Assortative mating and divergent male courtship behaviours between two cryptic species of nine-spined sticklebacks (genus Pungitius)

Abstract

Assortative mating based on ecologically divergent traits is a major driver of speciation among three-spined sticklebacks, however, little is known about reproductive isolation and variations in courtship behaviour among nine-spined sticklebacks. Here we demonstrate assortative mating and divergent courtship behaviours between two cryptic species of nine-spined sticklebacks using no-choice mate trials and kinematic analyses. Strong assortative mating was demonstrated in our tank experiments, highlighting the importance of prezygotic reproductive isolation in these species. Kinematic analyses showed that the freshwater type exhibited aggressive courtship behaviour with frequent ‘Attacking’ and spent more time exhibiting displacement activities. In contrast, the brackish-water type demonstrated a higher frequency of the ‘Zigzag-dance’ and ‘Nest-showing’. Our study highlights the value of nine-spined sticklebacks as a comparative system for the study of speciation and paves the way for future studies on the role of courtship behaviours and assortative mating in the evolution of sticklebacks.