(A) Amounts (μg per mL media) of AFB1 produced by A. flavus with different concentrations of D-glucose, D-glucal, or D-galactal (0, 2.5, 5, 10, 20 or 40 mg/mL). Data are presented as means ± S.D. (n = 3), from 3 independent experiments. (B) TLC analyses of AF production by A. flavus cultured in GMS media with different concentrations of D-glucal (0, 2.5, 5, 10, 20 or 40 mg/mL). (C) Growth curves of mycelia cultured in media with FG-4592 purchase 40 mg/mL D-glucose, D-glucal, or D-galactal for 5 d. (D) Numbers of spores produced per mL culture with D-glucose, D-glucal, or D-galactal. Data are presented as means ± S.D. (n = 3). We next examined if D-glucal or D-galactal inhibited mycelial growth, and found
that neither D-glucal nor D-galactal affected mycelial growth at the concentration of 40 mg/mL (Figure 2C). In contrast, additional D-glucose enhanced mycelial growth significantly, especially from the 3rd day onwards (Figure 2C). We next performed experiments on solid GMS
media with 40 mg/mL D-glucal or D-galactal to assess if these sugar analogs click here have any effect on sporulation, and observed that exogenous D-glucal inhibited sporulation significantly, while additional D-glucose enhanced sporulation (Figure 2D). No effect was observed for D-galactal. D-glucal promoted kojic acid biosynthesis, but inhibited fatty acid biosynthesis and glucose consumption We performed metabolomics analyses of mycelia of A. flavus A 3.2890 grown in media with or without 40 mg/mL D-glucal. The gas chromatography time-of-flight mass spectrometry (GC-TOF MS) based metabolomics technology developed in our lab has been shown to be a powerful
tool to elucidate metabolic changes in A. flavus[18]. For statistical analyses, we used nine replicates for each treatment. Partial least-squares (PLS) analyses of metabolite peak areas showed clustering of two distinct groups for mycelia grown in media with or without D-glucal, suggesting that exogenous D-glucal imposed significant PRKACG metabolic changes in mycelia (Figure 3). In particular, in the presence of D-glucal, the content of glucose, ribitol, glycerol and galactose were EVP4593 order increased significantly, while the content of TCA intermediates (succinic acid, malic acid and fumaric acid) and fatty acids (FAs) including palmitic acid, stearic acid, oleic acid and linoleic acid were decreased (Table 1). We also noticed that, in the presence of D-glucal, the content of two secondary metabolites, kojic acid and furanacetic acid, were increased by 2 and 159 fold, respectively. These results together suggest that D-glucal interferes with both primary and secondary metabolism. Figure 3 Mycelia grown in media with or without D-glucal showed significant differences in the accumulation of various metabolites. PLS analyses were performed using SIMCA-P V12.0. (A) Loadings plot obtained from PLS analyses of the entire GC-TOF MS dataset.
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