Fig. 2.

ATAF2 requirement for full expression of NIT2 but not for NIT1 and NIT3 upon IAN treatment. (A) Induction of ATAF2 upon IAN treatment was confirmed by qRT-PCR. (B-D) Expression pattern of NIT1, NIT2, and NIT3 in ataf2 knock-out mutant and Col-0 plants upon IAN treatment. Samples were harvested at the indicated time points following mock and 200 μM IAN treatments. Three independent biological replicate cDNA samples were tested for each experiment. Expression data was normalized with an endogenous Actin7 gene (internal reference). (E) Phenotypic abnormalities in 35S::ATAF2 plants. Six-week-old 35S::ATAF2 transgenic plants showed reduced size and height and lighter green leaf colors when compared with Col-0 plants. 35S::ATAF2 transgenic plants also exhibit stunted siliques and round-shaped cauline leaves. (F) RNA blot analysis of ATAF2 and NIT2 gene in wild type and 35S::ATAF2 transgenic plants. Twelve-day-old seedlings of wild type (WT), vector control (VT), 35S::ATAF2 transgenic (L8 and L16) plants were hydroponically cultured and harvested. The blot was hybridized with ATAF2 and NIT2 gene-specific probes. Ribosomal RNA (rRNA) was used as a loading control.