This study was carried out in the tissue culture laboratory, Faculty of Agriculture, Saba basha, Alexandria University, Egypt during the period from 2013 to 2015. An efficient and reliable protocol for in vitro propagation of Magnolia grandiflora, Linnwas optimized. However, nodal explants from field grown of magnolia were used during in vitro culture study for induction of multiple shoots. Nodal explants were effectively surface sterilized with 30% Clorox (sodium hypochlorite) as commercial bleaches for 20 min plus 1.5mg/l mercuric chloride for 5 min with few drops of Tween-20, also. Nodal explants were inoculated on various initiation or establishment media with different combinations of IBA and KIN and the neoformed shoots were cultured on proliferation (multiplication) media for the development of multiple shoots, and the elongation media to elongate of the neoformed shoot. The subsequent elongated shoots were rooted, successfully. The best medium for shoot initiation was Woody Plant Medium (WPM) supplemented with 2.0 mg/l KIN and 1.00 IBA. The favourable  medium for multiplication was the tested medium augmented with 5.0 mg/l KIN and 1.00 mg/l IBA.. Furthermore, the in vitro shoots showed healthy root development when the tested medium was supplemented with combination of 1.00 mg/l IBA and  NAA ,each in turn (rooting stage). The shoots of Magnolia grandiflora multiplication and rooted successfully when they cultured in WP medium supplemented with 1.00mg/l charcoal. The combination of sand: compost (1:3) was used as substratum for the hardening of the in vitro plantlets, as a potting mix, was the best suited mix for the acclimatization of plantlets.