Molecular-genetic characterization of three historically well-isolated cultivars of the Japanese Higogiku group of Chrysanthemum X morifolium Ramat. using 60 RAPD primers

Sixty RAPD primers of random amplified polymorphic DNA (RAPD) were used to investigate genetic difference, and to evaluate DNA finger printing patterns to develop molecular markers in three cultivars among the locally-isolated and the Higo-giku group in Chrysanthemum X morifolium such as ‘Kinkujaku', ‘Madonotsuki' and ‘Nowake' never allowed to be taken out of Kumamoto Prefecture, Japan. Different primers amplified varying numbers of DNA fragments, and altogether 353 reproducible DNA fragments were totally amplifi ed. The generated bands of DNA fragments were in the range of 300-8000 bp in ‘Kinkujaku', 200-8000 bp in ‘Madonotsuki', and 350-8000 bp in ‘Nowake'. Each primer of OPA-06, OPA-14, OPC-18 and OPC-20 revealed one band with no polymorphic DNA fragment among all cultivars studied here. Sixty-two and 48 bands generated by RAPD primers were specific to ‘Kinkujaku' and ‘Madonotsuki', respectively. In contrast, 26 bands were specific to ‘Nowake', and were generated by 21 primers. Pairwise test showed that the three cultivars of the Higo-giku group.