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. 2017 Oct 10;6:e27529. doi: 10.7554/eLife.27529

Figure 3. SAM, ACC, and ethylene contents in OsSAMS1 and S11 transgenic lines.

(A) SAM contents in the S11-overexpression lines and WT plants (using 40-d-old seedlings). (B) ACC contents in the S11-overexpression lines and WT plants (using 40-d-old seedlings). (C) Ethylene contents in the S11-overexpression lines and WT plants (using 40-d-old seedlings). (D) SAM contents in the OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). (E) ACC contents in the OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). (F) Ethylene contents in the OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). Tukey’s honestly significant difference post hoc test was performed for multiple comparisons. Letters indicate significant differences, p<0.05. Data are from three replicates. FW, fresh weight.

Figure 3.

Figure 3—figure supplement 1. Phenotype and identification of OsSAMS1 transgenic lines.

Figure 3—figure supplement 1.

(A) Expression levels of OsSAMS1 in WT, OX and RNAi transgenic lines. (14-d-old seedlings). The average (± standard deviation) values from three biological replicates are shown. Significant differences are indicated (*p<0.05, **p<0.01) based on Student’s t-test. (B) Northern blot analysis of OsSAMS1 mRNA expression level in WT, OX and RNAi transgenic lines. rRNAs were stained with ethidium bromide and served as loading controls. (C) Western blot analysis of OsSAMS1 protein expression level in transgenic lines. Actin was probed and served as a loading control. The black square indicates protein OsSAMS1 with flag tag, the black triangle indicates endogenous OsSAMS1 protein in rice. (D) Schematic of the OsSAMS1Cas9 construct. Key elements and the sgRNA sequence for specifically targeting OsSAMS1 are indicated. DNA sequencing chromatograms of two Ossams1 KO lines. The mutations, ‘T’ insertion in line Ossams1 KO#31 and ‘TTTACCTC’ deletion in line Ossams1 KO#39, which lead to premature termination in the amino acid sequence of OsSAMS1 are indicated. (E) Effect of SAM and ethylene on seed germination in the OsSAMS1 transgenic lines. SAM concentration was 1 mM, ethylene concentration was 10 ppm. Bar = 1 cm. (F) Phenotypic comparison of field-grown plants at the grain-filling stage. Bar = 15 cm (upper panel) and 10 cm (lower panel).
Figure 3—figure supplement 2. SAM, ACC and ethylene contents in OsSAMS1 transgenic lines.

Figure 3—figure supplement 2.

(A) SAM content in OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). (B) ACC content in OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). (C) Ethylene content in OsSAMS1 transgenic lines and WT plants (using 40-d-old seedlings). Tukey’s honestly significant difference post hoc tests were performed for multiple comparisons. Letters indicate significantly different results, p<0.05. Data are from three replicates. FW, fresh weight.