Adventitious rooting of cuttings is a complex developmental process in forest species, with several exogenous and endogenous factors influencing the outcome of the process. In this study we applied an in vitro working system, comprising two lines of microshoots with the same genotype but at a different ontogenetic stages, in two different tree species (chestnut and oak). We analyzed the expression of a gene encoding an AP2/ERF transcription factor from group VII in the initial hours of the adventitious rooting induction, both in rooting competent and incompetent microshoots. The analysis revealed that expression of this gene is related to wounding, ontogenetic stage and auxin in a complex and species-specific manner. Putative induction of the gene by auxin was also analyzed in the presence of naphthyl-phthalamic acid (NPA), an auxin transport inhibitor. In situ expression analysis in chestnut relates the gene activity to cambial divisions and root primordia in rooting competent microshoots, as well as in the root apex. The putative role of the gene during adventitious roots formation is discussed.