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Curculigo orchioides Gaertn is an endangered herb which is important ingredient of traditional medicine of Vietnam. In this study, six random primers were used for genetic diversity analysis of C. orchioides populations. Randomly Amplified Polymorphic DNA (RAPD) analysis of 92 samples showed the 65 polymorphic DNA fragments. The genetic diversity is high within the studied populations. Among these, Kon Tum has the highest diversity level (h=0.3889 and I=0.5636) while Lam Dong is lowest (h=0.1991 and I=3064). The degree of genetic differentiation among populations is also high (Gst=0.2505). Genetic identity between Quang Tri and Thua Thien Hue is highest (0.9407) while between Quang Tri and Lam Dong is lowest (0.7439). The dendrogram also showed that Quang Tri population is closely related to Thua Thien Hue population whereas Lam Dong population and Quang Tri, Thua Thien Hue populations have large distance. Present study indicated that C. orchioides genetic relationship and diversity levels are high. This is the first report on genetic diversity of C. orchioides in Vietnam and RAPD is a good molecular marker to study the genetic diversity of this species.