Phylogeography of the natal puddle frog Phrynobatrachus natalensis (Anura: Phrynobatrachidae) in Sub-Saharan Africa

The Family Phrynobatrachidae (Laurent, 1941) has one genus (Phrynobatrachus, Günther, 1862) containing 89 species. Commonly known as puddle frogs, they comprise one of the most species rich sub-Saharan amphibian groups. The objective of this study was to investigate P. natalensis from the Albertine Rift (AR) with systematic and phylogeographic methods. One mitochondrial DNA gene (16S = 555 base pairs [bp]) and one nuclear gene (RAG1 = 774 bp) were analyzed for 61 representatives of P. natalensis. Nine cryptic lineages were identified from Angola, Burundi, Côte d’Ivoire, Democratic Republic of the Congo, Ethiopia, Ghana, Mozambique, Namibia, Tanzania, Rwanda, and Uganda. Several of these lineages are likely new species. Genetic diversity among populations of P. natalensis for 16S was moderate to high (4–13%). Low genetic divergence was recovered in RAG1 (0–1%). Morphological characters were used to examine species boundaries in distinct lineages identified from the molecular phylogeny. No clear pattern was evident in these data. The results of this study suggest the diversity of P. natalensis has been severely underestimated. Additional research efforts should focus on the biogeographic patterns, reproductive isolation, acoustic calls, reproductive behavior, and ecology to determine species boundaries. Furthermore, it is imperative to sample more African sites, especially in the AR, to get a better understanding of African biodiversity