Induction and identification of bioactive compounds from callus extract of piper betle l. Var. nigra

Black betel (Piper betle L. var. Nigra) has been shown to have the potential for medicinal use. The black betel leaves contain 4.2% essential oil, alkaloids, flavonoids, saponins, tannins, eugenol, and chavicol. The application of plant tissue culture is needed to increase the production of secondary metabolites. This study aimed to investigate the impact of the combination of growth regulators (IBA, BAP, and kinetin) on the bioactive compounds of black betel callus extract. The experimental parts consisted of four treatments: I2B2, I1.5B1.5, I2K1.5, I1K1.5, and a control group. Callus induction time, fresh weight, dry weight, morphology, and bioactive compounds of black betel callus extract were observed. The findings showed that the combination of IBA and BAP, and IBA and kinetin influenced the growth of black betel leaves explants. The best concentration of growth regulators was the combination of 2 mg/L IBA and 2 mg/L BAP, which give the results of 9.33 days callus induction time, 650.68 mg fresh weight, and 54.22 mg dry weight. The grown callus had white to light brown colour with a compact texture. Furthermore, different treatments resulted in different bioactive compounds contained in the black betel leaves callus extract. Supplementation of I2B2 resulted in an extract containing 15 compounds, I1.5B1.5 supplementation yielded 8 compounds, I2K1.5 12 compounds, and I1K1.5 13 compounds. The bioactive compounds were predominated by methyl-β-D-Glucopyranoside (28.69%), 9,12-octadecadienoic acid (Z, Z; 15.07%), and hexadecanoic acid (11.03%).