Reproductive isolating mechanisms contributing to asymmetric hybridization in Killifishes (Fundulus spp.)

When species hybridize, one F1 hybrid cross type often predominates. Such asymmetry can arise from differences in a variety of reproductive barriers, but the relative roles and concordance of pre‐mating, post‐mating prezygotic, and post‐zygotic barriers in producing these biases in natural animal populations have not been widely investigated. Here, we study a population of predominantly F1 hybrids between two killifish species (Fundulus heteroclitus and F. diaphanus) in which >95% of F1 hybrids have F. diaphanus mothers and F. heteroclitus fathers (D♀ × H♂). To determine why F. heteroclitus × F. diaphanus F1 hybrids (H♀ × D♂) are so rare, we tested for asymmetry in pre‐mating reproductive barriers (female preference and male aggression) at a common salinity (10 ppt) and post‐mating, pre‐zygotic (fertilization success) and post‐zygotic (embryonic development time and hatching success) reproductive barriers at a range of ecologically relevant salinities (0, 5, 10, and 15 ppt). We found that F. heteroclitus females preferred conspecific males, whereas F. diaphanus females did not, matching the observed cross bias in the wild. Naturally rare H♀ × D♂ crosses also had lower fertilization success than all other cross types, and a lower hatching success than the prevalent D♀ × H♂ crosses at the salinity found in the hybrid zone centre (10 ppt). Furthermore, the naturally predominant D♀ × H♂ crosses had a higher hatching success than F. diaphanus crosses at 10 ppt, which may further increase their relative abundance. The present study suggests that a combination of incomplete mating, post‐mating pre‐zygotic and post‐zygotic reproductive isolating mechanisms act in concert to produce hybrid asymmetry in this system. Common Killifish (Fundulus heteroclitus; coloured blue) and Banded Killifish (Fundulus diaphanus; coloured orange) hybridize in areas of overlap; 95% hybrids between these species are F1s with F. diaphanus mothers. We examined reproductive isolation (RI) using all possible cross types across a range of ecologically relevant salinities (0‐15ppt, with 10ppt shown here) to examine how different barriers may contribute to the observed hybrid asymmetry.