Fig. 3
From: 4-Coumarate 3-hydroxylase in the lignin biosynthesis pathway is a cytosolic ascorbate peroxidase
Phenotypic characterization of Brachypodium c3h mutants. a Activation tagging construct pJJ2LBA and diagram of the T-DNA insertion in line JJ25124 (http://jgi.doe.gov). b Relative expression by qPCR, extractable activity and protein level by immunoblotting in c3h mutants compared with wild-type and T-DNA control line JJ22251 (apx3). The antibodies raised against C3H showed no cross-reactivity and detect a band of 29.5 kDa38. Line numbers are indicated in each lane. The full uncropped gel and blot shown are provided as a Source Data file. c Growth phenotype and transverse stem sections (UV-autofluorescence and phloroglucinol-HCl staining) of c3h mutants and apx3 and wild-type controls. d Total lignin (upper panel), S/G ratio (middle panel), and relative monolignol composition (lower panel) determined by thioacidolysis in c3h mutants and apx3 and wild-type controls. e Correlation plots for C3H activity with total lignin amount and individual lignin monomers. f Metabolite concentrations in mature stems of c3h mutants compared with apx3 and wild-type controls. Lignans were recognized from their fragmentation patterns. H/G lignan is 14.58 267 297 hydroxyphenyl guaiacyl lignan; G-lignan is 15.88 297 411 323 guaiacyl lignan, and S-lignan is 16.14 327 361 239 syringyl lignan glycoside (the first number is the retention time in min and the others are key mass-to-charge ratios, m/z). CWr cell wall residue. Error bars indicate mean ± SD, two-sided unpaired t-test. The Rsquared value (R2) was calculated from the linear regression model using Excel. Data points for all biological replicates are shown
