Discovery and Genomic Characterization of a Novel Phage P284 with Potential Lytic Ability Against Agrobacterium...

Agrobacterium tumefaciens (A. tumefaciens), the causal agent of crown gall disease, is a major threat to crop production worldwide. In this study, a novel lytic bacteriophage, designated P284, was identified and characterized for its antibacterial potential against A. tumefaciens. High-throughput sequencing revealed a 44,922 bp double-stranded DNA genome (G+C content 54.3%), with 66 predicted coding sequences, none associated with virulence, lysogeny, or antibiotic resistance. Genomic and phylogenetic analyses allocated P284 within the genus Atuphduovirus (subfamily Dunnvirinae), showing 94% nucleotide sequence identity and 100% query coverage with phage PAT1, representing a distinct species. Turbidity assays revealed that P284 (MOI = 1) strongly inhibits A. tumefaciens growth up to 48 h, achieving a 92% reduction in bacterial density. Transmission electron microscopy confirmed rapid adsorption and host cell lysis within 30 min. In silico predictions identified three putative depolymerases with properties suitable for recombinant applications. The phage exhibited stability across a wide pH range (3-9) and temperatures from -20 to 60 °C. These findings highlight the lytic activity and environmental resilience of P284, and whether it can control crown gall disease in planta remains to be evaluated.