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KMID : 0861020210360010009
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Korea Journal of Herbology
2021 Volume.36 No. 1 p.9 ~ p.17
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Development of ITS sequence based SCAR marker and multiplex-SCAR assay for the rapid authentication of Tetrapanacis Medulla and Akebiae Caulis
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Noh Pu-Reum
Kim Wook-jin
Park In-Kyu
Yang Sun-Gyu
Choi Go-Ya
Moon Byeong-Cheol
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Abstract
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Objectives : Tetrapanacis Medulla and Akebiae Caulis are one of the most frequently adulterated herbal medicines because of their confusability of terms in the ancient writings and the similarity of morphological features of dried herbal products. The major adulterant is Aristolochia manshuriensis (Guanmutong) which has a serious safety concern with its toxicity. To ensure the safety and quality of the two herbal medicines, it is necessary to discriminate the toxic adulterant from authentic species. The aim of this study is to develop SCAR markers and to establish the multiplex-SCAR assay for discrimination of four plant species related to Tetrapanacis Medulla and Akebiae Caulis.
Methods : ITS regions of fifteen samples of four species (Tetrapanax papyrifer, Fatsia japonica, Aristolochia manshuriensis, and Akebia quinata) collected from different sites were amplified and sequenced. Fifteen obtained ITS sequences were aligned and analysed for the detection of species-specific sequence variations. The SCAR markers were designed based on the sequence alignments and then, multiplex-SCAR assay enhancing rapidity was optimized.
Results : ITS sequences clearly distinguished the four species at the species level. The developed SCAR markers and multiplex-SCAR assay were successfully discriminated four species and detected the adulteration of commercial product samples by comparison of the amplified DNA fragment sizes.
Conclusions : These SCAR markers and multiplex-SCAR assay are a rapid, simple, and reliable method to identify the authentic Tetrapanacis Medulla and Akebiae Caulis from adulterants. These genetic tools will be useful to ensure the safety and to standardize the quality of the two herbal medicines.
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KEYWORD
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Tetrapanax papyrifer, Akebia quinata, Tetrapanacis Medulla, Akebiae Caulis, ITS, SCAR marker
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