Attempt was made to induce callus from different regions of seedling and to achieve high frequency shoot regeneration in sesame (Sesamum indicum L.) using local genotypes viz. GLW (Gulbarga local white), Western-II (W-II), DS-I, E-8 and Tumkur local (TL). Light green, healthy and moderately compact callus was obtained from the hypocotyls segments used as explants and callus was inoculated on to MS media supplemented with different combinations of growth regulators (viz., NAA, IAA, BAP, TDZ, AgNO3, ABA and Sucrose). The maximum number of shoot regeneration was in variety DS-I with 100% response achieved on MS media containing NAA 2.5mg/l, BAP 3.5mg/l and AgNO3 20µM followed by 58% response in variety W-II and 50% shoot response in variety TL and the mean shoot regeneration response was 54.9 and number of shoots pre callus were 1.85. And also the obtained callus were placed on the media containing TDZ of concentration 25 µM and IAA (2&3 µM). The maximum number of shoot regeneration was in variety DS-I with 83.3% & 75% response achieved on MS media containing TDZ 25µM and IAA 3, 2 µM respectively, followed by 58% shoot response in variety E8 and TL respectively on MS media containing TDZ 25µM and IAA 3, 2 µM respectively and the mean shoot regeneration percent was 52.35 and number of shoots pre callus were 1.59. The shoot regeneration will increase the chances of genetic improvement of sesame through biotechnological ways such as genetic transformation and in vitro cell line selections.