<p>Potato (<i>Solanum tuberosum</i> L.) is an important food crop that can help increase production to meet the growing population’s demand. However, the crop faces serious threats due to various stresses. An efficient in vitro regeneration protocol is a prerequisite for trait-specific modification using various biotechnological interventions. The present study investigates the role of putrescine in the shoot organogenesis of potato from leaf and internodal explants. Incorporating AgNO₃ and putrescine, either single or in combination, enhanced shoot organogenesis in both explants. Five out of six cultivars tested showed the best shoot organogenesis when cultured on MS2 medium (MS + 10 µM AgNO₃ + 15 µM GA₃ + 10 µM BA) containing 25 µM putrescine, thus establishing the efficacy of the developed protocol across the cultivars. In this study, beneficial role of putrescine on shoot organogenesis of potato in the presence of 15 µM GA₃ + 10 µM BA was documented. Using random amplified polymorphic DNA (RAPD) and the inter-simple sequence repeats (ISSR) markers, the data indicated that the regenerated shoot lines are genetically uniform and true-to-type to the mother plant. The developed efficient and reporoducible protocol effective for most of the economically important cultivars and can be utilized for various biotechnological interventions aiming at trait-specific improvement.</p>