Identification and expression analysis of twenty ARF genes in Populus

• Twenty Populus ARF genes were isolated and identified using RACE.
• The genomic structures and sequence similarity of the PeARF genes were examined.
• Dynamic expression patterns were profiled during adventitious root development.
• The corresponding PeARF proteins were localized to the nucleus in Populus cells.

The auxin response factor (ARF) family of transcription factors is a crucial component of auxin signaling and plays important roles regulating numerous growth and developmental processes in plants. We isolated and characterized 20 ARF genes involved in adventitious root development of Populus. Multiple protein sequence alignments revealed that the PeARF proteins contained a highly conserved region in their N-terminal portion corresponding to the DNA-binding domain of the Arabidopsis ARF family. Except for PeARF3.1, PeARF3.2, PeARF17.1 and PeARF17.2, the PeARF proteins contained a carboxyl-terminal domain related to the Arabidopsis domains III and IV, which are involved in homo- and heterodimerization. The exon–intron structures of the PeARF genes were determined by aligning cDNA and genomic sequences. As expected, most PeARF genes had a similar distribution of exon–intron structures. Temporal expression patterns of these genes were profiled during adventitious root development. All 20 PeARF genes were expressed in root, stem and leaf in a dynamic manner. Transient expression assays with Populus protoplasts demonstrated that these PeARFs were localized to the nucleus. These results suggest that PeARFs may play diverse regulatory roles in adventitious root development of Populus and contribute to improving our understanding of conserved and divergent aspects of auxin signaling in various species.