INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY, July 1983, p. 656-657 Vol. 33. No.

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0020-7713/83/030656-02$02.OO/O
Copyright 0 1983, International Union of Microbiological Societies

Pseudomonas fuscovaginae sp. nov., nom. rev.

KUNIYUKI MIYAJIMA,'" AKIO TANII,2 AND TADAHIKO AKITA3
Kitami Agricultural Experiment Station, Kunneppu, Hokkaido 099-14, Tokachi Agricultural Experiment
Station, Memuro, Hokkaido 082,2and Central Agricultural Experiment Station, Iwamizawa, Hokkaido
069-03,3Japan

The name Pseudomonas fuscovaginae Tanii, Miyajima and Akita 1976 was
omitted from the Approved Lists of Bacterial Names. Therefore, this name is here
revived for the organism to which it originally referred. The cells of strains of this
species are aerobic, gram negative, and rod shaped with polar flagella. They
oxidize glucose in oxidation-fermentation medium, and they produce a green
fluorescent pigment, oxidase, and arginine dihydrolase. Denitrification, p-glucosi-
dase, pit formation on polypectate gel and growth at 37°C are negative. Character-
istics that distinguish this species from other fluorescent pseudomonads which are
positive for arginine dihydrolase and oxidase are its ability to produce a
hypersensitivity reaction in tobacco plants and its inability to utilize 2-ketogluco-
nate or inositol. These bacteria were pathogenic to Oryza sativa and eight other
species of the Gramineae. The type strain is NCPPB 3085 (= PDDCC 5940).

The name Pseudomonas fuscovaginae was
validly published by Tanii et al. (7) for the
causative agent of sheath brown rot of rice
plants. This name was omitted from the Ap-
proved Lists of Bacterial Names (5). In accord-
ance with Rule 28a of the International Code of
Nomenclature of Bacteria (2), we propose to
revive the name according to the following de-
scription.
Pseudomonas fuscovaginae (ex Tanii et al.
1976) nom. rev. (fus. co. va. gi' nae, L. adj.
fuscus fuscous; L. fem. n. vagina vagina,
sheath; M. L. fem. n. fuscovaginae of a fuscous
vagina). Gram-negative , nonsporeforming, rod-
shaped cells with round ends, 0.5 to 0.8 by 2.0 to
3.5 km. Cells occur singly or in pairs and are
motile by means of one to four polar flagella.
After 4 to 5 days at 28°C on nutrient agar
moderate growth consisting of white to light
brown, smooth, glistening, raised, translucent,
circular, butyrous colonies 3 to 5 mm in diame-
starch are hydrolyzed, but esculin and arbutin
are not. Arginine dihydrolase and ammonia are
produced, but cytochrome oxidase, phenylala-
nine deaminase, urease, 2-ketogluconate, H2S,
and indole are not produced. Levan is not pro-
duced from sucrose.
No organic growth factors are required. Acid
is produced from glucose, arabinose, rhamnose,
and mannitol but not from maltose, sucrose,
raffinose, inulin, salicin, dextrin, adonitol,
erythritol, inositol, dulcitol, or a-methylgluco-
side. Citrate, malonate, succinate, urate, ace-
tate, p-alanine, L-valine, and L-lysine are uti-
lized, but tartrate, hippurate, 2-ketogluconate,
and polygalacturonic acid are not.
Aerobic. The optimal growth temperature is
approximately 28°C. No growth occurs at 37°C.
Characteristics that vary among strains of P .
fuscovaginae are shown in Table 1.

1 l(n%=:6/
TABLE 1. Characteristics in which 16 strains of P.
ter is produced. A green fluorescent, diffusible fuscovaginae differ from one another
pigment is produced on King's medium B. No
slime is produced on nutrient agar containing 5%
strains Strains that gave the
sucrose. Character
:t
f
S
O positive less common result
Metabolism of glucose is oxidative in Hugh-
Leifson oxidation-fermentation medium. Cata-
Tyrosinase 19 BA-1-1, 7101, 7106
lase and Kovacs oxidase tests are positive. Egg yolk reaction 81 BA-1-1, 705, 7101
Denitrification is negative; nitrate is not re- Acid from:
duced. The methyl red and Voges-Proskauer Xylose 94 6801
tests are negative. Lipolysis of margarine and Mannose 94 IK-4
Tween 80 is positive. No growth occurs in Lactose 81 BA-1-3, IK-2, 6801
nutrient broth supplemented with 5% NaC1. Trehalose 94 705
Peptonization and reduction of litmus milk with- Sorbitol 6 705
out coagulation are positive. Pits are not pro- Growth in KCN 19 BA-1-2, BM-1,
duced on polypectate gels at pH 8.5. Gelatin and broth 7103

