Extended Data Fig. 2: The afc1 and afc2 mutants contain mutations in CNGC2 and CNGC4.
From: A calmodulin-gated calcium channel links pathogen patterns to plant immunity
a–c, The flg22-triggered [Ca2+]cyt burst is reduced in afc1 and afc2 mutants. a, The [Ca2+]cyt burst was measured as relative luminescence units using a Varioskan Flash Spectral Scanning Multimode Reader as previously described9,30,31. The arrow indicates the time at which 500 nM flg22 was added. WT, wild type. b, Relative [Ca2+]cyt levels are depicted as luminescence counts per second relative to total luminescence counts remaining (L/Lmax)9,30,31. The remaining aequorin was discharged by the addition of 100 μl of 2 M CaCl2 with 20% ethanol per well. c, The bar graph represents the peak [Ca2+]cyt in the wild type and the two mutants after application of 500 nM flg22. d, NaCl (300 mM)-induced increases in [Ca2+]cyt in the afc1 and afc2 mutants are comparable to those of the wild type. For a–d, data are shown as mean ± s.e.m., n = 8 biologically independent leaf discs; experiments were repeated 3 times with similar results. P values are from two-sided Student’s t-tests. e, Genetic complementation group analysis indicates that AFC1 encodes CNGC2 and AFC2 encodes CNGC4. Scale bar, 2 cm. Plants of different genotypes were analysed at least twice independently and in all cases they show similar growth phenotypes. f, A schematic of CNGC with six-transmembrane domains and the cytosolic C-terminal region. The mutation in afc1 resulted in a truncated CNGC2 protein and the mutation in afc2 resulted in a G381E substitution in CNGC4.
