Rodent tuber is an anticancer plant. The natural genetic diversity of rodent tuber is low due to vegetative propagation. It is important to increase the plant's genetic diversity in order to obtain plants with a high amount of anticancer compounds. In vitro calli were irradiated with gamma rays to increase its genetic diversity. Seventeen clones of the first generation of vegetative mutants in a green house (MV1) were propagated until MV3. This research aimed to analyse the stability of mutation in MV3 based on morphology, RAPD and GC-MS markers. Clone 6-1-2 had the highest increase of shoots and leaf number than the control and the other MV3 clones while clone 6-1-3-4 had the highest fresh and dry weight. RAPD analysis using 15 primers produced 67 polymorphic DNA bands and showed four main clusters at the similarity coefficient cut-off of 0.87. The GC-MS showed that MV3 contained at least eight types of anticancer compound in the leaves and six types in the tubers; these were higher than in the control. MV3 leaves and tubers contained at least eight new anticancer compounds that were not found in the control. This research proved that rodent tuber MV3 clones were solid mutants and had a high potential for being developed into anticancer drugs.