Fig. 8

The influence of Ca²⁺ on the expression levels of some genes related to glycogen and trehalose regulation in the wild-type and Δpac-3 strains. Cells from the wild-type and Δpac-3 strains were cultured at pH 5.8 (0) for 24 h and shifted to 10 mM or 300 mM CaCl₂ for 15, 30 and 60 min. Mycelial samples were used to extract total RNA. Gene expression analysis was performed by RT-qPCR in the StepOnePlus™ Real-Time PCR system (Applied Biosystems) using the Power SYBR® Green and specific primers (Additional file 1: Table S1, qPCR). The actin gene was used as the reference gene, and the zero sample in the wild-type was used as the reference sample. At least four biological replicates were performed, and the data were analyzed using the relative quantification standard curve method. Bars indicate the standard deviation from the biological experiments. The asterisks indicate significant differences compared to the zero sample in the wild-type or mutant strains (Student’s t-test, P < 0.01)