Figure 6.

TIP4;3 affects ROS accumulation under cold stress. (a, b) Diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) histochemical staining of WT, TIP4;3‐OE and tip4;3 leaves under cold stress. The experiments were independently repeated three times. Representative photographs of leaf histochemical staining are shown. Scale bars, 1 mm. (c) Imaging of ROS levels in WT, TIP4;3‐OE and tip4;3 leaves under 25 °C and 4 °C conditions detected with H₂DCFDA. (d) Quantification of ROS levels in (c). Error bars represent mean ± SD (standard deviation) from three independent experiments (n = 10). ***P < 0.001, **P < 0.01, *P < 0.05 (two‐sided t‐test). (e) Exogenous application of glutathione (GSH) partially rescues the compromised cold sensitivity of tip4;3 mutants. Maize seedlings were treated with 10 mM GSH under 4 °C for 2 days. Representative photographs were taken after 2 days of recovery. (f) Leaf injury area for the seedlings in (e). Error bars represent mean ± SD (standard deviation) from 3 biological replicates (n = 5 plants for each replicate). Different letters represent significant differences (P < 0.05) determined by one‐way ANOVA with Tukey's multiple comparisons test.