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

Abstract

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 a plant’s body could be triggered by the occurrence of elicitor that is added to the culture’s medium, as biotic or abiotic elicitors. The addition of Ca²⁺ and Cu²⁺ ions in the culture’s medium as an abiotic elicitor would cause metal ion competition and interaction toward cells that are being cultured. Furthermore, it would influence ion transport from or to the cell cytoplasm. Finally, cytoplasm pH would be influenced so that both callus growth and secondary metabolite from the cultured cell will also be affected. This research used the in vitro callus culture method to obtain callus from explant (Morinda citrifolia leaf) and induced anthraquinone production. In vitro culture used in this research consisted of 3 stages. The first stage was the basal medium for sterilant objects. The second stage was the callus initiation medium to induce callus. The 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, dry callus weight, and anthraquinone content). The data were analyzed using ANOVA, followed by DMRT with a 5% confidence level. The result of the research indicated that the treatment with the addition of Ca²⁺ and Cu²⁺ ions on MS medium did not have any significant effect on callus growth and anthraquinone production.