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KMID : 0861020090240040115
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Korea Journal of Herbology
2009 Volume.24 No. 4 p.115 ~ p.120
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Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD
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Moon Byeong-Cheol
Lee Young-Mi
Chun Jin-Mi
Lee A-Young
Yoon Tae-Sook
Cheon Myeong-Sook
Choo Byung-Kil
Kim Ho-Kyoung
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Abstract
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Objectives: Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker.
Methods: To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and mutiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively.
Results: We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR.
Conclusions: These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.
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KEYWORD
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Rheum species, Rhei Radix et Rhizoma, Rhei Undulatai Rhizoma, Sequence Characterized Amplified Region(SCAR) marker, Random Amplified Polymorphic DNA(RAPD), Molecular authentication
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