COI metabarcoding of large benthic Foraminifera: Method validation for application in ecological studies

Monitoring community composition of Foraminifera (single-celled marine protists) pro-vides valuable insights into environmental conditions in marine ecosystems. Despitethe efficiency of environmental DNA (eDNA) and bulk-sample DNA (bulk-DNA) me-tabarcoding to assess the presence of multiple taxa, this has not been straightforwardfor Foraminifera partially due to the high genetic variability in widely used ribosomalmarkers. Here, we test the correctness in retrieving foraminiferal communities by me-tabarcoding of mock communities, bulk-DNA from coral reef sediment samples, andeDNA from their associated ethanol preservative using the recently sequenced cy-tochrome c oxidase subunit 1 (COI) marker. To assess the detection success, we com-pared our results with large benthic foraminiferal communities previously reportedfrom the same sampling sites. Results from our mock communities demonstrate thatall species were detected in two mock communities and all but one in the remainingfour. Technical replicates were highly similar in number of reads for each assigned ASVin both the mock communities and bulk-DNA samples. Bulk-DNA showed a signifi-cantly higher species richness than their associated eDNA samples, and also detectedadditional species to what was already reported at the specific sites. Our study con-firms that metabarcoding using the foraminiferal COI marker adequately retrieves thediversity and community composition of both the mock communities and the bulk-DNA samples. With its decreased variability compared with the commonly used nu-clear 18 S rRNA, the COI marker renders bulk-DNA metabarcoding a powerful tool toassess foraminiferal community composition under the condition that the referencedatabase is adequate to the target taxa.

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