Figure 6.

Induction of anaerobic genes requires ERF-VII genes but repression by extended night-induced starvation occurs downstream of ERF-VII stabilization by the N-end-rule. A, Wild-type plants (Col-0), the erf-vii mutant, and a transgenic line overexpressing a constitutively stable version of RAP.12 (35S:Δ-RAP2.12) were submerged (4 h) in the darkness or kept in the darkness in air after a pretreatment (6 h) either in the light or darkness. See Figure 5A for the experimental setup. The mRNA level was measured by RT-qPCR and data are expressed as relative to the 6hL+4hD sample of the Col-0 sample. Data are mean ± sd of three biological replicates (ANOVA, P < 0.05). B, same as in A, but using the 35S:MA-RAP2.3 line. C, Immunoblotting using an HA-tag antibody to measure the protein level of RAP2.3-HA. Plants of the 35S:RAP2.3-HA were treated as detailed in the figure. An α-actin antibody was used to verify the loading of protein on the electrophoresis.