Callus growth and anthraquinones production of Indian mulberry (Morinda citrifolia L.) in Murashige-Skoog’s medium (MS) supplemented with Ca2+ and Cu2+

Abstract

The objectives of the research were to study the effect of Ca2+ and Cu2+ ions in Murashige-Skoog’s medium on callus growth and anthraquinones production from Morinda citrifolia callus. The outline of the research was that the callus growth and secondary metabolite production from plant’s body could be triggered by the occurrence of elicitor that is added to culture’s medium, as biotic or abiotic elicitors. The addition of Ca2+ and Cu2+ ions in culture’s medium as abiotic elicitor would cause the occurrence of metal’s ion competition and interaction toward cells that being cultured, furthermore, it would influence ion transport from or to cell cytoplasm. Finally, cytoplasm pH would be influenced, so that both of callus growth and secondary metabolite from the cell that being cultured will also affected. In this research, in vitro callus culture method to obtain callus from explant (Morinda citrifolia leaf) and to induced anthraquinone production was used. In vitro culture used in this research consisted of 3 stages. First stage was the basal medium for sterilant object, second stage was the callus initiation medium to induce callus, and third stage was the treatment medium to induce anthraquinone production from callus. The research used factorial completely randomized design with 2 factors (Ca2+ ions: 0 mgl-1, 440 mgl-1, 880 mgl-1 and Cu2+ ions 0 mgl-1, 2,5 mgl-1, 5 mgl-1), with 3 replicates. Data collected were qualitative data (explant sterilization and callus morphology) and quantitative data (callus growth rate, callus dry weight, and anthraquinone content). The data were analyzed using ANOVA, followed by DMRT with 5% confidence level. The result of the research indicated that the treatment with addition of Ca2+ and Cu2+ ions on MS medium did not have any significant effect on callus growth and anthraquinone production.