Optimized Tissue Culture Protocol for Producing Virus-Free Potato Plants from Major Cultivars in Bangladesh

Background and Objective: Potato production in Bangladesh is hindered by viral infections that reduce yield and quality. Developing an optimized in vitro protocol for virus-free mini-tuber production is essential for improving seed quality. This study aimed to establish an efficient meristem-based tissue culture method using sprouts from five major potato cultivars-BARI Alu-7 (Diamant), BARI Alu-8 (Cardinal), BARI Alu-25 (Asterix), BARI Alu-29 (Courage), and BARI Alu-62- and to evaluate the influence of graded vitamin stock concentrations on plant regeneration performance. Materials and Methods: Meristem explants from each variety were cultured on media supplemented with five vitamin stock concentrations: T1 (90×), T₂ (95×), T3 (100×), T₄ (105×), and T₅ (110×), and a control (T₆). The vitamin mixtures differed in Glycine, Thiamine, Nicotinic acid, and Pyridoxine HCl content per 250 ml D₂O. Growth parameters, including plant height, root length, fresh and dry biomass of shoots and roots, number of nodes, and number of leaves, were recorded. Comparative performance across treatments was analyzed to determine the optimal formulation. All recorded data were analyzed using ANOVA under a Completely Randomized Design (CRD), and treatment means were compared by Duncan’s Multiple Range Test at p≤0.05 using Statistix 10. Results: The T₄ (105×) vitamin concentration showed the most consistent and superior response across the majority of varieties. In Diamant, T₄ produced the highest plant height, root length, shoot biomass, and node number. Cardinal responded best to T₁ for shoot and root biomass and leaf number, while T₃ recorded faster shoot initiation and greater height. In Asterix, T₄ led in early shoot initiation, dry biomass, and leaf count. For BARI Alu-62, T₄ produced the highest root length and biomass, whereas T₁ enhanced node and leaf formation. Overall, T₄ exhibited the strongest performance across multiple parameters and cultivars. Conclusion: The 105× vitamin formulation (T₄) demonstrates strong potential as an optimized medium for producing vigorous, virus-free potato plantlets suitable for mini-tuber production in major Bangladeshi cultivars. Its consistent performance across varieties highlights its applicability for large-scale clean seed programs. Future studies may validate this protocol at field-scale and refine it for additional local cultivars.