Studies have already shown that interlinking microbiomes with medicinal plants play a key role on plant growth processing. The plants-associated microorganisms may modulate the biosynthesis of bioactive secondary metabolites and stimulate their production. In the frame of the EU H2020 “MICROMETABOLITE” project, innovative technologies are developed by integrating microorganisms,including endophytic bacteria and fungi but also arbuscular mycorrhyzal fungi (AMF), on plant materials of Boraginaceae family.
As part of our study, eight different populations of Alkanna tinctoria were collected so far, from different locations close to Athens. The aerial part of the plant were separated from the roots, for each collection respectively and treated similarly. On the other hand, three other species of the genus Alkanna (A. graeca, A. hellenica and A. sfikasiana) were collected from Southern Greece, while a commercial sample of Alkanna tinctoria from Pakistan was used as a reference material.
Part of this study was focused on investigating and comparing metabolome variations between the four species in the aerial part, from different geographical regions of Greece, and the commercial sample. Some extracts were very rich in secondary metabolites. In addition pyrrolizidine alkaloids that are characteristic compounds of this family but also well known for their toxicity, were analyzed.
The extraction process of the plant material was performed sequentially with solvents of increasing polarity (c-Hexane, EtOAC and EtOH/H20 50:50) to get a broad metabolic profile. The extracts were analyzed with methods based on HPTLC, HPLC-PDA-ELSD, GC-MS and UPLC-HRMS. On those extracts a dereplication process combining the Dictionary of Natural Products, Molecular Networks, in silico MS/MS dereplication using customized libraries and algorithms using R statistical language were developed that will lead to a targeted isolation and identification of promising and new compounds.