Fig. 4

Gene structure and mutation analysis of the HvPTOX gene. The coding region of HvPTOX contains 10 exons (rectangles) and 9 introns (straight lines) (a). Various primers indicated with arrows (a) were used to analyze the mutated allele, and the primer sequences were included in Additional Table 2. Full-length of the coding region could not be obtained with the primer combination of F2 and R1 in mutants, but half of that at the 3′ was successfully amplified with primers F3 and R1 (b). The promoter (F4 and R2) and a region at the 2.5 kb upstream of the start codon (F5 and R3) were deleted altogether in mutants (c). RT-PCR analysis with 25, 28 and 30 PCR cycles failed to detect the expression of Hvptox in mutants. Actin was used as the internal control. The original gel images were included in Additional Figs. 3 and 4