Freshwater ecosystems provide a complex environment for microorganisms. In this study, we isolated diverse fungal strains from plant litter in freshwaters. These strains were identified using molecular phylogenetic analyses of rDNA and/or other gene sequences (TUB, GAPDH, and EF1). In addition, we examined their morphological characteristics by microscopy and cultural characteristics on several media. We identified six previously unrecorded Sordariomycetes species in Korea, i.e., Colletotrichum godetiae, Discosia rubi, Robillarda sessilis, Monochaetia dimorphospora, Idriella lunata, and Phialemoniopsis endophytica. Of these, D. rubi and M. dimorphospora exhibited high extracellular amylase, lipase, and protease activities, suggesting that these fungal isolates might play an important role as decomposers in freshwater ecosystems. Plant litter could thus be a good source for isolating and investigating previously undocumented fungal species in freshwater environments.