The Korea Journal of Herbology (대한본초학회지)

The Korea Association of Herbology (대한본초학회)
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Identification 4 kinds of Muxiang using Multiplex PCR

Multiplex PCR을 이용한 4 종류 목향(木香)의 감별

  • Doh, Eui Jeong (Department of Biological Sciences, Konkuk University) ;
  • Lee, Guemsan (Department of Herbology, College of Korean Medicine, Wonkwang University) ;
  • Ju, Young-Sung (Department of Herbology, College of Korean Medicine, Woosuk University) ;
  • Oh, Seung Eun (Department of Biological Sciences, Konkuk University)
  • 도의정 (건국대학교 생명특성화대학 생명과학특성학부 생명과학전공) ;
  • 이금산 (원광대학교 한의과대학 본초학교실) ;
  • 주영승 (우석대학교 한의과대학 본초학교실) ;
  • 오승은 (건국대학교 생명특성화대학 생명과학특성학부 생명과학전공)
  • Received : 2014.04.25
  • Accepted : 2014.05.14
  • Published : 2014.05.30
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Abstract

Objectives : Aucklandiae Radix (Muxiang) one of important herbal medicines in oriental medicine, is defined as the dried root of Aucklandia lappa (Asteraceae). Owing to the similarities in the morphology and name, Inulae Radix (Tu-Muxiang) and Vladimiriae Radix (Chuan-Muxiang) as well as Aristolochiae Radix (Qing-Muxiang) originated from other medicinal plants are often used as substitutes and/or adulterants of Aucklandiae Radix. Therefore, a reliable authentication of these herbal medicines is necessarily for the public health and prevention of misuse. Methods : 32 samples of medicinal plants supplying Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix were collected in Korea and China. The ITS (Internal transcribed spacer) nucleotide sequences of samples were determined. The PCR primers to amply DNA marker of each herbal medicine were designed basing on the specific ITS regions showing differences in the sequences among medicinal plants. Results : Primer set Al R/IS F designed in this work amplified 220 bp PCR product only in samples of Aucklandiae Radix. In contrast, primer set Ih F/IS R, Vs R/IS F, and AcR F1/Ac R amplified 250 bp product, 356 bp prouct, and 516 bp product respectively to identify Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. Conclusions : The primers designed basing on the nucleotide sequences of ITS regions appearing differenced in the sequences among medicinal plants amplified the DNA markers for the identification of Aucklandiae Radix, Inulae Radix, Vladimiriae Radix, and Aristolochiae Radix. These herbal medicines were more efficiently identified by multiplex PCR method using all primers in a single PCR process.

