Download Three New Methylobacterium Species

INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY, Jan. 1988, p. 124-127
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Copyright 0 1988, International Union of Microbiological Societies
Vol. 38, No. 1
Three New Methylobacterium Species: M . rhodesianum sp. nov.
M . zatmanii sp. nov., and M . fujisawaense sp. nov.
P. N. GREEN,l* I. J. BOUSFIELD,l AND D. HOOD2
National Collections of Industrial and Marine Bacteria Ltd., Torry Research Station, Aberdeen AB9 8DG, Scotland, and
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, England2
Three new Methylobacterium species ( M . zatmanii, M . rhodesianum, and M . fujisawaense) are proposed on
the basis of deoxyribonucleic acid-deoxyribonucleic acid homology. Strains are assigned to these and other
species of Methylobacterium on the basis of both the homology data and previously published phenotypic data.
Recent deoxyribonucleic acid (DNA)-DNA homology
studies on the genus Methylobacterium have shown that
there are at least five distinct homology groups (lA, lB, lC,
2B, and 2D) (9) within the genus that should be assigned
separate species status. Two of these groups correspond to
the species M . radiotolerans and M . extorquens, but the
others cannot be equated with any currently described
species of Methylobacterium. The type strains of M . orgar,iophilum, M . rhodinurn, and M . mesophilicum, the only
other species recognized at present, were shown to be
distinct from each other and from the five homology groups.
Thus, we consider the three anonymous homology groups to
represent as yet undescribed species of Methylobacterium.
The present paper presents formal proposals for and descriptions of these new species.
Strains assigned to the eight species of Methylobacterium
on the basis of both homology and phenotypic data are listed
in Table 1. Table 2 gives the phenotypic features that
differentiate among these species and among other probable
centers of variation within the genus.
Descriptions of the three new species are given below
(methodologies as previously described [S]).
Methylobacterium rhodesianum sp. nov. (rhbd’es.i.an.um
Id. L. neut. adj. rhodesianum named after the British
taxonomist Muriel Rhodes-Roberts for her contributions to
pseudomonad taxonomy). Mode of respiration, colony and
cell morphology, flagellar arrangement, Gram reaction, and
oxidase and catalase as previously described (6).
Grows at 15 and 30°C but not at 10°C. Most strains will
also grow at 37°C. Growth factors are not required. Casein,
cellulose, gelatin, lecithin, and DNA are not degraded.
Urease is produced, but P-galactosidase, indol, H,S, Lornithine and L-1 ysine decarboxylases, and L-arginine dihytlrolase are not. The methyl red and Voges-Proskauer tests
are negative. Some strains reduce nitrate to nitrite.
Sensitive to kanamycin, tetracyclines, gentamicin, alhamycin T, streptomycin, and framycetin but resistant to
tx-ythromycin, nalidixic acid, oleandomycin, spiramycin,
polymyxin B, colistin sulfate, penicillin G, and bacitracin.
The following compounds serve as sole carbon sources:
fructose, glycerol, malonate, succinate, fumarate, 2-oxoglutarate, lactate, malate, acetate, pyruvate, ethanol, propylene
glycol, methanol , monomethylamine, ethylamine, formate,
and betaine. Does not utilize monosaccharides other than
fructose, citrate, sebacate, tartrate, or trimethylamine.
The guanine-plus-cytosine content of the DNA ranges
from 69.8 to 71.2 moles percent (mol%) (9). We propose
* Corresponding author
NCIB 12249T (Pseudomonas strain 1 [16]), which was isolated from a fermentor operating with formaldehyde as a sole
carbon source, as the type strain. As a result of recent work,
including DNA-DNA homology studies (9), we also propose
that the following strains be considered as strains of M .
rhodesianum: Pseudomonas sp. strains NCIB 10598 and
10611, strain M159-1 (13), Rhodes-Roberts strain CS51, and
NCIB 12440 “Corynebacterium rubrum.” (It should be
noted that the National Collection of Industrial Bacteria
strain of “C. rubrum” is different from the strain deposited
in the DSM as DSM 1857). Other strains assigned to this
species solely on the basis of phenotypic data are listed in
Table 1.
