Skip to main content
SpringerLink
Log in

PCR Identification of Pathogenic Agrobacteria Detected in Vineyards of Krasnodar Krai Based on the Type of Ti Plasmids

  • Original Research
  • Published:
Moscow University Biological Sciences Bulletin Aims and scope Submit manuscript

Abstract—

Agrobacterium vitis and some strains of A. tumefaciens and A. rhizogenes cause a dangerous chronic disease on grape plants—crown gall. Molecular-genetic methods were used in the work to identify and study the diversity of pathogenic agrobacteria that cause crown gall in the vineyards of Krasnodar krai. The research material was 76 samples of young and lignified tumor-like outgrowths of crown gall of grapes collected from plants in 14 vineyards of Krasnodar krai with different origins of planting stock (Italy, France, Austria, Serbia, Bulgaria, and Russia). In all studied samples, the presence of A. vitis was identified using the PGF/PGR test-system for DNA analysis. When studying the diversity of agrobacteria by the type of Ti plasmids contained therein, four groups were identified: containing octopine-type Ti plasmids (59.5%), octopine/vitopine (13.5%), vitopine (13.5%), and nopaline type (13.5%). The obtained data indicate that crown gall of grapes in the vineyards of Krasnodar krai is caused by agrobacteria A. vitis, mainly of the octopine type.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Gelvin, S.B., Agrobacterium-mediated plant transformation: The biology behind the “gene-jockeying” tool, Microbiol. Mol. Biol. Rev., 2003, vol. 67, no. 1, pp. 16–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Tarbah, F.A. and Goodman, R.N., Rapid detection of Agrobacterium tumefaciens in grapevine propagating material and the basis for an efficient indexing system, Plant Dis., 1986, vol. 70, pp. 566–568.

    Article  Google Scholar 

  3. Burdinskaya, V.F. and Arestova, N.O., Latent infestation of grapes with bacterial cancer, Zashch. Karantin Rast., 2010, no. 10, pp. 38–39.

    Google Scholar 

  4. Kerr, A. and Panagopoulos, C.G., Biotypes of Agrobacterium radiobacter var. tumefaciens and their biological control, J. Phytopathol., 1977, vol. 90, no. 2, pp. 172–179.

    Article  Google Scholar 

  5. Ophel, K. and Kerr, A., Agrobacterium vitis sp. nov. for strains of Agrobacterium biovar 3 from grapevines, Int. J. Syst. Bacteriol., 1990, vol. 40, no. 3, pp. 236–241.

    Article  CAS  Google Scholar 

  6. Mousavi, S.A., Osterman, J., Wahlberg, N., Nesme, X., Lavire, C., Vial, L., Paulin, L., de Lajudie, P., and Lindstrom, K., Phylogeny of the Rhizobium-Allorhizobium-Agrobacterium clade supports the delineation of Neorhizobium gen. nov., Syst. Appl. Microbiol., 2014, vol. 37, no. 3, pp. 208–215.

    Article  CAS  PubMed  Google Scholar 

  7. Ramírez-Bahena, M.H., Vial, L., Lassalle, F., Diel, B., Chapulliot, D., Daubin, V., Nesme, X., and Muller, D., Single acquisition of protelomerase gave rise to speciation of a large and diverse clade within the Agrobacterium/Rhizobium supercluster characterized by the presence of a linear chromid, Mol. Phylogenet. Evol., 2014, vol. 73, pp. 202–207.

    Article  CAS  PubMed  Google Scholar 

  8. Slater, S.C., Goldman, B.S., Goodner, B., et al., Genome sequences of three agrobacterium biovars help elucidate the evolution of multichromosome genomes in bacteria, J. Bacteriol., 2009, vol. 191, no. 8, pp. 2501–2511.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Szegedi, E., Opines in naturally infected crown gall tumors, Vitis, 2003, vol. 42, no. 1, pp. 39–41.

    CAS  Google Scholar 

  10. Haas, J.H., Moore, L.W., Ream, W., and Manulis, S., Universal PCR primers for detection of phytopathogenic Agrobacterium strains, Appl. Environ. Microbiol., 1995, vol. 61, no. 8, pp. 2879–2884.

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Matveeva, T.V., Bogomaz, D.I., and Lutova, L.A., A method for the diagnostics of biomaterials for the presence of agrobacteria, RF Patent 2458142, Byull. Izobret., 2012, no. 22.

