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Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcoding.
Quaresma, Andreia; Brodschneider, Robert; Gratzer, Kristina; Gray, Alison; Keller, Alexander; Kilpinen, Ole; Rufino, José; van der Steen, Jozef; Vejsnæs, Flemming; Pinto, M Alice.
Afiliação
  • Quaresma A; Centro de Investigação de Montanha, Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253, Bragança, Portugal.
  • Brodschneider R; Institute of Biology, University of Graz, Universitätsplatz 2, 8010, Graz, Austria.
  • Gratzer K; Institute of Biology, University of Graz, Universitätsplatz 2, 8010, Graz, Austria.
  • Gray A; Department of Mathematics and Statistics, University of Strathclyde, Glasgow, UK.
  • Keller A; Center for Computational and Theoretical Biology, Hubland Nord, Würzburg, Germany.
  • Kilpinen O; Department of Bioinformatics, University of Würzburg, Am Hubland, BiocenterWürzburg, Germany.
  • Rufino J; Danish Beekeepers Association, Sorø, Denmark.
  • van der Steen J; Research Centre in Digitalization and Intelligent Robotics (CeDRI), Instituto Politécnico de Bragança, Bragança, Portugal.
  • Vejsnæs F; Alveus AB Consultancy, Oisterwijk, Netherlands.
  • Pinto MA; Danish Beekeepers Association, Sorø, Denmark.
Environ Monit Assess ; 193(12): 785, 2021 Nov 09.
Article em En | MEDLINE | ID: mdl-34755261
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 < 0.027). Statistical differences were also detected for DNA concentration, with EtOH samples producing lower yields than RT and FRZ samples in both countries and SG in Austria (P < 0.042). Yet, qualitative and quantitative assessments of floral composition obtained using high-throughput sequencing with the ITS2 barcode gave non-significant effects of preservation methods on richness, relative abundance and Shannon diversity, in both countries. While freezing and ethanol are commonly employed for archiving tissue for molecular applications, desiccation is cheaper and easier to use regarding both storage and transportation. Since SG is less dependent on ambient humidity and less prone to contamination than RT, we recommend SG for preserving pollen for metabarcoding. SG is straightforward for laymen to use and hence robust for widespread application in citizen science studies.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Código de Barras de DNA Taxonômico / Mel Tipo de estudo: Qualitative_research Limite: Animals Idioma: En Revista: Environ Monit Assess Assunto da revista: SAUDE AMBIENTAL Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Portugal

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Código de Barras de DNA Taxonômico / Mel Tipo de estudo: Qualitative_research Limite: Animals Idioma: En Revista: Environ Monit Assess Assunto da revista: SAUDE AMBIENTAL Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Portugal