Abstract
DNA barcoding coupled high resolution melting (Bar-HRM) is an emerging method for species discrimination based on DNA dissociation kinetics. The aim of this work was to evaluate the suitability of different primer sets, derived from selected DNA regions, for Bar-HRM analysis of species in Croton (Euphorbiaceae), one of the largest genera of plants with over 1,200 species. Seven primer pairs were evaluated (matK, rbcL1, rbcL2, rbcL3, rpoC, trnL and ITS1) from four plastid regions, matK, rbcL, rpoC, and trnL, and the nuclear ribosomal marker ITS1. The primer pair derived from the ITS1 region was the single most effective region for the identification of the tested species, whereas the rbcL1 primer pair gave the lowest resolution. It was observed that the ITS1 barcode was the most useful DNA barcoding region overall for species discrimination out of all of the regions and primers assessed. Our Bar-HRM results here also provide further support for the hypothesis that both sequence and base composition affect DNA duplex stability.
Highlights
Classification of Croton and uses in ethnomedicineCroton (Euphorbiaceae) is one of the largest genera of flowering plants, with between 1,200 and 1,300 species
The ITS1 was found to be optimal primer pair in this study, it is likely that other markers and/or combinations thereof might perform better in other plant groups as the results shown in this study are limited to the examined genus (Croton)
barcoding coupled high resolution melting (Bar-high resolution melting (HRM)) has proven to be a cost-effective and reliable method for the identification of the twelve Croton species tested in this study
Summary
Classification of Croton and uses in ethnomedicine. Croton (Euphorbiaceae) is one of the largest genera of flowering plants, with between 1,200 and 1,300 species. It is widespread in tropical areas, with habits ranging from large woody trees through climbing lianas to simple and prostrate weeds [1,2]. In Southeast Asia and Thailand there are at least 80 species and 30 species of Croton, respectively [3]. The complex subgeneric taxonomy of Croton relies on a provisional revision of the sections of the genus from the early 1990s [1]. Several studies, based on both classical taxonomy and phylogenetic analyses, have.
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