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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

Abstract

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.

Keywords

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

Acknowledgement

Grant : 동의보감 충부약재 활용기반 구축

Supported by : 한국한의학연구원

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