Application of randomly amplified polymorphic DNA (RAPD) markers and polyphenol oxidases (PPO) genes for distinguishing between the diploid (glaucum) and the tetraploid (leporinum) accessions in Hordeum murinum complex

Abstract

The genome of 12 wild barley accessions belonging to Hordeum murinum complex (eight out of them belong to H. m. glaucum and four belong to H. m. leporinum) was characterized using RAPD-PCR markers and polyphenol oxidases genes (PPO). A total of 123 RAPD markers with 22 arbitrary decamer primers were used. 56 out of the RAPD markers were common to all accessions, and the other 67 markers were polymorphic. 35 out of the polymorphic markers are distinguishable between the two taxa but not among all accessions, 15 RAPD markers are specific to H. m. glaucum and 17 are specific to H. m. leporinum. The PCR amplification of PPO genes revealed that amplified fragments from these genes are randomly distributed among the studied accessions regardless of their taxonomy. All data were analyzed using NT-SYS-pc (numerical taxonomy and multivariate analysis system) program to address the phylogenetic relationships between the studied taxa. The examined accessions were clustered into two main groups; the first one consists of 4 accessions representing the H. m. leporinum (diploid) and the other one consists of eight accessions representing the H. m. leporinum (tetraploid).

Key words: Hordeum, glaucum, murinum, leporinum, random amplified polymorphic DNA polymerase chainreaction (RAPD-PCR), polyphenol oxidases (PPO), numerical taxonomy and multivariate analysis system(NTSYS-pc).