Abstract
Pectobacterium atrosepticum is an economically important phytopathogen that is responsible for potato blackleg and soft rot, and for which current control strategies are limited. In this study, stem samples of potato crops exhibiting blackleg were taken from three farms in Co. Cork, Ireland, and they were found to be infected with P. atrosepticum. Three closely related bacteriophages (phages) that are specific to this phytopathogen were isolated and characterized, namely vB_PatP_CB1, vB_PatP_CB3, and vB_PatP_CB4 (abbreviated as CB1, CB3, and CB4). Both CB1 and CB3 were determined to infect 12 strains and CB4 10 strains of the 19 strains of P. atrosepticum tested. Morphology, latent periods, burst sizes, and their stability at various temperatures and pHs were also examined. Genome sequencing of the three phages revealed that they shared a minimum nucleotide identity of 93% with each other. Their genomes exhibited an Enquartavirinae genome organization, possessing several conserved proteins that were associated with phages of this group, like the type species Escherichia virus N4. Tandem electrospray ionization-mass spectrometry (ESI-MS/MS) allowed for the identification of ten structural proteins that form the virion of CB1, six that are conserved in phage N4. Biocontrol experiments demonstrated that the phages suppress soft rot formation upon co-inoculation with P. atrosepticum on whole tubers. The results of this study indicate that CB1 related phages could be good candidates for phage-based control.
Highlights
The genera of Pectobacterium and Dickeya, which are often referred to collectively as the soft rot Enterobacteriaceae (SRE), are phytopathogens that cause economically important losses in a wide range of arable and ornamental crops
Isolate identification was based on cavity formation on crystal violet pectate (CVP) medium, production of reducing substances from sucrose, acid production from α-methyl-glucoside, Pectobacterium genus-specific along with P. atrosepticum and P. carotovorum subsp. carotovorum species-specific polymerase chain reactions (PCRs), and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry
From which pectolytic isolates were found, fifteen were identified to be infected with P. atrosepticum with the remaining infection being identified to be caused by a mixture of P. atrosepticum with P. carotovorum subsp. carotovorum
Summary
The genera of Pectobacterium and Dickeya, which are often referred to collectively as the soft rot Enterobacteriaceae (SRE), are phytopathogens that cause economically important losses in a wide range of arable and ornamental crops. These bacteria are all Gram-negative, non-spore forming, facultative anaerobic rods that are typified by the production of extracellular pectinolytic enzymes during plant infection [1,2]. The predominant SRE species and subspecies that cause blackleg and soft rot of the potato crop in Europe are Pectobacterium atrosepticum, Pectobacterium carotovorum subsp. The success of these schemes vary and are highly weather dependent [8,9]
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