Comparative Genomics to Develop a Specific Multiplex PCR Assay for Detection of Clavibacter michiganensis

• DOI: 10.1094/phyto-10-19-0405-r
• Publication Date: 2019-12-04T14:35:35Z
• Authors: Shree P. Thapa; Michael O’Leary; Marie-Agnès Jacques; Robert L. Gilbertson; Gitta Coaker
• Keywords: Clavibacter michiganensis; Multiplex; Canker

Clavibacter michiganensis is a Gram-positive bacterial pathogen that proliferates in the xylem vessels of tomato, causing wilt and canker symptoms. Accurate detection crucial step confirming outbreaks developing management strategies. A major problem with existing methods are false-positive -negative results. Here, we report use comparative genomics 37 diverse strains, including 21 strains sequenced this study, to identify specific sequences C. targets. Genome-wide phylogenic analyses revealed additional diversity within genus Clavibacter. Pathogenic varied plasmid composition, highlighting need for based on chromosomal We utilized michiganensis-specific loci develop multiplex PCR-based diagnostic platform using two genes (rhuM tomA) an internal control amplifying both plant DNA (16s ribosomal RNA). The PCR assay specifically detected from panel 110 bacteria, other spp. pathogens tomato. was adapted detect presence seed tomato materials high sensitivity specificity. In conclusion, described method represents robust, tool infected plants.