In vitro propagation of AZ x M9 new apple rootstock resulted of breeding program of vegetative apple rootstock in Iran was investigated. Nodal explants were inoculated in Murashige and SKoog (MS) medium supplemented with various concentrations of 6-benzylaminon purine (BAP) (0.4, 0.8 and 1.2 mg/l) alone or with naphthalene acetic acid (NAA) (0.1 mg/l). We examined the effect of different concentrations of iron sequestrene (Fe-EDDHA) and thiamin vitamin in ½ MS and Linsmaier and Skoog (LS) media supplemented with indol-3-butric acid (IBA) (1 mg/l) on rooting of attained plantlets. According to our results, 0.4 BAP + 0.1 NAA treatment in MS medium was the best component which resulted in the highest proliferation rate (6.73). As well, we estimated the highest root number in ½ MS + 0.15 g/l Fe-EDDHA + 2.4 g/l thiamin.