Fig. 2.
L-methionine (L-Met) induces AtGLR3.5-dependent increases in [Ca2+]cyt in Arabidopsis. (A) Germination of wild-type (WT) and AtGLR3.5-RNAi seeds at different concentrations of L-Met. Seeds were sown on half-strength MS medium supplemented with L-Met and were scored for germination after incubation at 22 °C for 36 h. (B) Germination of WT and AtGLR3.5 T-DNA insertion lines (Atglr3.5-1 and Atglr3.5-2) at different concentrations of L-Met. Seeds were examined for germination after incubation at 22 °C for 36 h. (C) Germination of WT and AtGLR3.5-RNAi seeds in response to various treatments. Seeds were sown on modified MS media containing 0 mM CaCl2 (control), 0 mM CaCl2 and 10 µM L-Met, 1 mM CaCl2, 1 mM CaCl2 and 10 µM L-Met, 0 mM CaCl2 and 10 mM EGTA, or 0 mM CaCl2 supplemented with 10 µM L-Met and 10 mM EGTA, and were scored for germination after incubation at 22 °C for 36 h. Data in (A–C) are means (±SE) from three independent experiments. Significant differences were determined using Student’s t-test: **P<0.01. (D) [Ca2+]cyt-dependent FRET efficiency changes in WT and AtGLR3.5-RNAi plants in response to 1 mM L-Met. The primary roots of seedlings expressing yellow cameleon 3.60 (YC3.60) were used for the analysis and the time point when plants were treated with L-Met is indicated. Ten seedlings of each genotype were tested, and representative results are shown. (This figure is available in colour at JXB online.)