Untargeted In Silico Compound Classification—A Novel Metabolomics Method to Assess the Chemodiversity in Bryophytes

In plant ecology, biochemical analyses of bryophytes and vascular plants are often conducted on dried herbarium specimen as species typically grow in distant inaccessible locations. Here, we present an automated silico compound classification framework to annotate metabolites using untargeted data independent acquisition (DIA)–LC/MS–QToF-sequential windowed all theoretical fragment ion mass spectra (SWATH) ecometabolomics analytical method. We perform a comparative investigation the chemical diversity at global level composition metabolite families ten different fresh samples collected on-site stored for half year. Shannon Pielou’s indices, hierarchical clustering analysis (HCA), sparse partial least squares discriminant (sPLS-DA), distance-based redundancy (dbRDA), ANOVA with post-hoc Tukey honestly significant difference (HSD) test, Fisher’s exact test were used determine differences richness families, regard conditions, ecological characteristics, species. functionally annotated processes related structural integrity membranes cell walls (proto-lignin, glycerophospholipids, carbohydrates), defense (polyphenols, steroids), reactive oxygen (ROS) protection (alkaloids, amino acids, flavonoids), nutrition (nitrogen- phosphate-containing glycerophospholipids), photosynthesis. Changes also explained variance functioning like physiological adaptations dry environments (proteins, peptides, flavonoids, terpenes), light availability (flavonoids, terpenes, temperature (flavonoids), biotic interactions (steroids, terpenes). The results from this study allow construct traits that can be attributed biogeochemistry, habitat environmental changes interactions. Our accelerates complex annotation process metabolomics simplify patterns. show is powerful tool allows explore relationships both molecular biology by “zooming in” ecology out”. insights revealed our new research hypotheses enable detailed follow-up studies.