The species Xanthomonas oryzae includes two pathovars,
namely, oryzae and oryzicola (Swings et al., 1990). These
bacterial pathogens are closely related organisms and were
earlier named as pathovars of Xanthomonas campestris. Rice is
the main host for both pathogens, which are seed-borne and
seed-transmitted.
Bacterial leaf blight of rice (BLB) was first reported in
Fukuoka Prefecture, Japan, during 1884 in rice affected by X.
oryzae pv. oryzae. This disease is considered one of the most
serious rice diseases worldwide although it has declined in
incidence in Japan since the mid 1970s. Nevertheless it is still
prevalent worldwide. The disease was reported in South East
Asia in the early 1960s, where it is currently widespread, and it
still affects the rice crop in its severe form (Goto, 1992). It has
also been reported in several African countries, in Australia,
North America (Lousiana and Texas, US), Central and South
America (OEPP/EPPO, 2006a) but it is only of economic
importance in Asia. Other reported hosts include wild or minor
cultivated Poaceae: Brachiaria mutica, Cenchrus ciliaris,
Cyperus difformis, C. rotundus, Cynodon dactylon, Echinochloa
crus-galli, Leersia spp. (Leersia hexandra, L. oryzoides),
Leptochloa chinensis, Oryza spp., Panicum maximum, Paspalum
scrobiculatum, Zizania aquatica, Z. latifolia, Z. palustris,
and Zoysia japonica (Li et al., 1985; Bradbury, 1986; Valluva-
paridasan & Mariappan, 1989; EPPO/CABI, 1997; Saddler,
2002a). Cyperaceae (sedges) that are naturally infected include
Cyperus difformis and C. rotundus. Variation in virulence
has been observed between isolates, and many races have been
described; however, the different races have not been clearly
defined with specific reactions being assigned to each variety
(Mew, 1987). The diversity of the pathogen has been analysed
based on virulence and PCR-based DNA fingerprinting
(George et al., 1994). Low virulence strains have been reported
in the United States and India (Jones et al., 1989; Gnanamanickam
et al., 1993).
Bacterial leaf streak of rice (BLS) is caused by X. oryzae pv
oryzicola. The disease is present in Tropical Asia, West Afric
and Australia (OEPP/EPPO, 2006b). X. oryzae pv. oryzicol
has reached epidemic proportions in recent years in China
Other hosts affected by the pathogen are: Oryza spp., Leersi
spp., Leptochloa filiformis, Paspalum orbiculare, Zizani
aquatica, Z. palustris and Zoysia japonica. No races have bee
recorded, however, differences in virulence of strains have bee
observed in many countries (Vera Cruz et al., 1984; Adhikari &
Mew, 1985; Saddler, 2002b).
Further information on the biology and ecology of th
species can be found in the EPPO data sheet on X. oryza
(OEPP/EPPO, 1997)
Identity
Taxonomic position: Kingdom: Bacteria; Phylum: Proteobacteria
Class: Gammaproteobacteria; Order: Xanthomonodales; Family
Xanthomonodaceae; Genus: Xanthomonas (Dowson, 1939)
Name: Xanthomonas oryzae pv. oryzae (Ishiyama 1922
Swings et al. (1990)
Synonyms: Xanthomonas campestris pv. oryzae (Ishiyam
1922) Dye 1978; Xanthomonas oryzae (Ishiyama, 1921
Dowson 1943; Other synonyms are Xanthomonas itoana
Xanthomonas kresek, Xanthomonas translucens f. sp. oryzae
EPPO computer code: XANTOR
Phytosanitary categorization: EPPO A1 list no. 2, EU anne
II/A1 as Xanthomonas campestris pv. oryzae.
Xanthomonas oryzae pv. oryzicola
Name: Xanthomonas oryzae pv. oryzicola (Fang et al., 1956
Swings et al. (1990)
Fig. 1 Symptoms of X. oryzae pv. oryzae (bacterial leaf blight). (a) Rice field plants with X. oryzae pv. oryzae symptoms; (b) X. oryzae pv. oryzae symptoms
in rice nursery beds; (c) Rice seedlings showing X. oryzae pv. oryzae symptoms; (d) Wilting of transplanted seedlings; (e) Kresek symptoms in field plants;
(f) Progressive development of bacterial leaf blight in rice plants grown from infected seed.
Synonyms: Xanthomonas campestris pv. oryzicola (Fang
et al., 1956) Dye 1978; Xanthomonas oryzicola (Fang et al.
1956) Dowson 1943; Xanthomonas translucens f. sp. oryzicola
(Fang et al., 1956) Bradbury 1971
EPPO computer code: XANTTO
Phytosanitary categorization: EPPO A1 list no. 3, EU annex
II/A1 as Xanthomonas campestris pv. oryzicola.
Detection
Disease symptoms
X. oryzae pv. oryzae and X. oryzae pv. oryzicola can be clearly
distinguished by symptoms, which reflect the differences in
their modes of infection.
