In Vitro Shoot Proliferation and Plantlet Development of Musa acuminata (AAA) “Dwarf Cavendish” Using Bleach Sterilization and Biochar‐Supplemented Media

This study evaluated bleach‐based medium preparation and rice husk biochar (BC) supplementation for Musa acuminata (AAA) “Dwarf Cavendish” in vitro culture. The objectives were to assess the short‐term visible sterility of nonautoclaved half‐strength Murashige and Skoog (½ MS) medium supplemented with commercial bleach, identify a basal medium formulation supporting plantlet growth, and compare BC‐only media with BC combined with activated charcoal (AC). Blank ½ MS media supplemented with 0.5–0.7 mL L ⁻¹ Hygiene bleach (6% w/w NaOCl) remained visibly sterile for 2 weeks, with no visible contamination detected. However, this medium‐only assay did not assess endophytic contamination, residual‐chlorine phytotoxicity, or long‐term sterility during plantlet culture. In ½ MS medium, 1 mg L ⁻¹ BA, 0.1 mg L ⁻¹ NAA, and 0.1% (w/v) AC produced the highest numerical shoot response (1.66 ± 0.81 shoots per explant) and supported rooting after 8 weeks, indicating limited shoot proliferation and plantlet development rather than high‐frequency multiplication. BC alone at 0.01%–0.1% (w/v) suppressed plantlet growth, particularly root development; 0.01% BC produced no roots at 8 weeks. When BC was combined with 0.1% AC, 0.05% BC + 0.1% AC produced the highest numerical values for plant height (3.33 ± 0.13 cm), leaf number (5.16 ± 0.98), and root number (6.50 ± 1.04), whereas 0.01% BC + 0.1% AC produced the highest shoot number among BC‐containing treatments (1.50 ± 0.83 shoots per explant). Under the tested conditions, AC‐containing ½ MS medium supported plantlet growth and rooting. BC addition improved selected growth variables relative to BC‐only media but did not increase shoot proliferation beyond the AC‐only treatment. Further experiments should evaluate contamination, plantlet survival, and phytotoxicity throughout a complete culture cycle and optimize conditions for greater shoot proliferation.