Molecular and morphological discrimination of Chrysanthemum indicum using allele-specific PCR and T-shaped trichome
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ORIGINAL ARTICLE
Molecular and morphological discrimination of Chrysanthemum indicum using allele‑specific PCR and T‑shaped trichome Suleman Abid1 · Lalitha Kaliraj1 · Muhammad Huzaifa Arif1 · Joon Hurh1 · Jong Chan Ahn1 · Deok Chun Yang1,2 · Seok‑Kyu Jung1,2 Received: 14 June 2020 / Accepted: 11 September 2020 © Springer Nature B.V. 2020
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. Keywords Chrysanthemum indicum · SNP marker · Chloroplast DNA · rpoC2 · Multiplex allele-specific PCR · Trichomes
Introduction Medicinal plants are a source of several lifesaving drugs [1]. Chrysanthemum indicum L. is widespread in China and Korea and has been used as herbal medicine since ancient Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-020-05844-2) contains supplementary material, which is available to authorized users.
times. This plant produces small yellow-colored flowers useful for making tea, additives, and beverages [2]. Its blooms are also important in the fresh-cut flower industry [3]. The inflorescence of C. indicum, particularly the flower buds, possesses anti-inflammatory [4], hepatoprotective [5], and antioxidant properties [6]; hence, it has been used as an antiphlogistic to treat headache, relieve hypertension, conjunctivitis, furuncle, and carbuncle. The essential oils
* Deok Chun Yang [email protected]
Joon Hurh [email protected]
* Seok‑Kyu Jung [email protected]
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