Callus Induction and Bioactive Compounds from Piper betle L. var nigra

Black betel is a decoration plant which also have potential as source of medicine materials and alternative of safe antiseptic. In-vitro culture method can be applied to produce secondary metabolites using culture medium and optimal supplementation of growth regulators, in this case 2,4-dichlorophenoxyacetic acid (2,4-D). This study was aimed to determine the effect of 2,4-D concentration on callus induction time, percentage of explant forming callus, callus fresh weight, dry weight, morphology, and secondary metabolites and bioactive compounds profile. The study was designed as laboratory experiment using completely randomized design with 6 concentrations treatment and replicated 12 times. Culture medium used was MS medium supplemented with various concentration of 2,4-D (0.0; 0.5; 1.0; 1.5; 2.0; and 2.5 mg/L). After callus had grown, secondary metabolites content was analyzed using phytochemical screening. Result showed that 2,4-D growth regulator affected callus grown from black betel leaf explants. Growth regulator 2,4-D given at 2.5 mg/L was able to induce callus faster compared to other treatment, at mean induction time of 14 days. Explants supplemented with 2,4-D at 1.5 mg/L produced highest fresh and dry weight, at 0.8951 g and 0.0470 g respectively. Callus morphology with friable texture and yellowish white color was resulted from 1.5 mg/L 2,4-D treatment. Phytochemical analysis of secondary metabolites profile from 1.5 mg/L 2,4-D treatment indicated flavonoids content, while all 2,4-D concentration treatment (0.5; 1.0; 1.5; 2.0; and 2.5 mg/L) found to contain terpenoids. The main component is octadecanoic acid.