Fig. 2.
Expression levels of FUS3 and FUS3 target genes in FUS3 phosphomutants. (A) Seedling establishment (cotyledon expansion) assay of 3-month-old seeds showing rescue of fus3-3 desiccation intolerance by FUS3:FUS3-GFP, FUS3:FUS3S>A-GFP (lines #3 and #5), and FUS3:FUS3S>D-GFP (lines #11 and #13) phosphomutant lines. Seeds were cold stratified for 3 d and grown under constant light at 22 °C. The averages of three plates of 50 seeds ±SD are shown. (B) FUS3S>A–GFP and FUS3S>D–GFP were detected in the nuclei of almost all embryo proper cells in the cotyledon of walking stick/bent embryos. Scale bar=20 µm. (C) qPCR showing decreased FUS3S>A-GFP and slightly increased FUS3S>D-GFP transcript levels in transgenic lines. All plants were grown under long days (21 °C/18 °C) and siliques were collected at ~10 DAP (walking stick/bent cotyledon stages). (D) Transcript levels of FUS3 target genes measured by qPCR. 12S storage protein CRU2 (At1G03880), 2S storage protein 2S3 (At4G27160), fatty acid elongase FAE1/KCS18 (AT4G34520), and scorpion toxin proteinase/trypsin inhibitor SCT (At1g47540). FUS3S>A lines show lower transcript levels of most genes. Averages of three biological replicates ±SD are shown. Statistical significance against the wild type (WT) calculated by Welch’s t-test (*P<0.05, **P<0.01). All plants were grown under long days (21 °C/18 °C) and siliques were collected at ~10 DAP (walking stick/bent cotyledon stages). (E) Seed storage proteins extracted from 1 mg of dry seed visualized by Coomassie staining on a 15% SDS–polyacrylamide gel electrophoresis. (This figure is available in colour at JXB online.)