Development SCAR marker for the rapid authenticaton of Batryticatus Bombyx based on COI Sequences

COI 염기서열 기반 백강잠 신속 감별용 SCAR marker 개발 - 백강잠 유전자 감별 -

  • Kim, Wook Jin (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Yang, Sungyu (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Noh, Pureum (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Park, Inkyu (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Choi, Goya (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Song, Jun-Ho (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Moon, Byeong Cheol (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine)
  • 김욱진 (한국한의학연구원 한약자원연구센터) ;
  • 양선규 (한국한의학연구원 한약자원연구센터) ;
  • 노푸름 (한국한의학연구원 한약자원연구센터) ;
  • 박인규 (한국한의학연구원 한약자원연구센터) ;
  • 최고야 (한국한의학연구원 한약자원연구센터) ;
  • 송준호 (한국한의학연구원 한약자원연구센터) ;
  • 문병철 (한국한의학연구원 한약자원연구센터)
  • Received : 2019.08.19
  • Accepted : 2019.09.25
  • Published : 2019.09.30


Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.


Bombyx mori;Batryticatus Bombyx;COI;DNA barcoding;SCAR marker


Grant : 동의보감 충부약재 활용기반 구축

Supported by : 한국한의학연구원


  1. Kim WJ, Yang S, Choi G, Park I, Noh P, Seo CS, Moon BC. Development of conventional PCR and real-time PCR assays to discriminate the origins of Chinese pepper oil and herbal materials from Zanthoxylum. J Sci Food Agric. 2019 ; 99(4) : 2021-2029. doi: 10.1002/jsfa.9458.
  2. Xiong X, Yao L, Ying X, Lu L, Guardone L, Armani A, Guidi A, Xiong X. Multiple fish species identified from China's roasted Xue Yu fillet products using DNA and mini-DNA barcoding: Implications on human health and marine sustainability. Food Control. 2018 ; 88 : 123-30.
  3. Kim WJ, Yang S, Choi G, Moon BC. Identification of marker nucleotides for the molecular authentication of Araliae Continentalis Radix based on the analysis of universal DNA barcode, matK and rbcL, sequences. Kor J Herbol. 2016 ; 31(5) : 15-23.
  4. Ward RD, Zemlak TS, Innes BH, Last PR, Hebert PD. DNA barcoding Australias' fish species. Phil Trans R Soc. 2005 ; 360 : 1847-57.
  5. Kim WJ, Ji Y, Choi G, Kang YM, Yang S, Moon BC. Molecular identification and phylogenetic analysis of important medicinal plant species in genus Paeonia based on rDNA-ITS, mat K, and rbc L DNA barcode sequences. Genet Mol Res. 2016 ; 15(3) : gmr.15038472. doi: 0.4238/gmr.15038472.
  6. Ryu KS, Lee HS, Kim YS. Pharmaco-dynamic study of power in mice administered to maltose, sucrose and lactose. Korean J Seric Sci. 1999 ; 41 : 9-13.
  7. Oh TW, Kang SY, Kim KH, Song MY, Park YK. Antidiabetic effect of medicinal plants used for lower wasting-thirst in streptozotocin-induced diabetic rats. Kor J Herbology. 2013 ; 28(5) : 53-60.
  8. Yoon SH, Ha H. The effect of Bombycis corpus on Streptozotocin-induced diabetic rats. J Korean Soc Hygienic Sciences. 2006 ; 6(1-2) : 11-22.
  9. Korea Institute of Oriental Medicine. Defining Dictionary for Medicinal Herbs 'Hanyak Giwon Sajeon'. Retrived August 8, 2019, from
  10. Park KT, Lee JS, Lee SM, Shim SY. Distributional data on the superfamily Bombycoidea (Lepidoptera) in Korea. Inst of Agri Sci Kangwon Nat Univ. 1997 ; 8 : 47-81.
  11. CBOL Plant Working Group. A DNA barcode for land plants. PNAS. 2009 ; 106(31) : 12794-7.
  12. Kim WJ, Yang S, Choi G, Moon BC. Peptide nucleic acid based molecular authentication for identification of four medicinal Paeonia species using melting array analysis of the internal transcribed spacer 2 region. Molecules. 2017 ; 22 : 1922. doi:10.3390/molecules22111922.
  13. Moon BC, Kim WJ, Ji Y, Lee YM, Kang YM, Choi G. Molecular identification of the traditional herbal medicines, Arisaematis Rhizoma and Pinelliae Tuber, and common adulterants via universal DNA barcode sequences. Genet Mol Res. 2016 ; 15(1) : gmr.15017064. doi: 10.4238/gmr.15017064.
  14. Pincheira-Donoso D, Bauer AM, Meiri S, Uetz P. Global taxonomic diversity of living reptiles. PLoS ONE. 2013 ; 8(3) : e59741.
  15. Cho ES, Jung CG, Sohn SG, Kim CW, Han SJ. Population genetic structure of the ark shell Scapharca broughtonii Schrenck from Korea, China, and Russia based on COI Gene Sequences. Mar Biotechnol. 2007 ; 9 : 203-16.
  16. Guan S, Liu H, Zheng Y, Yu D, Nie A, Liu M. Inference of phylogenetics and evolution of Epinephelus septemfasciatus and 48 other species of Epinephelus genus using mitochondrial CO1 fragment sequences. Pakistan J Zool. 2014 ; 46(1) : 67-74.
  17. Ganie SH, Upadhyay P, Das S, Sharma MP. Authentication of medicinal plants by DNA markers. Plant Gene. 2015 ; 4 : 83-99.
  18. Moon BC, Kim WJ, Han KS, Yang S, Kang Y, Park I, Piao R. Differentiating authentic Adenophorae Radix from its adulterants in commercially-processed samples using multiplexed ITS sequence-based SCAR markers. Appl Sci. 2017 ; 7 : 660. doi:10.3390/app7070660.
  19. Noh P, Kim WJ, Yang S, Park I, Moon BC. Authentication of the herbal medicine Angelicae Dahuricae Radix using an ITS sequence-based multiplex SCAR assay. Molecules. 2018 ; 23(9) : 2134. doi: 10.3390/molecules23092134.
  20. Babaei S, Talebi M, Bahar M. Developing an SCAR and ITS reliable multiplex PCR-based assay for safflower adulterant detection in saffron samples. Food Control. 2014 ; 35 : 323-8.