Development of SCAR markers in olive (Olea europaea) by direct sequencing of RAPD products: applications in olive germplasm evaluation and mapping

RAPD markers generated by mixtures of two different 10-mer primers were developed for eight different olive cultivars used as parental lines in olive-breeding programs. Two RAPD bands were converted into dominant SCAR markers by direct sequencing of the RAPD products, avoiding the costly and time-consuming cloning step. The SCARs generated have maintained the original RAPD polymorphism among the cultivars and segregated according to Mendelian inheritance. Preliminary results suggest the use of the SCAR SCOeMS-2 for the marker-assisted selection of the high flesh/stone ratio. This strategy provides a rapid method for the characterization of RAPD markers and for the development of PCR-based markers with applications in olive mapping, paternal testing and germplasm characterization. The use of these markers in multiplexed PCRs, and the direct ethidium bromide detection of the PCR products in the test tube, facilitate their efficient and reliable breeding applications.