Fig. 2
Functional analysis of early mutations. a Partial alignment of Oryza MYB3 exon3s. Early mutations are by the asterisks. b Gene genealogy based on the entire coding regions of MYB3. The format follows Fig. 1. c Activation capacities by three MYB3s on the 1 Kb-long Nipponbare promoter of OsCHS (OsCHSpro1K). The promoter activity was measured by the ratio of two fluorescent luciferases (LUC/RUC) in three biological replicates (n = 3, but 5 for the negative control). A significant t-test (P < 0.001) on mean difference was shown by different letters. Data are represented as mean ± SE here and below. d Activations of MYB3s on the 1041 bp-long Nipponbare promoter of OsDFR (OsDFRpro1K). Each test had six biological replicates (four for negative control). Comparisons of averages by different letters were significant (t-test, P < 0.008 in both cases). e Impact of two early mutations (in square; comma refers to an indel) within 5′ region of OsDFR. Each test of transient expression had six biological replicates (four for negative control). Treatment comparisons in different letters were significant (t-tests, all P < 0.003). f Impact of one substitution (by the folded arrow and red letter) in the 5′ UTR of Hd3a. The lower panel shows the synthetic 5′UTR, with partial 5′UTR of OsDFR promoter (upper case and underlined) replaced by the 5′ UTR fragment (lower case) of Hd3a. The upper and italic sequences indicate restriction enzymes between the reporter and the tested 5′UTR region. The right panel shows reporter activities driven by OsC1/OsB2/OsTTG1 (TF complex) and the negative controls (n = 3). The mean difference is supported by a significant t-test (P < 0.001)