Fig. 3
Expression, subcellular localization, and enzyme activity of YGL3. (a) Schematic diagram of a rice seedling at the fourth leaf stage. SB, shoot base, corresponding to the stages P0–P3 of leaf development; L1–L4, the first leaf to fourth leaf. (b) Expression levels of YGL3 in WT and ygl3 plants at 32 °C and 22 °C. The RNA was extracted from two-week-old plants. The rice Actin gene was used as the internal control. (c) Expression of YGL3 in WT plants at the fourth leaf stage. (d) Protein abundance of YGL3 in WT and ygl3 plants at 32 °C and 22 °C. Total protein was extracted from leaves of two-week-old plants. β-Actin was used as a loading control. (e) qRT-PCR analysis of the expression levels of YGL3 in various tissues. R, root; S, stem; L, leaf; LS, leaf sheath; P, panicle; 6 d, 9 d, 12 d, 15 d, and 21 d correspond to the developing endosperm at 6, 9, 12, 15, and 21 d after fertilization, respectively. (f) Schematic illustration of the different lengths of YGL3 protein. (g) Subcellular location of YGL3, YGL31–31aa, and YGL332–459aa in rice protoplasts. GFP, green fluorescence; Red, chloroplast autofluorescence. Scale bars = 5 μm. (h) Enzyme activity of 4-hydroxy-3-methylbut-2-enyl diphosphate reductase in WT and ygl3 plants. Total protein was extracted from two-week-old plants. (i) IPP contents of WT and ygl3 plants. Total protein was extracted from two-week-old plants. (j) Immunoblot analysis of YGL3 protein. Protein samples were extracted from WT. β‐Actin was used as a loading control. TP, total protein; Chl, chloroplast; Thy, thylakoid membrane fraction; Str, stroma fraction. Data are shown as mean ± standard deviation from three biological replicates. Asterisks indicate statistical significance as determined by Student’s t-test (**P < 0.01; 0.01 < *P < 0.05)