Metabolomic Analysis of Extracellular Vesicles from the Cereal Fungal Pathogen Fusarium graminearum
Abstract
:1. Introduction
2. Materials and Methods
2.1. Microbial Cultures
2.2. Isolation of Extracellular Vesicles (EVs)
2.3. Preparation of Secretome (SEC) and Whole-Cell Lysate (WCL)
2.4. Detection of Zearalenone (ZEN) in EV Samples by ELISA
2.5. Cryo-Electron Microscopy
2.6. Sample Processing for Untargeted Metabolomic Analysis of Extracellular Vesicles (EVs)
2.7. Analysis of Untargeted Metabolomics Data
2.8. Statistical Analysis
2.9. Antimicrobial Assays
3. Results
3.1. Characterization of Extracellular Vesicles from Fusarium graminearum (F. graminearum)
3.2. Quantification of Zearalenone in Extracellular Vesicle (EV) Samples from Fusarium graminearum by ELISA
3.3. Extracellular Vesicle (EV) Samples from Fusarium graminearum Contain 2,4-Dihydroxybenzophenone (BP-1) and Cuminyl Alcohol
3.4. Metabolites Detected in F. graminearum EVs Reduce the Growth of Bacteria and F. graminearum
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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EV-Exclusive Metabolite | RT (min) | Adduct | Ref. m/z | Av. m/z | Error (ppm) | Formula | Ontology | ng/µL | RSD |
---|---|---|---|---|---|---|---|---|---|
Unknown (+) 40 | 0.606 | 261.13071 | 527.51 | 0.1107 | |||||
Unknown (+) 44 | 0.609 | 155.04849 | 23.12 | 0.1944 | |||||
Unknown (+) 54 | 0.622 | 233.04146 | 17.34 | 0.0486 | |||||
Unknown (+) 55 | 0.629 | 233.08037 | 158.07 | 0.0982 | |||||
Unknown (+) 56 | 0.632 | 289.10645 | 55.48 | 0.1317 | |||||
Unknown (+) 59 | 0.638 | 361.16351 | 18.22 | 0.1687 | |||||
Unknown (+) 60 | 0.641 | 236.05518 | 91.78 | 0.1371 | |||||
(5E,8Z)-4,7-dihydroxy-2-methyl-2,3,4,7-tetrahydrooxecin-10-one (Modiolide A) | 0.643 | [M+Na]+ | 237.05237 | 237.05177 | −2.531 | C10H14O4 | Oxocins | 386.11 | 0.1274 |
3-(4-chlorophenyl)-2-methyl-5,6,7,8-tetrahydropyrazolo[5,1-b]quinazolin-9(4H)-one | 0.646 | [M+Na]+ | 314.10532 | 314.10617 | 2.706 | C17H16ClN3O | Phenylpyrazoles | 473.18 | 0.0865 |
7,3′,4′-Trimethoxyflavone | 0.655 | [M+H]+ | 313.10999 | 313.11029 | 0.958 | C18H16O5 | 7-O-methylated flavonoids | 55.38 | 0.0849 |
Unknown (+) 65 | 0.656 | 336.08841 | 19.69 | 0.1516 | |||||
Unknown (+) 67 | 0.656 | 333.11313 | 59.93 | 0.1258 | |||||
Unknown (+) 68 | 0.657 | 191.03159 | 160.83 | 0.1826 | |||||
8-Acetylharpagide | 0.658 | [M+H]+ | 407.14999 | 407.14969 | −0.736 | C17H26O11 | Iridoid O-glycosides | 110.67 | 0.0974 |
Unknown (+) 70 | 0.659 | 406.15369 | 54.08 | 0.1324 | |||||
Unknown (+) 72 | 0.663 | 415.13553 | 39.69 | 0.1305 | |||||
Unknown (+) 76 | 0.677 | 199.02130 | 13.63 | 0.1758 | |||||
Hexaethylene glycol | 10.752 | [M+H]+ | 283.17511 | 283.17505 | −0.211 | C12H26O7 | Polyethylene glycols | 1123.70 | 0.1268 |
Unknown (+) 90 | 10.754 | 300.20163 | 734.73 | 0.2010 |
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Garcia-Ceron, D.; Truong, T.T.; Ratcliffe, J.; McKenna, J.A.; Bleackley, M.R.; Anderson, M.A. Metabolomic Analysis of Extracellular Vesicles from the Cereal Fungal Pathogen Fusarium graminearum. J. Fungi 2023, 9, 507. https://doi.org/10.3390/jof9050507
Garcia-Ceron D, Truong TT, Ratcliffe J, McKenna JA, Bleackley MR, Anderson MA. Metabolomic Analysis of Extracellular Vesicles from the Cereal Fungal Pathogen Fusarium graminearum. Journal of Fungi. 2023; 9(5):507. https://doi.org/10.3390/jof9050507
Chicago/Turabian StyleGarcia-Ceron, Donovan, Thy T. Truong, Julian Ratcliffe, James A. McKenna, Mark R. Bleackley, and Marilyn A. Anderson. 2023. "Metabolomic Analysis of Extracellular Vesicles from the Cereal Fungal Pathogen Fusarium graminearum" Journal of Fungi 9, no. 5: 507. https://doi.org/10.3390/jof9050507