In the Northeast, wild American ginseng (Panax quinquefolium L.) is typically found growing in the dense shade provided by deciduous hardwood tree species such as a sugar maple, in slightly acidic soils with relatively high calcium content. Woods cultivated ginseng is often grown in forest farming agroforestry systems under similar conditions. Supplemental calcium by soil incorporation of gypsum (CaSO4·2H2O) is often recommended for woods cultivated ginseng. The objective of this study was to investigate the effects of this practice on soil chemical properties, plant growth and quality of American ginseng. In a greenhouse pot culture experiment, 2-year-old seedlings were treated with 0, 2, 4, 8, or 16 Mt·ha–1 gypsum and grown for 12 weeks. Gypsum application decreased soil pH slightly, elevated soil electrical conductivity and increased available soil Ca and sulfate concentrations. Tissue calcium concentration was increased with by gypsum treatment, but shoot and root growth was reduced. HPLC analysis of root ginsenosides revealed that Re, Rb1, Rc, and Rb2, PT ginsenoside (sum of ginsenoside Rb1, Rc, Rb2, and Rd) and total ginsenoside concentration increased by gypsum soil amendment.