In the current research, an efficient protocol has been devised for production of calli and then regeneration for the important medicinal crop garlic (Allium sativum L.) via roots explants which were derived from the cloves on MS medium. Results showed that optimum calli production could be obtained on MS medium with 2, 4-D (0.5 mg/l) and Kin (0.5 mg/l). Calli were then regenerated into shoots after 4-6 weeks on regeneration medium i.e., MS medium with BA (0.5 mg/l) alone or BA (1.0 mg/l) along with 0.5 mg/l Kin. Also, optimum rooting could be observed on MS medium with growth hormones IAA (2.0 mg/l) and NAA (0.5 mg/l). The shoots obtained from calli developed into plantlets on plain ½ strength MS medium. This optimized protocol may be a step forward towards making in vitro garlic plantlets for various tissue culture studies and could be a useful tool in future for genetic transformation of this medicinally important everyday vegetable crop.