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On Genotyping Bacterial Strains of the Genera Pectobacterium and Pseudomonas: Pathogens of Bacterioses in Potatoes

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Abstract

Molecular-genetic research methods make it possible to identify phytopathogens with comprehensive characteristics of their hereditary material. We suggest a genotyping method based on the use of two restriction endonucleases for the cleavage of bacteria’s genomic DNA. Taq DNA polymerase added to the reaction mixture provides labeling of DNA fragments with biotinylated deoxycytosinetriphosphate (Bio-dCTP). The label is only incorporated into DNA having 3-prime recessed ends formed by the first enzyme. The second restriction endonuclease produces only blunt ends of fragments that are unable to incorporate the label. As a result of the DDSL (double digest selective label) reaction, 20–50 distinct DNA fragments are visualized on the filter, the amount and distribution of which are specific for each bacterial strain. This research was carried out using two pairs of restriction enzymes (XbaI/DraI and XbaI/Eco24I). The results indicate the same discriminatory capacity of these enzymes. Some advantage was noted for the XbaI/DraI combination, which makes it possible to detect differences in the region of longer DNA fragments in genetic profiles. The genetic profile generated by the BcuI/Eco32I pair of enzymes in Ps. fluorescens 894 was significantly different from that of Ps. xanthochlora, which could be anticipated while comparing these two species. In general, the results of genotyping agreed with the data on the origin of bacterial strains. In particular, microorganisms with the same genetic profile, D822 and D828, G399 and G400, as well as C960 and C966, were obtained from affected tubers of the same experimental field. This suggests the possibility of the contact spread of the pathogen. In some cases, the identity of strains was observed, despite their different geographic origin. This fact confirms contamination that occurred in the past during the use of the foreign seed potato.

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REFERENCES

  1. Nabhan, S., De Boer, S.H., Maiss, E., and Wydra, K., Taxonomic relatedness between Pectobacterium carotovorum subsp. carotovorum, Pectobacterium carotovorum subsp. odoriferum and Pectobacterium carotovorum subsp. brasiliense subsp. nov., J. Appl. Microbiol., 2012, vol. 113, no. 4, pp. 904–913. https://doi.org/10.1111/j.1365-2672.2012.05383.x

    Article  CAS  PubMed  Google Scholar 

  2. Vernon-Shirley, M. and Burns, R., The development and use of monoclonal antibodies for detection of Erwinia, J. Appl. Microbiolю, 1992, vol. 72, no. 2, pp. 97–102.

    Article  Google Scholar 

  3. Peltzer, S. and Sivasithamparam, K., Sero-groups of Ec associated with water, soil, tuber and stems of potato plants in Western Australia, N. Z. J. Agric., 1988, vol. 16, no. 3, pp. 265–270.

    Google Scholar 

  4. Faure, D. and Dessaux, Y., Quorum sensing as a target for developing control strategies for the plant pathogen Pectobacterium, Eur. J. Plant Pathol., 2007, vol. 119, pp. 353–365.

    Article  CAS  Google Scholar 

  5. Essarts, Y.R., Cigna, J., Quetu-Laurent, A., Caron, A., Munier, E., Beury-Cirou, A., Helias, V., and Faurea, D., Biocontrol of the potato blackleg and soft rot diseases caused by Dickeya dianthicola, Appl. Environ. Microbiol., 2016, vol. 82, no. 1, pp. 268–278. https://doi.org/10.1128/AEM.02525-15

    Article  CAS  Google Scholar 

  6. Pishchik, V.N., Chernyaeva, I.I., Vorobjev, N.I., and Lazarev, A.M., Characteristic features of virulent and avirulent strains of Erwinia carotovora, Microbiology (Moscow), 1996, vol. 65, no. 2, pp. 232–237.

    Google Scholar 

  7. Pishchik, V.N., Provorov, N.A., Vorobjov, N.I., Chizevskaya, E.P., Safronova, V.I., Kozhemyakov, A.P., and Tuev, A.N., Interactions between plants and associated bacteria in soils contaminated with heavy metals, Microbiology (Moscow), 2009, vol. 78, no. 6, pp. 785–793. https://doi.org/10.1134/S0026261709060162

    Article  CAS  Google Scholar 

  8. Xiu, J.H., Ji, G.H., Wang, M., Yang, Y.L., and Li, C.Y., Molecular identification and genetic diversity in Konnyaku’s soft rot bacteria, Acta Microbiol. Sin., 2006, vol. 46, no. 4, pp. 522–525.

