Azzollini, Antonio

Bicchi, Carlo

Boccard, Julien

Boggia, Lorenzo

Gindro, Katia

Lecoutre, Nicole

Rubiolo, Patrizia

Rudaz, Serge

Sgorbini, Barbara

Wolfender, Jean Luc

English

Institutional Research Information System University of Turin

 Detection and dynamics of volatile/non-volatile metabolite in-duction in fungal co-culture through a miniaturised MS-based metabolomic approach  Antonio Azzollini1, Lorenzo Boggia2, Julien Boccard1, Barbara Sgorbini2, Nicole Lecoutre3, Patrizia Rubiolo2,  Serge Rudaz1, Katia Gindro3, Carlo Bicchi2, Jean-Luc Wolfender1    1School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, Quai Ernest-Ansermet 30, CH-1211  Geneva 4, Switzerland; 2Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, 10125, Turin, Italy; 3Agroscope, Institute for Plant Production Sciences IPS, Mycology and Biotechnology, Route de Duiller 50, PO Box 1012, 1260 Nyon 1, Switzerland.  Microbial co-cultivation has emerged as a promising way for activating cryptic biosynthetic pathways and discovering novel antimicrobial metabolites [1]. For the success of such studies, a key element remains the development of standardized co-cultivation methods compatible with high throughput analytical procedures [2]. In our continuous studies on the dynamic stress induction of bioactive natural products in co-culture conditions, we have extended the investigations to modifications of the microbial volatile metabolome that might be of im-portance in the understanding of the interaction between microorganisms.    For this, an MS metabolomic strategy based on the analysis of volatile and non-volatile frac-tions of the fungal cultures directly grown in 20 ml vials was applied. The head space of each sample was profiled by SPME-GC-MS while the whole culture medium was analyzed by LC-HRMS after solvent extraction. This strategy was implemented for screening volatile/non-volatile metabolite inductions in an ecologically relevant fungal co-culture of Eutypa lata (Pers.) Tul. & C. Tul. (Diatrypaceae) and Botryospheria obtusa (Schwein.) Shoemaker (Botry-osphaeriaceae), two wood decaying fungi interacting in the context of esca disease of grape-vine [3]. Multivariate data analysis combining Analysis of Variance (ANOVA) and Orthogonal Partial Least Squares (OPLS) conducted to the selection of specific induced molecules related to each factor of the study.  A time-series study during nine days showed characteristic me-tabolite induction patterns. Interesting relationships between the dynamics of volatile/non-volatile metabolite de novo formation and up- or down-regulation could be highlighted. Some of the induced compounds (e.g., 2-nonanone) are bioactive and known to be produced by oth-er microorganisms. The developed strategy could help elucidating the microbial inter- and intra-species communication. Moreover it could be used for efficiently identifying signaling molecules with potential antimicrobial activity.   Keywords: volatile/non-volatile metabolite induction, microbial co-cultivation, SPME-GC-MS, LC-HRMS, metabolomics    References:  [1] Bertrand S, Bohni N, Schnee S, Schumpp O, Gindro K, Wolfender JL. Metabolite    induction via microorganism co-culture: A potential way to enhance chemical diversity for drug discovery. Biotechnol Adv 2014; 32: 1180?1204 [2] Bertrand S, Azzollini A, Schumpp O, Bohni N, Schrenzel J, Monod M, Gindro K, Wolfender JL. Multi-well fungal co-culture for de novo metabolite-induction in time-series studies based on untargeted metabolomics. Mol BioSyst 2014; 10: 2289?2298 [3] Glauser G, Gindro K, Fringeli J, De Joffrey JP, Rudaz S, Wolfender JL. Differential analysis of mycoalexins in confrontation zones of grapevine fungal pathogens by ultrahigh pressure liquid chromatography/time-of-flight mass spectrometry and capillary nuclear magnetic resonance. J Agric Food Chem 2009; 57: 1127?1134 

Detection and dynamics of volatile/non-volatile metabolite induction in a  fungal co-culture through a miniaturised MS-based metabolomic approach

https://iris.unito.it/bitstream/2318/1614803/1/Abstract_accepted.pdf

Detection and dynamics of volatile/non-volatile metabolite induction in a fungal co-culture through a miniaturised MS-based metabolomic approach

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