Fig. 8.

H₂O₂ accumulation and PCD detection in spl33. (A) DAB staining for H₂O₂ accumulation. (B) Trypan blue staining for cell death. (C, D) Aniline blue staining for callose accumulation under UV light; fluorescent regions indicate callose accumulation. (C) WT; (D) spl33. Scale bar: 100 μm. (E) DNA fragmentation detection in mesophyll cells by TUNEL assay. Red signal represents staining with propidium iodide, and yellow and green signals indicate TUNEL-positive nuclei of dead cells resulting from PCD. Scale bar: 100 μm. WT and spl33 leaf samples in (A–E) were analysed at 28 DAS.