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Development of molecular markers, based on chloroplast and ribosomal DNA regions, to discriminate three popular medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum

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Abstract

Identification of plant species is important for standardizing herbal medicine. Cynanchum wilfordii (Baekshuoh in Korean) and Polygonum multiflorum (Hashuoh in Korean) are important oriental medicinal herbs in Korea, Japan, and China. Cynanchum auriculatum is a faster growing and more productive plant than C. wilfordii; and, it is not recognized as a medicinal plant in the Korean Pharmacopoeia. C. wilfordii, P. multiflorum, and C. auriculatum are often misidentified in the Korean herbal medicine marketplace due to their morphological similarities and similar names. In this study, we investigated molecular authentication of these three medicinal plants using DNA sequences in the TrnL-F chloroplast intergenic region. Specific species identification was achieved by detecting allelic variations of single nucleotide polymorphisms (SNPs) using amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and high resolution melting curve analysis. Our results demonstrate that the intraspecific genetic distance between C. wilfordii and C. auriculatum is relatively low. We also developed a quantitative PCR assay using species-specific TrnL-F primers, which allowed us to estimate the ratio of C. wilfordii and C. auriculatum using varying ratios of mixed genomic DNA template from the two species. Additionally, to identify species in hybrid plants produced by cross-fertilization, we analyzed nuclear ribosomal DNA internal transcribed spacer regions in C. wilfordii and C. auriculatum by ARMS-PCR. Our results indicate that SNP-based molecular markers, usable to barcode tools could provide efficient and rapid authentication of these closely related medicinal plant species, and will be useful for preventing the distribution of products contaminated with adulterants.

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Acknowledgments

This research was supported by the Agro & Bio-industry Technology Development Program (Grant no. 314021-3-1-SB050), Ministry of Agriculture, Food and Rural Affairs, South Korea.

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Authors and Affiliations

  1. Department of Agonomy & Medicinal Plant Resources, Gyeongnam National University of Science & Technology, JinJu, 660-758, Republic of Korea

    Eun-Heui Han, Young-Wook Shin & Shin-Woo Lee

  2. Department of Food Science, Gyeongnam National University of Science & Technology, JinJu, Republic of Korea

    KyeMan Cho

  3. Gyeongnam Oriental Medicinal Herb Institute, Sancheong, Republic of Korea

    YoungMin Goo

  4. Gyeongsangnam-do Agricultural Research & Extension Services, JinJu, 660-985, South Korea

    ManBae Kim

  5. Department of Biology Education, College of Education, and IALS, Gyeongsang National University, JinJu, 660-701, Republic of Korea

    Yun-Hee Kim

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  1. Eun-Heui Han
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Correspondence to Yun-Hee Kim or Shin-Woo Lee.

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Han, EH., Cho, K., Goo, Y. et al. Development of molecular markers, based on chloroplast and ribosomal DNA regions, to discriminate three popular medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum . Mol Biol Rep 43, 323–332 (2016). https://doi.org/10.1007/s11033-016-3959-1

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  • DOI: https://doi.org/10.1007/s11033-016-3959-1

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