2.3 lite

Lipid droplets and glycogen staining mainly followed the method described previously [39 (link)]. The suspension of conidia (20 μL) was cultured on a hydrophobic membrane to induce formation of appressorium at 25 °C in the dark for 0 h, 2 h, 8 h, 16 h, and 24 h, respectively. Nile red staining was carried out by using the staining solution (final concentration 1 mM) for 30 min. Then the distribution of lipid droplets was observed under fluorescence microscopy (Zeiss LSM880, with a 40×objective). Similarly, glycogen was stained with I2/KI solution (1 mM) for 1 min. And the images were obtained with Zeiss microscopy (Zeiss A1). To analyze the glycogen, image processing and measurements were performed using ZEN software (Zeiss 2.3 lite). Images were selected using the ‘rectangle’ tool and the mean value of the selected region was calculated using the “Measure” plugin. The experiment was repeated 3 times, and 100 conidia were counted for each treatment.