Fungal Genome Extraction and Sequencing

Fungal mycelia were scraped from the colonies on PDA. The Biospin Fungus Genomic DNA Extraction Kit (Bioer Technology Co., Hangzhou, China) was used to extract total genomic DNA. The polymerase chain reaction (PCR) technique was utilized for the amplification of target DNA fragments. The primer pairs LR0R/LR5, NS1/NS4, ITS5/ITS4 and fRPB2-5F/fRPB2-7cR were used to amplify LSU, SSU, ITS and rpb2 [20 (link),21 (link),22 (link)]. The amplifications were carried out as detailed in Dong et al. [23 (link)]. The PCR thermal cycle program for the amplification of LSU, SSU and ITS was provided as initially 94 °C for 3 min, followed by 35 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 50 s, elongation at 72 °C for 90 s and a final extension at 72 °C for 10 min. The annealing was adjusted to 52 °C for rpb2. PCR products were checked on 1% agarose electrophoresis gels stained with Gel Red. The sequencing reactions were carried out by Shanghai Sangon Biological Engineering Technology and Services Co., Shanghai, China.