This study was intended to find out the well-defined protocol easily for in vitro propagation on different genotypes ofArganiaspinosa due to its difficulty in traditional propagation. In this respect, the highest survival percentage of shoot tips (100%) was found at 25% clorox for surface sterilization. While, genotype 3 was cultured on ¾ MS medium formed the highest number of leaves and number of shoots. For rooting, MS medium contained 3.0 mg/l NAA was suitable for genotype4. Plantlets after root development exhibited 79.66% survival in pots filled with mixture of peat moss: vermiculite (1:1, v/v) at genotype4. Inter simple sequence repeat (ISSR) analysis detected similarity to the mother tree in the in vitro regenerated plants from the shoot tips, that the direct and indirect regeneration protocol will be useful for Arganiaspinosatree production.