Fig. 4.

Preventing Pro biosynthesis results in a dramatic increase in callus formation in the msl2 msl3 background. (A) Production of apical callus in msl2 msl3 and msl2 msl3 p5cs1-4 seedlings in the presence and absence of exogenous Pro. The average of three biological replicates is presented, n≥25 seedlings each. Error bars represent s.d. *P<0.01, compared with msl2 msl3 seedlings of the same age (Student's t-test). (B) Pro content of the aerial tissue of mutant and wild-type seedlings grown as in A, measured as in Wilson et al. (2014). The average of two biological replicates (each performed in triplicate; n≥30 seedlings each) is presented. (C) Images of the meristematic region of seedlings stained with DAB or NBT. Scale bars: 1 mm. Quantification of H₂O₂ content by Amplex Red (D) and O₂⁻ production by NBT and formazan (E) in mutant and wild-type seedlings of indicated age. The average of three biological replicates (n≥20 seedlings each), two technical replicates each, is presented in both D and E. Statistical grouping in B,D,E was performed by ANOVA followed by Tukey's HSD test, P<0.01. Error bars indicate s.e.m. FW, fresh weight.