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Honey is one of the most frauded food products. Therefore, it is important to develop new analytical systems useful for its authentication. Honey contains intrinsic markers that can be used to identify and monitor its origin, including plant DNA mainly derived by pollen. In this study, we applied a next generation sequencing approach for honey authentication by detecting the prevalent botanical contribution and botanical composition of honeys of different origin. DNA was isolated from nine honeys (six monofloral honeys produced in Italy, two polyfloral honeys produced in East Europe and Chile respectively, and one honeydew honey) and PCR amplified for a chloroplast trnL barcoding fragment. Obtained amplicons were sequenced using the Ion Torrent sequencing platform. Sequence data was interpreted using a customized bioinformatic pipeline that used a reference plant sequence dataset derived by more than …