Abstract
As the most carcinogenic, toxic, and economically costly mycotoxins, aflatoxin B1 (AFB1) is primarily biosynthesized by Aspergillus flavus and Aspergillus parasiticus. Aflatoxin biosynthesis is related to oxidative stress and functions as a second line of defense from excessive reactive oxygen species. Here, we find that ethanol can inhibit fungal growth and AFB1 production by A. flavus in a dose-dependent manner. Then, the ethanol’s molecular mechanism of action on AFB1 biosynthesis was revealed using a comparative transcriptomic analysis. RNA-Seq data indicated that all the genes except for aflC in the aflatoxin gene cluster were down-regulated by 3.5% ethanol. The drastic repression of aflatoxin structural genes including the complete inhibition of aflK and aflLa may be correlated with the down-regulation of the transcription regulator genes aflR and aflS in the cluster. This may be due to the repression of several global regulator genes and the subsequent overexpression of some oxidative stress-related genes. The suppression of several key aflatoxin genes including aflR, aflD, aflM, and aflP may also be associated with the decreased expression of the global regulator gene veA. In particular, ethanol exposure caused the decreased expression of stress response transcription factor srrA and the overexpression of bZIP transcription factor ap-1, C2H2 transcription factors msnA and mtfA, together with the enhanced levels of anti-oxidant enzymatic genes including Cat, Cat1, Cat2, CatA, and Cu, Zn superoxide dismutase gene sod1. Taken together, these RNA-Seq data strongly suggest that ethanol inhibits AFB1 biosynthesis by A. flavus via enhancing fungal oxidative stress response. In conclusion, this study served to reveal the anti-aflatoxigenic mechanisms of ethanol in A. flavus and to provide solid evidence for its use in controlling AFB1 contamination.
Highlights
Aspergillus flavus is a saprophytic fungus being often found in mildewed grains, grain products, and other moldy organic matter, and causes the wastage of several important agricultural crops (Wild and Gong, 2010; Liang et al, 2015)
The diameters of A. flavus colonies appeared much smaller than the control after treatment with 2–6% ethanol in a dose-dependent manner, and the mycelia growth was completely inhibited by 6% ethanol when the initial concentration was ≤104 conidia/ml (Figure 1A)
These findings suggested that ethanol significantly inhibited fungal growth and aflatoxin B1 (AFB1) production by A. flavus
Summary
Aspergillus flavus is a saprophytic fungus being often found in mildewed grains, grain products, and other moldy organic matter, and causes the wastage of several important agricultural crops (Wild and Gong, 2010; Liang et al, 2015). This fungus is an opportunistic human and animal pathogen causing aspergillosis diseases (Amaike and Keller, 2010). It is more important to notice that this fungus can produce aflatoxins (AFs), the most potent natural carcinogen and. It is urgent to develop simple, economical, and effective ways to control A. flavus and subsequent AF contamination in food and feed, especially during storage and processing
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