Confirmation of bacterial leaf streak of rice caused by Xanthomonas oryzae pv. oryzicola in Vietnam

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ConfirmationofBacterialLeafStreakofRice
CausedbyXanthomonasoryzaepv.oryzicolain
Vietnam
ArticleinPlantDisease·December2015
DOI:10.1094/PDIS-03-15-0289-PDN

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Plant Disease Journal - 0(ja): - Abstract

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Confirmation*of*Bacterial*Leaf*Streak*of*Rice
Caused*by*Xanthomonas)oryzae*pv.*oryzicola
in*Vietnam

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Plant Disease Journal - 0(ja): - Abstract

27/05/2015 10:59

Environnement (IPME), Montpellier, France;

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The Xanthomonas oryzae species is one of the most important pathogens of rice (Oryza
sativa) (Niño-Liu et al. 2006). In early October of 2013, Vietnamese colleagues reported a
high prevalence of leaf bacterial diseases on irrigated rice fields of the Red River delta
(western coast of the Gulf of Tonkin). We performed an exploratory survey in provinces of
North Vietnam. In the Hanoi, Hung Yen and Nam Dinh provinces, we observed symptoms
reminiscent of bacteria leaf streak (BLS) with variable (<5 to 90%) incidenceprevalence.
While Tthe presence of BLS disease in Vietnam has been recorded in international databases
(Saddler 2002), but to our knowledge, the role of X. oryzae pv. oryzicola as the causal agent
has never been unambiguously established. Collected Field samples exhibited typical BLS
symptoms, such as water-soaked, linear lesions running along the veins and developing into
yellow to brown streaks with yellow droplets of exudates at the surface. Bacteria were
isolated by grinding 4 cm-long leaf pieces in sterile water. Tissue lysates were plated on
semi-selective PSA medium (Poulin et al. 2014) and incubated for 3-4 days at 28°C. Single,
yellow-pigmented, mucoid colonies were further purified on PSA medium. In parallel,
diagnostic multiplex PCR was performed on these colonies for microbial identification (Lang
et al. 2010). In total, isolates from 15 independent samples yielded a PCR profile
characteristic of X. oryzae pv. oryzicola. Six of these strains from the Nam Dinh and Hanoi
provinces were further analyzed. First, a portion of their gyrB gene was sequenced upon
following PCR amplification (Young et al. 2008). All six 780-bp quality-trimmed gyrB
sequences were deposited in GenBank under accession numbers KP872841 to KP872846.
They are identical to the gyrB sequence of strain BLS256 (GenBank CP003057), a reference
strain for the oryzicola pathovar, and exhibited ten polymorphic positions relative to the one
from PXO99A (GenBank CP000967), a oryzae pathovar reference strain. These strains were
also tested for pathogenicity by infiltrating suspensions at 1x108 CFU/ml in water into 4weeks-old O. sativa cv. Nipponbare leaves with a needleless syringe (3 infiltrations x 5 plants

per strain). Unlike water-infiltrated negative controls, all inoculated Lleaves inoculated with
Vietnamese isolates were indistinguishable from positive controls leaves challenged with
strain BLS256. In contrast to water-infiltrated negative controls,: they showed ttypical watersoaked lesions appeared after five days in the greenhouse (27°C, 12-h photoperiod, ~80%
humidity) and ultimately produced yellow exudates resembling those initially observed in rice
fields. For bacterial re-isolation, symptomatic tissue lysates were streaked on PSA medium.
Bacterial colonies showed a typical Xanthomonas morphology and were again assigned to the
oryzicola pathovar by multiplex PCR typing, thus fulfilling Koch's postulates. Theseis data
confirms the status of X. oryzae pv. oryzicola as the causing agent of BLS in two provinces of
Vietnam using modern molecular identification tools. To facilitate inquiries on the phylogeny
and virulence of Vietnamese X. oryzae pv. oryzicola, strains VXO32 and VXO39 have been
deposited in the Collection Française de Bactéries Phytopathogènes under accessions
CFBP8314 and CFBP8315, respectively. J.M. Lang et al. Plant Dis. 94:311, 2010 D.O. Niño-Liu
et al. Mol. Plant Pathol. 7:303, 2006 L. Poulin et al. Plant Dis. 98:1423, 2014. G.S. Saddler.
IMI Descriptions of Fungi and Bacteria 1458. CAB International, 2002. J.M. Young et al. Syst.
Appl. Microbiol. 31:366, 2008.

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