Fig. 5: Specific expression of ATG5-GFP in the integumentary tapetum of atg5-2 mutant promotes embryo pattern formation.

a Specific expression ATG5-GFP in the atg5-2 integumentary tapetum under the control of the integumentary tapetum-specific promoter NtTPE8. GFP GFP fluorescence, SR2200 SCRI Renaissance 2200. Scale bar: 20 µm. Observation of GFP in proTPE8:ATG5-GFP was performed three times with similar results. b, d mPS-PI staining showing representative integumentary tapetum (b) and embryos (d) in 4-DAP seeds from the WT, atg5-2 mutant and proTPE8:ATG5-GFP complementation plants. Scale bars: 50 µm (upper row) and 10 µm (lower row). c Quantitative analysis of the integumentary tapetum width. Values displayed in box and whisker plots are from three independent experiments, with ten seeds in each experiment (n = 30). Centerline, median values; top and bottom of boxes, 75th and 25th percentiles; top and bottom lines, maximum and minimum values. Tukey–Kramer multiple comparison test was used to determine the significant differences (one-way analysis of variance (ANOVA) between groups; P = 1.32E-15; F = 52.12). The labels a–c represent highly significant differences when compared at the 0.01 level. e Percentages of embryos identified in 4-DAP seeds from the WT, atg5-2 mutant, and proTPE8:ATG5-GFP complementation plants. Data are the mean ± SD from two independent experiments, with 50 embryos analyzed in each experiment (n = 100). f Representative images showing seeds from the WT, atg5-2 mutant, and proTPE8:ATG5-GFP complementation plants. Arrows indicate aborted seeds. Scale bars: 1 mm. g The number of normal and aborted seeds in the WT, atg5-2 mutant and proTPE8:ATG5-GFP complementation plants. Gray boxes indicate the number of normal seeds, while the white boxes represent the number of aborted seeds. Values are means ± SD (n = 4 independent ovaries), (two-tailed Student’s t test; ***P < 0.001).