High Resolution Melting (HRM) Genotyping in the Genus Origanum : Molecular Identification and Discrimination for Authent
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High Resolution Melting (HRM) Genotyping in the Genus Origanum: Molecular Identification and Discrimination for Authentication Purposes Charikleia Papaioannou1,2 · Konstantina Zeliou3 · Panayiotis Trigas4 · Vasileios Papasotiropoulos1 Received: 22 March 2020 / Accepted: 4 May 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Origanum L. (Lamiaceae) is an important genus of medicinal and aromatic plants used since ancient times as culinary herbs and remedies in traditional medicine. Although it is a relatively small genus, intra-generic species delineation, as well as its inter-generic relationships within tribe Mentheae, are still poorly understood. High resolution melting (HRM) analysis, coupled with microsatellite markers (SSRs), could facilitate the molecular identification and characterization of certain genotypes more efficiently and relatively faster when compared to other analytical methods. In this study, 38 Origanum samples corresponding to six Origanum taxa (O. dictamnus, O. majorana, O. onites, O. scabrum, O. sipyleum, and O. vulgare subsp. hirtum) were analyzed, using six microsatellite loci. Our goal was to molecularly identify and discriminate among the selected samples and to evaluate the ability of the HRM technique as an analytical tool for the discrimination of Origanum species from Greece. The temperature-shifted melting curves produced by the HRM analysis, resulted in 98 unique HRM profiles, which enabled the discrimination of the Origanum genotypes studied. According to the similarity dendrogram based on the HRM profiles, six unique clusters were formed, each one corresponding to a single taxon. In conclusion, HRM genotyping provided a fast, cost-effective method, well suited for the molecular characterization and identification of Origanum taxa and for the authentication of the original genetic material. Keywords High resolution melting analysis · Origanum · Microsatellites · Genotyping · Authentication
Electronic supplementary material The online version of this article (doi:https://doi.org/10.1007/ s10528-020-09970-1) contains supplementary material, which is available to authorized users. * Vasileios Papasotiropoulos [email protected] Extended author information available on the last page of the article
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Biochemical Genetics
Introduction Origanum L. (Lamiaceae) comprises 44 species (> 60 taxa, and > 18 hybrids) of herbaceous perennials or subshrubs, native to the Mediterranean and Eurasia, usually growing in rocky mountainous areas (Ietswaart 1980; Lukas et al. 2013). Within the genus, there are several culinary, fragrant, medicinal and ornamental plants, with great economic importance worldwide (Bakkali et al. 2008). Medicinally, oregano (Origanum vulgare L.) has been used for thousands of years as a stimulant, carminative, expectorant, and tonic, and to cure asthma, coughs, indigestion, rheumatism, toothache, headache, and spider bites (Vannozzi et al. 2018). Oregano, naturalized in almost all continents (Kokkini
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