The phylogenetic relationships of species and genera in the subfamily Nymphalinae from Myanmar were inferred using mtDNA sequence data from 608 bp of cytochrome oxidase subunit I (COI). A total of 20 species in 10 genera were sequenced and used to construct phylogenetic trees. The base composition of COI sequences was 38.1% T, 15.6% C, 31.6% A, 14.7% G, revealing strong AT bias (69.7%). The sequence distance of 20 species of Nymphalinae ranged from 1.5% to 15.5%. The transition of nucleotide substitution was more common than transversion. The transition between T and C were higher than transition between A and G, and the transversion between A and T was the highest amongst other types of transversion. The phylogenetic trees were constructed using the neighbor-joining (NJ) and maximum likelihood (ML) methods and showed almost identical topologies. The results indicated that the tribes Junoniini and Nymphalini (sensu Wahlberg et al., 2005) formed monophyletic groups but Kallimini was not monophyletic group. Rhinoplapa polynice formed sister group to Junoniini clade with moderate support in both trees. The relationship of species in Junoniini was ((Junonia + Yoma) + Hypolimnas) and the relationship in Nymphalini was (Symbrenthia + (Vanessa + (Kaniska + Polygonia))). The clustering results were almost identical to current morphological classification.