Rapid and Specific Detection of Burkholderia glumae in Rice Seed by Real-Time Bio-PCR Using Species-Specific Primers Based on an rhs Family Gene

In this study, we developed a reliable, quick, and accurate quantitative polymerase chain reaction (qPCR) assay to detect grain rot caused by Burkholderia glumae in rice seed. The control of bacterial grain rot is difficult, and the only practical methods for disease management rely on the use of pa...

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Publié dans:Plant disease. - 1997. - 96(2012), 4 vom: 01. Apr., Seite 577-580
Auteur principal: Kim, Byoung Kyu (Auteur)
Autres auteurs: Cho, Min Seok, Kim, Myeong Ho, Choi, Hyeon Jin, Kang, Man Jung, Shim, Hong Sik, Ahn, Tae-Young, Kim, Jaisoo, Park, Dong Suk
Format: Article en ligne
Langue:English
Publié: 2012
Accès à la collection:Plant disease
Sujets:Journal Article
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520 |a In this study, we developed a reliable, quick, and accurate quantitative polymerase chain reaction (qPCR) assay to detect grain rot caused by Burkholderia glumae in rice seed. The control of bacterial grain rot is difficult, and the only practical methods for disease management rely on the use of pathogen-free seed, appropriate culture practices, and resistant cultivars. Therefore, the specific detection of this pathogen in seed is essential for effective control of the disease. However, other Burkholderia spp. are also detected by currently available molecular and serological methods. In this study, we exploited the available genome sequence information in public databases to develop specific PCR primers for accurate diagnosis of B. glumae. An SYBR Green real-time PCR primer set was designed based on the rhs family gene (YD repeat protein) of B. glumae BGR1 because these genes are structurally diverse. The specificity of the primers was evaluated using purified DNA from 5 isolates of B. glumae, 6 different species of Burkholderia, and 18 other reference pathogenic bacteria. The assay was able to detect at least one genome equivalent of cloned amplified target DNA using purified DNA or 1 CFU per reaction when using calibrated cell suspension. This method is rapid and reliable and has great potential for analyzing large numbers of samples 
650 4 |a Journal Article 
700 1 |a Cho, Min Seok  |e verfasserin  |4 aut 
700 1 |a Kim, Myeong Ho  |e verfasserin  |4 aut 
700 1 |a Choi, Hyeon Jin  |e verfasserin  |4 aut 
700 1 |a Kang, Man Jung  |e verfasserin  |4 aut 
700 1 |a Shim, Hong Sik  |e verfasserin  |4 aut 
700 1 |a Ahn, Tae-Young  |e verfasserin  |4 aut 
700 1 |a Kim, Jaisoo  |e verfasserin  |4 aut 
700 1 |a Park, Dong Suk  |e verfasserin  |4 aut 
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