Jump to ContentJump to Main Navigation
Show Summary Details
More options …


More options …
Volume 69, Issue 1


Genetic diversity and hybridization of Pulsatilla tongkangensis based on the nrDNA ITS region sequence

Yan-Lin Sun / Wan-Geun Park / Hyun-Kyung Oh
  • National Institute of Biological Resources, Environmental Research Complex, Gyoungseo-dong, Seo-gu, Incheon, 404-708, Korea
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Soon-Kwan Hong
  • Department of Bio-Health Technology, Kangwon National University, Chuncheon, 200-701, Korea
  • Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, 200-701, Korea
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2013-11-15 | DOI: https://doi.org/10.2478/s11756-013-0284-1


Pulsatilla tongkangensis, a perennial medicinal herb growing on the cliffs of Tongkang, Kangwon Province, Korea, is accordingly named as P. tongkangensis as a new Pulsatilla species. Due to sharing similar morphological characters with other Pulsatilla species, how to accurately discriminate P. tongkangensis becomes a critical, urgently needed-solved problem. To understand the truth of P. tongkangensis derivation and the phylogenetic relationship in Pulsatilla, the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) as the most frequently used marker for distinguishing plant species was amplified from P. tongkangensis in this study. According to sequence alignment result, three P. tongkangensis-specific nucleotide sites were obtained to vary from other Pulsatilla species, particularly one at 417 bp showing completely homology among P. tongkangensis accessions and completely dissimilarity between P. tongkangensis and other Pulsatilla species. As known fungal contamination is common in herbal medicinal materials when improperly processed and stored, using universal primers of the ITS region could not only plant DNA but parasitic fungal DNA. Thus, to improve the amplification condition of the ITS region from P. tongkangensis and design plant-specific primer would the following focus of our study. This work will be useful for clearly understanding the phylogenetic relationship between P. tongkangensis and other species belonging to the genus Pulsatilla.

Keywords: Pulsatilla; Pulsatilla tongkangensis; molecular identification; nrDNA ITS

  • [1] Adams K.L. & Palmer J.D. 2003. Evolution of mitochondrial gene content: gene loss and transfer to the nucleus. Mol. Phylogenet. Evol. 29: 380–395. http://dx.doi.org/10.1016/S1055-7903(03)00194-5CrossrefGoogle Scholar

  • [2] But P.P.H. 1994. Herbal poisoning caused by adulterants or erroneous substitutes. J. Trop. Med. Hyg. 97: 371–374. Google Scholar

  • [3] Cho Y., Mower J.P., Qiu Y.L. & Palmer J.D. 2004. Mitochondrial substitution rates are extraordinarily elevated and variable in a genus of flowering plants. Proc. Natl. Acad. Sci. USA. 101: 17741–17746. http://dx.doi.org/10.1073/pnas.0408302101CrossrefGoogle Scholar

  • [4] Cullings K.W. & Vogler D.R. 1998. A 5.8S nulclear ribosomal RNA gene sequence database: applications to ecology and evolution. Mol. Ecol. 7: 913–923. http://dx.doi.org/10.1046/j.1365-294x.1998.00409.xCrossrefGoogle Scholar

  • [5] Doyle J.J. & Doyle J.L. 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull. 19: 11–15. Google Scholar

  • [6] Han C.K., Park Y.H., Jin D.Q., Hwang Y.K., Oh K.B. & Hand J.S. 2007. SK-PC-B70M from Pulsatilla koreana improves scopolamine-induced impairments of memory consolidation and spatial working memory. Brain Res. 1184: 254–259. http://dx.doi.org/10.1016/j.brainres.2007.09.084Web of ScienceCrossrefGoogle Scholar

  • [7] Hershkovitz M.A., Zimmer E.A. & Hahn W.J. 1999. Ribosomal DNA sequences and angiosperm systematic, pp. 268–326. In: Hollingsworth P.M., Bateman R.M. & Ornall R.J. (eds), Molecular Systematics and Plant Evolution, Taylor & Francis, London. http://dx.doi.org/10.1201/9781439833278.ch14CrossrefGoogle Scholar

  • [8] Huang S.Q., Takahashi Y. & Dafni A. 2002. Why dose the flower stalk of Pulsatilla cernua (Ranunculaceae) bend during anthesis? Am. J. Bot. 89: 1599–1603. http://dx.doi.org/10.3732/ajb.89.10.1599CrossrefGoogle Scholar

  • [9] Kaplan J., Chavan P., Warude D. & Patwardhan B. 2004. Molecular markers in herbal drug technology. Curr. Sci. 87: 159–165. Google Scholar

  • [10] Kim Z.S., Jo D.G., Jeong J.H., Kim Y.H., Yoo K.O. & Cheon K.S. 2010. Genetic diversity and structure of Pulsatilla tongkangensis as inferred from ISSR markers. Kor. J. Plant. Res. 23: 360–367. Google Scholar

  • [11] Lee A.K., Yuan T., Suh J.K., Choi D.S., Choi I.Y., Roh M.S., Joung Y.H., Ko J.Y. & Lee J.S. 2010. Pulsatilla tongkangensis, a natural hybrid swarm population hybridized with P. koreana based on RAPD and SNPs of chloroplast DNA. Hort. Environ. Biotechnol. 51: 409–421. Google Scholar

