Background: This study was conducted to investigate the effects of fertilization with the application of mixed organic matter after cultivating green manure crops on the growth and saponin content of Codonopsis lanceolata Trautv. Methods and Results: Five type of cultivation of green manure crops were done: hairy vetch as a single seedling crop (HV), hairy vetch and rye as a mixture of seedling (HV/R), additional fertilization with mixed organic matter after cultivation of a seedling mixture (HV/R/MO), chemical fertilizer (CF), and a non-treated control (NT). The total nitrogen content was the highest in the HV/R/ MO treatment, followed by the HV, CF, HV/R and NT treatments, respectively. Nitrogen fixation was higher in the single seedling treatment with hairy vetch (HV) than in the treatment with a mixture of hairy vetch and rye seedling (HV/R). Moreover, the growth of C. lanceolata was greatly increased by the application of additional fertilizer after treatment with the mixture of hairy vetch and rye seedlings. The HV/R/MO treatment produced the highest total lancemaside content among the treatments, followed by HV, HV/ R, CF, and NT, respectively. Conclusions: It was verified that C. lanceolata experienced superior growth with the application of mixed organic matter as additional fertilizer after the cultivation of green manure crops, and the content of its major saponins, lancemasides were also increased by this treatment.