Using morphological comparison method, the authors identified that the specimens collected in Binh Chau-Phuoc Buu Nature Reserve is Rothmannia wittii. Information on the distribution of this species in Vietnam was also reported. Moreover, based on molecular biology techniques, the matK and trnL-F regions of R. wittii were successfully amplified and sequenced. Also, the results showed that there were differences in matK and trnL-F sequences between R. wittii and R. daweishanensis that were similar in morphological characteristics.