Abstract: The morphological characters and karyotype of Rana tigrian rugulosa were analyzed using samples from Shiping and Yuanjiang of Yunnan. The results demonstrate that Rana tigrian rugulosa from Yuanjiang was bigger and got more weight than Shiping ones. Morphological characters such as length of hind limb, length of tibia, eardrum width and head width were significantly different between Yuanjiang and Shiping ones. Samples from different locations enjoyed the same karyotype: 2n=26(5L+8M+0s), NF=52. The chromosome No.3 of Rana tigrian rugulosa from Shiping was SM, while the rest chromosomes were M, and the secondary constriction located on 6p. The chromosome No.3, 6 and 8 from Yunjiang were SM, the rest were M, and the unobvious secondary constriction located on 3p and 6p. It was significant different at morphological characters and karyotype between Yuanjiang and Shiping samples. Yuanjiang and Shiping samples might be speculated as the geographical groups of Rana tigrian rugulosa.