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Sensitive and Specific Detection of Xanthomonas oryzae pv. oryzae by Real-Time Bio-PCR Using Pathovar-Specific Primers Based on an rhs Family Gene

May 2011 , Volume 95 , Number  5
Pages  589 - 594

Min Seok Cho, Man Jung Kang, Chang Kug Kim, Young-Joo Seol, Jang Ho Hahn, Soo Chul Park, and Duk Ju Hwang, National Academy of Agricultural Science, Rural Development Administration, 441-707 Suwon, Republic of Korea; Tae-Young Ahn, Department of Microbiology, Dankook University, Cheonan 330-714, Republic of Korea; and Duck Hwan Park and Chun Keun Lim, Division of Bio-resources Technology, College of Agriculture and Life Sciences, Kangwon National University, Chuncheon 200-701, Republic of Korea; Dong Suk Park, National Academy of Agricultural Science, Rural Development Administration, 441-707 Suwon, Republic of Korea



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Accepted for publication 14 January 2011.
Abstract

The present study describes bio-polymerase chain reaction (PCR) assays to detect bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae in rice. Successful control of X. oryzae. pv. oryzae requires a specific and reliable diagnostic tool. However, other X. oryzae pathovars are detected by currently available molecular and serological methods. In this study, SYBR Green real-time and conventional PCR primer sets were designed based on an rhs family gene of X. oryzae pv. oryzae KACC10331 because these genes are structurally diverse. The specificity of the primers was evaluated using purified DNA from 11 isolates of two X. oryzae pathovars, 21 other Xanthomonas species, and 4 other reference phytopathogenic bacteria and fungi. The assay was also able to detect at least two genome equivalents of cloned amplified target DNA using purified DNA. Thus, the SYBR Green real-time PCR-based method can be used for the rapid and specific detection of X. oryzae pv. oryzae and will potentially simplify and facilitate diagnosis and monitoring of this pathogen and guide plant disease management.



© 2011 The American Phytopathological Society