Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcoding

Quaresma, Andreia and Brodschneider, Robert and Gratzer, Kristina and Gray, Alison and Keller, Alexander and Kilpinen, Ole and Rufino, José and van der Steen, Jozef and Vejsnæs, Flemming and Pinto, M. Alice (2021) Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcoding. Environmental Monitoring and Assessment, 193 (12). 785. ISSN 0167-6369 (https://doi.org/10.1007/s10661-021-09563-4)

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Abstract

Pollen metabarcoding is emerging as a powerful tool for ecological research and offers unprecedented scale in citizen science projects for environmental monitoring via honey bees. Biases in metabarcoding can be introduced at any stage of sample processing and preservation is at the forefront of the pipeline. While in metabarcoding studies pollen has been preserved at - 20 °C (FRZ), this is not the best method for citizen scientists. Herein, we compared this method with ethanol (EtOH), silica gel (SG) and room temperature (RT) for preservation of pollen collected from hives in Austria and Denmark. After ~ 4 months of storage, DNAs were extracted with a food kit, and their quality and concentration measured. Most DNA extracts exhibited 260/280 absorbance ratios close to the optimal 1.8, with RT samples from Austria performing slightly worse than FRZ and SG samples (P