Figure 6.

Changes in 35S promoter methylation during vegetative development. A, Developmental stages of N. attenuata plants used for DNA isolation. The time of harvest is described as days post germination (dpg). Tissues were harvested from cotyledons and first true leaves of seedlings (15 dpg), leaves from rosette-stage plants (30 dpg), cauline leaves from elongating plants (45 dpg) and cauline leaves from flowering plants (60 dpg). B-F, Mean cytosine methylation of the 35S promoter among plant development, separately shown for the individual sites (CG, CHG and CHH). The lines ICE 4.4, PNA 1.2 and PNA 10.1 showed increasing methylation levels in the 35S promoter, whereas the stable expressing control lines (ICE 1.1 and PNA 8.6) showed low methylation levels (±SEM, n = 10-12 clones). For the detailed data from individual clones see Additional file 3.