Wheat leaf rust is caused by the obligate biotrophic fungus Puccinia triticina f. sp. tritici, which seriously affects wheat production. In this study, a novel mitovirus was identified in Puccinia triticina strain HN-1 and designated as "Puccinia triticina mitovirus 1" (PtMV1). The genome of PtMV1 consists of a single RNA molecule with a length of 2,380 nt and an A + U content of 54.7% that contains a single open reading frame (ORF). The ORF is predicted to encode a putative RNA-dependent RNA polymerase (RdRp) of 653 amino acids with a molecular mass of 74.77 kDa, containing six conserved motifs. The RdRp amino acid sequence of PtMV1 has a high degree of sequence similarity to the RdRps of unuamitoviruses. Phylogenetic analysis indicated that PtMV1 is a new member of the genus Unuamitovirus within the family Mitoviridae. To our knowledge, this is the first report of a fungal virus in Puccinia triticina.