Authentication of the Oriental Medicinal Plant Ligusticum tenuissimum (Nakai) Kitagawa (Korean Go-Bon) by Multiplex PCR

Abstract

The oriental medicinal plant Ligusticum tenuissimum (Korean name, Go-Bon) is widely used in Korea and China. L. tenuissimum (Go-Bon) has been employed in the treatment of headache and common cold, and as a fever remedy. The internal transcribed spacer (ITS) region was sequenced from thirty-four Go-Bon samples collected from botanical gardens and markets in Korea and China to identify and authenticate L. tenuissimum. Based on the ITS sequences, the thirty-four Go-Bon samples were classified into three groups: L. tenuissimum (Korean Go-Bon), L. jeholense (Chinese Go-Bon), and unknown Chinese Ligusticum species. Three specific primers were designed to identify the three groups of Ligusticum species using multiplex PCR. The established multiplex-PCR was proved to be effective for the differentiation of L. tenuissimum in commercial plant materials.

Key words

internal transcribed sequence (ITS) - Apiaceae - Ligusticum tenuissimum - Ligusticum jeholense - multiplex PCR - single nucleotide polymorphism (SNP)

References

• 1 Kitagawa M. Miscellaneous notes on Apiaceae (Umbelliferae) of Japan and Manchura.  Jpn J Bot. 1941;  17 557
• 2 Fading P, Watson M F. Ligusticum linnaeus, Sp.P1.1: 250. 1753.  Flora of China. 2005;  14 140-150
• 3 Lee H W, Choi J H, Park S Y, Choo B K, Chun J M, Lee A Y, Kim H K. Constituents comparison of component in native and cultivated species of Angelica tenuissima Nakai.  Korean J Med Crop Sci. 2008;  16 168-172
• 4 Han S H, Lee J H, Kil G J. A comparative study between cultivar and wild type of Angelica tenuissima in Korea.  Kor J Herbol. 2007;  22 1-7
• 5 Choi H K, Kim C K, Shin H C. Molecular reexamination of Korean Umbelliferae based on internal transcribed spacer sequences of rDNA: Ligusticum tenuissimum (Nakai) Kitagawa and Libanotis coreana (Wolff) Kitagawa.  J Plant Biol. 2000;  43 128-135
• 6 Ngan F, Shaw P, But P, Wang J. Molecular authentication of Panax species.  Phytochemistry. 1999;  50 787-791
• 7 Cui X M, Lo C K, Yip K L, Dong T T, Tsim K W K. Authentication of Panax notoginseng by 5S-rRNA spacer domain and random amplified polymorphic DNA (RAPD) analysis.  Planta Med. 2003;  69 584-586
• 8 Shim Y H, Choi J H, Park C D, Lim C J, Cho J H, Kim H J. Molecular differentiation of Panax species by RAPD analysis.  Arch Pharm Res. 2003;  26 601-605
• 9 Ha W Y, Shaw P C, Liu J, Yau F C, Wang J. Authentication of Panax ginseng and Panax quinquefolius using amplified fragment length polymorphism (AFLP) and directed amplification of minisatellite region DNA (DAMD).  J Agric Food Chem. 2002;  50 1871-1875
• 10 Choi Y E, Ahn C H, Kim B B, Yoon E S. Development of species specific AFLP-derived SCAR marker for authentication of Panax japonicus C. A. Meyer.  Biol Pharm Bull. 2008;  31 135-138
• 11 Wang J, Ha W Y, Ngan F N, But P P, Shaw P C. Application of sequence characterized amplified region (SCAR) analysis to authenticate Panax species and their adulterants.  Planta Med. 2001;  67 781-783
• 12 Cui G H, Tang X J, Huang L Q. Application of multiplex allele-specific PCR for authentication of Panax ginseng and P. quinquefolius.  China J Chin Mater Med. 2006;  31 1940-1943
• 13 Park M J, Kim M K, In J G, Yang D C. Molecular identification of Korean ginseng by amplification refractory mutation system-PCR.  Food Res Int. 2006;  39 568-574
• 14 Zhu S, Fushimi H, Cai S, Komatsu K. Species identification from ginseng drugs by multiplex amplification refractory mutation system (MARMS).  Planta Med. 2004;  70 189-192
• 15 Sasaki Y, Komatsu K, Nagumo S. Rapid detection of Panax ginseng by loop-mediated isothermal amplification and its application to authentication of ginseng.  Biol Pharm Bull. 2008;  31 1806-1808
• 16 Zhu S, Fushimi H, Komatsu K. Development of a DNA microarray for authentication of ginseng drugs based on 18S rRNA gene sequence.  J Agric Food Chem. 2008;  56 3953-3959
• 17 Hall T A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT.  Nucleic Acids Symp Ser. 1999;  41 95-98
• 18 Chen F, Chan H E, Wong K L, Wang J, Yu M T, But P P H, Shaw P C. Authentication of Saussurea lappa, an endangered medicinal material, by ITS DNA and 5S rRNA sequencing.  Planta Med. 2008;  74 889-892
• 19 Thompson J D, Gibson T J, Plewniak F, Jeanmougin F, Higgins D G. The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.  Nucleic Acids Res. 1997;  24 4876