The aim of this research was to know effect of methyl jasmonate addition in any concentration on the callus growth and alkaloid reserpine production from in vitro culture of pule pandak [Rauvolfia serpentina (L.) Bentham ex. Kurz.]. The research frame based on the advantage of pule pandak as medicine plants sources. The addition of methyl jasmonate on the callus medium would inhibit the callus growth. It also has effects for the synthesis stricthosidine synthase. This enzyme had essential role on the reserpine biosynthesis. The research was conducted of two steps. First, initiation step to induct the callus formation. Second, treatment step to induct reserpine production. The research was done by designing a factor random completed, that is methyl jasmonate concentration. Methyl jasmonate concentration which is 0 Î¼M, 10 Î¼M, 20 Î¼M, 40 Î¼M, 80 Î¼M, and 100 Î¼M. The result data was qualitative form, that was callus morphology and quantitative form, fresh weight and dried weight, and reserpine content. The analysis of data used the ANOVA and DMRT 5%. The result of this research indicated that increasing of methyl jasmonate addition reduced the callus growth. The more methyl jasmonate concentration (10, 20, 40, 80 Î¼M) was added, the higher reserpine was produced. Methyl jasmonate concentration in 80 Î¼M produced the highest reserpine content that is 0.75 mg/g.