Fig. 1: Phenotypes of astol1 mutant and MutMap-based cloning of OsASTOL1.
From: A molecular switch in sulfur metabolism to reduce arsenic and enrich selenium in rice grain
a The phenotype of WT and astol1 mutant treated with 0 or 20 μM As(III) for 2 days. Three plants for each genotype are shown. Scale bars, 20 mm. b Relative root elongation of WT and astol1 mutant treated with 0 or 20 μM As(III) for 2 days. Asterisks indicate significant difference by two-sided Student’s t test: ***P < 0.001. c Growth phenotype of 5-week-old WT, astol1(+/−) and astol1(+/+) mutant grown hydroponically. Two plants for each genotype are shown. Scale bars, 10 cm. d Biomass of 5-week-old WT, astol1(+/−) and astol1(+/+) mutant grown hydroponically. e Wild-type rice and astol1 mutants were grown in a paddy field until maturity. f Chromosomal location and gene structure of OsASTOL1. White and black boxes represent untranslated regions and exons, respectively, and the lines between them represent introns. Red line indicates the mutation site. g Validation of knock-out OsASTOL1/ OsASTOL1S189N protein in four independent CRISPR lines in the astol1(+/−) background using immunoblot analysis. OsASTOL1 protein (lower band) is detected by AtOAS-TL A1 antibody (anti-OAS-TLA) and the Rubisco large subunit is used as a loading control. The experiment was repeated three times and similar results were obtained. h Relative root elongation of WT, astol1(+/−), astol1(+/+), and four independent CRISPR lines in the astol1(+/+) background. Plants were treated with 20 μM As(III) for 2 days. i Biomass of 5-week-old WT, astol1(+/−), astol1(+/+), and CRISPR lines in astol1(+/−) or astol1(+/+) background. DW, dry weight. WT, wild type. astol1(+/−), astol1 heterozygote. astol1(+/+), astol1 homozygote. Data in b, d, h, and i are shown as means ± s.d., n = 10 for WT (control) and astol1 (As(III) treatment), 13 for astol1 (control) and WT (As(III) treatment) (b), 5 (d), 6 for astol1(+/+), 10 for WT and all CRISPR lines/astol1(+/+), 19 for astol1(+/−) (h) or 3 (i) biological replicates; each biological replicate represents an individual plant. Different letters in d, h, and i indicate significant differences (P < 0.05) using one-way ANOVA followed by Tukey’s test.
