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

Abstract

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. Using genomic data, we confirmed that the two species are genetically distinct, which can only be achieved by strong reproductive isolation through a prolonged time period. Through field surveys, we quantified microhabitat use, spatial distribution, and diets, and found extensive ecological overlap. Both species occupied similar substrates, elevations, and microclimates, and occurred in mixed-species aggregations. They only differ in their diets (inferred from stable isotopes), which might have some implications in reducing encounter rates. 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 almost complete sexual incompatibility between heterospecific pairs, with very rare interspecies 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 strong ecological divergence, providing rare empirical evidence that sexual barriers alone can maintain reproductive isolation in natural populations.