Methylobacterium zatmanii sp. nov. (zat.man’i.i M. L. m.
noun zatmanii, named after the British biochemist L. J.
Zatman for his work on the bacterial metabolism of onecarbon compounds.) Mode of respiration, colony and cell
morphology, flagellar arrangement, Gram reaction, and oxidase and catalase as previously described (6).
Grows at 20 and 37°C but not at 10°C. Growth factors are
not required. Casein, cellulose, gelatin, lecithin, Tweens,
and DNA are not degraded. Urease is produced, but pgalactosidase, indol, and H2S are not. The methyl red and
Voges-Proskauer tests are negative. Nitrate is not reduced,
Susceptible to kanamycin, tetracyclines, novobiocin,
neomycin, gentamycin, albamycin T, streptomycin, cephalothin, framycetin, and cephaloridine but resistant to nalidixic acid, oleandomycin, spiramycin, polymyxin B, colistin
sulfate, penicillin G, and bacitracin.
The following compounds serve as sole carbon sources:
fructose, glycerol, malonate, succinate, fumarate, 2-oxoglutarate, lactate, malate; acetate, pyruvate, ethanol, propylene
glycol, methanol, monomethylamine, ethylamine, and formate. Some strains utilize tartrate and trimethylamine. Does
not utilize monosaccharides other than fructose, citrate,
sebacate, aspartate, glutamate, or betaine.
The guanine-plus-cytosine content of the DNA ranges
from 69.4 to 70.3 mol% (9); for the type strain, this value is
70.3 mol%.
We propose NCIB 12243T(Pseudomonas strain 135 [16]),
which was isolated from a fermentor operating with formaldehyde as a sole carbon source, as the type strain. As a
result of recent work, including DNA-DNA homology studies (9), we also propose that the following strains be considered as strains of M . zatmanii: Pseudornonas spp. strains
NCIB 10604,10609, and 10610. Other strains assigned to this
species solely on the basis of phenotypic data are listed in
Table 1.
Methylobacterium fujisawaense sp. nov. (fu.ji.sa’wa.en.si?
N. L. neut. adj. fujisawaense coming from the Fujisawa
124
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VOL.38, 1988
NOTES
125
TABLE 1. Strains assigned to Methylobacterium species
Species
M . organophilum
M . extorquens
M . rhodinum
M . radiotolerans
M . mesophilicum
M . rhodesianum sp. nov.
M . zatmanii sp. nov.
M . fujisawaense sp. nov.
Strain or collection no.
XX (NCIB 1127gT)"
NCIB 9399Tu
Pseudomonas AM1 (NCIB 9133)"
Pseudomonas M27 (NCIB 9686)
"Protominobacter ruber" (NCIB 2879)"
453, 456, 465, 530, 539," 691, 724, 789, N-3, N-4, N-5,
N-10, N-12"
82A,b NCIB 10409
190B,' AM1 var. 471
"Pseudomonas methylica" strain 2
B020,d B026
111,' 270
19;f28, 50
NCIB 942lTU
0-1 (NCIB 10815T)"
Pseudomonas spp. (NCIB 9142" and 9143)
434," 532, 0-20
A47 (NCIB 11561T)"
Pseudomonas strain 1 (NCIB 12249T)"
Pseudomonas spp. (NCIB 10597'-10602, 10611")
"Corynebacterium rubrum""
CS51"*b
82, 83, 711, M159-1,"
B023,d B040,d 381B'
"Protaminobacter ruber"
"Protaminobacter ruber subsp. machidanus" (ATCC
21614)
Pseudomonas strain 135 (NCIB 12243=)"
Pseudomonas spp. (NCIB 10603,' 10604, 10606-10608,
10609-10610," 10612)
109,' 212, 216, 339, 390, 393, 463, 466, 471
590, N-11
0-31 (NCIB 12417T)u
044,' 115, 269"
35, 37, 45," 58, 420, 437, 470, 488 617," 670, 679, 696,
767, 781, 787, 788, 823, 0-6, 0-7, 0-46
BWd
"Mycoplana rubra" (NCIB 11272)h
D123"
8 p 9 , 26, 31, 36, 37, 39
Reference
Patt et al. (14)
Janota-Bassalik and Pedyk (11)
Peel and Quayle (15)
Anthony and Zatman (1)
Den Dooren de Jong (3)
Kouno and Ozaki (13)
Kirikova (12)
Heumann (8)
Ito and Iizuka (10)
Hayward (7)
Kouno and Ozaki (13)
Austin and Goodfellow (2)
Rock et al. (16)
Graf and Bauer (4)
Kouno and Ozaki (13)
Sat0 et al. (17)
Rock et al. (16)
Kouno and Ozaki (13)
Kouno and Ozaki (13)
Kouno and Ozaki (13)
" Strains assigned on the basis of DNA-DNA homology. Other strains are assigned on the basis of phenotypic characteristics only.
