Skip to main content
SpringerLink
Log in

Development of molecular markers for authentication of the medicinal plant species Patrinia by random amplified polymorphic DNA (RAPD) analysis and multiplex-PCR

  • Research Report
  • Published:
Horticulture, Environment, and Biotechnology Aims and scope Submit manuscript

Abstract

The authentication of components from plant species is vital for the quality control and standardization of herbal medicines. Due to morphological similarities when dried, it is very difficult to distinguish authentic medicinal plants Patrinia villosa and P. scabiosifolia from adulterants of P. rupestris and P. saniculifolia in the herbal medicine Patriniae Radix. To establish a reliable authentication tool distinguishing authentic Patriniae Radix species from adulterants, we used random amplified polymorphic DNA (RAPD) genomic profiling and obtained several species-specific DNA fragments for each of the four Patrinia species. Based on the sequences of these amplicons, we developed sequence characterized amplified region (SCAR) markers as stable molecular authentication tools. SCAR markers to identify the authentic Patriniae Radix species, P. villosa and P. scabiosifolia, were developed. To accurately identify adulterants, we also developed SCAR markers able to identify the inauthentic P. rupestris and P. saniculifolia species. Furthermore, we established multiplex-PCR SCAR assays to simultaneously distinguish all four species using combinations of species-specific SCAR marker primer sets. These efforts resulted in reliable DNA-based molecular markers to identify authentic Patriniae Radix species from their adulterants and a rapid authentication assay for the efficient identification of Patrinia species. These markers will facilitate the standardization of Patriniae Radix preparations and therefore prevent the distribution of adulterants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Literature Cited

  • Ahlawat A, Katoch M, Ram G, Ahuja A (2010) Genetic diversity in Acorus calamus L. as revealed by RAPD markers and its relationship with ß-asarone content and ploidy level. Sci Hortic 124:294–297

    CAS  Google Scholar 

  • Boleda MD, Briones P, Farres J, Tyfield L, Pi R (1996) Experimental design: A useful tool for PCR optimization. Biotechniques 21: 134–140

    CAS  PubMed  Google Scholar 

  • Cha JT, Lee JC, Lee YC (2012) Comparative study on anti-asthmatic activities of Patrinia scabisaefolia Fischer ex Link and Patrinia villosa Jussieu in a mouse model of asthma (Article in Korean). Korean J Herbol 27:75–82

    Article  Google Scholar 

  • Cheng JL, Long Y, Khan MA, Wei CL, Fu SL, Fu JJ (2015) Development and significance of RAPD-SCARs for the identification of Litchi chinensis Sonn. by improved RAPD amplification and molecular cloning. Electron J Biotechnol 18:35–39

    CAS  Google Scholar 

  • Choo BK., Moon BC, Ji Y, Kim BB, Choi G, Yoon T, Kim HK (2009) 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 32:24–30

    Article  CAS  PubMed  Google Scholar 

  • Coutinho Moraes DF, Still DW, Lum MR, Hirsch AM (2015) DNA-based authentication of botanicals and plant-derived dietary supplements: where have we been and where are we going? Planta Med 81:687–695

    Article  CAS  PubMed  Google Scholar 

  • Devaiah KM, Venkatasubramanian P (2008) Genetic Characterization and Authentication of Embelia ribes using RAPD-PCR and SCAR Marker. Planta Med 74:194–196

    Article  CAS  PubMed  Google Scholar 

  • Fushimi H, Komatsu K, Isobe M, Namba T (1997) Application of PCR-RFLP and MASA analyses on 18S ribosomal RNA gene sequence for the identification of three ginseng drugs. Biol Pharm Bull 20:765–769

    Article  CAS  PubMed  Google Scholar 

  • Horejsi T, Box JM, Staub JE (1999) Efficiency of randomly amplified polymorphic DNA to sequence characterized amplified region marker conversion and their comparative polymerase chain reaction sensitivity in cucumber. J Am Soc Hortic Sci 124:128–135

    CAS  Google Scholar 

  • Joshi K, Chavan P, Warude D, Patwardhan B (2004) Molecular markers in herbal drug technology. Curr Sci 87:159–165

    CAS  Google Scholar 

  • Kim JS, Kang SS (2013) Chemical constituents of plants from the genus Patrinia. Nat Prod Sci 19:77–119

    CAS  Google Scholar 

  • Kim WJ, Ji Y, Lee YM, Kang YM, Choi G, Kim HK, Moon BC (2014) Development of Molecular Marker for the authentication of Patriniae Radix by the analysis of DNA barcodes (Article in Korean). Korean J Herbol 29:45–53

    Google Scholar 

  • Korea Institute of Oriental Medicine (KIOM) (2016}) Defining Dictionary for Medicinal Herbs (Article in Korean). Available via http://boncho.kiom.re.kr/herbarium/codex.php?keyword=%ED%8 C%A8%EC%9E%A5&name_over=%ED%8C%A8%EC%9E%A5 %28%E6%95%97%E9%86%AC%29 Accessed 8 April 201

