Studying the diversity of viruses found in uncultivated fungi, including those forming mycorrhizal relationships, is essential. It’s equally important to explore viral communities in fungi that cause plant diseases or are economically significant. This dual approach helps us grasp the full scope of mycovirus diversity and evolution. Mycorrhizal fungi, in particular, host a wide range of viruses, shedding light on viral diversity and evolution. In this study, we present the discovery and complete genome characterization of a novel mitovirus infecting the hypogeous mycorrhizal fungus Tuber rufum. This virus, denominated “Tuber rufum mitovirus 1” (TrMV1) has a genome size of 2864 nucleotides with a G + C content of 37.53%. It contains a single open reading frame (ORF) responsible for encoding RNA dependent RNA polymerase (RdRp). Comparative analysis using BLASTp reveals that the protein encoded by TrMV1 shares significant sequence similarities with those found in the Triamitovirus genus. Specifically, TrMV1 shows the closest resemblance (43.35% identity) to Sopawar virus, a mitovirus previously detected in soil environments through metatranscriptomic analyses. Phylogenetic examination categorizes TrMV1 as a member of the Triamitovirus genus within the Mitoviridae family. This finding marks the first identification of a mitovirus within the hypogeous mycorrhizal fungus Tuber rufum. The discovery of TrMV1 expands our knowledge of Mitoviridae family diversity and evolution, contributing to the growing repository of mycovirus sequences.