The subspecies of Poephila acuticauda diverged roughly 0.5 MYA due to a temporary biogeographical barrier. Today, Poephila acuticauda hecki inhabits the eastern extreme of the species range in northern Australia and Poephila acuticauda acuticauda the western portion with a series of hybrid contact zones in the center. Phenotypic traits such as bill-color and sperm morphology are currently under investigation as potential reproductive barriers to determine whether speciation is still occurring. Because song is a precopulatory behavior in P. acuticauda, it is a candidate for both a pre- and post-zygotic reproductive barrier. The former would be facilitated by a clear subspecific difference in song and the latter by F1 hybrids either having similarities to parental lineage songs and reinforcing the subspecific divide in subsequent generations or reductions in song performance indicating hybrid breakdown. Here, we measure a suite of acoustic parameters in Raven Pro as well as frequency excursion and stereotypy to probe for differences in song characteristics and performance between subspecies and F1 hybrids. Principal component analyses revealed differences between the pure subspecies but grouped F1 hybrid song with P. a. hecki song. Both stereotypy scores and frequency excursion values were significantly higher in P. a. acuticauda song, with no significant difference between P. a. hecki and F1 hybrid song. Stereotypy scores were significantly higher for F1 hybrids with P. a. acuticauda fathers compared to those with P. a. hecki fathers. These data suggest that song could function as a prezygotic barrier via P. a. acuticauda song being acoustically different and more complex, thereby facilitating assortative mate choice. In turn, because F1 hybrids with physiological and vocal development deficits may struggle to learn the more complex song, hybrids may then struggle to obtain or breed with a mate making it possible for song to also function as a postzygotic barrier to gene flow.