Multiple nucleobase transporters contribute to boscalid sensitivity in Aspergillus nidulans

The development of fungicide-resistant fungal populations represents a major challenge for the agrochemical and agri-food sectors, which threatens food supply and security. The issue becomes complex for fungi that cause quantitative and qualitative losses due to mycotoxin biosynthesis. Nonetheless, currently, the molecular details underlying fungicide action and fungal resistance mechanisms are partially known. Here, we have investigated whether plasma membrane transporters contribute to specific fungicide uptake in the model fungus Aspergillus nidulans. Independent physiological tests and toxicity screening of selected fungicides provided evidence that the antifungal activity of Succinate Dehydrogenase Inhibitors (SDHIs) is associated with the expression of several nucleobase-related transporters. In particular, it was shown that a strain genetically inactivated in all seven nucleobase-related transporters is resistant to the fungicide boscalid, whereas none of the single null mutants exhibited significant resistance level. By constructing and testing isogenic strains that over-express each one of the seven transporters, we confirmed that five of them, namely, UapC, AzgA, FycB, CntA, and FurA, contribute to boscalid uptake. Additionally, by employing metabolomics we have examined the effect of boscalid on the metabolism of isogenic strains expressing or genetically lacking boscalid-related nucleobase transporters. The results confirmed the involvement of specific nucleobase transporters in fungicide uptake, leading to the discovery of corresponding metabolites-biomarkers. This work is the first report on the involvement of specific transporters in fungicide uptake and toxicity and their impact on fungal metabolism regulation and results might be further exploited towards the deeper understanding of fungal resistance to fungicides. © 2018 Elsevier Inc

Metabolic responses of two contrasting lentil genotypes to peg-induced drought stress

Among abiotic stresses, drought is undoubtedly one of the most severe environmental factors for a wide range of major crops, leading to considerable yield and economic losses. The adverse effects in crop yield reflect the result of a series of morphological and physiological changes but also changes in signaling pathways, transcriptional and post-transcriptional regulation of stress-responsive genes, and metabolic adaptations. Despite the exhausting studies elucidating plants’ metabolic response to drought, there is a knowledge gap in the biochemical mechanisms governing drought tolerance in lentil (Lens culinaris Medik.). The present study aimed to determine the fluctuations of the metabolite profiles of lentil genotypes with contrasting drought tolerance to discover possible biomarkers for screening tolerant genotypes at early growth stages. Lentil seedlings were subjected to osmotic drought stress, induced by polyethylene glycol, at two stress levels (2.5% and 5.0% PEG-6000) for a period of 20 days, while untreated plants were also included as controls. GC/EI/MS-mediated metabolic profiling was employed to monitor changes in response to osmotic drought stress. The data was subjected to OPLS-DA and OPLS-HCA for the discrimination between treatments and the discovery of trends and corresponding biomarkers. In total, the analysis yielded 150 metabolite features with highly reproducible patterns, of which the vast majority belonged to carbohydrates, carboxylic acids, and amino acids. Overall, findings highlight the differential accumulation of a series of compounds, and more importantly, the variable accumulation of certain metabolites, namely D-fructose, α,α-trehalose, myo-inositol, and L-tryptophan, in the contrasting genotypes, indicating that the adaptive metabolic responses to osmotic drought stress operate under strong genotypic dependency in lentil. Research findings provide insights into various aspects of lentil’s metabolism under drought and further offer the possibility of applying such knowledge towards effectively screening for drought-tolerant lentil germplasm at early growth stages. © 2021 by the authors. Licensee MDPI, Basel, Switzerland

Fusarium graminearum 1H NMR metabolomics

Raw 1H NMR spectra of Fusarium graminearum hyphae can be found at the website of the pesticide metabolomics group (PMG) of the Agricultural University of Athens at the address: http://www.aua.gr/pesticide-metabolomicsgroup/Resources/Fusarium_graminearum_NMR_spectra.html, accession number PMG-01–17. The data set support the research article “Implication of Fusarium graminearum Primary Metabolism in its Resistance to Benzimidazole Fungicides as revealed by 1H NMR Metabolomics” [1]

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

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: [Figure not available: see fulltext.]. © 2020, Springer Science+Business Media, LLC, part of Springer Nature