Honey bee-collected pollen for botanical identification via ITS2 metabarcoding: a comparison of preservation methods for citizen science
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DNA metabarcoding is emerging as a powerful method for botanical identification
of bee-collected pollen, allowing analysis of hundreds of samples in a single
high-throughput sequencing run, therefore offering unprecedented scale in
citizen science projects. Biases in metabarcoding can be introduced at any stage
of sample processing and preservation is the first step of the pipeline. Hence, it is
important to test whether the pollen preservation method influences
metabarcoding performance. While in metabarcoding studies pollen has typically
been preserved at −20°C, this is not the best method to be applied by citizen
scientists. Here, we compared the freezing method (FRZ) with ethanol (EtOH),
silica gel (SG) and room temperature (RT) in 87 pollen samples collected from
hives in Austria and Denmark.
While classical palynology has been the method of choice to assess botanical diversity of bee-collected pollen for multiple purposes, DNA metabarcoding is emerging as a powerful alternative being able to achieve high taxonomic identification accuracy. Moreover,DNA metabarcoding allows analysis of hundreds of samples in a single high-throughput sequencing run, therefore offering unprecedented scale in citizen science projects. Biases in metabarcoding can be introduced at any stage of sample processing and preservation
is at the forefront of the pipeline. Hence, it is important to test how sample preservation influences quality and quantitative performance of pollen metabarcoding. While inmetabarcoding studies pollen has typically been preserved at −20°C (FRZ), this is not the best method to be applied by citizen scientists.
