Capture by hybridization for full-length barcode-based eukaryotic and
prokaryotic biodiversity inventories for deep sea ecosystems
Abstract
Biodiversity inventory remains limited in marine systems due to
unbalanced access to the three ocean dimensions. The use of
environmental DNA (eDNA) for metabarcoding allows fast and effective
biodiversity inventory and is forecast as a future biodiversity research
and biomonitoring tool. However, in poorly understood ecosystems, eDNA
results remain difficult to interpret due to large gaps in reference
databases and PCR bias limiting the detection of some major phyla. Here,
we aimed to circumvent these limitations by avoiding PCR and
recollecting larger DNA fragments to improve assignment of detected taxa
through phylogenetic reconstruction. We applied capture by hybridization
(CBH) to enrich DNA from deep-sea sediment samples and compared the
results with those obtained through an up-to-date metabarcoding
PCR-based approach (MTB). Originally developed for bacterial communities
by targeting 16S rDNA, the CBH approach was applied to 18S rDNA to
improve the detection of species forming benthic communities of
eukaryotes, with particular focus on metazoans. The results confirmed
the possibility of extending CBH to metazoans with two major advantages:
i) CBH revealed a broader spectrum of prokaryotic, eukaryotic, and
particularly metazoan diversity, and ii) CBH allowed much more robust
phylogenetic reconstructions of full-length barcodes with up to 1900
base pairs. This is particularly important for taxa whose assignment is
hampered by gaps in reference databases. This study provides a database
and probes to apply 18S CBH to diverse marine systems, confirming this
promising new tool to improve biodiversity assessments in data-poor
ecosystems like those in the deep sea.