Abstract
Introduction
The production of high quality and safe food represents a main priority for the agri-food sector in the effort to sustain the exponentially growing human population. Nonetheless, there are major challenges that require the discovery of new, alternative, and improved plant protection products (PPPs). Focusing on fungal plant pathogens, the dissection of mechanisms that are essential for their survival provides insights that could be exploited towards the achievement of the aforementioned aim. In this context, the germination of fungal spores, which are essential structures for their dispersal, survival, and pathogenesis, represents a target of high potential for PPPs. To the best of our knowledge, no PPPs that target the germination of fungal spores currently exist.
Objectives
Within this context, we have mined for changes in the metabolite profiles of the model fungus Aspergillus nidulans FGSC A4 conidiospores during germination, in an effort to discover key metabolites and reactions that could potentially become targets of PPPs.
Methods
Untargeted GC/EI-TOF/MS metabolomics and multivariate analyses were employed to monitor time-resolved changes in the metabolomes of germinating A. nidulans conidiospores.
Results
Analyses revealed that trehalose hydrolysis plays a pivotal role in conidiospore germination and highlighted the osmoregulating role of the sugar alcohols, glycerol, and mannitol.
Conclusion
The ineffectiveness to introduce active ingredients that exhibit new mode(s)-of-action as fungicides, dictates the urge for the discovery of PPPs, which could be exploited to combat major plant protection issues. Based on the crucial role of trehalose hydrolysis in conidiospore dormancy breakage, and the subsequent involvement of glycerol in their germination, it is plausible to suggest their biosynthesis pathways as potential novel targets for the next-generation antifungal PPPs. Our study confirmed the applicability of untargeted metabolomics as a hypothesis-generation tool in PPPs’ research and discovery.
Graphic abstract
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source for glycerol that is biosynthesized immediately after 1 h and consumed in the course of germination. Similar changes of mannitol abundance indicate a possible osmoregulation role of polyols in early conidiospore germination. Sugars and Krebs cycle intermediates accumulate transiently during polarity establishment and germ tube emergence at 5–6 h of germination. Six biological replications were analyzed for each time point
source and c minimal media (MM) without carbon source. The arrows indicate representative germinated conidiospores. In the upper right corner, a magnified image of the conidiospores is displayed. Results indicate that A. nidulans conidiospores are able to germinate in sterilized H2O. However, in the absence of carbon and nitrogen sources the germination terminates. In the second row the corresponding treatments (d-f) with the addition of 20 μg mL−1 Validamycin A are shown. No visible differences were observed
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Data availability
“The metabolomics and metadata reported in this paper are available via Metabolights www.ebi.ac.uk/metabolights/MTBLS1565
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Acknowledgements
The authors acknowledge technical and laboratory support of the GC/EI-TOF/MS analyses by Ines Fehrle.
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This research is co-financed by Greece and the European Union (European Social Fund- ESF) through the Operational Program «Human Resources Development, Education and Lifelong Learning» in the context of the project “Strengthening Human Resources Research Potential via Doctorate Research” (MIS-5000432), implemented by the State Scholarships Foundation (ΙΚΥ).
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IFK, KAA, SA, and GD designed the study and performed literature research. IFK performed experimental research. IFK, KAA, AE, and JK analyzed data and performed statistical analysis. IFK, GD, JK, and KAA wrote the paper. All authors read and approved the manuscript.
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Kalampokis, I.F., Erban, A., Amillis, S. et al. Untargeted metabolomics as a hypothesis-generation tool in plant protection product discovery: Highlighting the potential of trehalose and glycerol metabolism of fungal conidiospores as novel targets. Metabolomics 16, 79 (2020). https://doi.org/10.1007/s11306-020-01699-7
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DOI: https://doi.org/10.1007/s11306-020-01699-7