Fig. 1

Transactivation activity and subcellular localization of ONAC066. a ONAC066 has transactivation activity. Full length and truncated mutants of ONAC066 were fused to GAL4-binding domain and transformed into yeasts. Yeast cells harboring each of the constructs, pGBKT7 empty vector (negative control) or pGBKT7-ONAC022 (positive control) were streaked on medium plates of SD/−Trp, SD/−Trp-His/3-AT and SD/−Trp-His/3-AT/x-α-gal for 3 days at 28 °C. b Schematic diagram of different constructs used in ONAC066-NACRS or ONAC066-JBS binding assays. c ONAC066 binds to NACRS and JBS in vivo. The Rec2-ONAC066 and Rec2 empty vectors were co-transformed with the reporter constructs pHis-NACRS or pHis-JBS into yeasts and co-transformed yeast cells were streaked by a series of 10-fold dilutions on plates of SD/−Trp-Leu, SD/−Trp-Leu-His, SD/−Trp-Leu-His/50 mM 3-AT and SD/−Trp-Leu-His/100 mM 3-AT. d ONAC066 is localized in nucleus. Agrobacteria harboring pCAMBIA1300-ONAC066-GFP or pCAMBIA1300-GFP were infiltrated into leaves of N. benthamiana plants expressing a red nucleus marker protein H2B-RFP. Leaf samples were collected at 48 h after agroinfiltration. Microscopic examination was performed under a confocal laser scanning microscope in dark field for green fluorescence (left), red fluorescence (middle left), white field for cell morphology (middle right) and in combination (right), respectively. Bar = 10 μm. Experiments in (a), (c) and (d) were repeated for three times with similar results