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Molecular and morphological discrimination of Chrysanthemum indicum using allele-specific PCR and T-shaped trichome

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Abstract

Chrysanthemum indicum L. is a traditional oriental medicinal herb prepared as a tea from flowers that have been used in China and South Korea since ancient times. It has a long history in the treatment of hypertension, inflammation, and respiratory diseases. Among Chrysanthemum species, C. indicum has more active chemical components as well as better therapeutic effects, and C. indicum is mostly used for medicinal purposes in South Korea. However, the usage of C. indicum has become problematic over the years due to the abundance of adulterated Chrysanthemum and confusion with morphologically related species such as C. morifolium, C. boreale, and Aster spathulifolius. Thus, here we developed a method for molecular authentication using chloroplast universal region rpoC2 and morphological authentication based on T-shaped trichomes of the adaxial leaf surface. By using a species-specific primer derived from the rpoC2 region, we established a multiplex allele-specific PCR for the discrimination of C. indicum. Amplicons of 675 bp for C. indicum and 1026 bp for other Chrysanthemum species were produced using both rpoC2-specific and common primers. These primers can be used to analyze dried samples of Chrysanthemum. Morphological discrimination was performed using T-shaped trichomes present only on the adaxial leaf surface of C. indicum species, and then molecular markers were utilized to authenticate C. indicum products from adulterant samples available in the market. Our results indicate that these molecular markers in combination with morphological differentiation can serve as an effective tool for identifying C. indicum.

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Funding

This study was supported by a grant from the Korea Institute of Planning & Evaluation for Technology in Food, Agriculture, Forestry & Fisheries (KIPET NO: 119111-01), Republic of Korea.

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Authors and Affiliations

  1. Graduate School of Biotechnology, College of Life Sciences, Kyung Hee University, Yongin si, Gyeonggi do, 17104, Republic of Korea

    Suleman Abid, Lalitha Kaliraj, Muhammad Huzaifa Arif, Joon Hurh, Jong Chan Ahn, Deok Chun Yang & Seok-Kyu Jung

  2. Department of Oriental Medicinal Biotechnology, College of Life Science, Kyung Hee University, Yongin si, Gyeonggi do, 17104, Republic of Korea

    Deok Chun Yang & Seok-Kyu Jung

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  1. Suleman Abid
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  2. Lalitha Kaliraj
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Correspondence to Deok Chun Yang or Seok-Kyu Jung.

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Abid, S., Kaliraj, L., Arif, M.H. et al. Molecular and morphological discrimination of Chrysanthemum indicum using allele-specific PCR and T-shaped trichome. Mol Biol Rep 47, 7699–7708 (2020). https://doi.org/10.1007/s11033-020-05844-2

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  • DOI: https://doi.org/10.1007/s11033-020-05844-2

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