Quantitative Real-Time Polymerase Chain Reaction Assay for Detection of Pectobacterium wasabiae Using YD Repeat Protein Gene-Based Primers

The aim of this study was to develop a quantitative polymerase chain reaction (qPCR) assay for specific detection of Pectobacterium wasabiae using a primer pair based on the YD repeat protein gene for amplification of a 140-bp DNA fragment from infected wasabi (Wasabia japonica), a member of the cru...

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Veröffentlicht in:Plant disease. - 1997. - 96(2012), 2 vom: 14. Feb., Seite 253-257
1. Verfasser: Kim, Myeong Ho (VerfasserIn)
Weitere Verfasser: Cho, Min Seok, Kim, Byoung Kyu, Choi, Hyeon Jin, Hahn, Jang Ho, Kim, ChangKug, Kang, Man Jung, Kim, Seong Hwan, Park, Dong Suk
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2012
Zugriff auf das übergeordnete Werk:Plant disease
Schlagworte:Journal Article
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245 1 0 |a Quantitative Real-Time Polymerase Chain Reaction Assay for Detection of Pectobacterium wasabiae Using YD Repeat Protein Gene-Based Primers 
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520 |a The aim of this study was to develop a quantitative polymerase chain reaction (qPCR) assay for specific detection of Pectobacterium wasabiae using a primer pair based on the YD repeat protein gene for amplification of a 140-bp DNA fragment from infected wasabi (Wasabia japonica), a member of the crucifer family. The soft rot caused by P. wasabiae is an emerging disease that is present in many wasabi-producing areas. However, specific and reliable methods for identifying the pathogen are not available. Therefore, a qPCR primer set for accurate diagnosis of P. wasabiae was developed from publically available genome sequences. A SYBR Green qPCR primer set was designed based on a YD repeat protein gene of P. wasabiae WPP163 because it is known that this gene is structurally diverse among species, pathovars, or subspecies. The specificity of the primer set was evaluated using genomic DNA from 5 isolates of P. wasabiae, 5 different species of Pectobacterium, and 16 other pathogenic reference bacteria. The primer set used in the PCR assay successfully amplified a 140-bp amplicon for all five P. wasabiae strains. No amplification was obtained from 29 other pathogenic bacteria. The assay was also able to detect at least two genomic DNA, or 3 CFU per reaction, when using calibrated cell suspension 
650 4 |a Journal Article 
700 1 |a Cho, Min Seok  |e verfasserin  |4 aut 
700 1 |a Kim, Byoung Kyu  |e verfasserin  |4 aut 
700 1 |a Choi, Hyeon Jin  |e verfasserin  |4 aut 
700 1 |a Hahn, Jang Ho  |e verfasserin  |4 aut 
700 1 |a Kim, ChangKug  |e verfasserin  |4 aut 
700 1 |a Kang, Man Jung  |e verfasserin  |4 aut 
700 1 |a Kim, Seong Hwan  |e verfasserin  |4 aut 
700 1 |a Park, Dong Suk  |e verfasserin  |4 aut 
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