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. 2021 Jul 8;24(8):102820. doi: 10.1016/j.isci.2021.102820

Figure 1.

Figure 1

Comparison of colony morphology, mycelial growth, DNA damage, and cell apoptosis in the wild-type (WT) and mutant strains (ΔAoras2, ΔAoras3, and ΔAorheb) of A. oligospora

(A) Colonies from the WT and mutant strains were cultured on PDA, TG, and TYGA plates for 7 days at 28°C.

(B) Colony diameters of the WT and mutant strains cultured on different media for 7 days. Error bars: Data are represented as mean ± SD. The asterisk indicates a significant difference between the mutants and the WT strain (n = 3 for the WT strain, n = 9 for each mutant strain; Tukey's HSD, p < 0.05).

(C) The proportions of deformed hyphal cells of the WT and mutant strains cultured on the PDA medium for 7 days.

(D) Mycelial morphology and TUNEL assay. The fungi were cultured on the PDA medium for 7 days, and the mycelia were then stained with 4ʹ,6-diamidino-2-phenylindole (DAPI), followed by the TUNEL analysis. a. The mycelia were stained with DAPI. b. Free DNA of the hyphae was re-stained with FITC-dUTP. c. The nuclei of the hyphae were stained with propidium iodide (PI). d. The mycelia were re-stained with Calcoflour White (CFW). e. The merged picture of (b)–(d). Samples were examined under a confocal laser scanning microscope. Scale bar, 10 μm.

(E) Analysis of DNA fragmentation and cell apoptosis in hyphal cells. The ratio of green to red fluorescence intensity was determined for at least 30 fields observed under a microscope, and the horizontal bars depict the median. The asterisk indicates a significant difference between the mutants and the WT strain (Tukey's HSD, p < 0.05).

(F) Relative transcription levels (RTLs) of Aoras2, Aoras3, and Aorheb genes in the WT strain at different time points. The red line indicates the standard (which has an RTL of 1) for the statistical analysis of the RTL of each gene in a deletion mutant compared with that in the WT strain under a given condition. Error bars: Data are represented as mean ± SD. An asterisk indicates significant difference in the expression of Aoras2, Aoras3, and Aorheb genes in the WT strain at days 3, 5, and 7 days compared with that at day 2 (n = 3 for each gene; Tukey's HSD, p < 0.05).