Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels
Fungal co-cultivation has emerged as a promising way for activating cryptic biosynthetic pathways and discovering novel antimicrobial metabolites. For the success of such studies, a key element remains the development of standardized co-cultivation methods compatible with high-throughput analytical...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-02-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2018.00072/full |
| _version_ | 1828482419388317696 |
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| author | Antonio Azzollini Lorenzo Boggia Julien Boccard Barbara Sgorbini Nicole Lecoultre Pierre-Marie Allard Patrizia Rubiolo Serge Rudaz Katia Gindro Carlo Bicchi Jean-Luc Wolfender |
| author_facet | Antonio Azzollini Lorenzo Boggia Julien Boccard Barbara Sgorbini Nicole Lecoultre Pierre-Marie Allard Patrizia Rubiolo Serge Rudaz Katia Gindro Carlo Bicchi Jean-Luc Wolfender |
| author_sort | Antonio Azzollini |
| collection | DOAJ |
| description | Fungal co-cultivation has emerged as a promising way for activating cryptic biosynthetic pathways and discovering novel antimicrobial metabolites. For the success of such studies, a key element remains the development of standardized co-cultivation methods compatible with high-throughput analytical procedures. To efficiently highlight induction processes, it is crucial to acquire a holistic view of intermicrobial communication at the molecular level. To tackle this issue, a strategy was developed based on the miniaturization of fungal cultures that allows for a concomitant survey of induction phenomena in volatile and non-volatile metabolomes. Fungi were directly grown in vials, and each sample was profiled by head space solid phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS), while the corresponding solid culture medium was analyzed by liquid chromatography high resolution mass spectrometry (LC-HRMS) after solvent extraction. This strategy was implemented for the screening of volatile and non-volatile metabolite inductions in an ecologically relevant fungal co-culture of Eutypa lata (Pers.) Tul. & C. Tul. (Diatrypaceae) and Botryosphaeria obtusa (Schwein.) Shoemaker (Botryosphaeriaceae), two wood-decaying fungi interacting in the context of esca disease of grapevine. For a comprehensive evaluation of the results, a multivariate data analysis combining Analysis of Variance and Partial Least Squares approaches, namely AMOPLS, was used to explore the complex LC-HRMS and GC-MS datasets and highlight dynamically induced compounds. A time-series study was carried out over 9 days, showing characteristic metabolite induction patterns in both volatile and non-volatile dimensions. Relevant links between the dynamics of expression of specific metabolite production were observed. In addition, the antifungal activity of 2-nonanone, a metabolite incrementally produced over time in the volatile fraction, was assessed against Eutypa lata and Botryosphaeria obtusa in an adapted bioassay set for volatile compounds. This compound has shown antifungal activity on both fungi and was found to be co-expressed with a known antifungal compound, O-methylmellein, induced in solid media. This strategy could help elucidate microbial inter- and intra-species cross-talk at various levels. Moreover, it supports the study of concerted defense/communication mechanisms for efficiently identifying original antimicrobials. |
| first_indexed | 2024-12-11T08:15:48Z |
| format | Article |
| id | doaj.art-2bf1625ef36642c2b4f2108a67c925a2 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-11T08:15:48Z |
| publishDate | 2018-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-2bf1625ef36642c2b4f2108a67c925a22022-12-22T01:14:47ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2018-02-01910.3389/fmicb.2018.00072326597Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile LevelsAntonio Azzollini0Lorenzo Boggia1Julien Boccard2Barbara Sgorbini3Nicole Lecoultre4Pierre-Marie Allard5Patrizia Rubiolo6Serge Rudaz7Katia Gindro8Carlo Bicchi9Jean-Luc Wolfender10School of Pharmaceutical Sciences (EPGL), University of Geneva, University of Lausanne, Centre Médical Universitaire (CMU), Geneva, SwitzerlandDepartment of Drug Science and Technology, University of Turin, Turin, ItalySchool of Pharmaceutical Sciences (EPGL), University of Geneva, University of Lausanne, Centre Médical Universitaire (CMU), Geneva, SwitzerlandDepartment of Drug Science and