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Characterization of a Lytic Bacteriophage against Pseudomonas syringae pv. actinidiae and Its Endolysin
Authors:Peien Ni  Lei Wang  Bohan Deng  Songtao Jiu  Chao Ma  Caixi Zhang  Adelaide Almeida  Dapeng Wang  Wenping Xu  Shiping Wang
Affiliation:1.School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; (P.N.); (L.W.); (B.D.); (S.J.); (C.M.); (C.Z.); (D.W.); (S.W.);2.Department of Biology and CESAM, University of Aveiro, Campus Universitário de Santiago, 3810-193 Aveiro, Portugal
Abstract:Pseudomonas syringae pv. actinidiae (Psa) is a phytopathogen that causes canker in kiwifruit. Few conventional control methods are effective against this bacterium. Therefore, alternative approaches, such as phage therapy are warranted. In this study, a lytic bacteriophage (PN09) of Psa was isolated from surface water collected from a river in Hangzhou, China in 2019. Morphologically, PN09 was classified into the Myoviridae family, and could lyse all 29 Psa biovar 3 strains. The optimal temperature and pH ranges for PN09 activity were determined as 25 to 35 °C and 6.0 to 9.0, respectively. The complete genome of PN09 was found to be composed of a linear 99,229 bp double-stranded DNA genome with a GC content of 48.16%. The PN09 endolysin (LysPN09) was expressed in vitro and characterized. LysPN09 was predicted to belong to the Muraidase superfamily domain and showed lytic activity against the outer-membrane-permeabilized Psa strains. The lytic activity of LysPN09 was optimal over temperature and pH ranges of 25 to 40 °C and 6.0 to 8.0, respectively. When recombinant endolysin LysPN09 was combined with EDTA, Psa strains were effectively damaged. All these characteristics demonstrate that the phage PN09 and its endolysin, LysPN09, are potential candidates for biocontrol of Psa in the kiwifruit industry.
Keywords:Pseudomonas syringae pv. actinidiae   kiwifruit   lytic bacteriophage   endolysin   phage therapy
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