Cold plasma treatment triggers antioxidative defense system and induces changes in hyphal surface and subcellular structures of Aspergillus flavus
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The cold atmospheric-pressure plasma (CAPP) has become one of the recent effective decontamination technologies, but CAPP interactions with biological material remain the subject of many studies. The CAPP generates numerous types of particles and radiations that synergistically affect cells and tissues differently depending on their structure. In this study, we investigated the effect of CAPP generated by diffuse coplanar surface barrier discharge on hyphae of Aspergillus flavus. Hyphae underwent massive structural changes after plasma treatment. Scanning electron microscopy showed drying hyphae that were forming creases on the hyphal surface. ATR-FTIR analysis demonstrated an increase of signal intensity for C=O and C-O stretching vibrations indicating chemical changes in molecular structures located on hyphal surface. The increase in membrane permeability was detected by the fluorescent dye, propidium iodide. Biomass dry weight determination and increase in permeability indicated leakage of cell content and subsequent death. Disintegration of nuclei and DNA degradation confirmed cell death after plasma treatment. Damage of plasma membrane was related to lipoperoxidation that was determined by higher levels of thiobarbituric acid reactive species after plasma treatment. The CAPP treatment led to rise of intracellular ROS levels detected by fluorescent microscopy using 2′,7′-dichlorodihydrofluorescein diacetate. At the same time, antioxidant enzyme activities increased, and level of reduced glutathione decreased. The results in this study indicated that the CAPP treatment in A. flavus targeted both cell surface structures, cell wall, and plasma membrane, inflicting injury on hyphal cells which led to subsequent oxidative stress and finally cell death at higher CAPP doses.
KeywordsAntioxidant defense system Aspergillus flavus Cold atmospheric pressure plasma FTIR Lipid peroxidation Oxidative stress
This work was supported by the Slovak Research and Development Agency under the contract no. APVV-16-0216 and by a project for the building of infrastructure for the modern research of civilization diseases, ITMS 26230120006.
Compliance with ethical standard
Conflict of interest
The authors declare they have no conflict of interest.
This paper does not contain any studies with human participants or animals performed by any of the authors.
- Amaike S, Keller NP (2011) Aspergillus flavus. Annu Rev Phytopathol 49:107–133. https://doi.org/10.1146/annurev-phyto-072910-095221 CrossRefPubMedGoogle Scholar
- van Bokhorst-van de Veen H, Xie H, Esveld E, Abee T, Mastwijk H, Nierop Groot M (2014) Inactivation of chemical and heat-resistant spores of Bacillus and Geobacillus by nitrogen cold atmospheric plasma evokes distinct changes in morphology and integrity of spores. Food Microbiol 45:26–33. https://doi.org/10.1016/j.fm.2014.03.018 CrossRefPubMedGoogle Scholar
- Conway GE, Casey A, Milosavljevic V, Liu Y, Howe O, Cullen PJ, Curtin JF (2016) Non-thermal atmospheric plasma induces ROS-independent cell death in U373MG glioma cells and augments the cytotoxicity of temozolomide. Br J Cancer 114:435–443. https://doi.org/10.1038/bjc.2016.12 CrossRefPubMedPubMedCentralGoogle Scholar
- Fridman A (2008) Plasma chemistry. Cambridge university pressGoogle Scholar
- Joshi SG, Cooper M, Yost A, Paff M, Ercan UK, Fridman G, Friedman G, Fridman A, Brooks AD (2011) Nonthermal dielectric-barrier discharge plasma-induced inactivation involves oxidative DNA damage and membrane lipid peroxidation in Escherichia coli. Antimicrob Agents Chemother 55:1053–1062. https://doi.org/10.1128/AAC.01002-10 CrossRefPubMedPubMedCentralGoogle Scholar
- Kiššová I, Deffieu M, Samokhvalov V, Velours G, Bessoule JJ, Manon S, Camougrand N (2006) Lipid oxidation and autophagy in yeast. Free Radic Biol Med 41:1655–1661. https://doi.org/10.1016/j.freeradbiomed.2006.08.012 CrossRefPubMedGoogle Scholar
- Lazovic S, Puac N, Radic N, Hoder T, Malovic G, Ráhel’ J, Černák M, Petrovic ZL (2008) Mass spectrometry of diffuse coplanar surface barrier discharge. Publ l’Observatoire Astron Beogr 84:401–404Google Scholar
- Mohamad R, Mohamed MS, Suhaili N, Salleh MM, Ariff A (2010) Kojic acid: applications and development of fermentation process for production. Biotechnol Mol Biol Rev 5:24–37Google Scholar
- Mošovská S, Medvecká V, Halászová N, Ďurina P, Valík Ľ, Mikulajová A, Zahoranová A (2018) Cold atmospheric pressure ambient air plasma inhibition of pathogenic bacteria on the surface of black pepper. Food Res Int 106:862–869. https://doi.org/10.1016/j.foodres.2018.01.066 CrossRefPubMedGoogle Scholar
- Panngom K, Baik KY, Nam MK, Han JH, Rhim H, Choi EH (2013) Preferential killing of human lung cancer cell lines with mitochondrial dysfunction by nonthermal dielectric barrier discharge plasma. Cell Death Dis 4:e642–e648. https://doi.org/10.1038/cddis.2013.168 CrossRefPubMedPubMedCentralGoogle Scholar
- Weiss M, Gümbel D, Hanschmann EM, Mandelkow R, Gelbrich N, Zimmermann U, Walther R, Ekkernkamp A, Sckell A, Kramer A, Burchardt M, Lillig CH, Stope MB (2015) Cold atmospheric plasma treatment induces anti-proliferative effects in prostate cancer cells by redox and apoptotic signaling pathways. PLoS One 10:e0130350. https://doi.org/10.1371/journal.pone.0130350 CrossRefPubMedPubMedCentralGoogle Scholar
- Zahoranová A, Henselová M, Hudecová D, Kaliňáková B, Kováčik D, Medvecká V, Černák M (2016) Effect of cold atmospheric pressure plasma on the wheat seedlings vigor and on the inactivation of microorganisms on the seeds surface. Plasma Chem Plasma Process 36:397–414. https://doi.org/10.1007/s11090-015-9684-z CrossRefGoogle Scholar