The Korea Journal of Herbology (대한본초학회지)

The Korea Association of Herbology (대한본초학회)
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Development of SCAR marker for the rapid assay of Paeng-hwal based on CO1 DNA barcode sequences

CO1 DNA 바코드 염기서열 기반 팽활(蟛螖) 신속 감별용 SCAR marker 개발

  • Wook Jin Kim (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Sumin Noh (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Goya Choi (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Woojong Jang (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine) ;
  • Byeong Cheol Moon (Herbal Medicine Resources Research Center, Korea Institute of Oriental Medicine)
  • 김욱진 (한국한의학연구원 한약자원연구센터) ;
  • 노수민 (한국한의학연구원 한약자원연구센터) ;
  • 최고야 (한국한의학연구원 한약자원연구센터) ;
  • 장우종 (한국한의학연구원 한약자원연구센터) ;
  • 문병철 (한국한의학연구원 한약자원연구센터)
  • Received : 2024.01.18
  • Accepted : 2024.03.25
  • Published : 2024.03.30
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Abstract

Objectives : Paeng-hwal is described as an insect herbal medicine used for digestive diseases in the Dong-ui-bo-gam. The origin of this herbal medicine is limited to several small crabs, such as Helice tridens. These crab species cohabitat in the same environment and share similar morphological characteristics, making it very difficult to distinguish and collect the individual species for use in dietary supplements or herbal medicines. This study was conducted to develop a genetic identification tool for discriminating among these closely related small crab species. Methods : CO1 DNA barcode regions of 15 samples from 6 species of small crabs were analyzed to obtain the individual sequences. To identify the correct species, comparative analyses were carried out using the database of the NCBI GenBank and the NIBR. SCAR primers were designed to develop simple and rapid assay methods using inter-species specific sequences. Optimal SCAR assay conditions were established through gradient PCR, and the limit of detection (LOD) was determined. Results : Six species of small crabs (Helicana tridens, Macrophthalmus abbreviatus, Helicana tientsinensis, Helicana wuana, Chiromantes dehaani, and Hemigrapsus penicillatus), which are distributed as Paeng-hwal, were identified through CO1 sequences analysis. We also developed SCAR markers to distinguish between six small crabs at the species level. Furthermore, we established the optimal PCR assay methods and the LOD of each individual species. Conclusions : The rapid and simple SCAR-PCR assay methods were developed to identify the species and control the quality of herbal medicines for Paeng-hwal based on the genetic analyses of CO1 DNA barcodes.

Keywords

Acknowledgement

본 연구는 한국한의학연구원 기본사업인 '지속가능한 한약표준자원 활용기술 개발' 과제(KSN1822320)의 지원으로 수행되었으며, 시료 확보에 도움을 주신 한약자원연구센터 연구자분들께 감사드립니다.

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