Strawberry leaves showing leaf blight symptoms were collected from six different farms in Ismailia and Beheira Governorates in Egypt during the 2020–2021 growing season. Eight bacterial isolates, i.e., Pa₁, Pa₂, Pa₃, Pa₄ (Ismailia farms) and Pa₅, Pa₆, Pa₇ and Pa₈ (Beheira farms) were isolated. A pathogenicity test of bacterial isolates was carried out using detached strawberry leaf technique. All bacterial isolates produced leaf blight disease symptoms. Isolates Pa₂ and Pa₆ showed the highest pathogenic characteristics with clear symptoms on detached strawberry leaves. The phenotypic, biochemical and physiological characters of the highest pathogenic isolates were confirmed by PCR analysis using 16S rRNA gene. The two bacterial isolates were identified as Pantoea ananatis with similarity of 97.05% with accession number MH_127816.1 (isolate Pa₂, Ismailia), while the isolate ( Pa₆, Beheira) with similarity of 97.03% with accession number NR_026045.1. The 16S rDNA sequences were deposited in the GenBank nucleotide databases under accession numbers OM258167 and OM279507, respectively. According to the pathogenicity test, morphological and physiological characteristics as well as molecular data (16S rRNA sequencing analysis), this finding is the first report of P. ananatis as a causal agent of strawberry leaf blight disease in Egypt.