Figure 5

Methylation levels across MYB10 in 'Honeycrisp' and 'Royal Gala'. Percent methylation in A) 'Honeycrisp' and B) 'Royal Gala' green and red stripes across the MYB10 locus (Genbank accession EU518249) (C), estimated using an assay combining McrBC digestions and real-time PCR amplification. Percent methylation indicates the proportion of copies cut by McrBC. Values on the X-axis indicate the location of the primers used relative to the ATG translation start site of MYB10. Panel C indicates the relative location of the TRIM element, the MYB10 promoter and three exons (1, 2, 3); this figure is not to scale. The blue triangles indicate the approximate positions of E-box motifs within the promoter region. The calculated % methylation for the -51 to 105 fragment in 'Honeycrisp' and the -2254 to -2098 in 'Royal Gala' were negative, therefore a value of 0 is indicated in the plot. Methylation in the -704 to -555 fragment in 'Royal Gala' could not be estimated given the extremely low template levels in the McrBC treated sample. The -1874 to -1681, -303 to -182, 146 to 257 and 494 to 710 fragments were not evaluated in 'Royal Gala' (N). Reactions were performed in triplicate and two or three independent digestions were used. Error bars are SE and stars indicate significant differences (p ≤ 0.05).