Abstract The study on the in vitro micropropagation of Robinia pseudoacacia L. shows the possibility of the rapid increase of the vegetative progeny using organogenesis. The culture system consists of sequential use of three media, namely, the initiation medium (MS medium supplemented with 0.6 mg · l−1 6-BA (6-benzylamino-purine), 0.1 mg · l−1 NAA (naphthalene acetic acid), proliferation medium (1/2 MS medium added with 1 mg · l−1 6-BA,) and root-induction medium (1/2 MS medium fortified with 0.2 mg · l−1 NAA). The resulting micropropagated plants, fully acclimatised to an in vivo environment, did not show any visually detectable morphological variation. In addition, we investigated the genetic stability (vegetative progeny to the donor plant) of a sample of 30 plants randomly taken from ca. 1067 micropropagated plants, by using three microsatellites loci Rops 15, Rops 16, Rops 18. Our results suggest that DNA sequence variations may occur in micropropagated plants.