Reproductive isolation between selfing and outcrossing species can arise through diverse mechanisms, some of which are directly associated with differences in mating system. We dissected cross incompatibility between the highly selfing morning glory Ipomoea lacunosa and its mixed-mating sister species I. cordatotriloba. We found that cross incompatibility is complex, with contributions acting both before and after fertilization. We then investigated whether the transition in mating system may have facilitated the evolution of these reproductive barrier components through mismatched floral morphology, differences in reproductive context, or both. We found evidence that morphological mismatch likely contributes to reproductive isolation in at least one cross direction and that other pollen-pistil interactions are present. We also identified hybrid seed inviability consistent with the predictions of the weak-inbreeder, strong-outbreeder hypotheses, suggesting endosperm misregulation plays an important role in cross incompatibility. In contrast, we did not find evidence consistent with the pre-zygotic weak-inbreeder, strong-outbreeder hypothesis. Our study highlights the complexity of reproductive isolation between outcrossing and selfing species and the extent to which evolutionary consequences of mating system transitions can facilitate speciation.

Please see the methods of our manuscript and the README files associated with each dataset for these details. 

The data files are a combination of .csvs and .txt files. Data analyses done using R version 4.0.3, and you can see the renv.lock file for all the R programs we used.