Optimization of Callus Induction Using Different Plant Hormone and Light Condition

Abstract

Clinacanthus nutans is a well-known source for pharmaceutical drugs with most of the active ingredients classified as secondary metabolites. The study objectives are to explore the effect addition of different plant hormones alone or by combinations in the MS plant growth medium and factor of light sources towards induction of callus via in vitro culture. In this study, nodal explant of C. nutans was cultured on Murashige and Skoog Medium (MS) supplemented with different concentrations of Naphthaleneacetic acid (NAA) (0.5, 1.0, 1.5, and 2.0 mg/L) and 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 1, 2, 3 and 4 mg/L) at dark and light conditions. In hormone combination treatments, study was conducted with addition on same concentration of NAA and BAP in MS medium. Study with single hormone in MS medium resulted that only NAA had successfully induced callus at both light and dark conditions. Highest friable callus produced is at MS + 1.5 mg/L NAA in dark as 100% callus induced with 1.470 ± 0.225 g fresh weight. Next, study on the effect of NAA + BAP combinations in MS medium resulted that the treatment induced compact callus formation in both light and dark condition. The highest number of callus produced is in treatment of MS + NAA (0.1 mg/L) + BAP (0.1 mg/L) with 0.569 ± 0.159 callus fresh weight. To conclude single treatment callus 1.5 mg/L NAA alone at dark condition was best to produce friable and proliferative callus.