Contribution of visual stimuli to mating and fighting behaviors of Drosophila prolongata

The importance of visual stimuli in a given behavior is often different, even between closely related species. Species of the genus Drosophila have been categorized into three classes according to their reproductive dependency on vision. Class I and II species reproduce in the darkness, whereas class III species never reproduce without vision. Therefore, in class III species, mutants at the white locus that are deprived of normal visual function are not able to be established as homozygous stocks. Drosophila prolongata is a species with a conspicuous body color pattern, suggesting that an increased importance of visual stimuli underlies the evolution of this species. In this study, to examine the behavioral dependency on visual stimuli in D. prolongata, we generated white gene-knockout (w⁻) mutants using the CRISPR/Cas9 system and examined their mating and fighting behaviors in light and dark conditions. The courtship and fighting activities of w⁻ males were extremely reduced regardless of the light condition. Furthermore, wildtype males did not perform any courtship or fighting behaviors in dark conditions. Our results showed that D. prolongata is a class III species that strongly relies on visual stimuli in both behaviors.