Fig. 2: Phenotypic analysis of the reproductive organs of osga3ox1 mutant rice.
a, b Pollen viability test of Nipponbare, OsGA3ox1+/− (heterozygous), and osga3ox1 mutant performed by conducting staining with the I2-KI solution. Pollen grains stained black are deemed viable; those stained red are deemed sterile (Bars = 0.5 mm) (a). Percentage of viable vs sterile (n = 5) (b). c Pollen viability test of GA-treated osga3ox1 mutant performed by conducting I2-KI staining, same as that for panel (b) (n = 3). P-values by Dunnett’s multiple comparison test are indicated. d Seed setting rate of Nipponbare, OsGA3ox1+/−, osga3ox1, and GA4-treated osga3ox1 mutant plants (n = 3). e In vitro pollen tube germination of Nipponbare and osga3ox1 mutant (n = 15). f Reciprocal crossing test between Nipponbare and osga3ox1 mutant plants. Arrowheads indicate pollen tubes. Bars = 0.5 mm. g, h Endogenous quantities of starch (g) and sucrose (h) contents in anthers of Nipponbare and osga3ox1 mutant (n = 3). i, j Enzyme activity of acid invertase in extractions from Nipponbare and osga3ox1 mutant pollen in the early binucleate stage. Summary of the extracted fractions (i) and their invertase activities (j) (these experiments were repeated three times with similar results). k Western blot analysis of OsCIN3 in extracted fractions in (I) and total extract of anthers using an OsCIN3 specific antibody. CBB staining of total extract is also shown. In all experiments, error bars = s.d. For panels (b) and (d): one-way ANOVA with Tukey’s multiple comparisons test. Different letters denote significant differences (p < 0.001). For panels (e), (g), and (h): ***p < 0.001, **p < 0.01, and *p < 0.05 by two-tailed paired t-tests.
