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Post-copulatory interactions between males and females involve highly coordinated, complex traits that are often rapidly evolving and divergent between species. Failure to produce and deposit eggs may be a common post-mating prezygotic barrier, yet little is known about what prevents the induction of egg-laying between species. The field crickets,  Gryllus firmus  and  G. pennsylvanicus  are isolated by a one-way reproductive incompatibility;  G. pennsylvanicus  males fail to fertilize  G. firmus  eggs or to induce normal egg-laying in  G. firmus  females. We use experimental crosses to elucidate the role of accessory gland-derived vs. testis-derived components of the  G. firmus  male ejaculate on egg-laying in conspecific and heterospecific crosses. Using surgical castrations to create ‘spermless’ males that transfer only seminal fluid proteins (SFPs) we test whether  G. firmus  male SFPs can induce egg-laying in conspecific crosses and rescue egg-laying in crosses between  G. pennsylvanicus  males and  G. firmus  females. We find  G. firmus  SFPs induce only a small short-term egg-laying response and that SFPs alone cannot explain the normal induction of egg-laying.  Gryllus firmus  SFPs also do not rescue the heterospecific cross. Testis-derived components, such as sperm or prostaglandins, most likely stimulate egg-laying or act as transporters for SFPs to targets in the female reproductive tract. These results highlight the utility of experimental approaches for investigating the phenotypes that act as barriers between species and suggest that future work on the molecular basis of the one-way incompatibility between  G. firmus  and  G. pennsylvanicus  should focus on divergent testis-derived compounds or proteins in addition to SFPs.

Influence of the Male Ejaculate on Post-Mating Prezygotic Barriers in Field Crickets