Identification of olive-tree cultivars with SCAR markers

Abstract

Summary There is an urgent need for the development of early identification techniques in olive-trees due to the economic importance of cultivar identification in periods of expansion like now. We have been able to identify 22 olivetree cultivars using only 10 different, specific, repeatable markers. These markers were designed by the cloning of significant RAPD bands obtained in PCR performed on bulked DNA to retain the genetic variability of each cultivar. Clones were partially or totally sequenced and new primers derived from these sequences were used to obtain Sequence Characterised Amplified Region (SCAR) fragments. We have demonstrated that the use of the 10 SCAR markers is enough to provide a simple, cheap, and reliable procedure to identify 22 geographically related olive-tree cultivars. Abbreviations: AFLP – amplification fragment length polymorphism; AP-PCR – arbitrarily primed PCR; ASAP – allele-specific associated primers; CAPS – cleaved amplified polymorphic sequence; CTAB – N-cetyl-N,N,Ntrimethyl-ammonium bromide; PCR – polymerase chain reaction; RAPD – random-amplified polymorphic DNA; RFLP – restriction fragment length polymorphism; SCAR – sequence-characterised amplified region; Ta – annealing temperature; UPGMA – unweighted pair group method with arithmetic averages