Keywords

References

  1. China Pharmacopoeia Committee. Pharmacopoeia of the People's Republic of China. Beijing : China Chemical Industry press. 2005 ; 1 : 41-2, Appendix X : 57.
  2. Korea Food and Drug Administration. The Korean Herbal Pharmacopoeia. Seoul : Korea Food and Drug Administration press. 2011 : 133-4, 362.
  3. Li AF, Sun AL, Liu RM. Preparative isolation and purification of costunolide and dehydrocostuslactone from Aucklandia lappa Decne by high-speed counter- current chromatography. J Chromatogr A. 2005 ; 1076 (1-2) : 193-7. https://doi.org/10.1016/j.chroma.2005.04.042
  4. Chang HM, But PPH. Pharmacology and applications of Chinese materia medica. Singapore : World Scientific. 1986 : 190-4.
  5. Taniguchi M, Kataoka T, Suzuki H, Uramoto M, Ando M, Arao K, Magae J, Nishimura T, Otake N, KNagai K. Costunolide and dehydrocostus lactone as inhibitors of killing function of cytotoxic T lymphocytes. Biosci Biotechnol Biochem. 1995 ; 59 : 2064-7. https://doi.org/10.1271/bbb.59.2064
  6. Damre AA, Damre AS, Saraf MN. Evaluation of sesquiterpene lactone fraction of Saussurea lappa on transudative, exudative and proliferative phases of inflammation. Phytother Res. 2003 ; 17 : 722-5. https://doi.org/10.1002/ptr.1152
  7. Choi HG, Lee DS, Li B, Choi YH, Lee SH, Kim YC. Santamarin, a sesquiterpene lactone isolated from Saussurea lappa, represses LPS-induced inflammatory responses via expression of heme oxygenase-1 in murine macrophage cells. Int Immunopharmacol. 2012 ; 13 : 271-9. https://doi.org/10.1016/j.intimp.2012.04.016
  8. Yang L, Chen X, Qiang Z, Jun Y, Ren LS, Xiang T. In vitro anti-Helicobacter pylori action of 30 Chinese herbal medicines used to treat ulcer diseases. J Ethnopharmacol. 2005 ; 98 : 329-33. https://doi.org/10.1016/j.jep.2005.01.020
  9. Chen CN, Huang HH, Wu CL. Isocostunolide, a sesquterpene lactone, induces mitochondrial membrane depolarization and caspase-dependent apoptosis in human melanoma cells. Cencer Lett. 2007 ; 246 : 237-52. https://doi.org/10.1016/j.canlet.2006.03.004
  10. Chen F, Chan HYE, Wong KL, Wang J, Yu MT, But PPH, Shaw PC. Authentication of Saussurea lappa, an endangered medicinal material, by ITS DNA and 5S rRNA Sequencing. Planta Med. 2008 ; 74 : 889-92. https://doi.org/10.1055/s-2008-1074551
  11. Shum KC, Chen F, Li SL, Wang J, But PPH, Shaw PC. Authentication of Radix Aucklandiae and its substitutes by GC-MS and hierarchical clustering analysis. J Sep Sci. 2007 ; 30 : 3233-9. https://doi.org/10.1002/jssc.200700232
  12. State Administration of Traditional Chinese Medicine of People's Republic of China. Chinese Materia Medica. Shanghai : Shanghai Scientific & Technical publishers. 1998 ; 7 : 722-7.
  13. Lord GM, Tagore R, Cook T, Gower P, Pusey CD. Nephropathy caused by Chinese herbs in the UK. Lancet. 1999 ; 354 : 481-2. https://doi.org/10.1016/S0140-6736(99)03380-2
  14. Nortier JL, Martinez M, Schmeiser H, Arlt V, Bieler C, Petein M. Urothelial carcinoma associated with the use of a Chinese herb (Aristolochia fangchi). N Engl J Med. 2000 ; 342 : 1686-92. https://doi.org/10.1056/NEJM200006083422301
  15. Korea Food and Drug Administration. The guideline on the visual and organoleptic examination of herbal medicine. Seoul : Korea Food and Drug Administration. vol 1. 2006 : 44, 99, 115.
  16. Vesela D, Saman D, Valterova I, Vanek T. Seasonal variations in the content of taxanes in the bark of Taxus baccata L. Phytochem Anal. 1999 ; 10 : 319-21. https://doi.org/10.1002/(SICI)1099-1565(199911/12)10:6<319::AID-PCA471>3.0.CO;2-W
  17. Bae SM, Jin YM, Jeong EH, Kim MB, Shin HY, Ro CW, Lee SC. Studies on proximate composition, Fatty acids and volatile compounds of Zanthoxylum schinifolium fruit according to harvesting time. Korea J Medicinal Crop Sci. 2011 ; 19 : 1-8. https://doi.org/10.7783/KJMCS.2011.19.1.001
