Binahong is a medicinal plant containing secondary metabolites with antibacterial and anti-inflammatory activities. The callus culture method is one of the commonly used methods to increase the content of secondary metabolites. The effect of plant growth regulators and light conditions on binahong internodes callus induction remains limited. The aimed of this study was to determine the optimal combination of concentrations of the growth regulators NAA and BAP to induce callus from binahong internode explants in vitro under different lighting conditions. This study employed a randomized block design (RBD) with two factors: light (light and dark) and NAA (0; 1; 2; 3) ppm and BAP (0; 0.5; 1; 1.5) ppm. The parameters observed included callus emergence time, percentage of callus-forming explants, fresh weight, dry weight, and callus morphology. The results showed a significant interaction between the plant growth regulators and lighting conditions for all parameters. The fastest callus formation time (6,8 DAP) occurred with BAP 0.5 ppm + NAA 2–3 ppm under light conditions, the highest percentage under dark conditions (95,25%) while under light conditions (92.94%), and the highest fresh and dry weights were observed with BAP 1.5 ppm + NAA 3 ppm (light) at 0.4128 g fresh weight and 0.0219 g dry weight. The callus was predominantly compact in texture, ranging in color from greenish-white to brown under light conditions and from white to brown under dark conditions. These findings indicate the role of NAA, BAP, and light in binahong internodes callus induction.
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