Nus-specific chemical information was expected to choose meaningful spectral matches, in spite of the significant improvement of annotation reliability via taxonomically informed metabolite annotation [26]. To highlight this limitation, as much as 4 spectral hits (ranked as described beneath) represented as their respective SMILES have been listed in Tables S5 14 (SI Sections three.4.four.1.1.4.four.1.ten). The 4 levels of accuracy reported within the Metabolomics Standard FCCP site Initiative were employed to describe the self-assurance level achieved in the function annotation (i.e., marker compounds = 1, all other Rapamycin Cancer putative annotations = two, Tables S5 14) [53]. Spectral matching of MS2 spectra against the GNPS spectral library led to 148 hits (four.0), whereas the taxonomically informed metabolite annotation against in silico ISDB-DNP afforded 2269 putative annotations (60.6 ; relative for the number of nodes in the FBMN) (SI Section three.4.6). Molecular networking research within the field of mycochemical analysis and as a result publicly readily available mass spectrometry data originating from fungal secondary metabolites could increase the GNPS-associated annotation hit rate as well because the all round annotation process. two.four.three. Specificity of Features and Phylogenetic Relevance Based on Figure five, the active clusters A, D, E, F, G, H, and I had been distinct for C. rubrophyllus and putatively identified as clusters comprising monomeric and dimeric anthraquinones (AQs), also as their chlorinated (confirmed by the isotopic pattern, SI Figures S32 35), glycosylated, methylated, and esterified derivates (SI Section three.four.four). Clusters F and H had been also partly precise for C. xanthophyllus. Cluster B was precise for C. rubrophyllus and C. xanthophyllus alike, pointing towards the possibility of a shared evolutionary history of these two species based on their secondary metabolite profiles. For many years, chemotaxonomy was viewed as the essential to resolving evolutionary relationships but was later replaced by molecular solutions (e.g., the investigation of sequences of the rDNA internal transcribed spacer (ITS) or the rDNA massive subunit (LSU)) [54]. Nonetheless, it was impossible to resolve basal phylogenetic relationships in the particularly species-rich genus Cortinarius, despite the fact that it represents one of the best-studied genera of Agaricomycetes (Basidiomycetes) and was addressed based on several multigene phylogenetic research [46,55]. By way of example, rDNA ITS sequences of C. claroflavus and C. xanthophyllus are identical [56], a reality highlighting the restricted resolution of molecular phylogenies primarily based on rDNA sequences only. These two species could be clearly distinguished primarily based on basidiome pigmentation and unique macrochemical color reactions.Metabolites 2021, 11,The C. glaucopus genome is comparatively massive (20k proteins (ncbi.nlm.nih.gov/genome/96691, accessed on 18 November 2021). Explora10 of 20 tive research like the full genome of all involved Cortinarius species are necessary to investigate pigment biosynthesis and its evolutionary history in these fungi in far more detail.Figure five. The FBMN with all the marked photoactive clusters (A) (top left corner) as well because the annotation final results for the clusters A . Options distinct for the active extracts are highlighted as colored nodes: C. rubrophyllus = red, C. xanthophyllus = purple. The node size depicts the features’ capacity to absorb visible light: significant = “VIS-Signal” present, small = “VIS-Signal” absent. The marker compounds (i.e., dermolutein (ID3), dermorubin (ID5), 5-c.