Ribosomal DNA의 internal transcribed spacer 염기서열과 RAPD 비교에 의한 작약류의 유연관계 분석

Phylogenetic analysis of herbaceous peony using ribosomal DNA partial sequencing and RAPDs

To analyse the genetic relationship among 14 peony accessions, ITS sequencing of rDNA and RAPD were exploited. We amplified 653bp regions including ITS(Ⅰ), 5.8S rDNA and ITS (Ⅱ). The amplified regions were sequenced and compared with GenBank database. The sequences of 5.8S rDNA region were identical among peony species and sub-species. Nucleotide differences at 16 different sites in ITS (Ⅰ) region were observed. In ITS (Ⅱ) region, 1 base pair deletions at one nucleotide site and nucleotide differences at 13 different sites were observed among sequences of 14 peony accessions and GenBank sequences. And at certain nucleotide sites that show nucleotide differences, two different nucleotides were observed in one nucleotide position. Results of ITS sequence analysis were consistent to the classification of peony species based on morphological characteristics. These results suggest that we can distinguish peony species by ITS sequence comparison. We also utilized RAPD analysis in order to study the phylogenetic relationship among peonies. Initially we screened 30 RAPD primers using genomic DNA and selected 11 primers showing distinct band patterns. Ninety eight reproducible bands were generated by 11 primers and the band patterns were analysed to generate dendrogram showing genetic diversity by UPGMA method. As a result, 14 peony accessions were classified into group A, B, C and D. Group D was subdivided into D-1, D-2 and D-3. This study shows that RAPD is a rapid and efficient tool for phylogenetic grouping of peony species and subspecies.