Bacterial leaf blight of rice (BLB)
X. oryzae pv. oryzae enters either through wounds or hydathodes
multiplies in the epitheme and moves to the xylem vessel
where active multiplication results in blight on the leaves. The
symptoms of the disease include leaf blight, wilting (kresek)
(Fig. 1) and pale yellow leaves. Leaf blight is characterized by
wavy elongated lesions, which develop along the leaf margins
They start as small water-soaked stripes from the tips where
water pores are found and rapidly enlarge in length and width,
orming a yellow lesion with a wavy margin along the lead
edges. Later on, diseased areas turn white to grey. These lesions
can develop on one or both sides of the leaf and occasionally
along the midribs, and leaf blight symptoms generally occur
rom maximum tillering stage and onwards. In young lesions,
drops of bacterial ooze can be observed early in the morning.
On panicles the disease causes grey to light brown lesions on
glumes that result in infertility and low quality of the grains.
Kresek is the result of systemic infection that is common in the
ropics in young plants and during the tillering stage of susceptible
cultivars. Leaves of infected plants wilt, roll up, turn grey-green
and whither, and entire plants finally die. Surviving plants look
tunted and yellowish. Yellow or pale yellow leaves are due to
ystemic infections that appear at tillering stage; the youngest
eaves become uniformly pale yellow or show a broad yellow
tripe, and bacteria are found in the internodes and crowns of
affected stems, but not in the leaf itself (Ou, 1985; Goto, 1992).
Bacterial leaf streak of rice (BLS)
X. oryzae pv. oryzicola is a foliar disease that appears at any
growth stage of the host. Cells of X. oryzae pv. oryzicola enter
through the stomata and multiply in the parenchyma tissues of
the leaves. X. oryzae pv. oryzicola infects mainly the parenchyma
of the cells of the leaves, but is not systemic. Initial symptoms
are small water-soaked, transparent interveinal streaks (Fig. 2),
which may elongate and darken. The transparent streaks
differentiate leaf streak lesions from those of X. oryzae pv.
oryzae that are opaque against the light. Bacterial exudates canbe observed as tiny yellow beads. The narrow, long, translucent
lesions may coalesce, forming large patches, and severely
affected fields appear burnt. It is at this stage that leaves wither,
turn brown and eventually die, and the disease can be difficult
to distinguish from bacterial leaf blight (Ou, 1985).
Isolation
Symptomatic samples are processed individually or in small
groups. Precautions are made to avoid cross contaminations
when collecting the samples and during the extraction process.
The samples should be processed as soon as possible after
collection and conserved at 4–8°C until use. Freshly prepared
sample extracts are necessary for a successful isolation. Several
procedures have been used for the isolation of X. oryzae pv.
oryzae from symptomatic and asymptomatic plant parts,
including seeds. Most of the procedures described for the
isolation of X. oryzae pv. oryzae from rice plants, can also be
applied for the isolation of X. oryzae pv. oryzicola. However, it
is known that both bacteria grow slowly on the isolating media
and can be overgrown by fast growing contaminants. Often these
contaminants are of yellow colour (e.g. Pantoea agglomerans
and Xanthomonas-like saprophytic bacteria) thus making
difficult the observations of colonies of the target organisms and
re often visually undistinguishable in colony morphology,
rowth, and colour from strains of both pathogens. Culturing
may fail from advanced stages of infection due to competition
r being overgrown by saprophytic organisms and also from
eeds with low levels of inoculum or under bad conditions of
orage. If disease symptoms are typical but isolation is negative,
he isolation step should be repeated.
irect plating from symptomatic and asymptomatic leaves
ollected from the field
acterial leaf blight in temperate regions can usually be
bserved during the latter part of the seed bed stage (Ou, 1985).
he bacteria can be detected from the upper part of infected
aves before symptoms appear (Goto, 1965; Misukami &
Wakimoto, 1969; Sakthivel et al., 2001). Tabei (1967) reported
hat symptomless rice seedlings may carry the pest.
The isolation of Xanthomonas from symptomatic material is
referable and can be performed using Peptone sucrose agar
PSA), Nutrient Broth Yeast Extract agar medium (NBY),
rowth Factor (GF) agar (Agarwal et al., 1989; Sakthivel et al.,
001). (Appendix 1). The bacteria can also be isolated on
utrient agar (NA) but with a very slow growth; semi-selective
gar substrates like modified XOS agar medium (mXOS)
Di et al., 1991; Gnanamanickam et al., 1994). X. oryzae pv.
ryzicola colonies are also isolated on modified Wakimoto’s
gar (Mew & Mistra, 1994) developed for the detection of X.
ryzae pv. oryzae, but with the omission of ferrous sulfate.
Surface disinfection with 70% alcohol for 15 s followed by
wo to three times rinsing with sterile water is used for isolation
om field plants. Plant parts showing fresh symptoms and with
xudates are selected whenever possible. Sections (2 mm ×
mm) from infected tissue and preferably from the advancing
ortion of lesions are selected. The exudates can be processed
eparately in 1 mL of sterile water. Narrow sections of infected
ssue are cut with a sterilized razor blade or scalpels, placed in
drop of sterile water and covered with a cover slip before the
bservation under the compound microscope for bacterial
reaming, an indication of bac
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