Previous studies on the Bufo viridis complex, which is distributed broadly across Iran, are incomplete and restricted to a few regions or a few samples. In this paper a new detailed study on the B. viridis complex in southern of Iran (from West to East) is presented. The analysis of 18 morphometric characters with univariate and multivariate methods reveals significant differences between three members of the B. viridis complex namely B. variabilis, B. luristanicus, and B. surdus distributed in southern part of Iran. Our result help to resolve an old taxonomic problem about B. surdus subgroup (taxa closely related to B. surdus) conifrming that B. luristanicus and B. surdus are distinct species. Moreover, for the ifrst time we report and describe karyotype details of B. luristanicus and B. surdus which conifrmed that they are diploid. Karyological studies demonstrate that all toads from three mentioned species have 2n=22 chromosomes. These chromosomes are arranged into two groups. First group has six large chromosomes and the second group is composed of five small chromosomes. These chromosomes are metacentric or submetacentric. The number of submetacentric chromosomes is different in three mentioned species of B. viridis complex. Neither sexual heteromorphism, nor secondary constriction was observed in any pairs of chromosomes.