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Optimation of Callus Induction from Piper betle L. var. Nigra Explants with Various Concentrations of Coconut Water and Addition of 2,4-D and BAP


Department of Biology, Faculty of Science and Technology, Universitas Airlangga, Surabaya, Indonesia
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Black betle [Piper betle (L.) var. Nigra] is one of the endemic species from Indonesia that has potential as medical plant because this plant can produce secondary metabolites such as alkaloids, flavonoids, tannins,
steroids, terpenoids and saponins. Secondary metabolites are isolated from callus culture with proper medium formulation to get optimal result. This study aimed at knowing the influence of variations in the concentration of coconut water and combination of 0.5 mg/l 2,4-D and 2.0 mg/l BAP towards induction of callus from P. betle L. var. Nigra’s leaf explants. The experiment consisted of five treatments which were repeated for six times. Observation was performed for eight weeks in parameters including percentage of callus formed from explants, callus induction time, callus fresh weight, dry weight and morphological features (texture and colour). Results showed that various concentrations of coconut water combined with addition of 0.5 mg/l 2,4 D and 2.0 mg/l BAP had effects on callus from Piper betle L. var. Nigra leaf explants. The best combination for callus induction from P. betle L. var. Nigra’s leaf explants was addition of 0.5 mg/L 2,4 D and 2.0 mg/l BAP combined with 5% coconut water, which resulted in the highest dry weight at 0.09 g and fastest induction time at 13.17 days. Callus of P. betle L. var. Nigra had compact texture in all treatments and dominant colour was brownish yellow but callus in eighth week mostly turned to black colour.
Key words : Callus, Piper betle L. var. Nigra, coconut