Genomic, proteomic and morphological characterization of two novel broad host lytic bacteriophages ΦPD10.3 and ΦPD23.1 infecting pectinolytic Pectobacterium spp. and Dickeya spp

• Published in: PloS one Vol. 10; no. 3; p. e0119812
• Main Authors: Czajkowski, Robert, Ozymko, Zofia, de Jager, Victor, Siwinska, Joanna, Smolarska, Anna, Ossowicki, Adam, Narajczyk, Magdalena, Lojkowska, Ewa
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 24.03.2015; Public Library of Science (PLoS)
• Subjects: Adsorption; Antibiotics; Bacteria; Bacteriophages - genetics; Bacteriophages - isolation & purification; Bacteriophages - physiology; Bacteriophages - ultrastructure; Bioinformatics; Biological control; Biotechnology; Burst size; Chromatography; Crop diseases; Crops; Disease control; Electrophoresis; Enterobacteriaceae; Enterobacteriaceae - virology; Erwinia carotovora; Fractionation; Gel electrophoresis; Gene sequencing; Genomes; Host Specificity; Infections; Laboratories; Latent period; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Morphology; Multiplicity of infection; Pathogens; Pectobacterium; Pectobacterium - virology; Pectobacterium carotovorum; Phages; Plant Diseases - microbiology; Plant Diseases - virology; Plant Tubers - microbiology; Plant Tubers - virology; Potatoes; Proteins; Proteomics; Pulsed-field gel electrophoresis; Random amplified polymorphic DNA; Seeds; Sodium lauryl sulfate; Solanum tuberosum - microbiology; Solanum tuberosum - virology; Stains; Poland; Canada; Netherlands; Europe
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0119812

Pectinolytic Pectobacterium spp. and Dickeya spp. are necrotrophic bacterial pathogens of many important crops, including potato, worldwide. This study reports on the isolation and characterization of broad host lytic bacteriophages able to infect the dominant Pectobacterium spp. and Dickeya spp. affecting potato in Europe viz. Pectobacterium carotovorum subsp. carotovorum (Pcc), P. wasabiae (Pwa) and Dickeya solani (Dso) with the objective to assess their potential as biological disease control agents. Two lytic bacteriophages infecting stains of Pcc, Pwa and Dso were isolated from potato samples collected from two potato fields in central Poland. The ΦPD10.3 and ΦPD23.1 phages have morphology similar to other members of the Myoviridae family and the Caudovirales order, with a head diameter of 85 and 86 nm and length of tails of 117 and 121 nm, respectively. They were characterized for optimal multiplicity of infection, the rate of adsorption to the Pcc, Pwa and Dso cells, the latent period and the burst size. The phages were genotypically characterized with RAPD-PCR and RFLP techniques. The structural proteomes of both phages were obtained by fractionation of phage proteins by SDS-PAGE. Phage protein identification was performed by liquid chromatography-mass spectrometry (LC-MS) analysis. Pulsed-field gel electrophoresis (PFGE), genome sequencing and comparative genome analysis were used to gain knowledge of the length, organization and function of the ΦPD10.3 and ΦPD23.1 genomes. The potential use of ΦPD10.3 and ΦPD23.1 phages for the biocontrol of Pectobacterium spp. and Dickeya spp. infections in potato is discussed.