Transcriptional Analysis of Fungal Mutants

Total RNA extraction and qRT-PCR analysis were performed as described by Guan et al. (55 (link)). Total RNA of the mycelia of the HN08 strain and mutants were extracted using the RNAprep Pure Plant Kit (Tiangen, Beijing, China). cDNA synthesis was performed with TransScript One-Step gDNA Removal and cDNA Synthesis SuperMix (TransGen Biotech, Beijing, China). The expression levels of the target genes were quantified by qRT-PCR performed with an ABI7500 sequence detection system (Applied Biosystems, Waltham, MA, USA). Reactions were performed in a total volume of 10 µL using the SYBR Premix Dimer Eraser Kit (Takara, Beijing, China). All of the reactions were repeated in at least three independent pools in three sets of biological replicates. The primer sequences were listed in Table S1.