A total of 1,159 Fusarium strains were isolated from sorghum grown in Danyang and Youngwol in 2017 and 2018. The isolates were analyzed to reveal genetic, toxigenic and pathogenic characteristics. Phylogenetic analysis using TEF-1α and RPB2 genes showed that the samples were contaminated with at least 17 Fusarium species. Among them, F. graminearum, F. proliferatum, F. thapsinum, F. incarnatum, and F. asiaticum were dominant species. In F. graminearum and F. asiaticum, F. graminearum–15-acetyl deoxynivalenol chemotype and F. asiaticum–nivalenol chemotype were frequent. Six Fusarium species tested produced one or more mycotoxins, except F. thapsinum and FTSC 11. F. proliferatum and F. fujikuroi had FUM1 gene (76.0% and 81.6%, respectively) and some isolates produced high level of fumonisin (over 1,000 μg/g). F. proliferatum and F. thapsinum were more virulent than other species on sorghum. These results indicate that Fusarium species in sorghum might produce multiple mycotoxins.