The fresh conidia were cultured stationarily on potato dextrose broth (PDB; PDA medium without agar) overnight to prepare the polarized growth conidia (Fig. S1). Then, polarized growth conidia were further used to evaluate micromorphological differences under the treatment with increased HHP. (1) At least 25 hyphal lengths were measured from the pressure of 0.1 MPa, 20 MPa, and 40 MPa in every 12 hours within 48 hours. (2) Mature mycelium was stained with Calcofluor white (CFW; Sigma-Aldrich, Shanghai), 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI; Sigma-Aldrich, Shanghai), or propidium iodide (PI; MKBio, Shanghai) as the previous report to determine the cell wall and nucleus of hypha under different HHP (38 (link), 39 (link)). Briefly, at least 6-day-old mycelium from the elevated HHP was stained with the fluorescent dye (1 g/L CFW; 0.5 g/mL DAPI; 1 mg/mL PI), followed by incubation in the dark for 5 min. Slides were washed with sterile water and observed using epifluorescence microscopy. Besides, dual staining with CFW-PI was also carried out to determine the viability following the previous reports (39 (link)). All staining experiments and groups were carried out at least in triplicate.
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