Skip to main content
Log in

Molecular, morphological and pathogenic characterization of Clavibacter michiganensis subsp. sepedonicus strains of different geographic origins in Georgia

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript


Potato ring rot and its causative pathogen Clavibacter michiganensis subsp. sepedonicus (Cms) have been studied in Georgia for the first time. The samples were collected in the fields and potato storage facilities in various potato producing regions of Georgia. Potato tubers were also collected at agrarian markets and the border checkpoints, from which potatoes are imported from neighboring countries. In 124 potato samples the detection of Cms was performed using specific PCR analysis and bioassay test. A total of 20 Cms isolates were purified and identified by subspecies-specific PSA-1 and PSA-R primers set. The isolates were characterized according to cell morphology, cultural-biochemical properties and virulence. The study of genetic diversity using molecular typing methods of PCR melting profile with different restriction enzymes and variable number tandem repeats PCR revealed significant differences between Cms isolates, which is not related to their geographical origin.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others


  • Abdelhai, M. H., Hassanin, H. A. M., & Sun, X. (2016). Comparative study of rapid DNA extraction methods of pathogenic Bacteria. American Journal of Bioscience and Bioengineering, 4(1), 1–8.

  • Anonymous. (1996). International standards for Phytosanitary measures. Part 1 - import regulations. Guidelines for pest risk analysis. Publication no. 2. FAO, Rome. 45. Plant Protect. Sci., 43, 35–46.

    Google Scholar 

  • Baer, D., & Gudmestad, C. N. (1993). Serological detection of nonmucoid strains of Clavibacter michiganensis subsp. sepedonicus in potato. Phytopathology, 83, 157–163.

    Article  Google Scholar 

  • Bergey, D. H., Holt, J. G., et al. (1994). Bergey's manual of determinative bacteriology (9th ed.). Baltimore: Williams & Wilkins.

    Google Scholar 

  • Bragard, C., Dehnen-Schmutz, K., Di Serio, F., Gonthier, P., Jaques Miret, J. A., Justesen, A. F., et al. (2019). Scientific opinion on the pest categorisation of Clavibacter sepedonicus. EFSA Journal, 17(4), 5670, 26 pp.

    Article  Google Scholar 

  • Dams, E. L., Hendriks, L., Van de Peer, Y., Neefs, J. M., Smits, G., Vandenbempt, I., & De Wachter, R. (1988). Compilation of small ribosomal subunit RNA sequences. Nucleic Acids Research, 16(Suppl), r87–r173.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • De Boer, S. H., & McCann, M. (1990). Detection of Corynebacterium sepedonicum in potato cultivars with different propensities to express ring rot symptoms. American Potato Journal, 67, 685–695.

    Article  Google Scholar 

  • De la Cruz, A. R., Wiese, M. V., & Schaad, N. W. (1992). A semiselective agar medium for isolation of Clavibacter michiganensis subsp. sepedonicus from potato tissues. Plant Disease, 76, 830–834.

    Article  Google Scholar 

  • Dinesen, I. G., & DeBoer, S. H. (1995). Extraction of Clavibacter michiganensis subsp. sepedonicus from composite samples of potato tubers. American Potato Journal, 72, 133–142.

    Article  Google Scholar 

  • Dziva, F., Muhairwa, A. P., Bisgaard, M., & Christensen, H. (2008). Diagnostic and typing options for investigating diseases associated with Pasteurella multocida. Veterinary Microbiology, 128, 1–22.

    Article  CAS  PubMed  Google Scholar 

  • Eichenlaub, R., & Gartemann, K. H. (2011). The Clavibacter michiganensis subspecies: Molecular investigation of gram-positive bacterial plant pathogens. Annual Review of Phytopathology, 49, 445–464.

    Article  CAS  PubMed  Google Scholar 

  • Elphinstone, J.G. (2010) Bacterial ring rot of potato - the facts (Clavibacter michiganensis subsp. sepedonicus). Summarized version. Available online:

  • Franc, G. D. (1999). Persistence and latency of Clavibacter michiganensis subsp. sepedonicus in field-grown seed potatoes. Plant Disiase, 83, 247–250.

    Article  Google Scholar 

  • Hayward, A. C., & Waterston, J. M. (1964). Corynebacterium michiganense. Descriptions of Pathogenic Fungi and Bacteria, No. 19. Wallingford: CAB International.

  • Heath, M. C. (2000). Hypersensitive response-related death. Plant Molecular Biology, 44, 321–334.

    Article  CAS  PubMed  Google Scholar 

  • Janse, J. D., & Wenneker, M. (2002). Possibilities of avoidance and control of bacterial plant diseases when using pathogen-tested (certified) or – Treated planting material. Plant Pathology, 51, 523–536.

    Article  Google Scholar 

  • Jansing, H., & Rudolph, K. (1998). Physiological capabilities of Clavibacter michiganensis subsp. sepedonicus and development of a semi-selective medium. Journal of Plant Pathology, 105, 590–601.

    CAS  Google Scholar 

  • Koneman, E. W., Allen, S. D., Janda, W. M., Schreckenberger, P. C., & Winn Jr., W. C. (1992). Colour atlas and textbook of diagnostic microbiology (4th ed.). Philadelphia: J. B. Lippinccott Co..

    Google Scholar 

  • Lelliott, R. A. & Stead, D. A. (1987). Methods for diagnosis of bacterial diseases of plants. Methods in plant pathology, vol. 2 T. F. Preece (Ed.), Oxford (GB): Blackwell scientific publications.

