Fig. 3

Analysis of the single bkn1 and bkn2 CRISPR mutants. a-e. Stage 13 pistils were pollinated with wild-type Col-0 pollen for 2 h and then fixed and stained with aniline blue. Brightfield images (left) show pollen grains adhered to the stigmatic papillae, and fluorescent images (right) show aniline blue stains of callose deposits in the pollen tubes. Representative images of pollinated Col-0 pistils (a) bkn1–2 (b) bkn1–3 (c), bkn2–1 (d) and bkn2–3 (e) show similar pollen adhesion and pollen tube penetration. Scale bar = 100 μm. f. Number of Col-0 pollen grains adhered to stigmas following aniline blue staining at 2-h post-pollination for Col-0 and the bkn1 and bkn2 mutants. During this staining procedure, ungerminated pollen grains are washed away and so the number of adhered pollen grains can be quantified. n = 10 stigmas per line. g. Seed set rates, following hand-pollination with Col-0 pollen. All bkn1 or bkn2 mutants display normal seed set, similar to Col-0. n = 10 siliques per line. Letters represent statistically significant groupings of p < 0.05 based on a one-way ANOVA with a Duncan post-hoc test. No significant differences were observed for (f) and (g)