Establishment of a stable and efficient in vitro regeneration system from leaf explants is a basic requirement for success in genetic transformation of peach. In this study, approximately 900 leaf explants of 'Guardian(A (R))' peach genotype were separated from in vitro shoots and cultured for shoot regeneration on A1/2 MS medium with different doses and combinations of thidiazuron (TDZ) and silver thiosulphate (STS). The results showed that callus bud formation and shoot regeneration occurred 4 and 6 weeks after culture initiation, respectively. All the regenerated shoots were from petioles of the leaf explants. Regeneration rates were significantly affected by different combinations of TDZ and STS (P < 0.05). Separation of means by Tukey test method revealed that on half strength MS media, high regeneration rates were obtained from the combinations of 6.81 A mu M TDZ + 10 A mu M STS, and of 9.08 A mu M TDZ + 10 A mu M STS (16 and 14 %, respectively). The number of shoots per regenerating explants was also significantly affected by growth regulator concentrations. The highest shoot number was obtained with the combination of 6.81 A mu M TDZ + 10 A mu M STS. Rooting percentage varied from 85 to 100 % depending on plant growth regulator used. On half strength MS medium containing 1.0 mg/l of IBA, an average of 7 roots per regenerating shoot was produced. The rooted plantlets were successfully acclimatized with 92 % survival rate under greenhouse conditions. This established regeneration system from leaf explants would facilitate the development of reliable peach genetic transformation procedures, helping accelerate peach genetic improvement using biotechnology approaches.