Apricot (Prunus armeniaca L) is widely considered as a recalcitrant species for shoot regeneration and genetic transformation. Previous reports indicated the possibility to obtain good regeneration from apricot hypocotyl slices: however, we report here an efficient and novel procedure to obtain direct shoot regeneration from the proximal zone of mature apricot cotyledons. Additionally, the system worked for all genotypes tested, which makes this protocol a very interesting and useful tool. Regeneration percentages were influenced by plant growth regulators and the initial dark incubation period, the former being critical to obtain high regeneration from stored apricot cotyledons. The highest regeneration percentages were achieved with TDZ (4 or 8 mu M) and 0.25 mu M IBA in combination with two weeks dark incubation period reaching up to 67.2%, 56.8%, 66.7%, 46.3% and 66.7% regeneration percentages for 'Canino', 'Dorada', 'Moniqui', 'Real Fino' and 'ansu Maxim' (Prunus armeniaca L var. ansu Maxim), respectively. Regeneration pattern appeared as multiple shoots per cotyledon. Histological studies showed that epidermal and subepidermal cells of the cotyledon were the cells from where direct organogenesis occurred. To the best of our knowledge, this is the first report of direct in vitro regeneration from mature apricot cotyledons. (C) 2013 Elsevier B.V. All rights reserved.