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Pollinating insects are of vital importance for the ecosystem and their drastic decline imposes severe consequences for the environment and humankind. The comprehension of their interaction networks is the first step in order to preserve these highly complex systems. For that purpose, the following study describes a protocol for the investigation of honey bee pollen samples from different agro-environmental areas by DNA extraction, PCR amplification and nanopore sequencing of the barcode regions rbcL and ITS. It was shown, that the most abundant species were classified consistently by both DNA barcodes, while species richness was enhanced by single-barcode detection of less abundant species. The analysis of the the different landscape variables exhibited a decline of species richness, Shannon diversity index, and species evenness with increasing organic crop area. However, sampling was only carried out in August and further investigations are suggested to display a more complete picture of honey bee foraging throughout the seasons.