Avoiding quantification bias in metabarcoding: Application of a cell biovolume correction factor in diatom molecular biomonitoring
Visualitza/Obre
Autor/a
Vasselon, Valentin
Bouchez, Agnès
Rimet, Frédéric
Jacquet, Stéphan
Corniquel, Méline
Topolczai, Kálmán
Domaizon, Isabelle
Data de publicació
2017-12-26ISSN
2041-210X
Resum
In recent years, remarkable progress has been made in developing environmental DNA metabarcoding. However, its ability to quantify species relative abundance remains uncertain, limiting its application for biomonitoring. In diatoms, although the rbcL gene appears to be a suitable barcode for diatoms, providing relevant qualitative data to describe taxonomic composition, improvement of species quantification is still required.
Here, we hypothesized that rbcL copy number is correlated with diatom cell biovolume (as previously described for the 18S gene) and that a correction factor (CF) based on cell biovolume should be applied to improve taxa quantification. We carried out a laboratory experiment using pure cultures of eight diatom species with contrasted cell biovolumes in order to (1) verify the relationship between rbcL copy numbers (estimated by qPCR) and diatom cell biovolumes and (2) define a potential CF. In order to evaluate CF efficiency, five mock communities were created by mixing different amounts of DNA from the eight species, and were sequenced using HTS and targeting the same rbcL barcode.
As expected, the correction of DNA reads proportions by the CF improved the congruence between morphological and molecular inventories. Final validation of the CF was obtained on environmental samples (metabarcoding data from 80 benthic biofilms) for which the application of CF allowed differences between molecular and morphological water quality indices to be reduced by 47%.
Overall, our results highlight the usefulness of applying a CF factor, which is effective in reducing over‐estimation of high biovolume species, correcting quantitative biases in diatom metabarcoding studies and improving final water quality assessment.
Tipus de document
Article
Versió del document
Versió acceptada
Llengua
Anglès
Matèries (CDU)
574 - Ecologia general i biodiversitat
Pàgines
37
Publicat per
Wiley
Publicat a
Methods in Ecology and Evolution
Citació
Vasselon, Valentin, Agnès Bouchez, Frédéric Rimet, Stéphan Jacquet, Rosa Trobajo, Méline Corniquel, Kálmán Tapolczai, and Isabelle Domaizon. 2018. "Avoiding Quantification Bias In Metabarcoding: Application Of A Cell Biovolume Correction Factor In Diatom Molecular Biomonitoring". Methods In Ecology And Evolution 9 (4): 1060-1069. Wiley. doi:10.1111/2041-210x.12960.
Número de l'acord de la subvenció
EC/COST/CA15219/EU/Developing new genetic tools for bioassessment of aquatic ecosystems in Europe/DNAqua-Net
Programa
Aigües Marines i Continentals
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