Development of a new approach based on NADH dehydrogenase subunit 1(ND1) marker and high resolution melting (HRM) analysis towards the authentication of the geographical origin of honey

In the present study, an approach for verifying the geographical origin of honey based on its entomological origin is proposed. The method was developed based on the polymerase chain reaction (PCR) amplification of a new markercontaining different single nucleotide polymorphisms characteristic of honeybees of different mitochondrial (mtDNA) lineages, therefore generating different fluorescent curves on HRM analysis. The method was successfully applied to honeys from Portugal and Italy, expected to be from lineages A and C, respectively, demonstrating their origin compliance.The authors are grateful to Programa Nacional Apícola 2020-2022 for funding the project “AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português”, to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO UIDB/00690/2020, to Fenapícola and Capemel for supplying the Portuguese honeys and to Dr. Antonio Nanetti (CREA-AA) for the Italian honeys. D. Henriques is supported by the project Bee Happy (POCI-01-0145-FEDER-029871) funded by FEDER, COMPETE 2020-POCIand FCT and A.R. Lopes by the PhD scholarship funded by the FCT SFRH/BD/143627/2019.