Inositol phosphorylceramide (IPC) participates in hyphal growth and serves as a signaling molecule that enables fungi to adapt to diverse environments. Here, a gene, encodes IPC synthase, was identified from the Aspergillus oryzae 3.042 genome and designated AoAur1. The characteristics, phylogenetic evolution, and resistance to aureobasidin A of AoAur1 were analyzed. The expression pattern of AoAur1 was markedly downregulated under temperature stress. Additionally, an RNAi-AoAur1 strain in which the AoAur1 expression was inhibited had mycelial that grew more quickly, had a higher frequency of hyphal fusion, and was more resistant to high-temperature stress than the control. Gene expression profiles showed that the genes related to IPC biosynthesis were obviously downregulated, while AoCerS, which participates in dihydroceramide biosynthesis, increased in the RNAi-AoAur1 strain at the three temperature treatments. A metabolomic analysis revealed that the intracellular IPC content decreased, and the accumulation of dihydroceramide and galactosylceramide increased significantly in the RNAi-AoAur1 strain. Thus, the inhibition of AoAur1 reduced IPC level followed by an increase in the contents of dihydroceramide and galactosylceramide that promote mycelial growth and the formation of spores in the RNAi-AoAur1 strain. Interestingly, the inhibition of AoAur1 also induced the expression of hyphal fusion-related genes, which promote hyphal fusion, thus, contributing to the transduction of stress signal to enhance the ability of cells to adapt to temperature stress. Our results demonstrated that the downregulation of AoAur1 and a decrease in the accumulation of IPC is one of the mechanisms that enables A. oryzae to adapt low- and high-temperature stress.
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Mannosyl inositol phosphoceramide
Potato Dextrose Agar
Quantitative real-time polymerase chain reaction
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We thank Dr. Van-Tuan Tran of the National Key Laboratory of Enzyme and Protein Technology, VNU University of Science for kindly providing the pEX1 vector.
This research was supported by grants from the National Natural Science Foundation of China (NSFC), Grant Number 31900063, and the Jiangxi Provincial Department of Education Science and Technology Project, Grant Number GJJ211103, and Natural Science Foundation of Jiangxi Province, grant number 20212BAB215005.
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Authors declare that they have no conflict of interests.
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Supplementary file1 Supplementary Figure 1. The schematic diagram of pEX1 plasmid used to construct the inhibition of AoAur1 expression (TIF 144 kb)
Supplementary file2 Supplementary Figure 2. Aspergillus oryzae was inoculated on PDA plates covered with cellophane at 22 °C, 30 °C and 42 °C for 3 days to analyze the gene expression level. PDA, potato dextrose agar (TIF 3160 kb)
Supplementary file3 Supplementary Figure 3. The phenotypes of Aspergillus oryzae inoculated on PDA plates covered with cellophane to analyze the dry biomass or the gene expression levels. (A) Control strain. (B) RNAi-AoAur1 strain 1. (C) RNAi-AoAur1 strain 2. PDA, potato dextrose agar (TIF 1720 kb)
Supplementary file4 Supplementary Table 1 Sequence IDs of the AUR protein or IPC synthase used to construct the phylogenetic NJ tree. IPC, inositol phosphorylceramide; NJ, neighbor-joining (DOCX 17 kb)
Supplementary file5 Supplementary Table 2 Primers of Aspergillus oryzae related-genes for qRT-PCR analysis. qRT-PCR, real-time quantitative reverse transcription PCR (DOCX 15 kb)
Supplementary file6: Supplementary material Construction details of the recombinant vector of RNAi- AoAur1 (DOCX 390 kb)
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Ge, J., Zhang, Z., Li, Y. et al. Inhibition of AoAur1 increases mycelial growth, hyphal fusion and improves physiological adaptation to high-temperature stress in Aspergillus oryzae. Arch Microbiol 204, 477 (2022). https://doi.org/10.1007/s00203-022-03075-6