  • Lee MY, Doh EJ, Kim YH, Ko BS, Oh SE (2008) Development of SCAR marker for discrimination of Artemisia princeps and A. argyi from other Artemisia herbs. Biol Pharm Bull 29:629–633

    Google Scholar 

  • Li G, Park YJ (2012) SCAR markers for discriminating species of two genera of medicinal plants, Liriope and Ophiopogon. Genet Mol Res 11:2987–2996

    Article  CAS  PubMed  Google Scholar 

  • Liu Y, Tan H, Qi Y, Ruan X, Liu N (2014) DNA markers in the authentication of Traditional Chinese Medicine. J Med Plants Res 8:121–127

    Google Scholar 

  • Moon BC, Choo BK, Cheon MS, Yoon T, Ji Y, Kim BB, Lee AY, Kim HK (2010) Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR. Plant Biotechnol Rep 4:1–7

    Article  Google Scholar 

  • Paran I, Michelmore RW (1993) Development of reliable PCR-based markers linked to downy resistance genes in lettuce. Theor Appl Genet 85:985–993

    Article  CAS  PubMed  Google Scholar 

  • Penner GA, Bush A, Wise R, Kim W, Domier L, Kasha K, Laroche A, Scoles G, Molinar SJ, et al (1993) Reproducibility of random amplified polymorphic DNA (RAPD) analysis among laboratories. PCR Meth Appl 2:341–345

    Article  CAS  Google Scholar 

  • Rafalski JA, Tingey SV (1993) Genetic diagnostics in plant breeding: RAPDs, microsatellites and machines. Trends Genet 9:275–280

    Article  CAS  PubMed  Google Scholar 

  • Sucher NJ, Carles MC (2008) Genome-based approaches to the authentication of medicinal plants. Planta Med 74:603–623

    Article  CAS  PubMed  Google Scholar 

  • Staub JE, Serquen F, Gupta M (1996) Genetic markers, map construction, and their application in plant breeding. Hortic Sci 31:729–741

    CAS  Google Scholar 

  • Sze SC, Song JX, Wong RN, Feng YB, Ng TB, Tong Y, Zhang KY (2008) Application of SCAR (sequence characterized amplified region) analysis to authenticate Lycium barbarum (wolfberry) and its adulterants. Biotechnol Appl Biochem 51:15–21

    Article  CAS  PubMed  Google Scholar 

  • Torelli A, Marieschi M, Bruni R (2014) Authentication of saffron (Crocus sativus L.) in different processed, retail products by means of SCAR markers. Food Control 36:126–131

    Article  CAS  Google Scholar 

  • Vos P, Hogers R, Bleeker M, Reijans M, Van de Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang J, Ha WY, Ngan FN, But PP, Shaw PC (2001) Application of sequence characterized amplified region (SCAR) analysis to authenticate Panax species and their adulterants. Planta Med 67:781–783

    Article  CAS  PubMed  Google Scholar 

  • World Health Organization (WHO)(2000) General guidelines for methodologies on research and evaluation of traditional medicines. Available via http://apps.who.int/iris/bitstream/10665/66783/1/WHO_ EDM_TRM_2000.1.pdf

    Google Scholar 

  • Williams JG, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wolff K, Schoen E, Rijn JPV (1993) Optimizing the generation of random amplified polymorphic DNAs in chrysanthemum. Theor Appl Genet 86:1033–1037

    Article  CAS  PubMed  Google Scholar 

  • Xie Y, Peng J, Fan G, Wu Y (2008) Chemical composition and antioxidant activity of volatiles from Patrinia villosa Juss obtained by optimized supercritical fluid extraction. J Pharm Biomed Anal 48:796–801

    Article  CAS  PubMed  Google Scholar 

  • Xin GZ, Lam YC, Maiwulanjiang M, Chan GKL, Zhu KY, Tang WL, Dong TTX, Shi ZQ, Li P, et al (2014) Authentication of Bulbus Fritillariae Cirrhosae by RAPD-derived DNA markers. Molecules 19:3450–3459

    Article  PubMed  Google Scholar 

  • Yip PY, Chau CF, Mak CY, Kwan HS (2007) DNA methods for identification of Chinese medicinal materials. Chin. Med 2:9

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. K-herb Research Center, Korea Institute of Oriental Medicine, Daejeon, 305-811, Korea

    Byeong Cheol Moon, Young Mi Lee, Wook Jin Kim, Yunui Ji, Young Min Kang & Goya Choi

Authors
  1. Byeong Cheol Moon
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Young Mi Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wook Jin Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yunui Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Young Min Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Goya Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Byeong Cheol Moon.

Electronic supplementary material

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Moon, B.C., Lee, Y.M., Kim, W.J. et al. Development of molecular markers for authentication of the medicinal plant species Patrinia by random amplified polymorphic DNA (RAPD) analysis and multiplex-PCR. Hortic. Environ. Biotechnol. 57, 182–190 (2016). https://doi.org/10.1007/s13580-016-0064-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13580-016-0064-2

Additional key words

Search

Navigation