Members of Cytospora encompass important plant pathogens, saprobes and endophytes on a wide range of woody hosts with a worldwide distribution. In the current study, we obtained seven representative isolates from six tree hosts of Betulaceae, Juglandaceae, Rosaceae, Tiliaceae and Ulmaceae in Mount Dongling of China. Based on morphological comparison and phylogenetic analyses using partial ITS, LSU, act, rpb2, tef1-α and tub2 gene sequences, we identified two known species (Cytospora leucostoma and C. pruinopsis) and two novel species (C. coryli and C. spiraeicola). These results represent the first study on Cytospora species associated with canker disease from Mount Dongling of China.