Molecular markers as a tool to verify sexual and apomictic off-spring of intraspecific crosses in Hypericum perforatum

The aim of this project was to establish RAPD markers to determine the percentage of sexual off-spring of the facultative apomictic plant species H. perforatum. We did reciprocal crosses between four different accessions (A x B, B x A, C x D, D x C) by mechanical emasculation and hand pollination. Genomic DNA of the parents and the off-spring was isolated and PCR conditions were optimized in order to obtain reproducible bands with RAPD markers. Of the 260 screened RAPD primers 127 revealed polymorphism between the parental lines of A and B, whereas 53 revealed no amplification products. Each progeny was tested for the presence of paternal bands with three primers. We found no sexual off-spring among the 22 progenies of A x B, the nine progenies of B x A and the ten progenies of D x C. However, we detected six sexual off-spring among the 45 progenies of C x D. We have proved that RAPD markers can be used to distinguish between sexual and apomictic off-spring in H. perforatum and that sexual off-spring can be obtained from intraspecific crosses. The percentage of sexual progeny might depend on the genotype of the parental lines.