Microsatellite high-resolution melting (SSR-HRM) analysis for genotyping and molecular characterization of an Olea europaea germplasm collection Aliki Xanthopoulou 1,2 , Ioannis Ganopoulos 1,2 , Georgios Koubouris 3 , Athanasios Tsaftaris 1,2 , Chrysa Sergendani 3 , Apostolos Kalivas 4 * and Panagiotis Madesis 1 * 1 Institute of Applied Biosciences, CERTH, Thermi, Thessaloniki 570 01, Greece, 2 Department of Genetics and Plant Breeding, School of Agriculture, Aristotle University of Thessaloniki, Thessaloniki 54 124, Greece, 3 Institute of Olive Tree and Subtropical Plants of Chania, Hellenic Agricultural Organization “Demeter”, Agrokipio 73100, Chania, Greece and 4 Cotton and Industrial Plants Institute, Hellenic Agricultural Organization “Demeter”, Thermi, Thessaloniki 57001, Greece Received 6 October 2013; Accepted 10 December 2013 Abstract Olea europaea L. has been cultivated in the Mediterranean region for thousands of years and is of major economic importance. The origin of olive cultivars remains as complex to trace as their identification. Thus, their molecular characterization and discrimination will enable olive germplasm management. In addition, it would be a useful tool for authentication of olive products. High-resolution melting (HRM) analysis, coupled with five microsatellite markers, was integrated to facilitate molecular identification and characterization of main O. europaea cultivars collected from the National Olive Tree Germplasm Collection established in Chania, Greece. The five microsatellite loci used were highly informative and generated a unique melting curve profile for each of the 47 cultivars and for each microsatellite tested. In particular, three microsatellite markers (DCA03, DCA09 and DCA17), which generated 29 HRM profiles, were sufficient to genotype all the olive cultivars studied, highlighting their potential use for cultivar identification. Furthermore, this assay provided a flexible, cost-effective and closed-tube microsatellite genotyping method well suited for molecular characterization of olive cultivars. Keywords: authentication; Gene Bank collection; genotyping; olive; SSR-HRM analysis Introduction Olive tree has been a well-known crop since ancient times. It is one of the most economically important crops and has highly valued healthy attributes. Olive tree classification is very difficult due to the number of varieties and the long life of the trees (Barranco et al., 2000). Complexity in the identification of olive cultivars is further increased by the different names under which they are known in different locations (Therios, 2009). As a result, the same variety is present with many different and variable names in different locations. Thus, highly accurate methods are needed to * Corresponding authors. E-mails: kalivasapostolis@yahoo.gr; pmadesis@certh.gr q NIAB 2014 ISSN 1479-2621 Plant Genetic Resources: Characterization and Utilization; 1– 5 doi:10.1017/S147926211400001X