Genetic relationship among acer species revealed by RAPD and PCR-RFLP

Ten different species of Acer including A. triflorum, A. saccharinum, A. pseudo-sieboldianum, A paimatum var. sanguineum, A. mono, A. ruburm, A. negundo, A. palmatum, A ginnala, and A. tegmentosum were compared with regard to their DNA variation based on RAPD profiles and PCR-RFLP. For RAPD study, only five species(A. triflorum, A. saccharinum, A pseudo-sieboldianum, A. palmatum var. sanguineum, and A. mono) consisting of more than 5 individuals were compared. For PCR-RFLP, 6 species that produced the target bands with the sequence specific primers were considered. Forty four loci were generated by RAPD with 4 arbitrary primers. Similarity between individuals was estimated by calculating the frequency of shared bands out of total bands. Cluster analysis based on the similarity produced two groups. While the first group was exclusively composed of A. pseudo-sieboldianum, the second comprised the remaining 4 species. When the primers specific to the gene sequences rpl2 and psbA in chloroplast DNA were used, A pseudo-sieboldianum again appeared to have different amplification pattern from the rest of Acerspecies examined. Analysis of the amplified psbA-rpl2 region(1.7 kb) by PCR-RFLP revealed that the species in the genus could be readily differentiated from each other by the fragmentation profile. We found that although RAPD worked on Acer species, it costed too much time and labor for the optimization of PCR conditions. On the other hand, the PCR-RFLP technique is very simple and straight forwarded to get reproducible results since it uses very stringent reaction conditions.