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. 2021 Jan 18;11:622513. doi: 10.3389/fpls.2020.622513

FIGURE 2.

FIGURE 2

Transient LsPDI1 expression in planta induces reactive oxygen species (ROS) burst and cell death. (A) Detection of ROS burst by 3,3’-diaminobenzidine (DAB) staining after LsPDI1-GFP expressed transiently for 12 h. GFP was the negative control. (B) Detection of relative electrolyte leakage (%) after LsPDI1-GFP transiently expressed for 96 h. GFP was the negative control (n = 3). (C) Nicotiana benthamiana leaves were infiltrated with Agrobacterium carrying GFP and LsPDI1-GFP and photographed after agroinfiltration for 96 h. GFP was the negative control (n = 3). (D) Detection of living rice protoplasts by fluorescein diacetate (FDA) staining and confocal laser-scanning microscopy after transiently transforming mCherry or LsPDI1-mCherry for 16 h (n = 5). The mCherry was the negative control. Bar = 50 μm. (E) Detection of dual-luciferase (LUC) activity in rice protoplasts after transiently co-expressing LUC and LsPDI1 or LUC and GFP (negative control) for 40 h using an LUC assay system (n = 5). “**” indicates significant differences between treatments (P ≤ 0.01; Student’s t-test).