Fig. 3

Ss-CAD-silenced transformants showing reduced superoxide accumulation. Exogenous H₂O₂ and NADPH treatments restored sclerotial development in Ss-CAD-silenced transformants. a NBT staining was used to detect superoxide accumulation at 3–4 dpi on PDA. Staining intensity was monitored under a light microscope, and blue staining represented superoxide accumulation. Ss-CAD-silenced transformants accumulated less blue formazan than the wild-type strain EP-1PNA367. b Treatment with exogenous H₂O₂ and NADPH recovered sclerotial development in Ss-CAD-silenced transformants. The effects of adding 6 mM H₂O₂ and 0.2 mM NADPH individually to PDA on S. sclerotiorum sclerotial development were evaluated. Photographs were taken at 10 dpi. c Weights of sclerotia formed under various culture conditions. Sclerotia produced on each plate were collected, air-dried, and weighed at 15 dpi. Three independent replicates were performed for each experiment. Values are means ± S.E. **P < 0.01; one-way ANOVA