Soytone

Protease secretion was determined after growing the strains on 1/5 TSA (15 g/L Tryptone (Roth, Karlsruhe, Germany), 5 g/L Soytone (Merck, Darmstadt, Germany), 5 g/L NaCl, 15 g/L Agar-Agar (Roth, Karlsruhe, Germany)) amended with 10% (w/v) powdered milk (Roth, Karlsruhe, Germany) at 28 °C in constant darkness or constant light (1800 lux). The mutant strains were inoculated in the center of the plate and colony diameter as well as halo diameter were measured every 24 hours for 2 days at 28 °C. A halo appearing around the inoculated area is indicative for exoprotease activity^{105 (link)}. Calculations are based on the ratio halo/mycelium. QM6aΔtmus53 was used as control for every set. At least three biological replicates were analyzed per strain.

To determine protease activity, strains were grown on TSA plates (3 g/l tryptone (Merck, Darmstadt, Germany), 1 g/l soytone (Merck, Darmstadt, Germany), 1 g/l NaCl, 20 g/l agar) supplemented with 1.5% milk powder (Roth, Karlsruhe, Germany) at 28°C in constant darkness or constant light (1,800 lux). Protease activity was manifested by halo formation. The sizes of the cleared zones and hyphal extension were recorded and the ratio of the diameter of the halo to the diameter of the colony was calculated. Three biological and two technical replicates were analyzed.