The production of dihydroxynaphthalene (DHN) melanin is known to be essential factor for pathogenicity in Colletotrichum lagenarium. However, the genetic diversity of melanin genes was not much known among Colletotrichum spp. To investigate the variability of melanin gene in Colletotrichum spp. that cause anthracnose on diverse crops including tomato, we cloned and sequenced partial sd, one of DHN melanin genes encoding for scytalone dehydratase, from eight strains of C. coccodes, C. acutatum, C. truncatum C. caricae, and C. musae. The size of PCR-amplified sd ranged 437 bp to 545 bp. The nucleotide sequence identity of sd among the Colletotrichum strains tested varied from 49% to 99%. All of the PCR-amplified sd from eight strains contain an intron and have two exons coding for 122 amino acids. Overall, the size and nucleotide sequence of sd varied among the five Colletotrichum spp. Sequence identity of the predicted scytalone dehydratase protein of 122 amino acids ranged 50 to 99%. Phylogentic analysis based on the sd nucleotide sequences revealed that the five Colletotrichum spp. could be genetically divided.