Abstract: 【Objective】Andrographis paniculata （Burm.F.） nees is one of the traditional Chinese medical materials. Establishing a technical system of ethyl methane sulfonate （EMS） mutation and creating a mutant resource library with rich phenotype variation will help to provide new materials for functional genomics research and variety breeding.【Method】The optimal conditions of A. paniculata mutant bank mutation were obtained by comparing and analyzing the effects of different EMS concentrations and treatment time on seed germination and seedling growth. The morphologies of mutants with different phenotypes after mutation were evaluated and identified.【Result】Among 5 treatments, the germination rate and seedling rate of seeds treated by EMS for 4 hours were significantly higher than those of the control. With the increase of EMS concentration and treatment time, the germination rate of A. paniculata seeds decreased and the mutagenesis rate increased. Among them, the highest mutagenesis rate （3.5%） was obtained under the treatment with 0.5% EMS concentration for 12 hours, followed by the treatment with 0.8% EMS concentration for 8 hours. Compared with the control, the induced mutant plants showed significant phenotypic differences. Furthermore, EMS induced mutations in a variety of A. paniculata plants, including leaf color, blade profile, leaf arrangement, plant height, branch and flower axis.【Conclusion】EMS mutagenesis method of A. paniculata seeds was established successfully. Many types of plants with specific phenotypes were obtained, which provided new materials for functional genomics research and new variety selection.