Licorice (Glycyrrhiza glabra ) is the rhizome and root known as sweet wood. Two varieties of Glycyrrhiza (var. glabra and var. glandulifera ) were studied. The major objective of this study was to establish callus induction on Glycyrrhiza and to assess the molecular genetic variation between the mother plants and their calli using ISSR markers. Calli were formed from leaves of Glycyrrhiza cultured on Murashige and Skoog’s medium supplemented with combinations of various concentrations of auxins and cytokinins. The highest callus induction and callus fresh weight of 2 varieties tested were obtained on a medium containing 0.5 mg/l 2,4-dichlorophenoxy acetic acid with 1mg/l 1-naphthalene acetic acid and 0.5 mg/l kinetin. Genomic DNA was isolated from plants and calli using cetyltriethyl ammonium bromide method and amplified by PCR using ISSR primers. The results showed the presence of genetic variations at the DNA level during in vitro culture between callus and mother plants.