A 33.4-kb fragment of the mitochondrial genome of Fusarium oxysporum has been sequenced. The fragment contains the complete gene sequences for 13 of the 14 proteins typically encoded by the mitochondrial genome of filamentous ascomycetes. Similarity searching revealed all encoded proteins to be most similar to those from other members of the Hypocreales. The fragment contains the complete small subunit rRNA gene, partial large rRNA subunit gene, and 12 tRNAs. Two introns were present, one in the nad5 gene and one in the large rRNA subunit gene, the latter containing a ribosomal protein gene.