Fig. 6

Overexpression of the ARGOS and ARL suppresses ethylene-dependent gene expression. a Gene expression was analyzed in dark-grown seedlings of the overexpression lines (35S::ARGOS #1 and 35S::ARL #2) grown in the absence and presence of 1 μL L⁻¹ ethylene. Expression of the indicated genes was determined by qRT-PCR, with the expression level of wild-type in the absence of ethylene set to 1. The left column shows genes whose expression is induced by ethylene, the right column genes whose expression is suppressed by ethylene. Data based on three biological replicates for each treatment. Statistical analysis was performed by unpaired two-tailed t-test with a Bonferroni correction (* p < 0.05; ** p < 0.01, *** p < 0.001), between the experimental samples and the corresponding wild-type control. b Overexpression of ARGOS or ARL reduces ethylene-dependent gene expression in a transient protoplast assay. Protoplasts were transfected with either of the ethylene-dependent luciferase reporters GCC-LUC or ERF1-LUC, as well as with a UBQ-GUS transgene for normalization of the transfection. Protoplasts were co-transfected with CsV::ARGOS or CsV::ARL constructs to determine their effect on the expression of the luciferase reporters. Protoplasts were treated in the absence or presence 10 μL L⁻¹ ethylene for 6 h prior to determining the relative LUC/GUS activity. Data from one experiment is shown from two independent experiments with similar results, each experiment including two biological replicates per sample treatment. Statistical analysis was performed by t-test with a Bonferroni correction (* p < 0.05; ** p < 0.01), between the experimental samples and the corresponding wild-type control