Вивчення життєздатних бактеріофагів за допомогою активації псевдолізогенного стану Pectobacterium carotovorum

Abstract

The rare occurrence of viable bacteriophages is an issue in studying bacterial-phage systems of the phytopathogenic bacterium Pectobacterium carotovorum subsp. carotovorum (Pcc). Aim. To obtain viable bacteriophages by activating pseudolysogeny of the Pcc strains. Methods. To obtain bacteriophage F66, isogenic clones of Nalr mutants of strains Pcc 66A and Pcc M2-4 were used. To study the properties of the phage, ultracentrifugation of viral particles, electron microscopy, electrophoretic separation, and comparative restriction analysis of virion DNA were used. Results. It was shown that some isogenic clones of Nalr -mutants of Pcc 66A and Pcc M2-4 can produce viable bacteriophages under conditions of intensive aeration. At the same time, induced phages are able to reproduce on the parent bacterial strains, which they were isolated from (pseudolysogenic response). A pure line was obtained for the phage isolate induced from pseudolysogenic Pcc 66A Nalr by consecutive single colony passages. This phage, named F66, was shown to be temperate and able to lyse and lysogenize strains of pectolytic bacteria isolated from soft rot affected potato tubers. Phage F66 has а virion with the A1 morphotype characterized by an isometric head with an average diameter of 52.0 ± 2.1 nm and a contractile tail with a length of 115.4 ± 3.2 nm. Showing low stability under environmental conditions, phage F66 differs significantly from the temperate phage ZF40 P. carotovorum, which has a similar virion morphology. Compared phages also differ in the restriction fragment patterns obtained using endonucleases HindIII, BamHI, and HpaI. Conclusions. Temperate phage F66 is a novel P. carotovorum virus. The method for activating the pseudolysogenic state proposed in the article is useful for obtaining viruses of phytopathogenic bacteria.