Three new species isolated from sugarcane rhizosphere in China, namely Conlarium baiseense sp. nov., C. nanningense sp. nov., and C. sacchari sp. nov., are described and illustrated. Molecular evidence (phylogenetic analysis of combined LSU, SSU, ITS and RPB2 sequence data) and phenotypical characters support their independent status from related and similar species. The new species, as dark spetate endophytes, inhabit sugarcane rhizosphere and can form a symbiosis with sugarcane.