Efficacy of Erwinia amylovora and Xanthomonas campestris pv campestris phages to control fire blight and black rot in vivo

Phytopathogens, such as Erwinia amylovora and Xanthomonas campestris, pose significant threats to agriculture, leading to substantial economic losses. Traditional chemical pesticides can harm soil fertility, contaminate water, and impact non-target organisms such as natural predators and pollinators, highlighting the need for sustainable pest control methods. This study explores the use of bacteriophages as biocontrol agents against E. amylovora, which causes fire blight, and X. campestris pv. campestris, responsible for black rot in cruciferous vegetables. Bacteriophages were isolated from urban wastewater and tested for their lytic activity against these pathogens. Three virulent phages were identified: ɸEF1 and ɸEF2 against E. amylovora and ɸXF1 against X. campestris pv. campestris. Genetic analysis confirmed the absence of known lysogeny-related genes, indicating that these phages are ideal candidates for biocontrol applications. In vitro assays demonstrated significant bacterial population reductions. Specifically, ɸEF1 killed 92.1% of the E. amylovora population at a multiplicity of infection (MOI) of 1 after 3 h, while ɸEF2 reduced the population by 98.1%. When combined in a 1:1 ratio, the two phages reduced E. amylovora populations by 99.7%, and no regrowth of resistant cells was observed, which was not the case when the phages were applied individually. ɸXF1 killed 99.9% of X. campestris pv. campestris populations at an MOI of 1 after 5 h. In vivo experiments using pears and kohlrabi as infection models further validated the phage effectiveness. Treated pears showed reduced fire blight symptoms, and kohlrabi plants exhibited markedly less necrosis from black rot compared to untreated controls.