  • de León, L., Rodríguez, A., López, M. M., & Siverio, F. (2008). Evaluation of the efficacy of immunomagnetic separation for the detection of Clavibacter michiganensis subsp. michiganensis in tomato seeds. J. Appl. Microbiol., 104(3), 776–786.

    Article  PubMed  Google Scholar 

  • Li, X., Tambong, J., Yuan, K. X., Chen, W. H., Lévesque, C. A., & De Boer, S. H. (2018). Re-classification of Clavibacter michiganensis subspecies on the basis of whole-genome and multi-locus sequence analyses. International Journal of Systematic and Evolutionary Microbiology, 68(1), 234–240.

    Article  CAS  PubMed  Google Scholar 

  • Lopez, M. M., Bertolini, E., Caruso, P., Penyalver, R., Marco-Noales, E., Gorris, M. T., Morente, C., Salcedo, C., Cambra, M., & Llop, P. (2005). Advantages of integrated approach for diagnosis of quarantine pathogenic bacteria in plant material. Phytopathologia Polonica, 35, 49–56.

    Google Scholar 

  • MacFaddin JF, editor. Biochemical Tests for Identification of Medical Bacteria. 3rd ed. Philadelphia:Lippincott Williams and Wilkins; 2000. p. 363–367.

  • Maciejewska, A. (2018). Analysis of variability of selected isolates of Clavibacter michiganensis subsp. sepedonicus (Spieckermann & Kotthoff) Davis et al., PhD thesis.

  • Malko, A., Frantsuzov, P., Nikitin, M., Statsyuk, N., Dzhavakhiya, V., & Golikov, A. (2019). Potato pathogens in Russia’s regions: An instrumental survey with the use of real-time PCR/RT-PCR in matrix format. Pathogens, 8(1).

  • OEPP/EPPO. (2006). EPPO standards PM 7 / 59 (1) diagnostic protocol – Clavibacter michiganensis subsp. sepedonicus. Bulletin OEPP / EPPO Bulletin, 36, 99–109.

    Article  Google Scholar 

  • Pastrik, K. H., & Rainey, F. A. (1999). Identification and differentiation of Clavibacter michiganensis subspecies by polymerase chain reaction-based techniques. Journal of Phytopathology, 147, 687–693.

    Article  CAS  Google Scholar 

  • Sambrook, J., Fritschi, E. F., & Maniatis, T. (1989). Molecular cloning: A laboratory manual. New York: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Schaad, N. W, Jones, J. B & Chu, W. (2001). Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed, pp. 218–235. American Phytopathological Society, St Paul (US.)

  • Slack, S. A. (1987). Biology and ecology of Corynebacterium sepedonicum. American Potato Journal, 64, 665–670.

    Article  Google Scholar 

  • Smith, J. M., Mc Namara D. G., Scott P R., Holderness M., Burger B. (1997). Clavibacter michiganensis subsp. sepedonicus. In CAB International (Ed), Quarantine pests for Europe (pp.986–990). University Press, Cambridge, UK.

  • Steel, K. J. (1961). The oxidase reaction as a taxonomic tool. J. Gen. MicroMol., 25, 297–806.

    Article  Google Scholar 

  • Van Vaerenbergh, J., De Paepe, B., Hoedekie, A., Van Malderghem, C., Zaluga, J., De Vos, P., & Maes, M. (2016). Natural infection of Clavibacter michiganensis subsp. sepedonicus in tomato (Solanum tuberosum). New Disease Reports, 33, 7.

    Article  Google Scholar 

  • Van der Wolf, J. M., Elphinstone, J. G., Stead, D. E., Metzler, M., Müller, P., Hukkanen, A. & Karjalainen, R. (2005). Epidemiology of Clavibacter michiganensis subsp. sepedonicus in relation to control of bacterial ring rot, Plant Research International B.V., Wageningen, February 2005, Report 95.

  • Węgierek-Maciejewska, A., Masny, A., Wołkowicz, T., Jagiełło, A., & Arseniuk, E. (2019). PCR MP method for differentiation of Clavibacter michiganensis subsp. sepedonicus isolates. American Journal Potato Research, 96(6), 588–604.

    Article  Google Scholar 

  • Żaczek, A., Struś, K., Sokołowska, A., Parniewski, P., Wojtasik, A., & Dziade, J. (2018). Differentiation of Clavibacter michiganensis subsp. sepedonicus using PCR melting profile and variable number of tandem repeat methods. Letters in Applied Microbiology, 68(1), 24–30.

    Article  PubMed  Google Scholar 

Download references


The Study was supported by the Regional Cooperative Research Grant project #60958, funded by the U.S. Defense Threat Reduction Agency through CRDF Global/GRDF.

We thank the State Laboratory of Agriculture of Georgia for kindly provided potato samples from Georgia’s border checkpoints.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Tinatin Sadunishvili.

Ethics declarations

Authors declare that the manuscript is entirely original and have not been, and will not be, published elsewhere if accepted for publishing in EJPP. The content and authorship of the submitted manuscript has been approved by all authors, and all prevailing local, national and international regulations and conventions, and normal scientific ethical practices have been respected. Authors declare no competing interests.

Conflict of interest

The authors TS, A.W-M., EA, DG, NA, NS, LA, SK and GK declare that they have no conflict of interest. Each authors provided the corresponding author with signed potential conflict of interest disclosure form.

Human and animal rights

This article does not contain any studies with human or animal subjects performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sadunishvili, T., Węgierek-Maciejewska, A., Arseniuk, E. et al. Molecular, morphological and pathogenic characterization of Clavibacter michiganensis subsp. sepedonicus strains of different geographic origins in Georgia. Eur J Plant Pathol 158, 195–209 (2020).

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: