|Lee, Ae Kyung -|
|Yuan, Tao -|
|Suh, Jeung Keun -|
|Choi, Dae Sung -|
|Choi, Ik-Young -|
|Joung, Young Hee -|
|Lee, Jong Suk -|
Submitted to: FLORA
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 1, 2011
Publication Date: June 30, 2011
Citation: Lee, A., Yuan, T., Suh, J., Choi, D., Choi, I., Roh, M.S., Joung, Y., Lee, J. 2011. Pulsatilla tongkanensis, a natural hybrid population hybridized with P. Koraiensis based on RAPD and SNPs of Chloroplast DNA. FLORA. 51;409-421. Interpretive Summary: A new Pulsatilla species growing on the cliffs of Donggang, Kangwon Province, Korea, was discovered and named P. tongkangensis. It is not known which forms of P. tongkangensis based on the orientation of flowers and varying flower colors represents the type specimen. Variation in flower color could result from hybridization with P. koreana or introgression in progeny of P. tongkangensis. There are no reports of hybridization with P. koreana, or genetic diversity studies of P. tongkangensis using molecular markers. Therefore, experiments were designed to investigate the genetic diversity of P. tongkangensis and the possibility of interspecific hybridization with P. koreana using RAPD and chloroplast SNP markers. A species-specific RAPD marker for each P. tongkangensis and P. koreana was identified, and both bands were present in some P. tongkangensis, indicating hybrid origin. Discrepancies in the identification of many P. tongkangensis accessions based on morphological characters, the presence of species-specific bands, and variations in nucleotide base differences suggest that significant genetic variation exists and that variation resulted possibly from cross pollination and introgression between these two closely related species. Based on the results of these studies, it is suggested that P. tongkangensis is a heterogeneous and evolving population resulting from genetic introgression from cross-pollination with P. koreana.
Technical Abstract: A new putative species of Pulsatilla, P. tongkangensis, growing in calcified rock cavities was discovered along the Donggang, Jeongseon Kun, Kangwon Province, Korea. Accessions of P. tongkangensis and P. koreana were characterized with randomly amplified polymorphic DNA (RAPD) markers and single nucleotide polymorphisms (SNPs) of the chloroplast trnQ, trnH, psbA gene and the atpB-rbcL intergenic spacer regions. Pulsatilla tongkangensis exhibits significant morphological variation in leaf morphology, flower color (white, purple blue, to light red), and flower bud orientation (lateral-facing to up-facing). RAPD and SNP analysis was performed on accessions collected from the native sites and with progeny from controlled pollination between P. tongkangensis and P. koreana. One P. tongkangensis accession with light-red flowers was clustered with P. koreana using RAPD markers, and three accessions of P. tongkangensis with pink-blue narrow petals, light-blue narrow petals, and white petals had the same SNPs as P. koreana. A species-specific RAPD band for both P. tongkangensis and P. koreana was identified, and both bands were present in some P. tongkangensis accessions, indicating these were of hybrid origin. The presence of species-specific bands and variations in SNPs suggest that significant genetic variation exists and that variation resulted possibly from cross pollination and introgression between these two closely related species. Based on the results of these studies, it is suggested that P. tongkangensis is a heterogeneous population resulting from introgression due to cross-pollination with P. koreana.