  18. Tetsuo OT, Takashi S, Hiroko M, Stefan W, Christoph N, Jin M. Molecular Phylogeny of Aristolochia sensu lato (Aristolochiaceae) based on Sequences of rbcL, matK, and phyA Genes, with Special Reference to Differentiation of Chromosome Numbers. System Bot. 2006 ; 31(3) : 481-92. https://doi.org/10.1600/036364406778388656
  19. White TJ, Bruns TD, Lee SB, Taylor JW. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, In PCR protocols: A guide to methods and applications. San Diego : Academic Press. 1990 : 315-322.
  20. Sonnante G, Galasso I, Pignone D. ITS sequence analysis and phylogenetic inference in the genus Lens Mill. Ann Bot. 2003 ; 91 : 49-54. https://doi.org/10.1093/aob/mcg007
  21. Gulbitti-Onarici S, Sancak C, Sumer S, Ozcan S. Phylogenetic relationships of some wild wheat species based on the internal transcribed spacer sequences of nrDNA. Curr Sci. 2009 ; 96 : 794-800.
  22. Pandey AK, Ali MA. Molecular markers in plant systematics I: nuclear sequences. In: Plant Sciences Research in India: Challenges and Prospects. India : Botanical Survey of India. 2006 : 21-36.
  23. Choo BK, Moon BC, Ji Y, Kim BB, Choi G, Yoon T, Kim HK. Development of SCAR markers for the discrimination of three species of medicinal plants, Angelica decursiva (Peucedanum decursivum) Peucedanum praeruptorum and Anthricus sylvestris, based on the internal transcribed spacer (ITS) sequence and random amplified polymorphic DNA (RAPD). Biol Pharm Bull. 2009 ; 32 : 24-30. https://doi.org/10.1248/bpb.32.24
  24. Mondini L, Noorani A, Pagnotta MA. Assessing plant genetic diversity by molecular tools. Diversity. 2009 ; 1 : 19-35. https://doi.org/10.3390/d1010019
  25. Haque I, Bandopadhyay R, Mukhopadhyay K. Intraspecific variation in Commiphora wightii populations based on internal transcribed spacer (ITS1-5.8S-ITS2) sequences of rDNA. Diversity. 2009 ; 1 : 89-101. https://doi.org/10.3390/d1020089
  26. Zuo Y, Chen Z, Kondo K, Funamoto T, Wen J, Zhou S. DNA barcoding of Panax species. Planta Med. 2010 ; 77 : 182-7.
  27. Zhang YB, Shaw PC, Sze CW, Wang ZT, Tong T. Molecular authentication of Chinese herbal materials. J Food Drug Anal. 2007 ; 15 : 1-9.
  28. Shiba M, Kondo K, Miki E, Yamaji H, Morota T, Terabayashi S, Takeda S, Sasaki H, Miyamoto K, Aburada M. Identification of medicinal Atractylodes based on ITS sequences of nrDNA. Biol Pharm Bull. 2006 ; 29 : 315-20. https://doi.org/10.1248/bpb.29.315
  29. Xie H, Huo K, Chao Z, Pan S. Identification of crude drugs from Chinese medicinal plants of the genus Bupleurum using ribosomal DNA ITS sequences. Planta Med. 2009 ; 75 : 89-93. https://doi.org/10.1055/s-0028-1088334
  30. Lee SKY, Li PT, Lau DTW, Yung PP, Kong RYC, Fong WF. Phylogeny of medicinal Phyllanthus species in China based on nuclear ITS and chloroplast atpB-rbcL sequences and multiplex PCR detection assay analysis. Planta Med. 2006 ; 72 : 721-6. https://doi.org/10.1055/s-2006-931580
  31. Holck A, Pedersen BO, Heir E. Detection of five novel GMO maize events by qualitative, multiplex PCR and fluorescence capillary gel electrophoresis. Eur Food Res Technol. 2010 ; 231 : 475-83. https://doi.org/10.1007/s00217-010-1302-y
  32. Mori K, Sakamoto Y, Mukojima N, Tamiya S, Nakao T, Ishii T, Hosaka T. Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato. Euphytica. 2011 ; 180 : 347-55. https://doi.org/10.1007/s10681-011-0381-6
  33. Kim MJ, Kim SH, Kim TS, Kee HY, Seo JJ, Kim ES, Park JT, Chung JK, Lee BK, Lee JE. Molecular Genetic Characterization of Shiga Toxin-producing E. coli Isolated from Diarrhea Patients and Cattle in Gwangju Area. Korea J Bacteriol Virol. 2009 ; 39 : 79-95. https://doi.org/10.4167/jbv.2009.39.2.79
  34. Lee MY, Doh EJ, Kim ES, Kim YH, Ko BS, Oh SE. Application of the Multiplex PCR Method for Discrimination of Artemisia iwayomogi from Other Artemisia Herbs Biol. Pharm Bull. 2008 ; 31 : 685-90. https://doi.org/10.1248/bpb.31.685
  35. Ajzenberg D, Collinet F, Mercier A, Vignoles P, Darde ML. Genotyping of Toxoplasma gondii Isolates with 15 Microsatellite Markers in a Single Multiplex PCR Assay. J Clin Microbiol. 2010 ; 48 : 4641-5. https://doi.org/10.1128/JCM.01152-10