This study represents the first published data of freshwater fungi from the River Nile in Egypt. Knowledge concerning the geographic distribution of freshwater ascomycetes and their asexual morphs in Egypt and in the Middle East is limited. Ninety-nine taxa representing 42 sexual ascomycetes, 55 asexual taxa and two basidiomycetes were identified from 959 fungal collections recorded from 400 submerged samples. Samples were randomly collected from the River Nile, in Sohag, Egypt in the winter and summer between December 2010 and August 2014. Fifty-eight taxa (22 sexual ascomycetes and 36 asexual taxa) were collected during winter, while 60 taxa (25 sexual ascomycetes, 33 asexual taxa and two basidiomycetes) were collected in summer season. Of the 99 taxa recorded, 50 are new records for Egypt, including five new genera and 30 new species., Three new genera and ten new species were described in previous articles. Fungi recorded from the two seasons were markedly different, with only 19 species common to both winter and summer collections. Asexual fungi dominated the fungal community during the two seasons. Taxonomical placements of 33 species were confirmed by molecular data based on LSU and SSU rDNA genes. Lolia aquatica (14.2%) was the dominant fungus in both winter and summer collections. Other dominant fungi were: Ceratorhiza sp. (19.5 %) and Limnoperdon sp. (13 %). These two basidiomycetes were the most common taxa in the summer, while they were absent in the winter. Common fungi were Coleophoma emperti (9.2 %), Zopfiella latipes (8 %), Discosporium tremuloides (5.5 %), Trematophoma lignicola (5.5 %) and Ophioceras commune (5 %). Specious genera recorded from the two seasons were Dictyosporium (6 species), Monodictys-like (3 species), Aniptodera (3 species), Lolia (3 species), Podospora (3 species), Zopfiella (3 species), and two species belong to each of the following genera: Achaetomium, Annulatascus, Lentithecium, Linocarpon, Cirrenalia, Ciliochora, Coleophoma, Colletogloeum, Clohesyomyces, Periconia, Pseudorobillarda and Stagonospora.