TSitologiya i Genetika 2019, vol. 53, no. 3, 38-46
Cytology and Genetics 2019, vol. 53, no. 3, 212–218, doi: https://www.doi.org/10.3103/S0095452719030058

To the question of genotyping strains of bacteria of the genera Pectobacterium and Pseudomonas – the pathogens of bacterioses of potatoes

Terletskiy V.P., Lazarev A.M.

  1. Федеральное государственное бюджетное научное учреждение «Всероссийский научно­исследовательский институт защиты растений», Санкт­Петербург, 196608, Россия
  2. Государственное автономное образовательное учреждение высшего образования Ленинградской области «Ленинградский государственный университет им. А.С. Пушкина», Санкт­Петербург, 196605, Россия

SUMMARY. Molecular genetic research methods make it possible to identify phytopathogens with comprehensive characteristics of their hereditary material. Our method of genotyping phytopathogenic bacteria is based on the use of two restriction endonucleases for digesting the genomic DNA (DDSL). The Taq DNA polymerase included in the reaction mixture provides labeling of DNA fragments with biotinylated deoxycytosinetriphosphate (Bio-dCTP). The label is incorporated only 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 a label. As a result of the DDSL reaction, 20–50 distinct DNA fragments are visualized on the filter, the amount and distribution of which is characteristic for each bacterial strain. The work was carried out using two pairs of restriction enzy-mes – XbaI/DraI and XbaI/Eco24I; the results indicate the same discriminatory ability of these combinations of enzymes. Some advantage is noted for the combination XbaI/DraI, which allows detecting differences in genetic profiles in the region of longer DNA fragments. The genetic profile generated by enzyme pair BcuI-Eco32I in Ps. fluorescens 894, was significantly different from that of Ps. xanthochlora, which could be anticipated considering that these are two different bacterial species. In general, genotyping data correlate with data concerning the origin of bacterial strains. In particular, strains with the same genetic profile, D822 and D828, G399 and G400, C960 and C966 were obtained from affected tubers of the same experimental fields, which allow the possibility of contact spread of the pathogen. In some cases, the identity of genetic profiles is demonstrated, despite their different geographic origins. This fact confirms the contamination that occurred in the past during the exchange of seed material.

Keywords: potato pathogens, strains, restriction endonucleases, genotyping

TSitologiya i Genetika
2019, vol. 53, no. 3, 38-46

Current Issue
Cytology and Genetics
2019, vol. 53, no. 3, 212–218,
doi: 10.3103/S0095452719030058

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