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 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.
Key words: Acalypha indica L., 2,4-D, callus culture, callus growth, flavonoid