    CAS  Google Scholar 

  9. Nabhan, S., Wydra, K., Linde, M., and Debener, T., The use of two complementary DNA assays, AFLP and MLSA, for epidemic and phylogenetic studies of pectolytic enterobacterial strains with focus on the heterogeneous species Pectobacterium carotovorum, Plant Pathol., 2012, vol. 61, pp. 498–508. https://doi.org/10.1111/j.1365-3059.2011.02546.x

  10. Terletskiy, V.P., Tyshchenko, V.I., Novikova, O.B., Borisenkova, A.N., Belash, D.E., and Yakovlev, A.F., The efficient molecular genetic technique for identification of Salmonella and Proteus strains, Russ. Agricul. Sci., 2013, no. 5, pp. 60–63.https://doi.org/10.3103/S1068367413060220

  11. Czajkowski, R., Perombelon, M.C.M., Jafra, S., Lojkowska, E., Potrykus, M., van der Wolf, J.M., and Sledz, W., Detection, identification and differentiation of Pectobacterium and Dickeya species causing potato blackleg and tuber soft rot: a review, Ann. Appl. Biol., 2015, vol. 166, pp. 18–38. https://doi.org/10.1111/aab.12166

    Article  CAS  PubMed  Google Scholar 

  12. Terletskiy, V.P., Tyshchenko, V.I., Novikova, I.I., Boikova, I.V., Tyulebaev, S.D., and Shakhtamirov, I.Y., An efficient method for genetic certification of bacillus subtilis strains, prospective producers of biopreparations, Microbiology (Moscow), 2016, vol. 85, no. 1, pp. 71–76. https://doi.org/10.1134/S0026261716010136

  13. Terletskiy, V., Kuhn, G., Francioli, P., and Blanc, D., Application and evaluation of double digest selective label (DDSL) typing technique for Pseudomonas aeruginosa hospital isolates, J. Microbiol. Methods, 2008, vol. 72, no. 3, pp. 283–287. https://doi.org/10.1016/j.mimet.2007.12.006

    Article  CAS  PubMed  Google Scholar 

  14. Vinogradova, S.V., Kyrova, E.I., and Ignatov, A.N., Complete genome sequencing of phytopathogenic bacteria, Potato Protection, 2014, no. 2, pp. 15–17.

  15. Khayi, S., Blin, P., Chong, T.M., Chan, K.G., and Faure, D., Complete genome anatomy of the emerging potato pathogen Dickeya solani type strain IPO 2222(T), Stand. Genomic Sci., 2016, vol. 11, pp. 87. https://doi.org/10.1186/s40793-016-0208-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ignatov, A.N., Lazarev, A.M., Panycheva, J.S., Provorov, N.A., and Chebotar, V.K., Potato phytopathogens of genus Dickeya—a mini review of systematic and etiology of diseases, Agricult. Biol., 2018, vol. 53, no. 1, pp. 123–131. https://doi.org/10.15389/agrobiology.2018.1.123eng

    Article  Google Scholar 

  17. Toth, I.K., van der Wolf, J.M., Saddler, G., Lojkowska, E., Helias, V., Pirhonen, M., and Elphinstone, J.G., Dickeya species: an emerging problem for potato production in Europe, Plant Pathol., 2011, vol. 60, no. 3, pp. 385–399.

    Article  Google Scholar 

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Authors and Affiliations

  1. All-Russia Research Institute of Plant Protection, 196608, St. Petersburg, Pushkin, Russia

    V. P. Terletskiy & A. M. Lazarev

  2. Pushkin Leningrad State University, 196605, St. Petersburg, Russia

    V. P. Terletskiy

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  1. V. P. Terletskiy
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  2. A. M. Lazarev
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Correspondence to V. P. Terletskiy or A. M. Lazarev.

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Translated by A. Panyushkina

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Terletskiy, V.P., Lazarev, A.M. On Genotyping Bacterial Strains of the Genera Pectobacterium and Pseudomonas: Pathogens of Bacterioses in Potatoes. Cytol. Genet. 53, 212–218 (2019). https://doi.org/10.3103/S0095452719030058

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