Fig. 3: Localization of PsAF5 and PsATG8 to mitochondria under different ROS-inducting conditions.

a Both H₂O₂ treatment and mitophagy-inducting conditions can increase the mitochondrial localization of PsAF5 and PsATG8. The treatment time for 1 mM H₂O₂ or 4 µg/mL ametoctradin was 1 h prior to observation. b The mitochondrial localization of PsAF5 and PsATG8 increased during infection. Prior to observation, the mycelia were treated with Mito Tracker Green FM for 1 h. c Mitophagy inducting conditions, aging, and N-starvation, can increase the mitochondrial localization of PsAF5. In (a–d), scale bars represent 5 μm. d Schematic diagram of the extraction of mitochondria from P. sojae using sucrose gradient centrifugation. e Verification of mitochondrial activity. The isolated mitochondria in (d) method were stained with Janus green B. f Mitochondrial integrity verification. The mitochondria isolated in (d) method were observed by transmission electron microscopy (TEM). g, h H₂O₂ treatment can promote the transfer of PsAF5 to mitochondria from the cytoplasm. Error bars in graphs represent means ± SEM (n = 3 biologically independent samples). Data in (a, b, c, d, g, h) represent at least 3 biological experiments, and data in (e, f) represent 2 biological experiments. Ordinary one-way ANOVA and Dunnett’s multiple comparisons test were used. Source data are provided as a Source Data file.