Biodiversity study on Puntius has been conducted in West Sumatera using a molecular technique. From the genetic analysis using the sequence of CO1 gene, the study discovered:(1) A new record species of Puntius in Diatas Lake, Batang Lembang, Batang Gumanti, Muara Pingai rivers (located in the eastern part of the Bukit Barisan mountain range) which is Barbodes binotatus banksi or B. banksi.(2). A new record of subspecies in Maninjau Lake and its tributary (located in the western part of the Bukit Barisan mountain range) which is Barbodes banksi maninjau.(3) A new record of subspecies in Batang Kuranji, Batang Katik, Batang Tarok and Lubuk Paraku rivers (located in the western part of the Bukit Barisan mountain range) which is Barbodes banksi kuranji. The results of this study add the evidence that the presence of Bukit Barisan mountain range in Sumatra Island contributed to genetic diversity, evolutionary process and speciation mechanism of freshwater fish in Sumatra. It is important to pay attention to the development of district or area in Sumatra.