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.

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

BOGGIA, LORENZO;SGORBINI, Barbara;RUBIOLO, Patrizia;BICCHI, Carlo;WOLFENDER, Jean Luc
2016

Abstract

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.
9th Joint Natural Products Conference 2016
Copenaghen
24-27 luglio
Abstract book
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volatile/non-volatile metabolite induction, microbial co-cultivation, SPME-GC-MS, LC-HRMS, metabolomics
Azzollini, Antonio; Boggia, Lorenzo; Boccard, Julien; Sgorbini, Barbara; Lecoutre, Nicole; Rubiolo, Patrizia; Rudaz, Serge; Gindro, Katia; Bicchi, Carlo; Wolfender, Jean-Luc
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2318/1614803
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