Romanian Biotechnological Letters Vol.19, No3, 2014 Copyright © 2014 University of Bucharest Printed in Romania. All rights reserved ORIGINAL PAPER Romanian Biotechnological Letters, Vol. 19, No. 3, 2014 9421 Barley cultivar discrimination and hybrid purity control using RAPD markers Received for publication, December 05, 2013 Accepted, February 27, 2014 IOANNIS G. MYLONAS 1 , ANDREAS GEORGIADIS 2 , APOSTOLOS P. APOSTOLIDIS 2* , KONSTANTINOS BLADENOPOULOS 3 AND METAXIA KOUTSIKA-SOTIRIOU 1 1 Department of Genetics and Plant Breeding, Faculty of Agriculture, Aristotle University of Thessaloniki, 541 24 Thessaloniki 2 Department of Animal Production, Faculty of Agriculture, Aristotle University of Thessaloniki 3 National Agricultural Research Foundation (N.AG.RE.F.), Institute of Cereals, 570 01 Thermi-Thessaloniki *Corresponding author: apaposto@agro.auth.gr Abstact In the present study, RAPD markers were used for cultivar discrimination and crosses certification in barley. Originally, ten arbitrary 10-mer primers were tested for the generation of amplification patterns in two successional reactions. From this patch a subset of four primers was selected to generate amplification patterns for five productive barley cultivars (Athinaida, Dimitra, Kos, Niki and Persephone) and their ten F 1 diallel hybrids. Additionally, RAPD analysis was performed to estimate the genetic distance between the five barley cultivars. Results showed that the four selected primers were able to discriminate all barley varieties and four out of ten F 1 diallel hybrids, and also to certify the success of all crosses. This findings show that RAPD markers could be employed as an additional tool in plant breeding programs, for the identification of barley cultivars and hybrids and the certification of crossings. Keywords: RAPD, genetic variation, cultivar identification, purity of F 1 seeds, Hordeum vulgare L. Introduction Barley (Hordeum vulgare L.) was domesticated in the Fertile Crescent about 10,000 years ago (A.A. JARADAT & al. [1]).Today is used as feed, malting and brewing grain and is one of the most important crops worldwide, as ranked 9 th in production in 2012 (www.fao.org, http://faostat.fao.org/site/339/default.aspx accessed on 10/2013) . Barley’s immense importance designates the necessity to develop methods for varietal identification, which is essential for breeders’ rights protection and for quality control in seed production. Furthermore, the application of simple discriminatory methods could be extended in malting and brewing industry, where the varietal identity and purity control of the supplied raw material is crucial in stabilizing production parameters and product features. Generally, phenotypic characterization has been extensively used in breeding programs to explore the genetic variation (K. HAMMER & al. [2]), differentiate cultivars and verify the crossing success. Nevertheless this approach has a lot of drawbacks, starting with the effect of