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KMID : 0364820090450040306
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Korean Journal of Microbiology
2009 Volume.45 No. 4 p.306 ~ p.311
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Development of a New PCR Method for Detection of Pectobacterium carotovorum
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No Ji-Na
Yoo Mi-Seon
Park Dong-Suk
Kim Jeong-Gu
Yoon Byoung-Su
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Abstract
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A new PCR method was developed to detect Pectobacterium carotovorum which is the causative agent of soft rot in Brassica pekinensis. A specific detection primer set based on Lytic murein transglycolase gene was designed and evaluated. Using ERB3_F (5’-TGC GAC ACC TCC TCA TCA CG-3’) and ERB3_R (5’-CTT ATC ACG CTG TAA CCA GC-3’) primers, 437 nucleotides long fragment was specifically amplified. The amplified products were observed in 52 out of 55 strains of P. carotovorum or Pectobacterium carotovorum subsp. carotovorum. On the other hand, no amplification was observed in 8 organisms including Chinese cabbage and potato. The optimal PCR condition for the ERB3_F/ERB3_R primer set was 58oC for annealing and 15 mM for MgCl2. With serially diluted templates, the specific PCR sensitivitie limit was 2×103 copies. Also, this method can be applied not only to DNA but also to field samples. This PCR method may be expected to be useful for specific detection of P. carotovorum.
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KEYWORD
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chinese cabbage, detection, diagnosis, P. carotovorum, soft rot
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