Complementation of Fungal Gene Mutants

The mutants were complemented with native gene copies of the wild-type strain 70-15. First, the pKO1B-HPH was built by the replacement of Ph3-GFP cassette with a HPH gene from pCB1003 [78] in pKO1B. The copies of the complementation genes were then cloned from the genomic DNA of the wild-type strain with the primers listed in Table S1 in Text S1 and were inserted into the XbaI/HindIII sites of pKO1B-HPH by the yeast recombinational cloning method. The constructed complementation plasmids were transformed into the mutants using the ATMT method, and the transformants were screened on selective medium containing 200 µg/ml hygromycin B. The gene-rescued transformants were identified by RT-PCR at the mRNA level (Figure S4C) with primers specific for the targeted genes (Table S1 in Text S1).