title: Genome, Proteome and Structure of a T7-Like Bacteriophage of the Kiwifruit Canker Phytopathogen Pseudomonas syringae pv. actinidiae.
abstract: Pseudomonas syringae pv. actinidiae is an economically significant pathogen responsible for severe bacterial canker of kiwifruit (Actinidia sp.). Bacteriophages infecting this phytopathogen have potential as biocontrol agents as part of an integrated approach to the management of bacterial canker, and for use as molecular tools to study this bacterium. A variety of bacteriophages were previously isolated that infect P. syringae pv. actinidiae, and their basic properties were characterized to provide a framework for formulation of these phages as biocontrol agents. Here, we have examined in more detail φPsa17, a phage with the capacity to infect a broad range of P. syringae pv. actinidiae strains and the only member of the Podoviridae in this collection. Particle morphology was visualized using cryo-electron microscopy, the genome was sequenced, and its structural proteins were analysed using shotgun proteomics. These studies demonstrated that φPsa17 has a 40,525 bp genome, is a member of the T7likevirus genus and is closely related to the pseudomonad phages φPSA2 and gh-1. Eleven structural proteins (one scaffolding) were detected by proteomics and φPsa17 has a capsid of approximately 60 nm in diameter. No genes indicative of a lysogenic lifecycle were identified, suggesting the phage is obligately lytic. These features indicate that φPsa17 may be suitable for formulation as a biocontrol agent of P. syringae pv. actinidiae.
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. andrew.pitman@plantandfood.co.nz.|New Zealand Institute for Plant and Food Research Limited, Private Bag 4704, Christchurch 8140, New Zealand. andrew.pitman@plantandfood.co.nz.
authors.forename: Rebekah A
authors.initials: RA
authors.lastname: Frampton
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. elpzacedo@gmail.com.|Departamento de Genetica, Universidad de Extremadura, Badajoz 06080, Spain. elpzacedo@gmail.com.
authors.forename: Elena Lopez
authors.initials: EL
authors.lastname: Acedo
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. vivienne.young@otago.ac.nz.
authors.forename: Vivienne L
authors.initials: VL
authors.lastname: Young
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. cheda856@student.otago.ac.nz.
authors.forename: Danni
authors.initials: D
authors.lastname: Chen
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. tonyu031@student.otago.ac.nz.
authors.forename: Brian
authors.initials: B
authors.lastname: Tong
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. corinda.taylor@otago.ac.nz.
authors.forename: Corinda
authors.initials: C
authors.lastname: Taylor
authors.affiliation: Otago Centre for Electron Microscopy, University of Otago, PO Box 56, Dunedin 9054, New Zealand. richard.easingwood@otago.ac.nz.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Easingwood
authors.affiliation: New Zealand Institute for Plant and Food Research Limited, Private Bag 4704, Christchurch 8140, New Zealand. andrew.pitman@plantandfood.co.nz.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Pitman
authors.affiliation: Centre for Protein Research, University of Otago, PO Box 56, Dunedin 9054, New Zealand. torsten.kleffman@otago.ac.nz.
authors.forename: Torsten
authors.initials: T
authors.lastname: Kleffmann
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. mihnea.bostina@otago.ac.nz.|Otago Centre for Electron Microscopy, University of Otago, PO Box 56, Dunedin 9054, New Zealand. mihnea.bostina@otago.ac.nz.
authors.forename: Mihnea
authors.initials: M
authors.lastname: Bostina
authors.affiliation: Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin 9054, New Zealand. peter.fineran@otago.ac.nz.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Fineran
mesh_terms: D029042:Actinidia / Q000382:microbiology*; D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005638:Fruit / Q000382:microbiology; D016679:Genome, Viral; D010935:Plant Diseases / Q000382:microbiology*; D017902:Podoviridae / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D020543:Proteome / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044224:Pseudomonas syringae / Q000502:physiology / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D020543:Proteome; D014764:Viral Proteins
keywords: Pseudomonas syringae pv. actinidiae; bacteriophage; biocontrol agent; canker; cryo-electron microscopy; genomics; kiwifruit; proteomics; single particle reconstruction; vB_PsyP_phiPsa17