Abstract: The present study was conducted to formulate single rapid in vitro callus induction and regeneration system for six different commercially important tomato cultivars of India. This study was done as a pre-requisite for the genetic manipulation studies to enrich vitamin E in tomato, which is ongoing research in the group. The varieties in this study include Sindhu and Shalimar (high altitude hybrid varieties), CO3, PKM, Vaishnavi and Ruchikar are hybrid varieties which are widely cultivated in the different agro climatic regions of India. Effect of the hormones, NAA (Naphthalene Acetic Acid), IAA (Indole Acetic Acid) in combination with BAP (Benzyl Amino Purine) and Kin (Kinetin) at varying concentrations were investigated on callus induction frequency. The varying concentrations of BAP were tested for regeneration capacity in all the cultivars. The highest frequency compact of callus formation was up to 90% for all the cultivars in 0.5 mg L-1 NAA+2 mg L-1 of BAP and regeneration was significantly higher with 3 mg L-1 BAP for all the cultivars. Hence, from this study, we could conclude that even though there are genotypic differences among the cultivars, common media and hormonal concentration can be used for the callus induction and for plant regeneration. In future, these results will be very useful for all the tomato genetic engineering studies.