M. E. Rhodes-Roberts, University College of Wales, Aberystwyth, United Kingdom.
T. S. Chandra, Indian Institute of Science, Bangalore, India.
T. G. Mitchell, British-American Tobacco Co., Southampton, United Kingdom.
ICI Ltd., Billingham, Cleveland, United Kingdom.
H. Stolp, University of Bayreuth, Bayreuth, Federal Republic of Germany.
B. Austin, Heriot-Watt University, Edinburgh, United Kingdom.
Boots Co. Ltd., Nottingham, United Kingdom.
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126
INT. J. SYST.BACTERIOL.
NOTES
TABLE 2. Features that differentiate among species and some unassigned Methylobacterium" strains
Substrates utilized as sole C sourceb
a,
*
5
U
Species or Strain
s
a,
*
0
U
*
1A
1B
1C
1E
1F
1G
1H
1J
2A
2B
2D
2E
2F
2G
2H
25
M . zatmanii
M . extorquens
M . rhodesianum
NCIB 9141 and
9145 (9)
N-6 (9)
M . rhodinum
M . orphanophilum
602 and 317 (9)
790 (9)
M . radiotoleruns
M.fujisawaense
M . mesophilicum
N-2 (9)
R14 (9)
D12 (9)
B46 (9)
7 and 35d
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a Owing to the slow growth of some strains on certain substrates, carbon utilization tests were read after 14 days of incubation at 30°C (5). Doubtful results
were checked by twice subculturing in liquid medium.
V, Variable result; W, weak growth.
Most strains which utilize sebacate can also utilize pimelate, suberate, azelate, and adipate.
Isolated by H. Stolp, University of Bayreuth, Bayreuth, Federal Republic of Germany.
region of Japan). Mode of respiration, colony and cell morphology, flagellar arrangement, Gram reaction, and oxidase
atnd catalase as for the type species.
Grows at 15 and 37°C. Some strains grow at 10°C. Growth
factors are not required. Gelatin, starch, cellulose, lecithin,
DNA, casein, and Tweens are usually not degraded. Urease
is produced, but indol, H,S, P-galactosidase, L-ornithine and
r.-lysine decarboxylases, and L-arginine dihydrolase are not.
Methyl red and Voges-Proskauer tests are negative. Nitrate
is not reduced by most strains.
Sensitive to kanamycin, tetracyclines, gentamicin, alhamycin T, streptomycin, and framycetin but resistant to
rialidixic acid, oleandomycin, polymyxin B, colistin sulfate,
penicillin G, and bacitracin.
The following compounds can serve as sole carbon
sources: D-glucose, L-arabinose, D-xylose, D-fucose, D-galactose, glycerol, malonate, succinate, fumarate, L-aspartate, L-glutamate, sebacate, 2-oxoglutarate, lactate, malate,
acetate, pyruvate, citrate, citraconate, ethanol, methanol,
and propylene glycol. Some strains can utilize fructose.
Monomethylamine, trimethylamine, and betaine are not
utilized. Grows on peptone-rich nutrient agar (Oxoid CM55).
The guanine-plus-cytosine content of the DNA ranges
from 70.8 to 71.8 mol% (9); for the type strain, this value is
71.8 mol%.
