The spatiotemporal distribution and dynamic changes of species composition of myxomycetes in humid subtropical forests of Houhe National Nature Reserve in southern China were studied by collecting samples of bark and ground litter within four seasons for isolation of myxomycetes in moist chamber cultures and by collecting fruiting bodies in the field from June to October. A total of 725 specimens of myxomycete fruiting bodies representing 77 species of 25 genera were registered, among them 302 records/51 species from field collections and 423 records obtained from 600 moist chamber cultures of ground litter and bark of living trees, which resulted in 35 and 39 species, respectively. Cornuvia serpula and Perichaena microspora were recorded for the first time in China. Both myxomycete species richness and diversity were significantly different in ground litter among sites and field collections among months. NMDS combined with Permanova indicated that the site had a greater impact on the myxomycete community of bark and litter than the season, while the month of field collections had a greater impact than the sampling sites. CCA and VPA analysis showed that the bark-inhabiting myxomycete species were mainly affected by spatial variables, while the litter-inhabiting and field collected species were more affected by climatic factors.