Agrobacterium tumefaciens (A. tumefaciens) is a plant pathogenic bacterium that causes crown gall disease, leading to significant economic losses in various agriculturally important crops, including apple, pear, peach, and almond. The difficulty of controlling A. tumefaciens with conventional pesticides underscores the need for alternative antibacterial agents such as bacteriophages. In this study, a novel lytic bacteriophage, named Agrobacterium phage PAT1 (PAT1), with high lysis potential against A. tumefaciens, was isolated from wastewater. The interaction between PAT1 and A. tumefaciens cells, investigated using Transmission electron microscopy, revealed that PAT1 efficiently adsorb, infect, and replicate on A. tumefaciens in a very short time period (i.e., ≤30 min for a complete infectious cycle). Furthermore, the turbidity assay showed that PAT1 (MOI = 1) exerted a significant inhibitory effect on A. tumefaciens for 48 hours, resulting in an 82% reduction in bacterial growth. Additionally, stability tests showed that PAT1 was highly resistant to a broad range of pH (4-10) and temperatures (4-60°C). Bioinformatics analyses of the genomic sequence suggested that PAT1 is a new phage closely related to Agrobacterium phages of the genus Atuphduovirus within the family Autographiviridae. The genome size was 45,040 base pairs with a G+C content of 54.5%, consisting of 54 coding sequences (CDS), of which the functions of 22 CDS were predicted, including two endolysins genes which could be used as antimicrobials against A. tumefaciens. Additionally, no lysogenic mediated genes or genes encoding virulence factors, antibiotic resistance, or toxins were detected in PAT1 genome. Overall, PAT1 demonstrated suitability as a potential biocontrol agent for combating A. tumefaciens infections, expanding the limited catalog of such lytic A. tumefaciens phages. However, its efficacy in controlling crown gall disease in planta remains to be evaluated.