Production of interspecific Campanula hybrids by ovule culture: exploring the effect of ovule isolation time

The Campanula genus comprises several economically important ornamental plants species. Wide hybridisation is a method to increase phenotypic variability, but is limited due to interspecies hybridisation barriers. In this study we investigated whether ovule culture could be used to increase the success rate of interspecific hybridisation between C. portenschlagiana × C. poscharskyana and C. medium × C. formanekiana . The effect of different ovule isolation times on ovule germination in vitro was examined. In general, the number of collectible ovules and ovule germination was low. Interspecific hybrids between C. medium and C. formanekiana exhibited an increased number of viable ovules with later isolation time, but with different ovule germination rates. A parent-of-origin effect on both the number of collectible ovules and ovule germination was observed for C. medium × C. formanekiana . Histological analysis of embryo and endosperm development in collectible ovules isolated at different time points from interspecific crosses showed that the vast majority of ovules did not contain an embryo. When present, embryo development only progressed with ovule collection time in the C. medium and C. formanekiana crosses. The occurrence of miscoloured seedlings in interspecific crosses indicated incompatibilities between the parental lines that could not be prevented by reciprocal crossing. The low number of collectible ovules and germination rates might be inhibited due to fertilisation barriers. With this study, a protocol for ovule culture was established and the usefulness of ovule culture to obtain interspecific hybrids of selected Campanula species was demonstrated.