The effect of 2,4-dichlorophenoxyacetic acid (2,4-D) on callus growth and production flavonoid content on culture callus Acalypha indica L.

Abstract

The Objectives of the research were to study the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) on callus initiation and growth, as well as on flavonoid content on Acalypha indica callus. The outline of the research was that Acalypha indica as a medicinal plant potentially to be developed by in vitro culture method. The addition of 2,4-D in culture medium could induce cell proliferation, meanwhile, it also affects callus growth and production. Completely randomized design with three replicates was used in this study. Data collected were analyzed using ANOVA and followed by DMRT with 5% confidence level. The research consists of two experiments. Firstly, to study the effect of medium Murashige and Skoog’s composition on callus production. The media used in this experiment were medium MS and medium MS with 2,4-D 0,5 mg/L and kinetin 0,5 mg/L. Secondly, to study the effect of 2,4-D on callus growth with 4 concentrations (0 mg/L, 1 mg/L, 2 mg/L and 3 mg/L). The research variables were initiation time, weight change, texture and color of callus. Chlorophyll content was analyzed spectrophotometrically, but flavonoid content was analyzed using thin layer chromatography (TLC). The research on callus production stage indicated that 0,5 mg/L 2,4-D and 0,5 mg/L kinetin were better than 0 mg/L 2,4-D and 0 mg/L kinetin, and resulted in highest weight change at eight weeks (920 mg). Additional amount of 3 mg/L of 2,4-D resulted in the best growth of callus indicated by highest addition of weight (10160 mg) at eight weeks. The chlorophyll analysis showed that 0 mg/L 2,4-D produced highest chlorophyll content of 3,74 µmol. The callus chlorophyll content would lower with rising 2,4-D concentration. The flavonoid analysis using TLC was negative on callus.