Freshwater fungi were collected from rivers and streams, including freshwater, sediment, and plant litter samples. While surveying unrecorded freshwater fungi in Yeongsan-river, three fungal strains, NNIBRFG37669, NNIBRFG34665, NNIBRFG37668 and NNIBRFG34669 were isolated from the freshwater environment. Cosmospora obscura NNIBRFG37669 was isolated from freshwater in Keumgye-reservoir. NNIBRFG37669 grew fully by 2.3cm on PDA at 25℃ for 7 days. The colony on PDA was white. Conidia was ovoid to ellipsoidal. Ophiobolus cirsii NNIBRFG37668 was isolated from freshwater in Keumgye-reservoir. NNIBRFG37668 grew fully by 1.6cm on PDA at 25℃ for 7 days. The colony on PDA, white to yellowish. Conidia was overlapping or lying parallel or spiral, greenish-yellow, without sheath or appendages. Didymella macrophylla NNIBRFG34665 was isolated from freshwater in Dogap-reservoir. NNIBRFG34665 grew fully by 6.0cm on PDA at 25℃ for 7 days. The colony on PDA, greyish brown. Conidia was aseptate, hyaline, smooth-walled, and thin-walled, ranging from ovoid to cylindrical shape. Phyllachora phyllostachydis NNIBRFG34669 was isolated from freshwater sediment in Dogap-reservoir. NNIBRFG34669 grew fully by 3.1cm on PDA at 25℃ for 7 days. The colony on PDA, as greenish grey to dark green at the center. Conidia was aseptate, hyaline, smooth, obovoid to ellipsoidal. The stains were identified using molecular phylogenetic analyses of rDNA. We identified three species previously unrecorded in Korea.