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Over the past two decades, numerous novel species have been identified within <i>Dictyosporiaceae</i>, primarily in <i>Dictyocheirospora</i> and <i>Dictyosporium</i>. A recent monograph has revealed that these two genera exhibit a distinct preference for freshwater habitats, particularly in southern China. However, further investigation into the distribution and diversity of the two genera in Guangdong and Guizhou Provinces remains insufficient. In this study, we conducted an analysis of four intriguing cheiroid hyphomycetes collected from flowing rivers in these two regions. Through morphological and phylogenetic analyses incorporating combined LSU, SSU, ITS, and <i>tef1-α</i> sequence data, we have identified them as a novel species in <i>Dictyocheirospora</i> (<i>Dictyoc. submersa</i> sp. nov.), two novel species in <i>Dictyosporium</i> (<i>Dictyos. guangdongense</i> sp. nov. and <i>Dictyos. variabilisporum</i> sp. nov.), and one previously documented species (<i>Dictyos. digitatum</i>). Specifically, the identification of <i>Dictyos. guangdongense</i> is primarily based on its distinct morphology, characterized by complanate, cheiroid, and brown to dark brown conidia, with a hyaline, short, and atrophied appendage arising from the apical cell of the outer row. In addition, the morphological distinctions between <i>Dictyocheirospora</i> and <i>Dictyosporium</i> are further clarified based on our new data. This study also highlights a few phylogenetic matters regarding <i>Dictyosporiaceae</i>.