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VOL. 33, 1983 NOTES 657

TABLE 2. Characters of diagnostic value in distinguishing P.fuscovaginae from other fluorescent

pseudomonads that are positive for arginine dihydrolase and oxidase
P. fuscovaginae P. Juorescens biotype:b
Character type strain P. marginalis' P . putidab
A B C F G
Denitrification - c' + - + + + -
Levan formation from sucrose - +
P-Glucosidase - +
Gelatin liquefaction + +
Utilization of
L- Arabinose +
++ + - +
+ d+ d
Trehalose + + + -
2- Ke togluconate - + + + + + +
Inositol - + + + + + -
Sorbitol - + + d + d -
Adonitol - + d + - d -
Polygalacturonic acid - - - - _ _ -
Pits on polypectate gel (pH 8.5) -
Hypersensitivity reaction in tobacco +
Data from Hildebrand and Schroth (1) and Lelliott et al. (3).
Data from Sands et al. (4)and Stanier et al. (6).
+, >90% of strains positive; -, 4 0 % of strains positive; d, 10 to 90% of strains positive.

A hypersensitive reaction is produced when
cells are inoculated into tobacco leaves, but
potato soft rot is not produced.
Pathogenicity was demonstrated on Oryza sa-
tiva, Hordeum vulgare, Triticum aestivum,
Avena sativa, Zea mays, Lolium perenne, Bro-
mus marginatus, Phleum pratense, and Phalaris
arundinacea but was not apparent on two spe-
cies of the Solanaceae, six species of the Legu-
minosae, Petroselinum crispum, Brassica olera-
cea, Lactuca sativa, or Citrus limon. On 0.
sativa, water-soaked dark green spots appear
first; then these spots become brown to dark
brown blotches on the flag leaf sheaths. Infected
young panicles show a brown to dark brown
discoloration. On the other eight species of the
Gramineae tested, water-soaked lesions appear
and then become greyish white or reddish brown
spots on the leaves.
P. fuscovaginae is similar to other fluorescent
pseudomonads that are positive for arginine
dihydrolase and oxidase (1, 3, 4, 6). Table 2 lists
the characters which have diagnostic value for
differentiating P . fuscovaginae from other spe-
cies of this group. Table 2 shows that P.fusco-
vaginae is most similar to the saprophytes Pseu-
domonas putida and Pseudomonas fluorescens
biotype G (6). There are consistent differences
between P . fuscovaginae and P . putida in six
characters (gelatin liquefaction; utilization of
trehalose, 2-ketogluconate, and hippurate;
starch hydrolysis; and hypersensitive reaction).
P . fluorescens biotype G also differs in six
characters (utilization of arabinose, 2-ketogluco-
nate, inositol, sorbitol, and adonitol; and hyper-
sensitive reaction).
Strains were isolated from diseased leaf
sheaths of 0. sativa in Japan.
The type strain of this species is 6801
(= NCPPB 3085 [National Collection of Plant
Pathogenic Bacteria, Harpenden, England]
= PDDCC 5940 [Plant Diseases Division Culture
Collection, Auckland, New Zealand]).
We are most grateful to M. Goto (University of Shizuoka,
Shizuoka, Japan) for invaluable help during this investigation.
We also thank T. Narita (Hokkaido Plant Protection Associa-
tion, Sapporo, Japan) for constructive criticism and comments
and T. Ui (University of Hokkaido, Sapporo, Japan) and T.
Baba (Hokkaido Central Agricultural Experiment Station,
Naganuma, Japan) for advice and encouragement.
LITERATURE CITED
1. Hildebrand, D. C., and M. N. Schroth. 1971. Identification
of the fluorescent pseudomonads, 281-287. In H. P. Maas
Geesteranus (ed.), 3rd International Conference on Plant
Pathogenic Bacteria, Wageningen. Centre for Agricultural
Publishing and Documentation, Wageningen, The Nether-
lands.
2. Lapage, S. P., P. H. A. Sneath, E. F. Lessel, V. B. D. Sker-
man, H. P. R. Seeliger, and W. A. Clark (ed.). 1975. Inter-
national code of nomenclature of bacteria. 1975 Revision.
American Society for Microbiology, Washington, D.C.
3. Lelliott, R. A., E. Billing, and A. C. Hayward. 1966. A
determinative scheme for the fluorescent plant pathogenic
pseudomonads. J. Appl. Bacteriol. 29:470-489.
4. Sands, D. C., M. N. Schroth, and D. C. Hildebrand. 1970.
Taxonomy of phytopathogenic pseudomonads. J. Bacteri-
01. 101~9-23.
5. Skerman, V. B. D., V. McGowan, and P. H. A. Sneath
(ed.). 1980. Approved lists of bacterial names. Int. J. Syst.
Bacteriol. 30:225-420.
6. Stanier, R. Y., N. J. Palleroni, and M. Doudoroff. 1966.
The aerobic pseudomonads: a taxonomic study. J. Gen.
Microbiol. 43:159-271.
7. Tanii, A., K. Miyajima, and T. Akita. 1976. The sheath
brown rot disease of rice plant and its causal bacterium,
Pseudomonas fuscovaginae A. Tanii, K . Miyajima et T.
Akita sp. nov. Ann. Phytopathol. SOC.Jpn. 42540-548.

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