Fig. 3: The C-terminal truncated gc1 confers low glume coverage by accumulating higher protein level.

a Immunoblot analysis of Myc tag fused GC1 and gc1 proteins extracted from the young panicles of transgenic GC1-OE and gc1-OE sorghum plants, respectively. The relative expression of GC1 and gc1 in transgenic plants compared to Wheatland were detected by qPCR. b Glume length evaluation of five GC1 haplotypes among 482 sorghum accessions. Data are mean ± s.e.m. c Comparison of GC1 expression level among the five GC1 haplotypes. The two ends of the box plot and the upper, middle, and lower box lines represent the upper edge, lower edge, median and two quartiles of values in each group. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test in b and c. d Schematic diagram of peptide structure of natural or synthetic GC1, gc1-a, gc1-b, gc1-c, gc1-d, SiGC1, SiGC1¹⁰⁴, and SiGC1¹²⁴ proteins. G, G protein γ subunit domain. T, Predicted transmembrane domain. The gray color shows the out-frame amino acids. e GFP tag fused with various deletion-based GC1 and SiGC1 proteins showed in d were transiently expressed in N. benthamiana leaves. Protein levels were subsequently analyzed by western blotting with anti-GFP antibody after 30 μM CHX treatment for 0 and 6 h. The expression of GFP tag from tobacco leaves were detected by RT-PCR. f The protein accumulation of GFP-GC1 and GFP-gc1-a extracted from tobacco leaves after treatments with CHX (30 μM) or CHX (30 μM) + MG132 (50 μM) for 0, 6, 12, and 24 h were analyzed by western blotting with anti-GFP antibody. Plant actin was used as the loading control. CHX cycloheximide. The numbers on the blot bands indicate the relative protein quantification by determination of grayscale value. Three independent experiments were performed in e and f. Source data are provided as a Source Data file.