Fig. 5: gnom mutants highly resemble praf mutants.
a 4-day-old seedlings of wild type, praf4t;5c;8c;9c, and loss-of-function gnom (segregated from gnomB/E) mutants. b 4-week-old plants of wild type, praf triple mutants (praf4t;8c;9c and praf5c;8c;9c), and gnomB/E (trans-heterozygously complementing B4049 and emb30-1 alleles). c Confocal images show stomatal phenotypes of 5-day-old adaxial cotyledon epidermis of the designated genotypes. Stomatal lineage and guard cells are manually traced and highlighted. Three independent experiments were performed. Scale bar = 10 μm. d Box plots show quantification of stomata lineage index for (c). Box plots show first and third quartile (box), median (line) and mean (cross). n, # cotyledons counted. Student’s unpaired t tests were used for comparison. Two-sided P values are 0.4439 (for gnom vs. praf4t;5c;8c;9c), 0.0258 (for gnom vs. gnom;praf4t;5c;8c;9c), and 0.2922 (for basl-2;gnom vs. gnom). n.s. not significant; *P < 0.05. e Box plots show quantification of divisional asymmetry in the stomatal lineage for (c). Box plots show first and third quartile (box), median (line) and mean (cross). n, # daughter cell pairs. Student’s unpaired t tests were used for comparison. Two-sided P values are 0.1707 (for gnom vs. praf4t;5c;8c;9c), 0.9274 (for gnom vs. gnom;praf4t;5c;8c;9c), and 0.6190 (for basl-2;gnom vs. gnom). n.s., not significant. f GFP-BASL (green) lost polarization in gnom. Magenta, cell outlines stained by PI. Data represent the results of three independent experiments. g Confocal images show GFP-BASL (green) merged with 8 μM FM4-64 staining (red) (top) or GFP-channel only (bottom). Three-day old seedlings were pre-incubated with FM4-64 for 40-min (left), then treated with 70 μM BFA for 2 h (middle), followed by 2 h wash-out (right). Yellow arrowheads indicate BFA-induced endomembrane aggregations. White arrows, protein polarization. Note no BASL polarization observed after BFA treatment (middle). Three independent experiments were performed. Scale bar, 5 μm.
