Chilli Pepper is the most important vegetable and widely consumed spice around the world. For genetic transformation or genome editing it is very important and prerequisite to establish an in vitro regeneration protocol. Callus formation is the essential step for regeneration. Pepper is highly recalcitrant plant. The present study was conducted to observe the high-quality callus induction from the leaf explant of hot pepper cultivar BARI Morich-2 under different hormonal concentration of auxin and cytokinin. Leaf from 10 -15 days grown in vitro grown seedlings was cultured on MS medium supplemented with 2,4-D (1.5 mg/l) and different combinations of BAP (0.2 mg/l, 0.4 mg/l and 0.6mg/l) or Zeatin (0.2 mg/l, 0.4 mg/l and 0.6mg/l). All combinations of phytohormones showed callus induction with variations of appearance and frequency.The leaf explants cultured on MS medium with 30% sucrose and 0.6 mg/l BAP + 1 .5mg/l 2,4-D were produced friable greenish calli and healthy protoplast from the calli were successfully isolated. Our findings speculated that leaf grown calli can be subjected to regenerate whole plant, genetic transformation, transient expression of gene of interest and application of genome editing tools.
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