Fig 4.

Reduced expression of rom2 results in an increased susceptibility to cell wall-perturbing agents and heat associated with cytosolic leakage. (A) In a series of 10-fold dilutions derived from a starting suspension of 5 × 10⁶ conidia ml⁻¹ of the indicated strains, aliquots of 3 μl were spotted onto AMM agar or, when indicated, onto YG or potato dextrose agar. When indicated, AMM or YG was supplemented with Congo red (CR) (8 μg ml⁻¹), calcofluor white (CFW) (40 μg ml⁻¹), sorbitol (Sorb) (1.2 M), or doxycycline (Doxy). AMM agar plates were incubated at 37°C for 36 h or, when indicated, at 48°C for 48 h. YG and potato dextrose agar plates were incubated at 37°C for 24 h. (B) A total of 1.5 × 10³ conidia were spotted on AMM agar and incubated at 37°C. After 36 h, the agar plates were shifted to 48°C. After 6 h of incubation at 48°C, the agar plates were overlaid with soft agar supplemented with 5-bromo-4-chloro-3-indolylphosphate and incubated for another 30 min at 48°C. Cell lysis (enzymatic activity) is visualized by blue coloring.