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Pathogens 2018, 7(4), 90; https://doi.org/10.3390/pathogens7040090

Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels

1
Institute of Graduate Studies in Sciences, Programme of Molecular Biology and Genetics, Istanbul University, Suleymaniye, Istanbul 34116, Turkey
2
Faculty of Arts and Sciences, Department of Molecular Biology and Genetics, Istanbul Yeni Yuzyil University, Cevizlibag, Istanbul 34010, Turkey
3
Faculty of Sciences, Department of Molecular Biology and Genetics, Istanbul University, Vezneciler, Istanbul 34134, Turkey
*
Author to whom correspondence should be addressed.
Received: 16 October 2018 / Revised: 9 November 2018 / Accepted: 19 November 2018 / Published: 22 November 2018
(This article belongs to the Special Issue Fusarium)
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Abstract

Fusarium graminearum and F. culmorum are phytopathogens, which cause destructive diseases in cereals. Epidemics of these phytopathogens are caused by mycotoxin contamination and the reduction of crop quality. In this study, the alteration due to in vitro camphor treatment on F. culmorum 9F and F. graminearum H11 isolates was investigated in terms of epigenetic, cellular, and transcription levels. Camphor with different concentrations (0.2, 0.4, 0.8, 1, 2, and 4 µg/µL) was applied to potato dextrose agar (PDA) growth media. The minimum inhibitory concentration (MIC) and the half maximal inhibitory concentration (IC50) were calculated as 2 and 1 µg/µL, respectively. hog1, mst20, CAT, POD, mgv1, stuA, and tri5 genes, which are related to various cellular processes and pathogenesis, were examined by qPCR assay. qPCR analysis showed that camphor treatment leads to the downregulation of tri5 expression but the upregulation of the remaining genes. Apoptosis and oxidative stress were confirmed via acridine orange/ethidium bromide (AO/EB) and dichlorofluorescin diacetate (DCF-DA) staining, respectively. Moreover, coupled restriction enzyme digestion-random amplification (CRED-RA) assay, used for DNA methylation analysis, was carried out to evaluate epigenetic alterations. The decrease in genomic template stability (GTS) values, which resulted due to the alterations in random amplified polymorphic DNA (RAPD) profiles caused by camphor treatment, were detected as 97.60% in F. culmorum 9F and 66.27% in F. graminearum H-11. The outer and inner methylated cytosine profiles are determined by CRED-RA assay as type I–IV epigenetic alterations. The outcomes indicated that camphor could lead to alterations at several molecular levels of F. graminearum and F. culmorum. View Full-Text
Keywords: apoptosis; camphor; epigenetics; gene expression; oxidative stress apoptosis; camphor; epigenetics; gene expression; oxidative stress
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Gazdağlı, A.; Sefer, Ö.; Yörük, E.; Varol, G.İ.; Teker, T.; Albayrak, G. Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels. Pathogens 2018, 7, 90.

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