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. 2019 Jul 30;7(8):225.
doi: 10.3390/microorganisms7080225.

Oxidative Stress Response of Aspergillus oryzae Induced by Hydrogen Peroxide and Menadione Sodium Bisulfite

Huanhuan Shao  1   2 , Yayi Tu  1 , Yijing Wang  1 , Chunmiao Jiang  1 , Long Ma  1 , Zhihong Hu  1 , Jiangfan Wang  1 , Bin Zeng  3 , Bin He  4
Affiliations

Oxidative Stress Response of Aspergillus oryzae Induced by Hydrogen Peroxide and Menadione Sodium Bisulfite

Huanhuan Shao et al. Microorganisms. .

Abstract

Oxidative stress response protects organisms from deleterious effects of reactive oxygen species (ROS), which can damage cellular components and cause disturbance of the cellular homeostasis. Although the defensive biochemical mechanisms have been extensively studied in yeast and other filamentous fungi, little information is available about Aspergillus oryzae. We investigated the effect of two oxidant agents (menadione sodium bisulfite, MSB, and hydrogen peroxide, H2O2) on cellular growth and antioxidant enzyme induction in A. oryzae. Results indicated severe inhibition of biomass and conidia production when high concentration of oxidants was used. Transcriptomic analysis showed an up-regulated expression of genes involved in oxidoreduction, such as catalase, glutathione peroxidase, and superoxide dismutase. In addition, it was observed that oxidative stress stimuli enhanced the expression of Yap1 and Skn7 transcription factors. Further, metabolomic analysis showed that glutathione content was increased in the oxidative treatments when compared with the control. Moreover, the content of unsaturated fatty acid decreased with oxidative treatment accompanying with the down-regulated expression of genes involved in linoleic acid biosynthesis. This study provided a global transcriptome characterization of oxidative stress response in A. oryzae, and can offer multiple target genes for oxidative tolerance improvement via genetic engineering.

Keywords: Aspergillus oryzae; antioxidant enzymes; fatty acids; glutathione; oxidative stress.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The density of spores, dry biomass and phenotype as affected by oxidative treatment. (A) The density of spores in the oxidative treatments (H2O2 and MSB) and the control; (B) The dry biomass in the oxidative treatments and the control. The mycelia were collected by peeling them off from the plates and dried overnight for the determination of biomass; The bars represent the average (±SE) of biological repeats. (C) The phenotype of A. oryzae in the control and the oxidative treatment at 72 h. The colony growth was retarded and the conidia production was inhibited. WT: the control; H2O2: the H2O2 treatments (3 and 6 mM); MSB: the MSB treatments (0.28 and 0.42 mM).
Figure 2
Figure 2
Distribution of differentially expressed genes (DEGs) in the five samples. (A) DEGs’ distribution between control and oxidative treatment; (B) Venn diagram exhibiting the DEGs’ distribution in five samples. WT: the control; H2O2: the H2O2 treatments (3 and 6 mM); MSB: the MSB treatments (0.28 and 0.42 mM).
Figure 3
Figure 3
The expression profile of catalase (CAT), GPX1 (glutathione peroxidase) and superoxide dismutase (SOD) in the five samples. WT: the control; H2O2: the H2O2 treatments (3 and 6 mM); MSB: the MSB treatments (0.28 and 0.42 mM).
Figure 4
Figure 4
The expression profile of Yap1 and Skn7 transcription factors in the five samples. WT: the control; H2O2: the H2O2 treatments (3 and 6 mM); MSB: the MSB treatments (0.28 and 0.42 mM).
Figure 5
Figure 5
The content of glutathione (GSH) and expression profile of genes involved in glutathione biosynthesis in the oxidative treatments (H2O2 and MSB) and the control. (A) The standard curve of GSH concentration. (B) The changes of GSH level in response to oxidative stress. The bars represent the average (±SE) of biological repeats. The content of GSH increased with the increasing concentration of H2O2 and MSB. (C) The expression pattern of DEGs involved in the linoleic acid biosynthesis pathways (from left to right: the control, 3 mM and 6 mM H2O2, 0.28 mM and 0.42 mM MSB).
Figure 6
Figure 6
DEGs involved in the linoleic acid biosynthesis pathways and the changes of C18 fatty acid in response to oxidative stress. (A) The expression pattern of DEGs involved in the linoleic acid biosynthesis pathways (from left to right: the control, 3 mM and 6 mM H2O2, 0.28 mM and 0.42 mM MSB); (B) The changes of C18 fatty acid in response to oxidative stress. The bars represent the average (±SE) of biological repeats. WT: the control; H2O2: the H2O2 treatments (3 and 6 mM); MSB: the MSB treatments (0.28 and 0.42 mM).
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