Answer: A.Reproductive isolation occurred through behavioral isolation.
Speciation is a process of development of new species from the previously existing species due to reproductive barriers occur among the members of the population of species. The reproductive barrier arises due to behavior and physical differences. Behavior isolation occurs when the two populations of the same species develop some differences in behavior, to attract their mates for mating, which can lead to reproductive isolation among members of same species or related species.
In the given situation, the related species of leopard frogs have developed specific courtship call, this will lead to reproductive isolation due to behavior isolation. This will help in identifying mates of the same species and this will act as a reproductive barrier among the other related species.