Visualizing AhERF and AhDof Gene Expression in Arabidopsis

The coding sequences of the AhERF and AhDof genes, were fused with the 5’ region of the green fluorescent protein (GFP) driven by the CaMV35 promoter. The constructions were subsequently employed to transform A. thaliana plants. The above procedures were performed as described previously [14 (link)]. For GFP analysis, roots were observed under an inverted LSM510 confocal laser scanning microscope (Zeiss, Oberkochen, Germany). For visualization, seedlings were stained and mounted in 10 μg/ ml propidium iodide (PI) solution (Sigma). The green fluorescent protein (GFP) was excited with the 488 nm laser line of an argon laser, whereas PI was excited with the 514 nm laser line. The resulting images were acquired using the multi-channel mode. For GFP analysis of whole seedlings, a Lumar V.12 stereoscopic microscope with a GFP filter (Carl Zeiss Microscopy GmbH, Jena, Germany) was used.