Figure 4.
cml38 mutants are insensitive to the inhibitory activity of AtRALF1 on root growth, and sensitivity is restored by exogenous treatment with CML38 or by introgression of the PROCML38:CML38-GFP gene. A, Arabidopsis seedlings (2 days old) were treated with AtRALF1 for 48 h. Values are the means ± S.D. (error bars) of at least 25 seedlings. ***, p < 0.001 (Student's t test). ns, not significant. B, 2-day-old seedlings were transferred to medium containing 5 μm of AtRALF1, and the root length was measured after 6 days of treatment. Data represent mean values ± S.D. (n = 15). Student's t tests were used to compare treated with AtRALF1 and water control. ***, p < 0.001. C, Arabidopsis seedlings (2 days old) were treated with AtRALF1 (R1), CML38, or CML39 proteins for 48 h. Values are means ± S.D. (error bars) of 35 seedlings. *, p < 0.05; **, p < 0.01; ***, p < 0.001 (Student's t test). D, introgression of the PROCML38:CML38-GFP gene into the cml38-1 mutant restores the root growth inhibition caused by AtRALF1. Two-day-old parental lines cml38-1 and PROCML38:CML38-GFP or F3 progeny PROCML38:CML38-GFP/cml38-1 were treated with water or AtRALF1 for 3 days. Values are the means ± S.D. of at least 30 seedlings. ***, p < 0.01 (Student's t test). E, Arabidopsis seedlings (2 days old) were treated with water, AtRALF1 (R1), 12 μg of anti-CML38 (anti-CML) antibody, 12 μg of preimmune serum, or combinations of the peptide with anti-CML or preimmune serum for 48 h. Values are means ± S.D. of 30 seedlings. ***, p < 0.001 (Student's t test).
