- A Quantitative and Direct PCR Assay for the Subspecies-Specific Detection of Clavibacter michiganensis subsp. michiganensis Based on a Ferredoxin Reductase Gene
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Min Seok Cho , Jang Ha Lee , Nam Han Her , ChangKug Kim , Young-Joo Seol , Jang Ho Hahn , Ji Hyoun Baeg , Hong Gi Kim , Dong Suk Park
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J. Microbiol. 2012;50(3):496-501. Published online June 30, 2012
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DOI: https://doi.org/10.1007/s12275-012-1611-x
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The Gram-positive bacterium Clavibacter michiganensis subsp. michiganensis is the causal agent of canker disease in tomato. Because it is very important to control newly introduced inoculum sources from commercial materials, the specific detection of this pathogen in seeds and seedlings is essential for effective disease control. In this study, a novel and efficient assay for the detection and quantitation of C. michiganensis subsp. michiganensis in symptomless tomato and red pepper seeds was developed. A pair of polymerase chain reaction (PCR) primers (Cmm141F/R) was designed to amplify a specific 141 bp fragment on the basis of a ferredoxin reductase gene of C. michiganensis subsp. michiganensis NCPPB 382. The specificity of the primer set was evaluated using purified DNA from 16 isolates of five C. michiganensis subspecies, one other Clavibacter species, and 17 other reference bacteria. The primer set amplified a single band of expected size from the genomic DNA obtained from the C. michiganensis subsp. michiganensis strains but not from the other C. michiganensis subspecies or from other Clavibacter species. The detection limit was a single cloned copy of the ferredoxin reductase gene of C. michiganensis subsp. michiganensis. In conclusion, this quantitative direct PCR assay can be applied as a practical diagnostic method for epidemiological research and the sanitary management of seeds and seedlings with a low level or latent infection of C. michiganensis subsp. michiganensis.
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Citations
Citations to this article as recorded by  - Development and Validation of a Field-Based Colorimetric LAMP Assay for the Detection of Clavibacter michiganensis in Tomato Plants
Glykeria Mermigka, Maria Megariti, Dimitris Malliarakis, Marianthi G. Pagoulatou, Electra Gizeli, Dimitrios E. Goumas
Plants.2026; 15(3): 372. CrossRef - Clavibacter michiganensis Reframed: The Story of How the Genomics Era Made a New Face for an Old Enemy
Ebrahim Osdaghi, Hamid Abachi, Marie‐Agnes Jacques
Molecular Plant Pathology.2025;[Epub] CrossRef - Advances in the detection technology of vegetable soil borne fungi and bacteria
Lida Chen, Guiyun Lü, Songhan Yang, Binbin Gong, Yusong Lu, Xiaolei Wu, Jingrui Li, Hongbo Gao
Frontiers in Microbiology.2024;[Epub] CrossRef - Bacterial wilt and canker of tomato: fundamentals of a complex biological system
Emmanuel Martínez-Castro, Ramón Jarquin-Gálvez, Ángel Gabriel Alpuche-Solís, Moisés Roberto Vallejo-Pérez, Juan Gualberto Colli-Mull, José Pablo Lara-Ávila
Euphytica.2018;[Epub] CrossRef - Characterization and Comparison of Clavibacter michiganensis subsp. nebraskensis Strains Recovered from Epiphytic and Symptomatic Infections of Maize in Iowa
Azeem Ahmad, Gladys Y. Mbofung, Jyotsna Acharya, Clarice L. Schmidt, Alison E. Robertson, Richard A Wilson
PLOS ONE.2015; 10(11): e0143553. CrossRef
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