Anthracnose caused by Colletotrichum is one of the most severe diseases of Hevea brasiliensis. However, research on the diversity and geographical distribution of Colletotrichum remains limited in China. In this study, we investigated the phylogenetic diversity of Colletotrichum isolates associated with symptomatic tissues of H.brasiliensis from four provinces of China (Hainan, Guangdong, Guangxi, and Yunnan). Based on multi-locus phylogenetic analyses and phenotypic characteristics, five species were distinguished, including two known species (C. fructicola, C. siamense), one novel species of C. gloeosporioides species complex (C. ledongense), and two novel species of C. acutatum species complex (C. bannanense and C. australisinense). Of these, C. siamense and C. australisinense have been recognized as major causative agents of anthracnose of H. brasiliensis.