Callose Staining for Magnaporthe Interaction

Callose staining was performed as Zhang et al. [4 (link)] described. Briefly, 6 fresh leaf sheaths at 0, 16, 24 and 36 h after of Magnaporthe oryzae 95234I-1b inoculation, respectively. After cleaning with 95% ethanol, plant leaves were incubated in ethanol-emulsifiable solution (phenol: glycerol: lactic acid: water: ethanol = 1:1:1:1:8) at 65°C until the green color was completely removed, washed with 50% ethanol and sterilized water. After staining in 0.1% aniline blue for 1 h, callose was observed and photographed under DMI4000B fluorescence inverted microscope (Leica Camera AG, Wetzlar, Germany), and the number of callose was counted in each treatment according to 20 field of vision.