Sperm ultrastructure of six Australian hylid frogs from two genera (Litoria and Cyclorana): phylogenetic implications

The spermatozoa of four fossorial (Litoria alboguttata, Cyclorana brevipes, Cyclorana novaehollandiae and Cyclorana cryptotis) and two non-fossorial australian hylid frogs (Litoria aurea and Litoria moorei) together with previously examined Litoria (Hylidae: Anura) are compared. In spermatozoal ultrastructure (in particular the structure of the sperm tail) Cyclorana includes species which appear derived (apomorphic) relative to non-fossorial species of Litoria while the fossorial L. alboguttata groups with Cyclorana. All hylid species examined here are united by the bufonoid synapomorphy of a conical subacrosomal cone consisting of separate sheaves and the eubufonoid synapomorphy of a mitochondrial sheath or collar separated by a cytoplasmic canal from the centriolar region and tail. Spermatozoal symplesiomorphies for the Eubufonoidea, seen in Litoria (with the exception of L. alboguttata), are the well developed thin undulating membrane with juxta-axonemal and axial fibre. L. alboguttata, C. novaehollandiae and C. brevipes appear monophyletic in the apomorphic modification of the undulating membrane as a thick, dense structure. In L. alboguttata and C. novaehollandiae this structure retains a swelling, at the free edge, homologized with the axial rod. C. brevipes has a further apomorphy as the undulating membrane forms a parallel-sided dense structure with no separate differentiation of an axial fibre. C. cryptotis, however, retains the plesiomorphic sperm tail with a thin undulating membrane, juxta-axonemal and axial fibre. That these differences in spermatozoal ultrastructure have phylogenetic significance is endorsed by the similarity of the fertilization biology of the species examined. On the basis of sperm ultrastructure three separate lineages are discerned within Cyclorana s. lat.: 1) C. cryptotis; 2) L. alboguttata and C. novaehollandiae; and 3) C. brevipes. The evidence of sperm ultrastructure, supported by previously published molecular, morphological and karyological data, clearly places Litoria alboguttata within the genus Cyclorana.