Improving DNA isolation from honey for the botanical origin identification

Honey is a natural product highly consumed due its known association with health benefits. Monofloral honeys are perceived as better quality products, being the most appreciated by consumers, thus attaining higher market values. Therefore efficient tools are needed as alternatives to the classical microscopic analysis presently used for the botanical origin identification of honey. In the present work, the use of DNA-based methods for the botanical species identification of honey is proposed. For this purpose, five DNA extraction methods (the kits NucleoSpin Plant (methods A and B) and DNeasy Plant Mini Kit, and the in-house CTAB-based and Wizard methods) combined with three different sample pre-treatments were applied to four honey samples (3 monofloral honeys of Calluna vulgaris, Lavandula spp. and Eucalyptus spp. and one multifloral honey). The 15 DNA extraction protocols were compared in terms of DNA integrity, yield and purity, as well as capacity of amplification targeting universal and adh1 specific genes of C. vulgaris. The results demonstrated the superior efficacy of the Wizard method in terms of DNA quality and amplification capacity, when combined with the sample preparation treatment with a mechanical disruption step of pollen to improve DNA yield. Although with considerable lower DNA yields, the CTAB and DNeasy methods were also successful because both were able to clearly amplify heather DNA from the monofloral heather honey.This work has been supported by FCT through grants PEst-C/ EQB/LA0006/2013 and NORTE-07-0124-FEDER-000069-Food Science. S. Soares is grateful to FCT PhD grant (SFRH/BD/75091/2010) financed by POPH-QREN (subsidised by FSE and MCTES).