  • [12] Lee W.K., Choi H.W. & Bang J.W. 2004. Karyotype analysis of five species of genus Pulsatilla. Kor. J. Med. Crop Sci. 12: 490–493. Google Scholar

  • [13] Lee Y.N. 2000. Genus Pulsatilla in Korea. Bull. Korea Plant Res. 1: 9–15. Google Scholar

  • [14] Li D.Z., Liu J.Q., Chen Z.D., Wang H., Ge X.J., Zhou S.L., Gao L.M., Fu C.X., Chen S.L. 2011. Plant DNA barcoding in China. J. Syst. Evol. 49: 165–168. http://dx.doi.org/10.1111/j.1759-6831.2011.00137.xCrossrefWeb of ScienceGoogle Scholar

  • [15] Seo J.S., Yun J.H., Baek I.S., Leem Y.H., Kang H.W., Cho H.K., Lyu Y.S., Son H.J. & Han P.L. 2010. Oriental medicine Jangwonhwan reduces Abeta(1–42) level and beta-amyloid deposition in the brain of Tg-APPswe/PS1dE9 mouse model of Alzheimer disease. J. Ethnopharmacol. 128: 206–212. http://dx.doi.org/10.1016/j.jep.2010.01.014Web of ScienceCrossrefGoogle Scholar

  • [16] Sparg S.G., Light M.E. & Van Staden J. 2004. Biological activities and distribution of plant saponins. J. Ethnopharmacol. 94: 219–243. http://dx.doi.org/10.1016/j.jep.2004.05.016CrossrefGoogle Scholar

  • [17] Tomlinson B., Chan T.Y., Chan J.C., Critchley J.A. & But P.P. 2000. Toxicity of complementary therapies: an eastern perspective. J. Clin. Pharmacol. 40: 451–456. http://dx.doi.org/10.1177/00912700022009206CrossrefGoogle Scholar

  • [18] White T.J., Bruns T., Lee S. & Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, pp. 315–322. In: Innis M.A., Gelfand D.H., Sninsky J.J. & White T.J. (eds), PCR protocols-a guide to methods and applications, Academic Press, San Diego, Calif. Google Scholar

  • [19] World Health Organization 2008. Traditional Medicine. Fact Sheet N134. Geneva, World Health Organization. Google Scholar

  • [20] Xu H.J., Shi X.W., Ji X., Du Y.F., Zhu H. & Zhang L.T. 2011. Qualitative and quantitative determination of nine main active constituents in Pulsatilla cernua by high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. J. Separ. Sci. 34: 308–316. http://dx.doi.org/10.1002/jssc.201000660CrossrefGoogle Scholar

  • [21] Xu Y.J., Bai L., Liu Y.H., Liu Y., Xu T.H., Xie S.X., Si Y.S., Zhou H.O., Liu T.H. & Xu D.M. 2010. A new triterpenoid saponin from Pulsatilla cernua. Molecules 15: 1891–1897. http://dx.doi.org/10.3390/molecules15031891Web of ScienceCrossrefGoogle Scholar

  • [22] Yoo K.O., Cheon K.S. & Jang S.K. 2009. Environmental and ecological characteristics of Pulsatilla tongkangensis habitats. Kor. J. Env. Eco. 23: 439–446. Google Scholar

  • [23] Yu S., Fu J., Zhou R., Liu B. & Yan X. 2008. The first report of windflower leaf spot caused by Ascochyta anemones in China. Plant Prot. 34: 147–148. Google Scholar

  • [24] Zhang W., Wendel J.F. & Clark L.G. 1997. Bamboozed again! Inadvertent isolation of fungal rDNA sequences from bamboos (Poaceae: Bambusoideae). Mol. Phylogenet. Evol. 8: 205–217. http://dx.doi.org/10.1006/mpev.1997.0422CrossrefGoogle Scholar

About the article

Published Online: 2013-11-15

Published in Print: 2014-01-01

Citation Information: Biologia, Volume 69, Issue 1, Pages 24–31, ISSN (Online) 1336-9563, DOI: https://doi.org/10.2478/s11756-013-0284-1.

Export Citation

© 2013 Slovak Academy of Sciences. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

Yuhua Shi, Mingming Zhao, Hui Yao, Pei Yang, Tianyi Xin, Bin Li, Wei Sun, and Shilin Chen
Scientific Reports, 2017, Volume 7, Number 1
Gábor Sramkó, Levente Laczkó, Polina A. Volkova, Richard M. Bateman, and Jelena Mlinarec
Molecular Phylogenetics and Evolution, 2019, Volume 135, Page 45
Monika SZCZECIŃSKA, Grzegorz ŁAZARSKI, Katarzyna BILSKA, and Jakub SAWICKI
TURKISH JOURNAL OF BOTANY, 2017, Volume 41, Page 329
Monika Szczecińska and Jakub Sawicki
International Journal of Molecular Sciences, 2015, Volume 16, Number 9, Page 22258

Comments (0)

Please log in or register to comment.
Log in