Development of species specific RAPD markers using populus cpDNA

Chloroplast DNAs(cpDNA) from eight different species of Populus(P. alba, P. glandulosa, P. alba×P. glandulosa, P. davidiana, P. maximowiczii, P. nigra×P. maximowiczii, P. nigra, and P. Koreana×P. nigra), Salix pseudo-lasiogyne and Nicotiana tabacum were compared by PCR amplification. The cpDNAs were amplified at relatively low annealing temperature so that many bands could be observed. Amplification of a spacer between 16S γRNA gene and 23S γRNA gene located in inverted repeats in cpDNA resulted in a major band in size of 2.3 Kb and several minor bands of various sizes. A band specific to Populus was found when the amplification was done at lower annealing temperature. Amplification with primers deduced from γpoC1 and γpoC2 gene produced the major target band as well as several additional (minor) bands that could be used for the differentiation of each Populus species. The minor bands appeared repeatedly even when different thermocycles were employed. With the amplification profile, it was possible to differentiate Populus species.