy agar at 28 uC for 24 h. Cellular fatty acids of P.
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P. Schumann and others
Fig. 4. Polar lipids of P. marina DSM 21750T and K. algae DSM
22126T after two-dimensional TLC and detection with molybdophosphoric acid and heating at 180 6C for 5 min. DPG,
Diphosphatidylglycerol; PG, phosphatidylglycerol; PI, phosphatidylinositol; PIMs, phosphatidylinositol mannosides.
marina DSM 21750T and K. algae DSM 22126 were
analysed by GC using an Agilent 6850 chromatograph with
the MIDI Microbial Identification System (library TSBA40,
4.10; Sherlock software package, version 6.1). The fatty acid
profiles of the type strains of K. algae and P. marina agreed
in displaying the fatty acids anteiso-C15 : 0, C18 : 0, C16 : 0 and
anteiso-C17 : 0 as major components, but the quantitative
ratios of the cellular fatty acids differed (Table S2).
For reinvestigation of the base composition, genomic DNA
from the two type strains was degraded to nucleosides by
using P1 nuclease and bovine intestinal mucosa alkaline
phosphatase, as described by Mesbah et al. (1989). The
nucleosides were separated by reversed-phase HPLC
(Shimadzu LC 20A) according to the method described
by Tamaoka & Komagata (1984). A DNA G+C content of
70.8±0.3 mol% for both type strains was calculated from
the ratio of deoxyguanosine to thymidine.
On the basis of the agreement in 16S rRNA gene sequences,
MALDI-TOF mass spectra, peptidoglycan type, major
menaquinone, DNA base composition, polar lipids and
cellular fatty acids, as well as in morphological, physiological and biochemical characteristics, and because of the
observed DNA–DNA binding of 94 %, the reclassification
of Koreibacter algae as a later heterotypic synonym of
Paraoerskovia marina is proposed according to Rules 38
and 42 of the Bacteriological Code (Lapage et al., 1992). As a
result of the new taxonomic information obtained in this
study, emended descriptions of the genus Paraoerskovia
and of Paraoerskovia marina are given.
Emended description of Paraoerskovia Khan et al.
2009
non-motile, facultatively anaerobic rods. Catalase-positive
and oxidase-negative. Do not produce aerial mycelium.
The peptidoglycan type is A4a L-Lys–L-Ser–D-Glu
(A11.48). The cell wall contains acetylated muramic acid
residues and galactose. The major quinone is menaquinone
MK-9(H4). The polar lipids are phosphatidylglycerol,
diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides. Major fatty acids are anteisoC15 : 0, C18 : 0, C16 : 0 and anteiso-C17 : 0. The G+C content of
the DNA is 71 mol%. Phylogenetically, the genus belongs
to the family Cellulomonadaceae (suborder Micrococcineae,
order Actinomycetales). The type species is Paraoerskovia
marina.
Emended description of Paraoerskovia marina
Khan et al. 2009
Paraoerskovia marina (ma.ri9na. L. fem. adj. marina of the
sea, marine).
The description is as given by Khan et al. (2009) with the
following modifications. Cells are 0.4–0.6 mm wide and
1.0–1.6 mm long. Forms creamy yellow-coloured colonies,
1–2 mm in diameter on HSMA or ISP 2 plates after 3–
5 days of incubation at 28 uC. Carotenoid-type pigments
are present. Growth occurs at 10–35 uC (optimum at
28 uC), at pH 6.0–10.0 (optimum pH 7.0–8.0) and at 0–
8 % (w/v) NaCl. According to the API 50CH system,
positive for acid production from starch, glycogen,
cellobiose, sucrose, trehalose, gentiobiose, maltose, Larabinose, D-fructose, D-galactose, D-glucose, D-mannose,
D-xylose, aesculin ferric citrate, glycerol and turanose (type
strain) and negative for the 33 other tests of the panel.
