The present research was carried out to study the effect of some growth regulators such as 6- benzyl amino purine (BAP) at 0.5, 1.0, 2.0 and 5.0 mg/L, Naphthalene acetic acid (NAA) at 0.1 and 0.5 mg/L and their combinations in the initiation media. Besides studding the influence of such growth regulators (BAP and NAA) at concentration of 0.2, 1.0, 2.0 or 4.0 mg/L BAP and 0.1 or 0.2 mg/L NAA in the multiplication media. In addition, different auxins {i.e., Indole-3-butyric acid (IBA), Naphthalene acetic acid (NAA) and Indole-3 acetic acid (IAA)} at 0.1, 0.3, 0.5 and 1.0 mg/L were applied separately in the rooting media on the in vitro micropropagation and acclimatization of Dendranthema grandiflorum plants. The chrysanthemum cultivar used in this research was namely flyer. To evaluate the adaptation degree of chrysanthemum cultivar to the culture medium composition, the single nodal explant (0.5 – 1.0 cm) were surface sterilized and cultured on MS medium supplemented with sucrose (30 g/L), agar (7 g/L). The initiated shoots were excised and cultured on the multiplication and rooting media. The obtained results indicated that explants grown on medium containing 1.0 mg/L BAP + 0.5 mg/L NAA produced the highest shoots number. While, explants grown on medium containing 2.0 mg/L BAP + 0.1 mg/L NAA produced the tallest shoots and internodes. Moreover, explants grown on medium fortified with 1.0 mg/L BAP + 0.1 mg/L NAA produced the highest leaves number. The obtained results also indicated that, the explants grown on medium supplemented with 1.0 mg/LBAP+ 0.2mg/L NAA produced the highest shoots number. While, those grown on the multiplication medium containing 1.0 mg/L BAP + 0.1 mg/L NAA produced the longest shoots and internodes. Finally, shoot tips grown on medium containing 2.0 mg/l BAP + 0.1 mg/L NAA formed the highest  leaves number. Moreover, rooting of  the multiplied shoots were achieved on the same medium used in the multiplication stage except for the macro-elements of MS medium which used at half strength, sucrose at 20 g/L and 0.5 mg/L IBA which formed the roots initiation within a few days, followed by the medium which contained1.0 mg/L IBA which produced the highest roots number. While, medium containing 0.1 mg/L IBA resulted in the tallest roots per explants. For the acclimatization process, the combination of peat moss + perlite + vermiculite (1:1:1 v/v) showed its superiority over all the other growing media used, as it gave 100% survival percentage (%), increase in leaves number per shoots, shoots length (cm), and internodes length.