Talinum paniculatum is a herbaceous plant known for its high medicinal value. GmCHI transgenic T. paniculatum in the T1 generation, that contain 4.24 mg/g and 2.74 mg/g of flavonoid, respectively, which reflect increases of 7.4-fold and 4.8-fold, respectively, compared to that in wild-type plants. In order to increase flavonoid acquisition from the biomass of GmCHI transgenic T. paniculatum in the T1 generation with high flavonoid content, it is necessary to study the effect of multiple shoots and rooting in transgenic plants. In this paper, we present the results of studying the effects of a combination of plant growth regulator on multiple shoots and rooting in GmCHI transgenic T. paniculatum. The cotyledons are suitable materials to create multiple shoot for GmCHI transgenic T. paniculatum when using the MS medium supplemented with kinetin.  The addition of 2.0 mg/l of BAP gave a higher multi-shooting effect than the addition of 1.5 mg/l of kinetin or the combination of 1.5 mg/l of kinetin with 0.8 mg/l of IBA. Using 0.5mg/l IAA had better rooting ability than combining 0.5mg/l IAA with kinetin.