Micropropagation of Nopal Cactus (Opuntia ficus-indica) Via Areole Proliferation: Development of an Efficient In Vitro Protocol

Opuntia ficus-indica (nopal cactus) is an economically and ecologically important succulent with applications in food, medicine, and sustainable materials, however conventional propagation methods suffer from low multiplication rates, genetic variability, and disease susceptibility, limiting commercial cultivation potential. This study aimed to develop an efficient micropropagation protocol for O. ficus-indica using areole explants through optimization of hormone concentrations for shoot initiation, elongation, rooting, and acclimatization. Areole explants from disease-free cladodes were surface-sterilized and cultured on Murashige and Skoog (MS) medium, with shoot proliferation evaluated using different combinations of 6-benzylaminopurine (BAP) and 1-naphthaleneacetic acid (NAA), while rooting efficiency was assessed using indole-3-butyric acid (IBA) at varying concentrations. A two-phase acclimatization protocol was implemented to ensure successful transplantation. Optimal shoot proliferation (14 shoots per explant, 4-7 cm length) was achieved on MS medium supplemented with 1.5 mg/L BAP and 0.3 mg/L NAA, while maximum rooting efficiency (85%) with healthy root development occurred at 1.0 mg/L IBA. The two-phase acclimatization protocol resulted in 80-95% survival rates under controlled hardening conditions. This protocol provides a reliable, scalable method for mass propagation of disease-free O. ficus-indica plantlets, supporting commercial cultivation, conservation efforts, and biotechnological applications.