According to the API ZYM system, positive for leucine
arylamidase, a- and b-glucosidases, lipase, N-acetyl-bglucosaminidase and esterase (type strain) and negative
for the 13 other enzymes included in the test panel. Able to
degrade starch and carboxymethyl-cellulose but unable to
degrade casein, cellulose, gelatin or chitin. For the type
strain, menaquinones MK-9 and MK-9(H2) are detected,
in addition to the major menaquinone MK-9(H4). Traces
of xylose occur in addition to the major cell-wall sugar
galactose. The type strain is sensitive to vancomycin
(50 mg) and resistant to bacitracin (10 mg), gentamicin
(30 mg), kanamycin (30 mg), nalidixic acid (30 mg),
nitrofurantoin (300 mg), nystatin (100 IU) and streptomycin (10 mg).
The type strain is CTT-37T (5NBRC 104352T 5DSM
21750T), isolated from a sediment sample collected from a
beach on the coast of Tottori city, Japan.
References
Paraoerskovia (Pa.ra.oer.sko9vi.a. Gr. prep. para beside;
N.L. fem. n. Oerskovia a bacterial genus name; N.L. fem. n.
Paraoerskovia beside or close to Oerskovia).
Barney, M., Volgyi, A., Navarro, A. & Ryder, D. (2001). Riboprinting
The description is as given by Khan et al. (2009) with the
following modifications. Cells stain Gram-positive and are
Bruce, J. (1996). Automated system rapidly identifies and charac-
222
and 16S rRNA gene sequencing for identification of brewery
Pediococcus isolates. Appl Environ Microbiol 67, 553–560.
terizes microorganisms in food. Food Technol 50, 77–81.
Downloaded from www.microbiologyresearch.org by
International Journal of Systematic and Evolutionary Microbiology 63
IP: 88.99.165.207
On: Sun, 18 Jun 2017 09:43:13
Koreibacter algae is a synonym of Paraoerskovia marina
Cashion, P., Holder-Franklin, M. A., McCully, J. & Franklin, M. (1977).
Schleifer, K. H. (1985). Analysis of the chemical composition and
A rapid method for the base ratio determination of bacterial DNA.
Anal Biochem 81, 461–466.
Schleifer, K. H. & Kandler, O. (1972). Peptidoglycan types of bacterial
De Ley, J., Cattoir, H. & Reynaerts, A. (1970). The quantitative
measurement of DNA hybridization from renaturation rates. Eur
J Biochem 12, 133–142.
Felsenstein, J. (1981). Evolutionary trees from DNA sequences: a
maximum likelihood approach. J Mol Evol 17, 368–376.
Felsenstein, J. (1985). Confidence limits on phylogenies: an approach
using the bootstrap. Evolution 39, 783–789.
Gordon, R. E., Haynes, W. C. & Pang, C. H.-N. (1973). The Genus
Bacillus. Agriculture Handbook no. 427. Washington, DC: Agricultural
Research Service, United States Department of Agriculture.
Hall, T. A. (1999). BioEdit: a user-friendly biological sequence
alignment editor and analysis program for Windows 95/98/NT.
Nucleic Acids Symp Ser 41, 95–98.
Huss, V. A. R., Festl, H. & Schleifer, K. H. (1983). Studies on the
spectrophotometric determination of DNA hybridization from
renaturation rates. Syst Appl Microbiol 4, 184–192.
Khan, S. T., Harayama, S., Tamura, T., Ando, K., Takagi, M. & Kazuo,
S. Y. (2009). Paraoerskovia marina gen. nov., sp. nov., an
actinobacterium isolated from marine sediment. Int J Syst Evol
Microbiol 59, 2094–2098.
Kim, O. S., Cho, Y. J., Lee, K., Yoon, S. H., Kim, M., Na, H., Park, S. C.,
Jeon, Y. S., Lee, J. H. & other authors (2012). Introducing EzTaxon-e:
a prokaryotic 16S rRNA gene sequence database with phylotypes that
represent uncultured species. Int J Syst Evol Microbiol 62, 716–721.
