Use of microbiome technologies and bioassays for the search for new bioactive compounds in plants and symbionts: implications for metabolic responses, molecular diversity and bioactivity

• Grant number: 21/09693-5
• Support type: Regular Research Grants
• Duration: March 01, 2022 - February 29, 2024
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal researcher: Marcos José Salvador
• Grantee: Marcos José Salvador
• Home Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Assoc. researchers: Candida Aparecida Leite Kassuya ; Derlene Attili de Angelis ; Fabiana Fantinatti Garboggini

Abstract

This research aims to carry out integrated investigations with plants (Amaranthaceae, Gesneriaceae and Annonaceae) and microorganisms from the microbiome of their leaf endosphere to evaluate the effect of plant/endophyte interactions on metabolic responses, molecular diversity and biosynthesis of bioactive substances. The main focus will be the search for substances with photoinduced biological effect for application in antimicrobial and antitumor PDT and/or protein Kinase C (PKC) inhibitors in plants and symbionts, using chemical and biological assays to direct the study. Therefore, the development of procedures for monitored study by bioassays will be sought, by analysis of absorption in the visible region of interest in PDT, evaluation of photochemical efficiency and by UHPLC-UV/DAD-ESI-MS, MS/ MS, NMR and the GNPS platform ("Global Natural Products Social Molecular Networking") in dereplication approaches. Plant species will be collected, identified and their extracts prepared, being then submitted to screening tests, being selected two plant species with better results in terms of chemical profile and bioactivity to continue the study. For the selected in natura plants, the aim will be to describe the diversity of microorganisms (bacteria and fungi) associated with the endosphere of their leaf blades and carry out the isolation and identification of cultivable microorganisms using culture media, microbiological analysis and molecular techniques. At first, for each of the plant species selected in the screening, a symbiotic microorganism with stable culture and higher biomass yield will be chosen to continue the integrated chemical, biological and biotechnological study. For the two plant species and microorganisms selected in the previous steps, we will seek to establish conditions for in vitro axenic cultivation and establish protocols for experimentation with pure cultures and synthetic co-cultures axenic/endophyte plant for proof of concept and for evaluating the effect of plant/endophyte interactions on plant metabolic responses, molecular diversity and bioactivity. Using genome mining tools, an analysis of the molecular genetics of the microorganisms selected in the study will be carried out to determine the biosynthetic gene clusters (BCG) of some of the identified molecules in order to explore the biosynthetic potential of endophytes. Promising extracts for bioactivity and with constituents that could not be identified in the dereplication step will be fractionated and, for active metabolites purified by the use of different chromatographic methods, their molecular structures will be determined using different methods analysis (IR, UV-vis, 1D and 2D NMR, mass spectrometry, specific rotation and circular dichroism). Therefore, with this study, investigations will be carried out in order to better understand the metabolic interactions between plant and its microbiota for prospecting (isolation and characterization) of bioactive molecules. This project also aims to train human resources (16 in guidance at the moment: 2PD, 9 Dr, 3 MS and 2 IC, all with CNPq, CAPES and FUNCAMP grants) able to carry out research of excellence for the generation of high-value technologies aggregate, combined with the conservation and sustainable use of biodiversity. (AU)