방풍류의 감별을 위한 분자마커의 탐색과 활용
Development amd Application of PCR-based Markers for the Discrimination of Bang-Poong and Related Species
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초록
한약재로 사용되는 방풍류는 절단되어 유통되므로 외부 형 태적인 특징만으로 구분하기가 어려워 방풍류로 사용되는 방 풍, 식방풍, 석방풍, 갯방풍 등 4종에 대해 PCR에 기초한 RAPD 마커를 이용하여 SCAR 마커를 개발하고자 하였다. RAPD 분석결과 밴드의 패턴은 다양하게 나타났으며 다형성 의 밴드 수는 총 215개로 전체 밴드수의 98%였다. RAPD 분 석에서 각 방풍류를 구별 할 수 있는 특이적인 밴드를 나타 내는 primer는 방풍에서 4개의 primer, 식방풍은 6개의 primer, 석방풍은 4개의 primer, 갯방풍은 6개의 primer를 선 발하였고, 그 중 특히 primer 425는 4종의 방풍류의 감별에 유용하였고, 이를 이용하여 SCAR 마커로 전환하는데 이용하 였다. 특이적인 단편을 클로닝하여 염기서열 분석으로 특이 primer를 제작하고 제작된 primer로 방풍류 시료 16개에 적 용하였을 때, 국내의 야생에서 주로 자생하는 석방풍은 215 bp, 그리고 국내에서 가장 많이 재배 또는 생산되는 갯방풍은 177 bp와 300 bp에서 뚜렷하게 나타났다. 따라서 갯방풍과 석 방풍의 감별 가능성을 제시할 수 있으며 개발된 SCAR 마커 를 이용하여 시중에 유통되고 있는 방풍류 건조약재의 감별 에 유용한 마커로 활용될 수 있을 것이다.
Abstract
Bang-Poong and related species are an important herbal medicine. However, it is difficult to determinethe commercial dry material through anatomical and chemotaxonomical characteristics. Here, we used a PCR-based technique for an accurate discrimination of Bang-Poong and related species. With the RAPD primers, 215RAPDs (random amplified polymorphic DNAs) were obtained, and 98% of them showed polymorphic patterns.RAPDs from the four primers were appropriate for the discrimination of S. divaricata(TURCZ.) SCHISKIN, those from thesix primers for P. japonicum THUNBERG, those from the four primers for P. terebinthaceumFISHER, and those from thesix primers for G. littoralisFr. SCHMIDT. The specific bands from the primer 425 were obtained and used to developSCAR (sequence characterized amplified region) markers, based on the sequence information of the RAPDmarkers. The SCAR primers generated a 215 bp fragment specific to Peucedanum terebinthaceum FISHER, and a177 bp and a 300 bp fragment specific to G. littoralisFr. SCHMIDT. As a result, the three SCAR markers were able todiscriminate from two Bang-Poong related species.
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