Since the Modern Synthesis, many species concepts have considered characters mediating mate recognition essential to both the identification and definition of species. To explore divergence in mate recognition systems, calls were recorded and toads measured from all three species of the monophyletic Bufo microscaphus species complex including five populations of Bufo californicus, three populations of B. microscaphus, and one population of B. mexicanus. Call duration, dominant frequency, and pulse rate were significantly related to temperature, but not snout-vent length. When adjusted for temperature, calls of B. californicus had a significantly longer call duration, higher dominant frequency, and slower pulse rate than B. mexicanus and B. microscaphus, which did not differ from one another. However, the magnitude of variation among populations of B. californicus was similar to that between species. Discriminant analysis using call variables provided some separation of B. californicus and B. mexicanus from B. microscaphus, and cross-validation analysis correctly classified approximately 75% of B. californicus and B. mexicanus. Given only slight divergence in mate recognition systems between B. californicus and the other two taxa, the biological significance of this difference remains ambiguous. Comparisons among B. americanus group members indicate that hybridizing taxa may or may not exhibit divergence in advertisement calls. Maintenance of independently evolving lineages may be driven by other evolutionary mechanisms. These results support the notion that species recognition is best viewed as an effect of mate recognition.