oryzicola in Vietnam Article in Plant Disease · December 2015 DOI: 10.1094/PDIS-03-15-0289-PDN CITATIONS 0 READS 159 9 authors , including: Some of the authors of this publication are al
Trang 1Confirmation of Bacterial Leaf Streak of Rice Caused by Xanthomonas oryzae pv oryzicola in Vietnam
Article in Plant Disease · December 2015
DOI: 10.1094/PDIS-03-15-0289-PDN
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Editor-in-Chief: Mark L Gleason Published by The American Phytopathological Society Home > Plant Disease > Table of Contents > Abstract
Previous Article | Next Article Accepted for publication
http://dx.doi.org/10.1094/PDIS-03-15-0289-PDN
Confirmation*of*Bacterial*Leaf*Streak*of*Rice
Caused*by*Xanthomonas)oryzae*pv.*oryzicola
in*Vietnam
Dr Tuan Tu Tran, PhD
Institut de Recherche pour le Développement (IRD), UMR 186 Interaction Plantes Microorganismes Environnement (IPME), Montpellier, France; tuan-tu.tran@ird.fr
Miss Nga Nguyen
Vietnam National University of Agriculture, Centre for Plant tropical disease, Hanoi, Viet Nam; vietnga54t@gmail.com
Miss Ngân Phan
Vietnam National University of Agriculture, Dept Molecular Biology and Applied biotech, Hanoi, Hanoi, Viet Nam, 10000;
Agriculture Genetics Institute, Pham Van Dong street, Hanoi, Viet Nam, 10000, 084 1642
615 877; ptngancnsh.tbf@gmail.com
Miss Hong Thi Nguyen
Agricultural Genetics Institute, Mutation and Heterosis, Pham Van Dong, Hanoi, Tuliem, Viet Nam, 10000; nguyenhongdhnn@gmail.com
Dr Boris Szurek
Institut de Recherche pour le Développement (IRD), UMR 186 Interaction Plantes Microorganismes Environnement (IPME), Montpellier, France; boris.szurek@ird.fr
Dr Ralf Koebnik
Institut de Recherche pour le Développement (IRD), UMR 186 Interaction Plantes Microorganismes Environnement (IPME), Montpellier, France; ralf.koebnik@ird.fr
Dr Ham Le Huy
Agricultural Genetics Institute, Hanoi, Viet Nam; lhham@agi.ac.vn
Dr Cuong Ha Viet
Vietnam National University of Agriculture, Centre for Plant tropical disease, Hanoi, Viet Nam; cuongvietha@gmail.com
Dr Sebastien Cunnac, PhD
Institut de Recherche pour le Développement, UMR 186 Interaction Plantes Microorganismes
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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
4-weeks-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 infiltrated negative controls,: they showed ttypical water-soaked 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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