Authentication of ‘Cereza del Jerte’ cherry cultivars using real time PCR

The purpose of this work was to develop a real-time polymerase chain reaction method for the determination of sweet cherry cultivars for the authentication of “Cereza del Jerte”. Three autochthonous cultivars of the “Picota” type sweet cherry, ‘Ambrunés’, ‘Pico Negro’, and ‘Pico Colorado’, and the foreign cultivar ‘Sweetheart’ were used in the study. Three molecular markers were obtained through analysis of RAPD profiles with OPD11 primer, after testing 27 primers, one of 822 bp for the ‘Pico Negro’ cultivar and two markers of 763 bp and 635 bp for the ‘Ambrunés’ cultivar. Those markers were then used to design new primer pairs for real-time PCR with SYBR® Green, one of which allowed the Picota type cherry cultivars to be distinguished from the Sweetheart cultivar since the amplification products had distinct melting temperatures.