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To comprehensively understand the effect of selenium (Se) supplementation on Codonopsis pilosula, effects of sodium selenite levels on Se accumulation, growth, physiology and quality were explored by using hydroponic experiments. The results were as follows: (1)Treatment with 0.2 mg·L-1 Se promoted leaf size, plant height and biomass of C. pilosula, while treatment with 10 mg·L-1 Se suppressed them. (2)Both increasing Se level and exposure time to C. pilosula seedlings could raise its Se contents in root and leaf, and Se was distributed in organs with an order of root>leaf>stem. (3)Treatment with 0.2 mg·L-1 Se increased photosynthetic pigment content and root activity, and reduced malondialdehyde, proline and hydrogen peroxide contents, while treatment with 10 mg·L-1 Se had opposite effects. (4)Treatment with 0.2 mg·L-1 Se had positive effects on accumulations of lobetyolin, polysaccharide, and soluble protein, while treatment with 10 mg·L-1 Se had negative effects on the accumulations of polysaccharide and soluble protein. In summary, sodium selenite has double effect on C. pilosula; application of appropriate Se is beneficial to its growth, plant physiology and quality, and 0.2 mg·L-1 Se level is the optimum. This study may improve us to understand the effect of selenite level on the biology effect of C. pilosula, which will be useful for the basic research and cultivation of Se-enriched C. pilosula.