转化方法：

DNA mediated transformation was done as follows;

 a young mycelium was grown overnight at 30 ◦ C or 37 ◦ C 180 rpm in minimal medium with supplements, harvested by filtration (Whatman filter paper),

1）washed with 0.6 M MgSO4 ,

2）suspended in 5 ml DSPS (1.1 M KCl, 0.1 M citric acid【柠檬酸】, pH 5.8) with 100 mg of lysing enzymes from Tricho- derma harzianum (Sigma L1412), 100 mg of lysozyme from chicken egg white (Sigma L7651) and 100 mg of albumin bovine fraction V (Sigma A4503). The slurry was incu- bated at 30 ◦ C, 100 rpm for 1–2 h and protoplasts harvested by filtration through a one layer Miracloth,

3） washed by centrifugation 4500 × g, 4 ◦ C, 10 min, twice with 50 ml STC (1.2 M Sorbitol, 50 mM CaCl2 , 50 mM Tris pH 7.5).

4) The final pellet was suspended in 1ml STC and stored at 4◦C until further use. In a falcon tube  of pEXPYR plasmid DNA was added onto 50 μl STC (final volume) along with addi- tional 150μl of protoplast suspension (∼108 ), incubated at RT for 15–20 min prior to the addition of 2 ml of 60% PEG solution (60% PEG4000 in STC). The transforming mixture was mixed carefully by swirling and incubated at room temperature for 10–15 min, 8 ml of STC was added and 1 ml poured onto protoplast-recovery (1.2 M sorbitol) and transformant-selection (no uracil, uridine or 5-FOA) basic medium plates (medium without yeast extract or vitamins). Plates were incubated at 30 ◦ C or 37 ◦ C for one day and then inverted.