Validating a multi-locus metabarcoding approach for characterizing mixed-pollen samples

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dc.contributor.authorWizenberg, Sydney B.
dc.contributor.authorNewburn, Laura R.
dc.contributor.authorPepinelli, Mateus
dc.contributor.authorConflitti, Ida M.
dc.contributor.authorRichardson, Rodney T.
dc.contributor.authorHoover, Shelley E. R.
dc.contributor.authorCurrie, Robert W.
dc.contributor.authorGiovenazzo, Pierre
dc.contributor.authorZayed, Amro
dc.date.accessioned2023-12-12T17:46:30Z
dc.date.available2023-12-12T17:46:30Z
dc.date.issued2023-11-04
dc.date.updated2023-12-01T04:27:00Z
dc.description.abstractBackground The mutualistic interaction between entomophilous plants and pollinators is fundamental to the structure of most terrestrial ecosystems. The sensitive nature of this relationship has been disrupted by anthropogenic modifications to natural landscapes, warranting development of new methods for exploring this trophic interaction. Characterizing the composition of pollen collected by pollinators, e.g. Apis mellifera, is a common means of exploring this relationship, but traditional methods of microscopic pollen assessment are laborious and limited in their scope. The development of pollen metabarcoding as a method of rapidly characterizing the abundance and diversity of pollen within mixed samples presents a new frontier for this type of work, but metabarcoding may have limitations, and validation is warranted before any suite of primers can be confidently used in a research program. We set out to evaluate the utility of an integrative approach, using a set of established primers (ITS2 and rbcL) versus melissopalynological analysis for characterizing 27 mixed-pollen samples from agricultural sites across Canada. Results Both individual markers performed well relative to melissopalynology at the family level with decreases in the strength of correlation and linear model fits at the genus level. Integrating data from both markers together via a multi-locus approach provided the best rank-based correlation between metagenetic and melissopalynological data at both the genus (ρ = 0.659; p < 0.001) and family level (ρ = 0.830; p < 0.001). Species accumulation curves indicated that, after controlling for sampling effort, melissopalynological characterization provides similar or higher species richness estimates than either marker. The higher number of plant species discovered via the metabarcoding approach simply reflects the vastly greater sampling effort in comparison to melissopalynology. Conclusions Pollen metabarcoding performed well at characterizing the composition of mixed pollen samples relative to a traditional melissopalynological approach. Limitations to the quantitative application of this method can be addressed by adopting a multi-locus approach that integrates information from multiple markers.
dc.identifier.citationPlant Methods. 2023 Nov 04;19(1):120
dc.identifier.doi10.1186/s13007-023-01097-9
dc.identifier.urihttp://hdl.handle.net/1993/37864
dc.language.isoeng
dc.language.rfc3066en
dc.publisherBMC
dc.rightsopen accessen_US
dc.rights.holderThe Author(s)
dc.subjectpollen
dc.subjectpollination
dc.subjectmetabarcoding
dc.subjectmetagenetic
dc.subjectpalynology
dc.subjectMelissopalynology
dc.titleValidating a multi-locus metabarcoding approach for characterizing mixed-pollen samples
dc.typeJournal Article
local.author.affiliationFaculty of Agricultural and Food Sciences::Department of Entomology
oaire.citation.issue120
oaire.citation.titlePlant Methods
oaire.citation.volume19
project.funder.identifierhttp://dx.doi.org/10.13039/100008762
project.funder.nameGenome Canada
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