Fig. 8

Comparison of ROS accumulation in root tips of HyPRP1-silenced cotton seedlings inoculated with V. dahliae. a Representative microphotographs of NBT staining. The roots were inoculated with NBT for 40 min and were then subjected to microscopic observation to determine O₂⁻ production. Scale bar = 1 mm. b Representative microphotographs of DAB staining. The roots were placed in DAB for 10 h and were then subjected to microscopic observation to determine H₂O₂ production. Scale bar = 1 mm. c Representative confocal images of DCFH-DA staining. The roots were placed in DCFH-DA for 30 min and were then examined to determine ROS production. Scale bar = 100 μm. At each time point of each treatment, nine roots from three separate plants were examined. Data represent the means ± SE from three biological replicates; asterisks indicate statistically significant differences according to Tukey’s multiple comparisons test (*P < 0.05, **P < 0.01, ***P < 0.001). 10-day-old seedlings with two fully expanded cotyledons were used for VIGS. After two weeks later HyPRP1-silenced plants were inoculated with V. dahliae