    Google Scholar 

  12. Pulawska, J., Willems, A., and Sobiczewski, P., Rapid and specific identification of four Agrobacterium species and biovars using multiplex PCR, Syst. Appl. Microbiol., 2006, vol. 29, no. 6, pp. 470–479.

    Article  CAS  PubMed  Google Scholar 

  13. Sawada, H., Ieki, H., and Matsuda, I., PCR detection of Ti and Ri plasmid from phytopathogenic Agrobacte-rium strains, Appl. Environ. Microbiol., 1995, vol. 61, no. 2, pp. 828–831.

    Google Scholar 

  14. Suzaki, K., Yoshida, K., and Sawada, H., Detection of tumorigenic Agrobacterium strains from infected apple saplings by colony PCR with improved PCR primers, J. Genet. Plant Pathol., 2004, vol. 70, no. 6, pp. 342–347.

    Article  CAS  Google Scholar 

  15. Bini, F., Kuczmog, A., and Putnoky, P., Novel pathogen-specific primers for the detection of Agrobacterium vitis and Agrobacterium tumefaciens, Vitis, 2008, vol. 47, no. 3, pp. 181–190.

    CAS  Google Scholar 

  16. Eastwell, K.C., Kenneth, C., Leslie Willis, G., and Cavileer, D.T., A rapid and sensitive method to detect Agrobacterium vitis in grapevine cuttings using the polymerase chain reaction, Plant Dis., 1995, vol. 79, no. 8, pp. 822–827.

    Article  CAS  Google Scholar 

  17. Herlache, T.C., Hotchkiss, A.T., Burr, T.J., and Collmer, A., Characterization of the Agrobacterium vitis pehA gene and comparison of the encoded polygalac-turonase with the homologous enzymes from Erwinia carotovora and Ralstonia solanacearum, Appl. Environ. Microbiol., 1997, vol. 63, no. 1, pp. 338–346.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Kawaguchi, A., Sawada, H., Inoue, K., and Nasu, H., Multiplex PCR for the identification of Agrobacterium biovar 3 strains, J. Genet. Plant Pathol., 2005, vol. 71, no. 1, pp. 54–59.

    Article  CAS  Google Scholar 

  19. Lim, S.H., Kim, J.G., and Kang, H.W., Novel SCAR primers for specific and sensitive detection of Agrobacterium vitis strains, Microbiol. Res., 2009, vol. 164, no. 4, pp. 451–460.

    Article  CAS  PubMed  Google Scholar 

  20. Canaday, J., Gerard, J.C., Crouzet, P., and Otten, L., Organization and functional analysis of three T-DNAs from the vitopine Ti plasmid pTiS4, Mol. Gen. Genet., 1992, vol. 235, nos. 2–3, pp. 292–303.

    Article  CAS  PubMed  Google Scholar 

  21. Szegedi, E. and Bottka, S., Detection of Agrobacterium vitis by polymerase chain reaction in grapevine bleeding sap after isolation on a semiselective medium, Vitis, 2002, vol. 41, no. 1, pp. 37–42.

    CAS  Google Scholar 

  22. Szegedi, E., Bottka, S., Mikulas, J., Otten, L., and Sule, S., Characterization of Agrobacterium tumefaciens strains isolated from grapevine, Vitis, 2005, vol. 44, no. 1, pp. 49–54.

    CAS  Google Scholar 

  23. Bini, F., Geider, K., and Bazzi, C., Detection of Agrobacterium vitis by PCR using novel virD2 gene-specific primers that discriminate two subgroups, Eur. J. Plant Pathol., 2008, vol. 122, no. 3, pp. 403–411.

    Article  CAS  Google Scholar 

  24. Argun, N., Momol, M.T., Maden, S., Momol, E.A., Reid, C.L., Celek, H., and Burr, T.J., Characterization of Agrobacterium vitis strains isolated from Turkish grape cultivars in the Central Anatolia region, Plant Dis., 2002, vol. 86, no. 2, pp. 162–166.

    Article  CAS  PubMed  Google Scholar 

  25. Lamovšek, J., Zidaric, I., Pleško, I.M., Urek, G., and Trdan, S., Comparative study of diagnostic methods used for monitoring of common grape vine (Vitis vinifera L.) crown gall (Agrobacterium vitis Ophel & Kerr) in Slovenia, Acta Agric. Slov., 2015, vol. 103, no. 2, pp. 313–321.

