Genetic stability in tissue-cultured tomato plants was examined by randomly amplified polymorphic DNA (RAPD) analysis. Picloram was used for the first time as alternative auxin, along with benzyladenine (BA) for callus induction in tomato. Calli were induced from leaf explants on Murashige and Skoog's (MS) medium supplemented with 8.88 μM BA and 4.13 μM picloram. Regeneration was obtained after culturing freshly-induced calli on MS medium containing 17.7 μM BA alone. Microshoots were rooted in the presence of 10 μM indole-butyric acid (IBA) on MS medium. DNA samples from the mother plant and 11 randomly selected regenerated plants, obtained from a single callus, were subjected to RAPD analysis for the detection of putative somaclones. Six arbitrary decamer primers produced polymorphic amplification products. In this set of experiments, fifteen non-parental bands were observed, of which three were shared and twelve unique. The estimation of genetic similarity coefficient based on RAPD band-sharing data indicated that ten regenerated plants were more than 95% similar to the mother plant, except one, LS5, which was found to be distinctly different. This report demonstrates the feasibility of easy induction of regenerative calli by using combination of picloram and BA and the possibility of detecting genetic variation through RAPD analysis among callus-regenerated plants in tomato at an early stage of growth.