Fesem s 4800 scanning electron microscope

To characterize the colonization of “Pingli-5” (HR, Highly Resistant) and “Red Globe” (HS, Highly Susceptible) by B. cinerea, 2–3 cm² leaf pieces were collected at 4, 8, 12, 18, 24, 36, 48, 72, and 96 hpi, fixed, and decolorized in ethanol/trichloromethane (3:1, v/v) containing 0.15% (w/v) trichloroacetic acid, before clearing in saturated chloral hydrate, and were then stored in 20% glycerol. Samples were subsequently stained with aniline blue solution (for staining fungal tissues a blue color) and examined with an Olympus BX-51 microscope (Olympus Corporation, Japan). For each sample, fungal germination, and infection percentages were examined. For scanning electron microscopy (SEM), leaf tissues were cut into small pieces (0.5–1 cm²), fixed in 4% (v/v) glutaraldehyde in phosphate buffer (0.1 M, pH 6.8) for 12 h at 4°C, and rinsed in the same buffer four times for 10–15 min. After dehydration in a graded ethanol series (30, 50, 70, 80, 90, 100%, v/v), the samples were then critical-point dried, coated with gold in a sputter coater, and examined with a JEOL FESEM S-4800 scanning electron microscope at 15 kV (Cheng et al., 2012 (link)).

The structural growth of B. cinerea on leaves and berries of one representative genotype, “Ju mei gui” or “Summer black,” was observed using SEM (JEOL FESEM S-4800 scanning electron microscope, JEOL, Tokyo, Japan). Infected leaf and berry skins were cut into 1.0–1.5 cm² pieces and immersed in 4% glutaraldehyde. After vacuum infiltration for 30 min, the infected leaf and berry skins were rinsed five times for 5, 10, 15, 20, 20 min, respectively, with 0.1 M sodium phosphate buffer (PBS) (pH 6.8). The segments were dehydrated in an ethanol gradient: 30%, 50%, and 70% for 15 min each; 80% and 90% for 20 min each; and 100% alcohols twice for 30 min. Finally, the samples were incubated in acetone for 30 min and isoamyl acetate twice for 15 and 30 min in three biological replicates. The segments were desiccated by CO₂, coated with gold in a sputter coater, and then observed under a scanning electron microscope at 15 kV [43 ].