    Article  Google Scholar 

  26. Genov, I., Atanassov, I., Tsvetkov, I., and Atanassov, A., Isolation and characterization of Agrobacterium strains from grapevines in Bulgarian vineyards and wild grapes, V. vinifera ssp. silvestris, Vitis, 2015, vol. 45, no. 2, pp. 97–101.

    Google Scholar 

  27. Winans, S.C., Allenza, P., Stachel, S.E., McBride, K.E., and Nester, E.W., Characterization of the virE operon of the Agrobacterium Ti plasmid pTiA6, Nucleic Acid Res., 1987, vol. 15, no. 2, pp. 825–837.

    Article  CAS  PubMed  Google Scholar 

  28. Tolba, I.H. and Zaki, M.F., Characterization of Agrobacterium vitis isolates obtained from galled grapevine plants in Egypt, Ann. Agric. Sci., 2011, vol. 56, no. 2, pp. 113–119.

    Article  Google Scholar 

  29. Kuzmanović, N., Vanović, M., Prokić, A., Gašic, K., Blagojević, N., Puławska, J., and Obradović, A., Identification and characterization of Agrobacterium spp. isolated from apricot in Serbia, Eur. J. Plant Pathol., 2013, vol. 137, no. 1, pp. 11–16.

    Article  CAS  Google Scholar 

  30. Rouhrazi, K. and Rahimian, H., Characterization of Iranian grapevine isolates of Rhizobium (Agrobacterium) spp., J. Plant Pathol., 2012, vol. 94, no. 3, pp. 555–560.

    Google Scholar 

  31. Filo, A., Sabbatini, P., Sundin, G.W., Zabadal, T.J., Safe, G.R., and Cousins, P.S., Grapevine crown gall suppression using biological control and genetic engineering: A review of recent research, Am. J. Enol. Vitic., 2013, vol. 64, no. 1. doi 10.5344/ajev.2012.12038

    Google Scholar 

  32. Doyle, J.J. and Doyle, J.L., A rapid DNA isolation procedure for small quantities of fresh tissue, Phytochem. Bull., 1987, vol. 19, pp. 11–15.

    Google Scholar 

  33. Makarkina, M.V., Ilnitskaya, E.T., and Stepanov, I.V., Identification of Agrobacteria on grape plants with symptoms of crown gall lesions in ampelocenosises of Krasnodar Territory using the PCR method, Russ. Agric. Sci., 2017, vol. 43, no. 5, pp. 402–405.

    Article  Google Scholar 

  34. Makarkina, M.V. and Il’nitskaya, E.T., Identification of the latent stage of bacterial cancer of grapes by the method of polymerase chain reaction, Nauchn. Tr. Sev.-Kavk. Fed. Nauchn. Tsentra Sadovod. Vinograd. Vinodel., 2017, vol. 13, pp. 100–104.

    Google Scholar 

  35. Ignatov, A.N., Khodykina, M.V., Vinogradova, S.V., Polityko, V.A., Kornev, K.P., and Mazurin, E.S., First report of Agrobacterium vitis causing crown galls of wine grape in Russia, Plant Dis., 2016, vol. 100, no. 4, p. 853.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. North Caucasian Federal Scientific Center of Horticulture, Viticulture, and Winemaking, Krasnodar, 350901, Russia

    M. V. Makarkina, M. V. Makarkina, E. T. Ilnitskaya & S. V. Tokmakov

Authors
  1. M. V. Makarkina
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. T. Ilnitskaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. V. Tokmakov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. V. Makarkina.

Additional information

Russian Text © The Author(s), 2019, published in Vestnik Moskovskogo Universiteta, Seriya 16: Biologiya, 2019, Vol. 74, No. 1, pp. 50–58.

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Makarkina, M.V., Ilnitskaya, E.T. & Tokmakov, S.V. PCR Identification of Pathogenic Agrobacteria Detected in Vineyards of Krasnodar Krai Based on the Type of Ti Plasmids. Moscow Univ. Biol.Sci. Bull. 74, 40–47 (2019). https://doi.org/10.3103/S0096392519010048

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.3103/S0096392519010048

Keywords

Search

Navigation