Recently, ambrosia beetles have become very important pest of 2~5 year old apple trees with M9 dwarf rootstocks in South Korea. The beetles have killed the branches and stems of the young trees, especially, frozen damage trees in winter or drought stressed tree in spring. By the increase in planting area and weaken property in winter of M9 dwarf rootstock, ambrosia beetles are becoming a key pest in Korean apple orchards using M9 rootstock. According to the survey of damaged apple trees by ambrosia beetles, Xylosandrus germanus Blandford, Xyleborus apicalis Blandford and Xyleborinus saxeseni (Ratzeburg). These insects are hosts of the ambrosia fungi. ITS region of rDNA has shown to be a useful source for phylogenetic studies and identifying speices in previous published articles. We analyzed the nucleotide sequences of ITS1, 5.8S and ITS2 region of ambrosia fungi isolated from three ambrosia beetles, in order to observe molecular variation among the fungi strains and to reveal phylogenetic relationships.