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| Main Authors: | , , , |
|---|---|
| Format: | Artículo científico |
| Language: | en |
| Published: |
ISME communications
2026
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| Online Access: | https://pubmed.ncbi.nlm.nih.gov/42116847/ |
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Table of Contents:
- Converting relative amplicon abundances to absolute abundances via flow cytometry: metagenomic validation and application to long ocean transects. Williams, Nathan L R Bei, Qicheng Raut, Yubin Fuhrman, Jed A With microbes critical for ocean ecological and biogeochemical processes, we need to understand their abundance and diversity distributions. While traditional amplicon sequencing provides only relative abundance data, and the strongly preferred absolute abundances can be determined from samples spiked with internal standards, few oceanographic studies with absolute abundances exist. However, many have flow cytometry (FCM) data that should allow us to retrospectively "anchor" the relative abundances into absolute abundances. We tested this hypothesis with data from the 29th Atlantic Meridional Transect (AMT29) cruise where we had FCM of , amplicons corrected with internal standards, and absolute cell count estimates from single copy and metagenomics. Anchoring the AMT29 amplicon data with FCM (used because phycoerythrin in is reliably detected by FCM in surface waters) yielded results strongly correlated with amplicon data corrected with internal standards (Pearson's = 0.94, slope = 0.73), FCM ( = 0.80, slope = 0.43), and -based genome counts (Pearson's = 0.94, slope = 0.62). Seeing this method worked reasonably well, we then generated estimates of absolute rRNA gene abundances from the Global rRNA Universal Metabarcoding of Plankton (GRUMP) transects that had FCM data (Pacific ~65 N to ~40S). These FCM-anchored gene copy estimates also showed strong correlations to FCM data (i.e. anchor with and predict ), with values ranging from 0.48-0.86. While the results are clearly only reasonable estimates, we believe the approach has the potential to significantly enhance the value of amplicon data which have accompanying FCM data.