Quantitative property of metabarcoding
Metabarcoding on the 18S rRNA region of DNA is a common strategy to study zooplankton. Various sections of this region have been used as barcodes (e.g. , V9 or V4 the standard markers for planktonic eukaryotes since Tara Oceans studies). In this study, we choose to amplify and sequence the V1V2 region, originally used for meiofaunal zoobenthos (Fonseca et al., 2014) and specific to Metazoan (Lejzerowicz et al., 2021). It appears to be one of the best markers to assess marine community changes (Cordier et al., 2018). However, it is important to consider that some taxa are not well characterized, e.g. ,Pleuromamma sp. or Chaetognatha . This may be due to a secondary structure on their ribosomal region which might make their DNA harder to amplify as the primer regions show high fidelity in the available sequences in the NCBI. This could impact our results sincePleuromamma sp., which is highly abundant at BATS, is known to rapidly react to the new primary production and to contribute significantly to the ecosystem carbon flux (between 4% and 70%, Steinberg et al., 2000). Chaetognaths are also common in this study region. They are thought to be linked to the bloom community, since their abundance usually follows copepod density maxima and decreases with increasing stratification of the water column in the Sargasso Sea (Ivory et al., 2019) and their morphology matches this cluster’s characteristics. Therefore, due to chaetognath’s sequences low amplification we might have missed potential interspecific interactions. Generally, correlations are found between the number of reads per ASV and the corresponding taxon biomass or abundance (Bucklin et al., 2019). Significant relationship also exists between the proportion of input material and the proportion of sequences obtained per species from metabarcoding, however large uncertainties remain (Lamb et al., 2019). Here, positive correlations were found for some coarse taxonomic groups. A negative correlation was found for ostracods abundances versus sequence reads proportions. It was not the case in Matthews et al. (2021) in which they found a positive correlation between ostracods’ relative abundances and proportion of sequences for both 18S V4 and COI primers. It might be due to a technical bias from the metabarcoding process. On the contrary, some groups appeared to be well suited for a quantitative metabarcoding use. Positive correlations were found for abundance and biomass of Cyclopoida, Doliolida, Harpacticoida and Larvacea. For biomass, similar trends were observed for Cyclopoida, Doliolida and Larvacea (Matthews et al., 2021). The proportion of reads of Chaetognatha and Cladocera well correlated with their abundance but not their biomass. Chaetognatha are not well identified by 18S V1V2 (Matthews et al., 2021) which might explain the absence of correlation between relative number of sequences and biomass. On the contrary, relative number of sequences and biomass was positively correlated for Cephalopoda despite their low abundance. For the remaining taxa, the absence of significant correlations might be due to not-specific or inaccurate conversion factors from biovolume to dry weight. Hence, more studies such as the one by Maas et al. (2021) should be done before assuming a quantitative nature.