Sexual isolation maintains species boundaries between Hawaiian crickets in sympatry despite weak habitat isolation

Understanding the relative importance of different reproductive isolating barriers in maintaining species boundaries is fundamental to understanding how closely related species coexist without hybridizing. Habitat isolation and sexual isolation are often the first barriers between closely related species, yet they are rarely studied together, making their relative contributions to reproductive isolation unclear. We conducted the first comprehensive study of both barriers in Hawaiian swordtail crickets ( Laupala ), focusing on the recently diverged, sympatric species pair Laupala kona and Laupala hualalai . Through field surveys, we quantified microhabitat use, spatial distribution, and diel singing activity, and found extensive ecological overlap: both species occupied similar substrates, elevations, and microclimates, and occurred in mixed-species aggregations. Despite this weak habitat isolation, they have divergent sexual traits and exhibit strong sexual isolation in lab assays. The species differ significantly in male pulse rate and cuticular hydrocarbon profiles, and females consistently preferred conspecific song stimuli. Mating trials showed complete sexual incompatibility between heterospecific pairs, with no successful copulations. Together, these results demonstrate that sexual isolation, rather than habitat partitioning, maintains species boundaries between L. hualalai and L. kona in sympatry. Our findings highlight the sufficiency of sexual isolation to prevent hybridization even in the absence of ecological divergence, providing rare empirical evidence that sexual barriers alone can maintain reproductive isolation in natural populations.