A method for the micropropagation of pepino (Solanum muricatum Aiton) from nodal segments was developed. Detailed surface disinfection methods of leaves and nodes were tested. Via establishment of initial culture using nodal segments, effect of Murashige and Skoog Basal Medium (MS) with N-6-benzylaminopurine (BAP) (1, 2, 3, 4, 5, 6 and 7 mg/l) or MS with alpha-naphthalene acetic acid (NAA) (2, 4, 6 and 8 mg/l) on some morphologic and vital parameters were noted. To obtain the best shoot multiplication medium from shoot tips of in vitro growth plants; MS with BAP (1, 2, 3, 4, 5 and 6 mg/l) and control medium without any plant growth regulators were tested. In both of the two stages, proliferation rate (%), number of shoots (shoot number/explant), length of shoots (cm), callusing rate (%) and callus size (cm) were determined. In addition, at initial culture, rooting rate (%), number of roots (root number/explant) and length of roots (cm) were also evaluated. Fully expanded shoots were transferred rooting medium MS with or without indole-butyric acid (IBA) (0, 0.5, 1, 2 and 3 mg/l) and rooting rate (%), classification of root quality, callusing rate (%), callus size (cm), and length of shoots (cm) were determined. Initiation time of rooting according to treatments were also observed in every day and fully in vitro rooted plants transferred two different acclimatization media under two different climatic conditions step by step and for each step survival rate (%) were evaluated.