For RNA sequencing (RNA-seq) analysis, 1 × 108 conidia of the gemA deletion mutant and its parental wild-type (WT) strains were incubated in liquid MM for 18 h at 37°C. After incubation, mycelium was filtered and immediately harvested in liquid nitrogen. The samples were then sent to Personal Biotechnology (Nanjing, China) for the following sequencing. The raw sequence reads have been submitted to SRA (https://www.ncbi.nlm.nih.gov/sra ) at NCBI with six accession numbers SRR19440562–SRR19440564 (WT-1, WT-2, and WT-3, three biological replicate samples of WT) and SRR19440565–SRR19440567 (ΔgemA-1, ΔgemA-2, and ΔgemA-3, three biological replicate samples of ΔgemA).
For RNA isolation, the sample preparation is similar to that described in RNA-seq analysis. The total RNA was obtained using the purification kit (UNIQ-10 column, Sangon). Digestion and reverse transcription were performed using the HiScriptII Q RT SuperMix for the qPCR (gDNA wiper) kit from Vazyme to obtain cDNA. qPCR was then performed using the AceQ qPCR SYBR green master mix (Vazyme). All procedures were performed according to the manufacturer's instructions.
For RNA isolation, the sample preparation is similar to that described in RNA-seq analysis. The total RNA was obtained using the purification kit (UNIQ-10 column, Sangon). Digestion and reverse transcription were performed using the HiScriptII Q RT SuperMix for the qPCR (gDNA wiper) kit from Vazyme to obtain cDNA. qPCR was then performed using the AceQ qPCR SYBR green master mix (Vazyme). All procedures were performed according to the manufacturer's instructions.