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

2021Journal article Open access
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dc.contributor.authorQuaresma, Andreia
dc.contributor.authorBrodschneider, Robert
dc.contributor.authorGratzer, Kristina
dc.contributor.authorGray, Alison
dc.contributor.authorKeller, Alexander
dc.contributor.authorKilpinen, Ole
dc.contributor.authorRufino, José
dc.contributor.authorSteen, Jozef van der
dc.contributor.authorVejsnæs, Flemming
dc.contributor.authorPinto, M. Alice
dc.date.accessioned2021-11-17T15:53:01Z
dc.date.available2021-11-17T15:53:01Z
dc.date.issued2021
dc.description.abstractPollen 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.pt_PT
dc.description.sponsorshipWe are deeply indebted to Susana Lopes and Maria Magalhães, from CIBIO—Research Centre in Biodiversity and Genetic Resources—InBIO Associate Laboratory, for their time devoted to library preparation and sequencing in the MiSeq. AQ acknowledges the PhD scholarship (DFA/BD/5155/2020) funded by FCT. This work was funded by the Health and Food Safety Directorate General, European Commission through the project INSIGNIA—Environmental monitoring of pesticide use through honeybees SANTE/E4/SI2.788418-SI2.788452- INSIGINIA-PP-1–1-2018. Fundação para a Ciência e a Tecnologia (FCT) provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020).pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationQuaresma, Andreia; Brodschneider, Robert; Gratzer, Kristina; Gray, Alison; Keller, Alexander; Kilpinen, Ole; Rufino, José; Steen, Jozef van der; Vejsnæs, Flemming; Pinto, M. Alice (2021). Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcoding. Environmental Monitoring and Assessment. ISSN 0167-6369. 193:12, p. 1-20pt_PT
dc.identifier.doi10.1007/s10661-021-09563-4pt_PT
dc.identifier.issn0167-6369
dc.identifier.urihttp://hdl.handle.net/10198/24219
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectDNA metabarcodingpt_PT
dc.subjectPollenpt_PT
dc.subjectDNA barcodingpt_PT
dc.subjectPreservation biaspt_PT
dc.subjectSilica gelpt_PT
dc.subjectPreservationpt_PT
dc.subjectCitizen sciencept_PT
dc.titlePreservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcodingpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.issue12pt_PT
oaire.citation.titleEnvironmental Monitoring and Assessmentpt_PT
oaire.citation.volume193pt_PT
person.familyNameQuaresma
person.familyNameRufino
person.familyNamePinto
person.givenNameAndreia
person.givenNameJosé
person.givenNameM. Alice
person.identifier.ciencia-id4F1A-4E4A-3F23
person.identifier.ciencia-idC414-F47F-6323
person.identifier.ciencia-idF814-A1D0-8318
person.identifier.orcid0000-0002-8678-5800
person.identifier.orcid0000-0002-1344-8264
person.identifier.orcid0000-0001-9663-8399
person.identifier.scopus-author-id57119742600
person.identifier.scopus-author-id55947199100
person.identifier.scopus-author-id8085507800
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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relation.isAuthorOfPublication1e24d2ce-a354-442a-bef8-eebadd94b385
relation.isAuthorOfPublication0667fe04-7078-483d-9198-56d167b19bc5
relation.isAuthorOfPublication.latestForDiscovery0667fe04-7078-483d-9198-56d167b19bc5

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