A rapid method for species identification and interspecies introgression based on a PCR-RFLP: application to the closely related Neotropical toads Rhinella marina and R-schneideri (Anura: Bufonidae)

Toad species Rhinella marina and R. schneideri (Bufonidae, Anura) are widely distributed in Amazon and Cerrado/Caatinga, respectively. Recent data has hypothesized that these two toad species share a history of a massive introgressive hybridization. Therefore, the aim of the present work was the precise identification of organisms with signs of past introgression or actual ongoing hybridization. In this paper we conducted a method based on polymerase chain reaction-restriction fragment length polymorphism, through the use of the CfoI enzyme. The individuals of R. marina have two specific digested bands, which is not digested in R. schneideri, possessing the hybrids three bands. The method was very efficient and allowed fast identification of these species.