During our investigation of saprophytic fungi in Guizhou and Hainan provinces, China, three hyphomycetes were collected from terrestrial and freshwater habitats. Based on morphological characteristics and phylogenetic analyses of combined ITS, LSU, tef 1-α, and rpb 2 sequence data, two new species are introduced: Distoseptisporahainanensis and D.lanceolatispora . Additionally, one known species, D.tectonae , previously unreported from Edgeworthiachrysantha , is newly reported. Detailed descriptions, illustrations, and a phylogenetic tree to show the two new species and the new host record of Distoseptispora are provided. In addition, a checklist of Distoseptispora species with their locations, lifestyles, habitats, and hosts is provided., Competing Interests: The authors have declared that no competing interests exist., (Xue-Mei Chen, Xia Tang, Jian Ma, Ning-Guo Liu, Saowaluck Tibpromma, Samantha C. Karunarathna, Yuan-Pin Xiao, Yong-Zhong Lu.)