We propose NCIB 12417T (= strain 0-31 [13]) as the type
s,train. As a result of recent work, including DNA-DNA
homology studies (9), we also propose that strains 45, 269,
and 617 (13) be considered as strains of M . fujisawaense.
Other strains assigned to this species solely on the basis of
phenotypic data are listed in Table 1.
LITERATURE CITED
1. Anthony, C., and L. J. Zatman. 1964. The microbial oxidation of
methanol. I. Isolation and properties of Pseudomonas M27.
Biochem. J. 92:609-614.
2. Austin, B., and M. Goodfellow. 1979. Pseudomonas mesophilica: a new species of pink bacteria isolated from leaf surfaces.
Int. J. Syst. Bacteriol. 29:373-378,
3. Den Dooren de Jong, L. E. 1927. Uber protaminophage Bakterien. Zentralbl. Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt.
2 71:193-232.
4. Graf, W., and L. Bauer. 1973. Red bacterial growth (Corynebacterium rubrum n. spec.) in tap water systems. Zentralbl.
Bakteriol. Parasitenkd. Infektionskr. Hyg. Abt. 1Orig. Reihe A
236:513-530.
5. Green, P. N., and I. J. Bousfield. 1982. A taxonomic study of
some gram-negative facultatively methylotrophic bacteria. J.
Gen. Microbiol. 12tk623-638.
6. Green, P. N., and I. J. Bousfield. 1983. Emendation of Methyfobacterium Patt, Cole, and Hanson 1976; Methylobacterium
rhodinum (Heumann 1962) comb. nov. corrig. ; Methylobacterium radiotoleruns (It0 and Iizuka 1971) comb. nov. corrig.; and
Methylobacterium mesophilicum (Austin and Goodfellow 1979)
comb. nov. Int. J. Syst. Bacteriol. 335375477.
7. Hayward, A. C. 1960. Relationship between Protaminobacter
ruber and some red pigmented pseudomonads. J. Appl. Bacterial. 23:ii.
8. Heumann, W. 1962. Die Methodik der Kreuzung sternbildener
Downloaded from www.microbiologyresearch.org by
IP: 88.99.165.207
On: Sun, 14 May 2017 03:41:38
NOTES
VOL.38, 1988
Bakterien. Biol. Zentralbl. 81:341-354.
9. Hood, D., C. S. DOW,and P. N. Green. 1987. DNA:DNA
hybridization studies on the pink pigmented facultative methylotrophs. J. Gen. Microbiol. 133:709-720.
10. Ito, H., and H. Iizuka. 1971. Taxonomic studies on a radioresistant Pseudornonas. XII. Studies of the micro-organisms of
cereal grain. Agric. Biol. Chem. 3515661571.
11. Janota-Bassalik, L., and D. Pedyk. 1961. Ability of Fiavobacterium extorquens Bassalik to utilise various sources of carbon
with particular reference to glucose. Acta Microbiol. Pol.
10~225-238.
12. Kirikova, N. N. 1970. Properties of two strains of Pseudornonas
utilizing one-carbon compounds. Microbiology 39: 12-16. (English translation of Mikrobiologiya.)
13. Kouno, K., and A. Ozaki. 1975. Distribution and identification of
14.
15.
16.
17.
127
methanol-utilizing bacteria, p. 11-21. In Proceedings of the
International Symposium on Microbial Growth on C, Compounds. Society of Fermentation Technology, Osaka, Japan.
Patt, T. E., G. C. Cole, and R. S. Hanson. 1976. Methylobacterium, a new genus of facultatively methylotrophic bacteria. Int.
J. Syst. Bacteriol. 26:226229.
Peel, D., and R. Quayle. 1961. Microbial growth on C, compounds. I. Isolation and characterization of Pseudomonas AMI.
Biochem. J. 81:465469.
Rock, J. S., I. Goldberg, A. Ben-Bassat, and R. I. Mateles. 1976.
Isolation and characterization of two methanol utilizing bacteria. Agric. Biol. Chem. 40:2129-2135.
Sato, K., S. Ueda, and S. Shimizu. 1977. Form of vitamin B,, and
its role in a methanol-utilizing bacterium, Protarninobacter
ruber. Appl. Environ. Microbiol. 33515-521.
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