Assessment of the botanical origin of Bulgarian honey samples using melissopalynological, DNA barcoding and NMR analyses

Polyfloral honey samples from Bulgaria were subject to parallel analyses of their botanical origin and composition using traditional melissopalynology, DNA barcoding based on the plastid rbcL gene and NMR analysis. The obtained datasets were compared with each other to evaluate the information capacity of the applied experimental methods. The results from the melissopalynological and DNA-barcoding studies demonstrated a significantly higher resolution of the latter, revealing the presence of pollen from a total of 17 plant families, 21 plant genera and 5 plant species in comparison to pollen from only 7 plant families, 3 plant genera and 4 plant species identified by melissopalynology. The higher resolution of DNA barcoding allows a more detailed characterisation of the diet and foraging preferences of honey bees, including foraging on plant species growing in lower abundance in the area. The comparison of the quantitative data on floral honey composition for several plant genera and species reveals significant differences between the relative abundance of the pollen grains estimated by melissopalynological analysis and the relative abundance of rbcL clones in rbcL libraries determined after DNA barcoding. All three applied methods confirm the polyfloral botanical origin of the analysed samples and support routine NMR use for the assessment of the floral origin of honey.