Kimura, M. (1980). A simple method for estimating evolutionary rates
of base substitutions through comparative studies of nucleotide
sequences. J Mol Evol 16, 111–120.
Lapage, S. P., Sneath, P. H. A., Lessel, E. F., Skerman, V. B. D.,
Seeliger, H. P. R. & Clark, W. A. (editors) (1992). International Code of
Nomenclature of Bacteria (1990 Revision). Bacteriological Code.
Washington, DC: American Society for Microbiology.
Lee, D. W. & Lee, S. D. (2010). Koreibacter algae gen. nov., sp. nov.,
isolated from seaweed. Int J Syst Evol Microbiol 60, 1510–1515.
Mesbah, M., Premachandran, U. & Whitman, W. B. (1989). Precise
measurement of the G+C content of deoxyribonucleic acid by highperformance liquid chromatography. Int J Syst Bacteriol 39, 159–167.
Saitou, N. & Nei, M. (1987). The neighbor-joining method: a new
method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425.
http://ijs.sgmjournals.org
primary structure of murein. Methods Microbiol 18, 123–156.
cell walls and their taxonomic implications. Bacteriol Rev 36, 407–
477.
Schumann, P. (2011). Peptidoglycan structure. Methods Microbiol 38,
101–129.
Schumann, P. & Gvozdyak, O. R. (2012). International Committee on
Systematics of Prokaryotes Subcommittee on the taxonomy of the
suborder Micrococcineae. Minutes of the meeting, 9 September 2011,
Sapporo, Japan. Int J Syst Evol Microbiol 62, 755.
Smibert, R. M. & Krieg, N. R. (1994). Phenotypic characterization. In
Methods for General and Molecular Bacteriology, pp. 607–654. Edited
by P. Gerhardt, R. G. E. Murray, W. A. Wood & N. R. Krieg.
Washington, DC: American Society for Microbiology.
Staneck, J. L. & Roberts, G. D. (1974). Simplified approach to
identification of aerobic actinomycetes by thin-layer chromatography.
Appl Microbiol 28, 226–231.
Tamaoka, J. & Komagata, K. (1984). Determination of DNA base
composition by reversed phase high-performance liquid chromatography. FEMS Microbiol Lett 25, 125–128.
Thompson, J. D., Gibson, T. J., Plewniak, F., Jeanmougin, F. &
Higgins, D. G. (1997). The CLUSTAL_X windows interface: flexible
strategies for multiple sequence alignment aided by quality analysis
tools. Nucleic Acids Res 25, 4876–4882.
Tindall, B. J., Sikorski, J., Smibert, R. M. & Krieg, N. R. (2007).
Phenotypic characterization and the principles of comparative
systematics. In Methods for General and Molecular Microbiology, pp.
332–393. Edited by C. A. Reddy, T. J. Beveridge, J. A. Breznak,
G. A. Marzluf, T. M. Schmidt & L. R. Synder. Washington, DC:
American Society for Microbiology.
Tóth, E. M., Schumann, P., Borsodi, A. K., Kéki, Z., Kovács, A. L. &
Márialigeti, K. (2008). Wohlfahrtiimonas chitiniclastica gen. nov., sp.
nov., a new gammaproteobacterium isolated from Wohlfahrtia
magnifica (Diptera: Sarcophagidae). Int J Syst Evol Microbiol 58,
976–981.
Wayne, L. G., Brenner, D. J., Colwell, R. R., Grimont, P. A. D., Kandler,
O., Krichevsky, M. I., Moore, L. H., Moore, W. E. C., Murray, R. G. E. &
other authors (1987). Report of the ad hoc committee on
reconciliation of approaches to bacterial systematics. Int J Syst
Bacteriol 37, 463–464.
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Sun, 18 Jun 2017 09:43:13
223