Dataset Information

Effect of Water Activity on Conidia Germination in Aspergillus flavus.

ABSTRACT: In this study, we explored the mechanism underlying Aspergillus flavus conidia germination inhibited by decreased water activity. The impact of low water activity was analyzed at 4 h, 8 h and 12 h. Additionally, we demonstrated that low water activity affected cell shape and decreased cell sizes. Transcriptomics found numerous differentially expressed genes (DEGs) during the first 12 h of germination, with 654 DEGs observed among 4 h, 8 h and 12 h. In particular, more DEGs were detected at 8 h of germinating. Therefore, proteomics was performed at 8 h, and 209 differentially expressed proteins (DEPs) were speculated, with 94 up-regulated and 115 down-regulated. Combined analysis of KEGG of transcriptomics and proteomics demonstrated that the dominant pathways were nutrient metabolism and translation. We also found several DEGs and DEPs in the Mitogen Activated Protein Kinase (MAPK) pathway. Therefore, we concluded that low water activity inhibited conidia germination, causing unregular morphology. In addition, low water activity influenced expression of creA, TreB in carbohydrate metabolism, Clr4, RmtA in amino acid metabolism and RPL37, RPL3 in translation in Aspergillus flavus.

SUBMITTER: Jia S

PROVIDER: S-EPMC9504883 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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Effect of Water Activity on Conidia Germination in <i>Aspergillus flavus</i>.

Jia Sifan S Li Chong C Wu Kuntan K Qi Desheng D Wang Shuai S

Microorganisms 20220829 9

In this study, we explored the mechanism underlying <i>Aspergillus flavus</i> conidia germination inhibited by decreased water activity. The impact of low water activity was analyzed at 4 h, 8 h and 12 h. Additionally, we demonstrated that low water activity affected cell shape and decreased cell sizes. Transcriptomics found numerous differentially expressed genes (DEGs) during the first 12 h of germination, with 654 DEGs observed among 4 h, 8 h and 12 h. In particular, more DEGs were detected a ...[more]

PMID: 36144346

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Project description:Aspergillus flavus conidia are widespread in air; they attach to food and feed crops and secrete aflatoxins, which results in serious contamination. Germination of A. flavus conidia is the most critical step in contamination of food by A. flavus. This study aims to gain an insight into A. flavus conidia through dormancy to germination to provide a theoretical basis for inhibition of A. flavus conidia germination. The morphological changes and regulation mechanism of A. flavus conidia germination at 0, 4, 8, and 12 hours were observed. Transcriptomic and metabolomic analyses showed that conidia became active from dormancy (0 hour) to the initial stage of germination (4 hours), cellular respiration and energy metabolism increased, and amino acids and lipids were synthesized rapidly. The number of differentially expressed genes and differential metabolites was highest at this stage. Besides, we found that conidia germination had selectivity for different carbon and nitrogen sources. Compared with monosaccharides, disaccharides, as the only carbon source, significantly promoted the germination of conidia. Moreover, MepA, one of genes in the ammonium transporter family was studied. The gene deletion mutant ΔMepA had a significant growth defect, and the expression of MeaA was significantly upregulated in ΔMepA compared with the wild-type, indicating that both MepA and MeaA played an important role in transporting ammonium ions.IMPORTANCEThis is the first study to use combined transcriptomic and metabolomics analyses to explore the biological changes during germination of Aspergillus flavus conidia. The biological process with the highest changes occurred in 0-4 hours at the initial stage of germination. Compared with polysaccharides, monosaccharides significantly increased the size of conidia, while significantly decreasing the germination rate of conidia. Both MeaA and MepA were involved in ammonia transport and metabolism during conidia germination.

Project description:BackgroundAspergillus flavus, one of the causative agents of human fungal keratitis, can be phagocytosed by human corneal epithelial (HCE) cells and the conidia containing phagosomes mature into phagolysosomes. But the immunological responses of human corneal epithelial cells interacting with A. flavus are not clear. In this study, we report the expression of immune response related genes of HCE cells exposed to A. flavus spores using targeted transcriptomics.MethodsHuman corneal epithelial cell line and primary cultures were grown in a six-well plate and used for coculture experiments. Internalization of the conidia was confirmed by immunofluorescence microscopy of the colocalized endosomal markers CD71 and LAMP1. Total RNA was isolated, and the quantity and quality of the isolated RNA were assessed using Qubit and Bioanalyzer. NanoString nCounter platform was used for the analysis of mRNA abundance using the Human Immunology panel. R-package and nSolver software were used for data analysis. KEGG and FunRich 3.1.3 tools were used to analyze the differentially expressed genes.ResultsDifferent morphotypes of conidia were observed after 6 h of coculture with human corneal epithelial cells and found to be internalized by epithelial cells. NanoString profiling showed more than 20 differentially expressed genes in immortalized human corneal epithelial cell line and more than ten differentially expressed genes in primary corneal epithelial cells. Distinct set of genes were altered in their expression in cell line and primary corneal epithelial cells. KEGG pathway analysis revealed that genes associated with TNF signaling, NF-KB signaling, and Th17 signaling were up-regulated, and genes associated with chemokine signaling and B cell receptor signaling were down regulated. FunRich pathway analysis showed that pathways such as CDC42 signaling, PI3K signaling, and Arf6 trafficking events were activated by the clinical isolates CI1123 and CI1698 in both type of cells.ConclusionsCombining the transcript analysis data from cell lines and primary cultures, we showed the up regulation of immune defense genes in A. flavus infected cells. At the same time, chemokine signaling and B cell signaling pathways are downregulated. The variability in the expression levels in the immortalized cell line and the primary cultures is likely due to the variable epigenetic reprogramming in the immortalized cells and primary cultures in the absence of any changes in the genome. It highlights the importance of using both cell types in host-pathogen interaction studies.

Dataset Information

Effect of Water Activity on Conidia Germination in Aspergillus flavus.

Publications

Effect of Water Activity on Conidia Germination in <i>Aspergillus flavus</i>.

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