Fig. 1. Gene editing of OsRAL5 gene and phenotypic characterization of grain size and rice quality. A, Schematic diagram of sgRNA target sites (TS) and homozygous mutations in TS1 and TS2. B, Wild type NJJG and the OsRAL5-edited mutant seedlings (dOsRAL5-TS) at the grain-filling stage. Scale bar, 15 cm. C, Phenotypic differences in grain length and grain width between NJJG and dOsRAL5-TS mutants. Scale bar, 5 mm. D‒H, Comparisons of grain length (D), width (E), thickness (F), 1000-grain weight (G), and grain yield per plant (H) of NJJG and dOsRAL5-TS mutants. I, Comparison of the appearance of brown rice and starch granules between NJJG and dOsRAL5-TS mutants. Scale bars, 10 μm. J‒L, Comparisons of chalkiness degree (J), Rapid Visco Analyser profiles (K), and amylopectin chain length distribution (L) between NJJG and dOsRAL5-TS mutants. NJJG, Nanjing Jinggu. Data are Mean ± SD (n = 4). * and ** indicate P < 0.05 and P < 0.01 by the Student’s t-test, respectively.

Fig. 2. Comparisons of contents of seed storage substances and expression analysis of grain size- and storage substance-related genes in Nanjing Jinggu (NJJG), dOsRAL5-TS mutants, and OsRAL5-OE plants. A, Total starch content. B, Total protein content. C, Total lipid content. D, Amylose content. E, Glutelin content. F, Albumin content. G, Globulin content. H, Prolamin content. I, Comparison of expression levels of genes related to grain size (GS3, GS5, GL6, and GL7), storage proteins (GluA, GluB, GluD, Prol14, and RP10), and starch synthesis (OsAGPS2b, OsAGPL2, GBSSI, SSI, SSIIa, and SSIIIa). Data are Mean ± SD (n = 4). * and ** indicate P < 0.05 and P < 0.01 by the Student’s t-test, respectively.