Planta Med 2006; 72(13): 1223-1226
DOI: 10.1055/s-2006-951695
Original Paper
Biochemistry and Molecular Biology
© Georg Thieme Verlag KG Stuttgart · New York

Molecular Authentication of the Traditional Tibetan Medicinal Plant Swertia mussotii

Chun-Ying Xue1 , De-Zhu Li1 , Jin-Mei Lu1 , Jun-Bo Yang1 , Jian-Quan Liu2
  • 1Laboratory of Plant Biodiversity and Biogeography, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan P. R. China
  • 2Northwest Plateau Institute of Biology, Chinese Academy of Sciences, Xining, QingHai, P. R. China
Further Information

Publication History

Received: December 5, 2005

Accepted: August 25, 2006

Publication Date:
18 October 2006 (online)

Abstract

Swertia mussotii is an important species in Tibetan folk medicine. However, it is quite expensive and frequently adulterated, so reliable methods for authentication of putative specimens and preparations of the species are needed to protect consumers and to support conservation measures. We show here that the chloroplast (cp) DNA rpl16 intron has limited utility for differentiating S. mussotii from closely related species, since the cpDNA rpl16 sequences are identical in S. mussotii and two other species of Swertia. However, the rDNA internal transcribed spacer (ITS) sequences differ significantly between S. mussotii and all of 13 tested potential adulterants. Thus, the ITS region provides a robust molecular marker for differentiating the medicinal S. mussotii from related adulterants. Therefore, a pair of allele-specific diagnostic primers based on the divergent ITS region was designed to distinguish S. mussotii from the other species. Authentication by allele-specific diagnostic PCR using these primers is convenient, effective and both simpler and less time-consuming than sequencing the ITS region.

References

  • 1 Yang Y C. Tibetan Medicines. Qinghai. Qinghai People Press 1991: p 111
  • 2 Ding J Y, Sun H F. Effective components antihepatitis of Swertia mussotii I, separate and identify mangiferin and olinolic acid.  Chin Tradit Herb Drugs. 1980;  9 391-2
  • 3 Sun H F, Ding J Y. Separate and identify xanthones of the Swertia mussotii .  Acta Bot Sin. 1981;  23 464-9
  • 4 Sun H F, Hu B L, Ding J Y. Glucoside components of the Swertia mussotii .  Acta Bot Sin. 1991;  33 31-7
  • 5 Yang H L, Liu J Q. Seed germination of Swertia mussotii, an important application in Tibetan folk medicine.  Acta Bot Yunnanica. 2005;  27 295-300
  • 6 Baldwin B G, Sanderson M J, Porter J M, Wojciechowski M F, Campbell C S, Donoghue M J. The ITS region of nuclear ribosomal DNA: a valuable source of evidence on angiosperm phylogeny.  Ann Mo Bot Gard. 1995;  82 247-77
  • 7 Lau D TW, Shaw P C, Wang J, But P PH. Authentication of medicinal Dendrobium species by the internal transcribed spacer of ribosomal DNA.  Planta Med. 2001;  67 456-60
  • 8 Ding X Y, Xu L S, Wang Z T, Zhou K Y, Xu H, Wang Y Q. Authentication of stems of Dendrobium ficinale by rDNA ITS region sequences.  Planta Med. 2002;  68 191-2
  • 9 Sun Y, Fung K P, Leung P C. Characterization of medicinal Epimedium species by 5S rRNA gene spacer sequencing.  Planta Med. 2004;  70 287-8
  • 10 Ding X Y, Wang Z T, Zhou K Y, Xu L S, Xu H, Wang Y Q. Allele-specific primers for diagnostic PCR authentication of Dendrobium officinale .  Planta Med. 2003;  69 587-8
  • 11 Chassot P, Nemomissa S, Yuan Y M, Küpfer P. High paraphyly of Swertia L. (Gentianaceae) in the Gentianella-lineage as revealed by nuclear and chloroplast DNA sequence variation.  Plant Syst Evol. 2001;  229 1-21
  • 12 Doyle J J, Doyle J L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue.  Phytochem Bull. 1987;  19 11-5
  • 13 White T J, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, White T PCR protocols: A guide to methods and application. San Diego; Academic Press 1990: p 315-22
  • 14 Jordan W C, Courtney M W, Neigel J E. Low levels of intraspecific genetic variation at a rapidly evolving chloroplast DNA locus in North American duckweeds (Lemnaceae).  Am J Bot. 1996;  83 430-9
  • 15 Ho T N, Xue C Y, Wang W. The origin, dispersal and formation of the distribution pattern of Swertia L. (Gentianaceae).  Acta Phytotax Sin. 1994;  32 525-37
  • 16 Liu J Q, Chen Z D, Liao Z X, Lu A M. Comparison of the ITS sequences of the Tibetan medicine ”zang yin chen”-Swertia mussotti and its adulterant species.  Acta Pharm Sin. 2001;  36 67-70

Dr. Chun-Ying Xue

Laboratory of Plant Biodiversity and Biogeography

Kunming Institute of Botany

Chinese Academy of Sciences

Kunming 650204

Yunnan

People’s Republic of China

Phone: +86-871-522-3508

Fax: +86-871-521-7791

Email: chyxue@mail.kib.ac.cn