The Korea Journal of Herbology (대한본초학회지)

The Korea Association of Herbology (대한본초학회)
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Molecular Authentication and Phylogenetic Relationship of Bupleurum Species by the rDNA-ITS Sequences

rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석

  • Moon, Byeong-Cheol (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Choo, Byeong-Kil (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Ji, Yun-I (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Yoon, Tae-Sook (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Lee, A-Young (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Cheon, Myeong-Sook (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Kim, Bo-Bae (Center of Herbal Resources Research, Korea Institute of Oriental Medicine) ;
  • Kim, Ho-Kyoung (Center of Herbal Resources Research, Korea Institute of Oriental Medicine)
  • 문병철 (한국한의학연구원 한약자원연구센터) ;
  • 추병길 (한국한의학연구원 한약자원연구센터) ;
  • 지윤의 (한국한의학연구원 한약자원연구센터) ;
  • 윤태숙 (한국한의학연구원 한약자원연구센터) ;
  • 이아영 (한국한의학연구원 한약자원연구센터) ;
  • 전명숙 (한국한의학연구원 한약자원연구센터) ;
  • 김보배 (한국한의학연구원 한약자원연구센터) ;
  • 김호경 (한국한의학연구원 한약자원연구센터)
  • Published : 2009.09.30
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Abstract

Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.

Keywords

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