Abstract
The democratization of sequencing technologies fostered a leap in our knowledge of the diversity of marine phytoplanktonic microalgae, revealing many previously unknown species and lineages. The evolutionary history of the diversification of microalgae can be inferred from the analysis of their genome sequences. However, the link between the DNA sequence and the associated phenotype is notoriously difficult to assess, all the more so for marine phytoplanktonic microalgae for which the lab culture and, thus, biological experimentation is very tedious. Here, we explore the potential of a high-throughput untargeted metabolomic approach to explore the phenotypic–genotypic gap in 12 marine microalgae encompassing 1.2 billion years of evolution. We identified species- and lineage-specific metabolites. We also provide evidence of a very good correlation between the molecular divergence, inferred from the DNA sequences, and the metabolomic divergence, inferred from the complete metabolomic profiles. These results provide novel insights into the potential of chemotaxonomy in marine phytoplankton and support the hypothesis of a metabolomic clock, suggesting that DNA and metabolomic profiles co-evolve.
Highlights
Phytoplanktonic eukaryotes are phylogenetically highly diverse, as they have many representatives in most super-groups of the eukaryotic tree of life [1,2]
We explored the potential of an untargeted metabolomic approach including pigments, lipids, and other uncharacterized metabolites to investigate chemotaxonomic markers in 12 marine microalgal strains from 11 species, including 9 microalgae from the green lineage; the Mamiellales Ostreococcus tauri [20], O. mediterraneus [21], Bathycoccus prasinos [22], Micromonas commoda [23], and Mantoniella sp., the Chlorellales Picochlorum costavermella [24], and strains from basal groups Nephroselmis sp. and Pyramimonas sp
Algal ethyl acetate extracts were analyzed by UHPLC-ESI+ -HRMS2, and acquired ion chromatograms were processed through an untargeted metabolomic workflow in Compound Discoverer 2.1 software (Thermo Scientific) to generate extracted ion chromatograms across samples, detect and quantify corresponding metabolites, and generate the observations/variables matrix used for further statistical analyses
Summary
Phytoplanktonic eukaryotes are phylogenetically highly diverse, as they have many representatives in most super-groups of the eukaryotic tree of life [1,2]. Our knowledge on the diversity of phytoplanktonic green microalgae has greatly increased with the democratization of DNA sequencing and genomics, but it is likely to stay behind that of their terrestrial relatives, including land plants, whose estimated species number exceeds 400,000 [5]. We explored the potential of an untargeted metabolomic approach including pigments, lipids, and other uncharacterized metabolites to investigate chemotaxonomic markers in 12 marine microalgal strains from 11 species, including 9 microalgae from the green lineage; the Mamiellales Ostreococcus tauri [20], O. mediterraneus [21], Bathycoccus prasinos [22], Micromonas commoda [23], and Mantoniella sp., the Chlorellales Picochlorum costavermella [24], and strains from basal groups Nephroselmis sp. This leads us to discuss the hypothesis of a metabolomic corollary of the molecular clock, a central tenet of molecular evolution
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