Technology, University of Turin, Turin, ItalyPlant Protection, Mycology and Biotechnology, Agroscope, Nyon, SwitzerlandSchool of Pharmaceutical Sciences (EPGL), University of Geneva, University of Lausanne, Centre Médical Universitaire (CMU), Geneva, SwitzerlandDepartment of Drug Science and Technology, University of Turin, Turin, ItalySchool of Pharmaceutical Sciences (EPGL), University of Geneva, University of Lausanne, Centre Médical Universitaire (CMU), Geneva, SwitzerlandPlant Protection, Mycology and Biotechnology, Agroscope, Nyon, SwitzerlandDepartment of Drug Science and Technology, University of Turin, Turin, ItalySchool of Pharmaceutical Sciences (EPGL), University of Geneva, University of Lausanne, Centre Médical Universitaire (CMU), Geneva, SwitzerlandFungal co-cultivation has emerged as a promising way for activating cryptic biosynthetic pathways and discovering novel antimicrobial metabolites. For the success of such studies, a key element remains the development of standardized co-cultivation methods compatible with high-throughput analytical procedures. To efficiently highlight induction processes, it is crucial to acquire a holistic view of intermicrobial communication at the molecular level. To tackle this issue, a strategy was developed based on the miniaturization of fungal cultures that allows for a concomitant survey of induction phenomena in volatile and non-volatile metabolomes. Fungi were directly grown in vials, and each sample was profiled by head space solid phase microextraction gas chromatography mass spectrometry (HS-SPME-GC-MS), while the corresponding solid culture medium was analyzed by liquid chromatography high resolution mass spectrometry (LC-HRMS) after solvent extraction. This strategy was implemented for the screening of volatile and non-volatile metabolite inductions in an ecologically relevant fungal co-culture of Eutypa lata (Pers.) Tul. & C. Tul. (Diatrypaceae) and Botryosphaeria obtusa (Schwein.) Shoemaker (Botryosphaeriaceae), two wood-decaying fungi interacting in the context of esca disease of grapevine. For a comprehensive evaluation of the results, a multivariate data analysis combining Analysis of Variance and Partial Least Squares approaches, namely AMOPLS, was used to explore the complex LC-HRMS and GC-MS datasets and highlight dynamically induced compounds. A time-series study was carried out over 9 days, showing characteristic metabolite induction patterns in both volatile and non-volatile dimensions. Relevant links between the dynamics of expression of specific metabolite production were observed. In addition, the antifungal activity of 2-nonanone, a metabolite incrementally produced over time in the volatile fraction, was assessed against Eutypa lata and Botryosphaeria obtusa in an adapted bioassay set for volatile compounds. This compound has shown antifungal activity on both fungi and was found to be co-expressed with a known antifungal compound, O-methylmellein, induced in solid media. This strategy could help elucidate microbial inter- and intra-species cross-talk at various levels. Moreover, it supports the study of concerted defense/communication mechanisms for efficiently identifying original antimicrobials.http://journal.frontiersin.org/article/10.3389/fmicb.2018.00072/fullfungal co-culturemetabolomicsinduced metabolitesantifungalvolatilenon-volatile |
| spellingShingle | Antonio Azzollini Lorenzo Boggia Julien Boccard Barbara Sgorbini Nicole Lecoultre Pierre-Marie Allard Patrizia Rubiolo Serge Rudaz Katia Gindro Carlo Bicchi Jean-Luc Wolfender Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels Frontiers in Microbiology fungal co-culture metabolomics induced metabolites antifungal volatile non-volatile |
| title | Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels |
| title_full | Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels |
| title_fullStr | Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels |
| title_full_unstemmed | Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels |
| title_short | Dynamics of Metabolite Induction in Fungal Co-cultures by Metabolomics at Both Volatile and Non-volatile Levels |
| title_sort | dynamics of metabolite induction in fungal co cultures by metabolomics at both volatile and non volatile levels |
| topic | fungal co-culture metabolomics induced metabolites antifungal volatile non-volatile |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2018.00072/full |
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