Molecular dissection of the first Staphylococcus cohnii temperate phage IME1354_01

•IME1354_01 is the first isolated temperate phage of Staphylococcus cohnii, expand our knowledge of the diversity of the Staphylococcal phage contingent.•IME1354_01 is a novel temperate phage with only 10% homology coverage and 83.73% nucleotide identity, which is sufficient to be a new genus.•Comparative genomic and phylogenetic analysis showed that IME1354_01 shows distant evolutionary relationships with Bacillus phage and Listeria phage. In staphylococcal phage research, studies specific to coagulase-negative staphylococci (CoNS) remain severely under-represented, and the number of temperate bacteriophages is limited. This investigation identifies a novel temperate phage IME1354_01 from the strain Staphylococcus cohnii IME1354, which was isolated from the skin of a patient with foot ulcer disease. The phage IME1354_01 is the first isolated temperate phage of S. cohnii, and was determined to have a long-tail morphology using TEM. Its genome was found to be a 42,706-bp linear dsDNA molecule with a GC content of 34%. The integration of IME1354_01 occurred using a tRNA-Ser coding gene, and it did not affect tRNA-Ser function. The genome of IME1354_01 is most closely related to that of the temperate Staphylococcus arlettae phage vB_Sars_BM31 with 10% homology coverage and 83.73% nucleotide identity. In addition, they showed similarities mainly in the DNA replication, DNA packaging and partial morphogenesis modules. We propose that a new genus should be created for IME1354_01 based on the intergenomic similarities (maximum is 23%) obtained from the VIRIDIC calculations. The isolation and in-depth study of the novel phage, IME1354_01, will improve our understanding of the evolutionary relationship between temperate phages and their hosts.