Feature-Based Molecular Networking-An Exciting Tool to Spot Species of the Genus with Hidden Photosensitizers.

Fungi have developed a wide array of defense strategies to overcome mechanical injuries and pathogen infections. Recently, photoactivity has been discovered by showing that pigments isolated from produce singlet oxygen under irradiation. To test if this phenomenon is limited to dermocyboid Cortinarii, six colourful species belonging to different classical subgenera (i.e., , , , , and ) were investigated. Fungal extracts were explored by the combination of in vitro photobiological methods, UHPLC coupled to high-resolution tandem mass spectrometry (UHPLC-HRMS), feature-based molecular networking (FBMN), and metabolite dereplication techniques. The fungi   () and   () exhibited promising photobiological activity in a low concentration range (1-7 µg/mL). Using UHPLC-HRMS-based metabolomic tools, the underlying photoactive principle was investigated. Several monomeric and dimeric anthraquinones were annotated as compounds responsible for the photoactivity. Furthermore, the results showed that light-induced activity is not restricted to a single subgenus, but rather is a trait of species of different phylogenetic lineages and is linked to the presence of fungal anthraquinones. This study highlights the genus as a promising source for novel photopharmaceuticals. Additionally, we showed that putative dereplication of natural photosensitizers can be done by FBMN.